Der Leitfähigkeitstest — Ein Testsystem zur Ermittlung der membrantoxischen Eigenschaften von Substanzen

The Conductivity Test — Determining the Membrane-toxic Properties of Substances A new biotest system is introduced which is specific in terms of the measured effect but not in terms of the applied organisms. The system is able to detect the effect of membrane-toxic compounds e.g. detergents, organic solvents, and radical formers. Released into the environment, these substances damage biological membranes and therefore cause ion leakage into a surrounding medium. Ion leakage is taken as a measure for membrane damage and is determined as an increase of the electrical conductivity in the medium (conductivity test). The results presented for the tensides benzalkonium chloride, LAS (linear alkylbenzenesulfonate), and APG (alkylpolyglycoside) demonstrate the capability of the conductivity test to distinguish between the differing toxicological potential of these membrane-toxic substances. In contrast to most of the conventional biotests, the endpoint of the conductivity test is independent of a specific organism. This independence is demonstrated by the results obtained with benzalkonium chloride treatment of Elodea canadensis, Lemna minor, and Salvinia natans. The conductivity test represents a versatile, easy to handle biotest system for the assessment of ecotoxicological effects caused by membrane-toxic substances.     

Fate of Benzalkonium Chloride in a Sewage Sludge Low Temperature Conversion Process Investigated by LC‐LC/ESI‐MS/MS

Thermocatalytic low temperature conversion (LTC) is a new method for the disposal of sewage sludge. Using this method, sludge is converted into a residual solid (coal) along with reaction water, oil, and non‐condensable gases. The oil can be used as an energy source and the coal as a substitute for charcoal. To this end, it is important to determine whether there are any easily available contaminants present in the coal generated by the process. Contaminants that can be strongly sorbed by sewage sludge solids are, e. g., pharmaceuticals and disinfectants. As an example the fate of the persistent and strong adsorbing disinfectant benzalkonium chloride (BAC) has been investigated within the LTC process. The sewage sludge was spiked with BAC and then subjected to the LTC process. The resulting coal was extracted and analyzed using LC‐LC/ESI‐MS/MS (ion trap). BAC could not be detected in the LTC coal, although it could be extracted from the spiked sludge before the LTC treatment. It can thus be concluded that the investigated compound is not easily available in the coal, and hence that its use does not present a risk.     

Scum in Activated Sludge Plants: Impact of Non‐filamentous and Filamentous Bacteria

Excessive scum production is a widespread phenomenon in present activated sludge wastewater treatment. The question how foaming is initiated and stabilized is still unanswered. Hydrophobic wastewater ingredients and surface active material such as synthetic surfactants are discussed among others as major causative agents for scum production. Focusing on biological impacts non‐filamentous bacteria isolated from scum turned out to contribute to flotation by both cell surface hydrophobicity and emulsification activity, depending on the prevailing substrate and milieu conditions. The biological characterization of scum based on microscopic sludge investigation of conspicuous microorganisms resulted in a significant shift of filamentous and non‐filamentous organism populations with Gram‐positive bacteria prevailing in present nutrient removal plants as compared to the situation ten years ago. Their hydrophobic cell surface is supposed to support adherence and stabilization of interfaces and thus promote sludge flotation. In scum six types of filamentous bacteria turned out to be numerous: Microthrix parvicella and nocardioform actinomycetes, both of them being enriched in the scum fraction, moreover Nostocoida limicola and Eikelboom types 0041/0675, 1851, and 0092. Possible interactive mechanisms between non‐filamentous and filamentous scum bacteria and their selection factors are discussed in order to contribute to a better understanding of scum formation and to provide efficient troubleshooting measures.     

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.     

Assessment of the Comparative Toxicity of Sewage Effluent from 10 Sewage Treatment Plants in the Area of Sydney, Australia using an Amphipod and Two Sea Urchin Bioassays

In 1995, Sydney Water Corporation undertook an ecological and human health risk assessment for 10 sewage treatment plants (STPs) that discharge primary (6 STPs) and secondary (4 STPs) treated effluents into coastal waters in the Sydney and Illawarra regions, NSW, Australia. A program of toxicity testing of effluent from the 10 STPs was undertaken to determine the toxic effects of the effluents and as a weight-of-evidence study for the risk assessment.

Three types of bioassays were used to test multiple samples of effluent from each STP. Tests used were the sea urchin (Heliocidaris tuberculata) fertilization and larval development bioassays and the amphipod (Allorchestes compressa) survival bioassay.

Comparing between STPs, primary treated effluent was slightly more toxic than secondary treated effluent for all three tests. The highest toxicity was recorded for those STPs that use primary treatment and discharge to the shoreline. Comparing between tests, the sea urchin fertilization bioassay was slightly more sensitive than the sea urchin larval development bioassay, with the amphipod survival bioassay being the least sensitive.

This paper will describe the results of this toxicity testing program and discuss the application of the study results as weight-of-evidence for the risk assessment study.     

Drug-resistance transfer in Escherichia coli in New York Bight sediment

Sediment from the sewage sludge dump site area in the New York Bight contains bacteria resistant to antibiotics and heavy metals. The purpose of this study was to determine if antibiotic resistance could be transferred from donor to recipient Escherichia coli strains inoculated into glass vessels containing sediment and seawater obtained near the dump site. Temperature was maintained at 10°C, the mean winter temperature of benthic water at the dump site. Transconjugants (recipients which inherit donor genes for tetracycline resistance) were isolated from the seawater and sediment within one hour after inoculation and were found in all subsequent sediment samples for one month. Donor and recipient E. coli remained viable in the sediment for at least one month. Our results indicate that sewage sludge polluted sediment may serve as an environment conducive to conjugal transfer of antibiotic resistance genes.     

The Growth of Micromycetes in Activated Sludge Media

Today already numerous micromycetes are a recognized part of the biocenosis of growth in receiving waters strongly polluted by organically rich waste waters and of growth on biofilter media surfaces. The principal representatives are some Deuteromycetes, Mucorales and Saccharomycetaceae, whereas the so-called true aquatic fungi (Oomycetes) with the exception of Leptomitus lacteus are practically absent. The growth of micromycetes in activated sludge remains still an unanswered question. Generally, the filaments in the activated sludge are ascribed to filamentous bacteria and as long as fungi were isolated from activated sludge their presence has been reported only in the form of spores. The nitrogen deficiency with regard to carbon present in the waste water is considered as one of the factors enhancing the development of fungi in the activated sludge. In a series of 13 tests carried out on laboratory-scale batch models using synthetic waste water, where the carbon source was glucose and the nitrogen source was (a) sodium nitrate, (b) ammonium sulphate, and (c) peptone, the fungal growth was studied in different models with the BOD:N ratio varying from 20 to 1000. The models were seeded with thickened activated sludge from the municipal sewage works. The sludge did not contain any microscopically detectable forms of fungoid growth. It was found that the fungi in activated-sludge medium grow well and both in the forms of single cells, occuring independently and in chains, as well as in the form of long branched filaments. In some instances, the micromycetes formed up to 80 to 100 per cent of the biocenosis of the activated-sludge flocs, but the dependency of their development on the increasing BOD:N ratios in the waste water has not been ascertained. Fifteen species of micromycetes were isolated and identified, and in addition several representatives of Saccharomycetaceae, which were not identified.     

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.     

Kinetic Studies and Anaerobic Co‐digestion of Vegetable Market Waste and Sewage Sludge

The effect of anaerobic co‐digestion of vegetable market waste and sewage sludge was studied extensively. The effects of co‐digestion were compared with the separate digestion of vegetable market waste and sewage sludge. The batch studies were carried out using three bench scale reactors having 1.5 L working volume. The cumulative biogas production shows that the organic waste available from the vegetable waste contains easily biodegradable organic matter compared with the sewage sludge. First order reaction kinetics is maintained throughout the methanation fermentation. The reductions in volatile solids (VS) in the three reactors were in the range of 63–65 %. The specific gas production for vegetable waste was higher (0.75 L biogas/g VS_in and 1.17 L biogas/g VS_des) than for the sewage sludge (0.43 L biogas/g VS_in and 0.68 L biogas/g VS_des). Consequently, the specific gas production for the co‐digestion of the mixture of vegetable waste and sewage sludge (0.68 L biogas/g VS_in and 1.04 L biogas/g VS_des) was considerably higher than for the sewage sludge only. Batch kinetics of anaerobic digestion is useful in predicting the performance of digesters and for the design of appropriate digesters.     

Wachstum gentechnisch veränderter E. coli K—12-Zellen in Klärschlamm aus zwei Kläranlagen Growth of Genetically Modified E. coli K - 12 Cells in Sewage Sludge of Two Different Sewage Treatment Plants

The release of genetically engineered microorganisms may occur from research laboratories of production plants. The most important pathway of release is via the sewage system and the sewage treatment plants into surface waters. The growth of these organisms might represent a risk to the environment. Therefore the growth of a genetically engineered strain of E. coli K-12 was investigated. Aqueous supernatant from sewage sludge of two domestic sewage plants was used as medium. This medium was treated with sterile filtration and amended with nutrients. The test strain E. coli K-12 W3110i^qM15Nal^r(pBR322) did not show permanent growth under the conditions employed. The result is interpreted by the presence of bacteriophages and the poor nutritional conditions in the medium.     

Laboratory Testing of Sulfamethoxazole and its Metabolite Acetyl‐Sulfamethoxazole in Soil

The fate of ¹⁴C‐labeled sulfamethoxazole and acetyl‐sulfamethoxazole in soil has been investigated with special respect to possible entry routes of human and veterinary pharmaceuticals into soil environments. Therefore, the stability of the test substances was monitored first in sewage sludge and bovine manure. Within the incubation period of 72 d, 1% at maximum of the initially applied radiotracers was released as ¹⁴C‐carbon dioxide while ?75% was transferred to non‐extractable residues that were operationally defined by the ethyl acetate extraction. Test‐sludge and test‐manure samples with defined aged residues were prepared and, supplementary to standard solutions, applied to silty‐clay soil samples. After standard and test‐sludge application, soil/water distribution coefficients of K_d < 5 L kg^–1 were determined revealing both test substances as potential leachers. In contrast, the sorption of sulfamethoxazole increased after test‐manure application (K_d > 10 L kg^–1). In the long‐term degradability tests, the metabolic fate of both test substances was characterized by the continuous decrease of extractable residues, resulting in disappearance times of DT₉₀ ? 33 d, and the increase of non‐extractable residues. Mineralization reached 11% at maximum. Thereby, the dynamics of these processes differed whether the test substances were applied via standard, test‐sludge or test‐manure application. This fact emphasized the relevance of entry route specific matrix effects on the fate of both test substances in soil.     

Effect of Co‐culture and Nutrients Supplementation on Bioremediation of Crude Petroleum Sludge

Ex‐situ bioremediation of real‐field crude petroleum sludge was evaluated to elucidate the role of co‐culture (bioaugmentation) and external nutrients supplementation (biostimulation) under anaerobic microenvironment. Maximum removal of total petroleum hydrocarbons (TPH) was observed by integrating biostimulation with bioaugmentation (R5, 44.01%) followed by bioaugmentation alone (R4, 34.47%), co‐substrate supplemented operations [R6, 23.36%; R3, 16.5%; R2, 9.88%] and control (R1, 4.36%). Aromatics fraction showed higher degradation in all the conditions studied. Fate of six selected polycyclic aromatic hydrocarbons (PAHs) was evaluated during bioremediation. Among these, four ring PAHs compounds showed good degradation by integration of biostimulation with bioaugmentation (R5) while bioaugmentation alone (R4) documented good degradation of three ring PAHs. Lower ring PAHs compounds showed good degradation with the application of biostimulation (R6). Fluorescent in situ hybridization (FISH) detected the presence of known PAHs degrading microorganisms viz., Bacillus, Pseudomonas, Acido bacteria, Sulphur reducing bacteria Firmicutes, etc. Application of biostimulation and bioaugmentation strategies alone or in combinations documented noticeable influence on the degradation of petroleum sludge.     

Assessment of Cyto‐ and Genotoxic Effects of a Variety of Chemicals Using Saccharomyces cerevisiae

Saccharomyces cerevisiae as the most simple eukaryotic organism is broadly accepted as a laboratory model organism. For the detection of potential toxic effects of pure compounds and complex composed samples like wastewater a miniaturised short‐term in vitro cyto‐ and genotoxicity screening assay was developed. The assay based on genetically modified S. cerevisiae cells deleted in the prominent drug efflux transporters Pdr5, Snq2, and Yor1 that facilitate pleiotropic drug resistance. The yeast strain devoid of these proteins that mediate the efflux of structurally diverse hydrophobic compounds exhibited an increased sensitivity to a variety of organic compounds. The DNA damage inducible RAD54 promoter fused to a yeast optimized derivative of the GFP (green fluorescent protein) gene from the jelly fish Aequorea victoria served as an indicator of DNA damage in this strain. Various pure compounds including the direct‐acting genotoxins methyl‐N‐nitro‐N‐nitrosoguanidine (MNNG), 4‐nitroquinoline‐N‐oxide (4‐NQO), methyl methanesulfonate (MMS) and hydroxyurea as well as the heavy metals cadmium and chromium(VI), the insecticide lindane and the central nervous system stimulant caffeine were tested exhibiting dose dependent induction of green fluorescence. All compounds were in parallel examined for chronic toxicity. A bioassay detecting simultaneously geno‐ and cytotoxic effects of potential toxicants in a single assay can be an important tool with a variety of applications in environmental monitoring and aquatic ecotoxicology. By partial automation and miniaturisation to microtitration scale this bioassay enables sensitive and fast biomonitoring for a multitude of samples.     

Biodegradation and transport of benzene,toluene, and xylenes in a simulated aquifer: comparison of modelled and experimental results

Both laboratory experiments and numerical modelling were conducted to study the biodegradation and transport of benzene–toluene–xylenes (BTX) in a simulated semi‐confined aquifer. The factors incorporated into the numerical model include advection, hydrodynamic dispersion, adsorption, and biodegradation. The various physico‐chemical parameters required by the numerical model were measured experimentally. In the experimental portion of the study, BTX compounds were introduced into the aquifer sand. After the contaminants had been transported through the system, BTX concentrations were measured at 12 equally spaced wells. Subsequently, microorganisms obtained from the activated sludge of a sewage treatment plant and cultured in BTX mixtures were introduced into the aquifer through the 12 sampling wells. The distribution data for BTX adsorption by the aquifer sand form a nonlinear isotherm. The degree of adsorption by the sand varies, depending on the composition of the solute. The degradation time, measured from the time since the bacteria were added to the aquifer until a specific contaminant was no longer detectable, was 35–42 h for BTX. The dissolved oxygen, after degradation by BTX compounds and bacteria, was consumed by about 40–60% in the entire simulated aquifer; thus the aerobic conditions were maintained. This study provides insights for the biodegradation and transport of BTX in aquifers by numerical modelling and laboratory experiments. Experimental and numerical comparisons indicate that the results by Monod degradation kinetics are more accurate than those by the first‐order degradation kinetics. Copyright © 2002 John Wiley & Sons, Ltd.     

Scum in Nutrient Removal Plants: The Role of Carbon Sources in “Microthrix parvicella” Growth

Scum formation is a widespread problem in activated sludge nutrient removal plants. It often comes along with an excessive development of the filamentous bacterium “Microthrix parvicella” stabilizing the flotation process. As “M. parvicella” was found to depend on long‐chain fatty acids (LCFA) as sole carbon source not only in vitro but also in situ, some options of in‐situ substrate supply are discussed. Wastewater concentrations of fatty acids in the range of 2 to 15 mg L^‐1 and homologue concentrations from synthetic surfactant degradation below 10 mg L^‐1 rule out these substrates as source for excessive biomass production. They might, however, well be suitable for start‐up of a “M. parvicella” population. Build‐up of excessive biomass might rely on fatty acid supply originating in cell walls of lysed stationary phase bacteria of long residence time sludge fractions such as scum layers. Moreover, biogenic surfactants such as rhamnolipids have been proved to be an excellent carbon source for excessive biomass production in vitro.     

Untersuchungen zum Abbau von Pharmaka in kommunalen Kläranlagen mit HPLC-Electrospray-Massenspektrometrie

Examination of the Degradation of Drugs in Municipal Sewage Plants Using Liquid Chromatography-Electrospray Mass Spectrometry Numerous drugs can be identified in the secondary effluent of municipal sewage plants. In order to obtain information about the degree of elimination (adsorption, aerobic degradation) of these compounds, a batch reactor containing different drugs in environmentally relevant concentrations and a suspension of activated sludge was coupled to a HPLC-MS-MS system. During a testing period of three days concentration-time-curves were recorded. For most of the examined drugs (acetamidoantipyrine, crotamiton, diclofenac, primidone, propyphenazone) solely a strong decrease of the initial concentration within the first 15 minutes was observed, which was interpreted primarily as adsorption to the activated sludge. For acetaminophenol and pentoxifyllin an additional slower decrease in concentration within several hours was observed. This slower elimination was interpreted to be caused mainly by primary degradation. It could be shown that dihydrocodeine is oxidized to hydrocodone in the batch reactor. The conjugate acetaminophen glucuronide was cleaved.     

Subsurface Injection of Treated Sewage into a Saline-Water Aquifer at St. Petersburg, Florida — Water-Quality Changes and Potential for Recovery of Injected Sewage

The city of St. Petersburg is testing subsurface injection of treated sewage into the Floridan aquifer as a means of eliminating discharge of sewage to surface waters and as a means of storing treated sewage for future nonpotable reuse. The injection zone at the test site at the start of injection contained saline water with chloride concentrations ranging from 14,000 to 20,000 milligrams per liter (mg/1). Treated sewage with a mean chloride concentration of 170 mg/1 was injected through a single well for 12 months at a mean rate of 4.7 × 10⁵ cubic feet per day. The volume of water injected during the year was 1.7 × 10⁸ cubic feet. Dissolved oxygen was contained in the sewage prior to injection. Water removed from the injection zone during injection was essentially free of oxygen. Probable growth of denitrifying bacteria and, thus, microbial denitri-fication, was suggested by bacterial counts in water from two observation wells that were close to the injection well. The volume fraction of treated sewage in water from wells located 35 feet and 733 feet from the injection well and open to the upper part of the injection zone stabilized at about 0.9 and 0.75, respectively. Chloride concentrations stabilized at about 1,900 mg/1 in water from the well that was 35 feet from the injection well and stabilized at about 4,000 mg/1 in water from the well that was 733 feet from the injection well. These and other data suggest that very little near injection-quality treated sewage would be recoverable from storage in the injection zone.     

Natural oligotrophy vs. pollution-induced eutrophy on the SE Mediterranean shallow shelf (Israel): Environmental parameters and benthic foraminifera

The eastern Mediterranean is naturally highly oligotrophic, but urbanization along the Levant coast has led to raised organic and nutrient loads. This study tracks living foraminiferal assemblages at two sites near an activated sewage sludge outfall from 11/2003 to 5/2004. Oligotrophic site PL29 shows seasonal variations in O_2, chlorophyll a, and organic carbon, and has an abundant, diverse benthic foraminiferal assemblage living at various in-sediment depths. At eutrophic site PL3, ∼16 years of sludge injection favor a depleted assemblage primarily of opportunist foraminifera. This site shows less seasonality, is subjected to organic matter overload, O₂-stress, and periodic anoxia, foraminifera are less abundant and diverse, and live at shallower depths. The assemblages at both sites represent a common pool of species, with Ammonia tepida highly dominant. Benthic foraminifera were therefore found to be sensitive to trophic trajectories, respond on sub-seasonal time-scales, and track injection and dispersal of organic loads on the shelf.