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.     

Products and Kinetics of the OH‐radical‐induced Dealkylation of Atrazine

Hydroxyl radicals, generated radiolytically in N₂O/O₂‐saturated solutions, yield in their reaction with atrazine equal amounts of deethylatrazine and acetaldehyde (40% of OH radical yield) and deisopropylatrazine and acetone (16%), respectively. The precursors of deethylatrazine and acetaldehyde is their Schiff base which hydrolyzes slowly (OH^–‐catalyzed: k = 5.2 dm³ mol^–1 s^–1). The hydrolysis of the Schiff base of deisopropylatrazine and acetone is too fast to be detected. In a pulse radiolysis experiment, the intermediate formed upon OH‐radical attack (k = 3·10⁹ dm³ mol^–1 s^–1) has a strong absorption at 440 nm. It decays in the presence of oxygen (k = 1.3·10⁹ dm³ mol^–1 s^–1), and upon deprotonation [pK_a(peroxyl radicals) ≈ 10.5] the peroxyl radicals thus‐formed eliminate superoxide radicals (k = 2.9·10⁵ s^–1). s‐Triazine itself reacts much more slowly with OH radicals (k = 9.7·10⁷ dm³ mol^–1 s^–1). This can explain, why in the case of atrazine in comparison to other aromatic compounds, e.g. toluene, the addition of the OH radical to the ring (estimated at ca. 40%) is of relatively little importance as compared to an H‐abstraction from (activated) positions of the side groups.     

Gene Expression Analysis for Exposure to Estrogenic Substances

Zebrafish (Danio rerio) were exposed with 17β‐estradiol (E2) and nonylphenol (NP) in different concentrations. Gene expression analysis was carried out by two different approaches. First, RT‐PCR experiments were performed for the examination of expression levels of the two marker genes vitellogenin and aromatase. This approach showed a significant increase in the expression of the vitellogenin gene in exposed male fish (500 ng/L 17β‐estradiol and 250 μg/L nonylphenol). This egg yolk protein is usually not synthesized in male vertebrates. A slight decrease of expression of the aromatase gene was observed in exposed female zebrafish. Aromatase is known to catalyze the conversion of androgens to estrogens. Second, DNA microarray experiments were carried out, which allow the simultaneous examination of the expression levels of a great number of marker genes. The microarray experiments resulted in an up‐regulation of vitellogenin up to 850‐fold. In addition, several other genes were identified to be up‐regulated by estrogens, for example the high mobility group box protein ssrp1 (78‐fold) or the chaperonin containing t‐complex polypeptide 1, beta subunit cctb (22‐fold).     

The effect of conservation tillage on runoff erosivity and soil erodibility during concentrated flow

Crop residues in conservation tillage systems are known to cause both a reduction in the erosive runoff power and an increase in the topsoil erosion resistance. In this study, the relative importance of both mechanisms in reducing soil loss by concentrated flow erosion is examined. Therefore, a method to calculate the effective flow shear stress responsible for soil detachment in the presence of a residue cover is applied. The determination of effective flow shear stress is based on the recalculation of the hydraulic radius for residue treatments. The method was tested in a laboratory flume by comparing soil detachment rates of identical pairs of soil samples that only differ in the presence or absence of crop residues. This shear stress partitioning approach and a soil detachment correction were then applied to a dataset of soil detachment measurements on undisturbed topsoil samples from a no‐till field plot on a loess‐derived soil, sampled during one growing season. Results indicate that only a small fraction (10% on average) of the difference in soil detachment rate between conventional and conservation tillage can be attributed to the dissipation of shear forces on the residues. The remaining decrease in soil detachment during concentrated runoff after a two‐year application of conservation tillage can be explained by the increased dry bulk density and root and crop residue content in the topsoil that reduces soil erodibility. After correcting for the presence of residues, the temporal variability in soil detachment rates (D_r) during concentrated flow for a given flow shear stress (τ) for both treatments can be predicted fairly well (R² = 0·87) from dry soil bulk density (DBD, representing consolidation effects), soil moisture content (SMC, representing antecedent rainfall conditions), the dry mass of organic material (OM, representing root growth and residue decomposition) and saturated soil shear strength σ_{s, sat} using an equation of the form: This study is the first to show that the effect of conservation tillage on soil detachment rates is a result of soil property modifications affecting soil erodibility, rather than a result of the surface residue decreasing flow erosivity. Copyright © 2007 John Wiley & Sons, Ltd.     

Retardation of Fin Regeneration in Freshwater Fish Oreochromis mossambicusby Zinc

Regeneration of partially amputated caudal fin was studied in freshwater fish Oreochromis mossambicus exposed to sublethal concentrations of zinc (5.0 mg L^—1 and 10.0 mg L^—1) under ambient laboratory conditions over a period of 20 days. Caudal fin regeneration was measured on 5th, 10th, 15th and 20th day of exposure and after amputation. Significant ( p < 0.05) retardation in fin regeneration was observed on day 5th and 10th in fish exposed to the nominal concentration of 5.0 mg L^—1 Zn, while retardation was found highly significant ( p < 0.01) at all the observations in 10.0 mg L^—1. The maximum inhibition in caudal fin regeneration (20.8 % and 24.3 %) was found during the initial observation at both of the concentrations (5.0 mg L^—1 and 10.0 mg L^—1) of zinc exposure. Later on the regeneration rate was almost as good as in the control group. Thus in this study fin regeneration was significantly inhibited at all time points following Zn exposure as a detrimental effect of Zn to fish. This study demonstrates that fish caudal fin regeneration is a simple assay, sensitive and easy to perform, and can serve as a model to determine the toxicity of pollutants in aquatic environment.     

Tidal mixing in sill regions: influence of sill depth and aspect ratio

A non-hydrostatic model in cross-sectional form with an idealized sill is used to examine the influence of sill depth (h _s) and aspect ratio upon internal motion. The model is forced with a barotropic tide and internal waves and mixing occurs at the sill. Calculations using a wide sill and quantifying the response using power spectra show that for a given tidal forcing namely Froude number F _r as the sill depth (h _s) increases the lee wave response and vertical mixing decrease. This is because of a reduction in across sill velocity U _s due to increased depth. Calculations show that the sill Froude number F _s based on sill depth and across sill velocity is one parameter that controls the response at the sill. At low F _s (namely F _s ≪ 1) in the wide sill case, there is little lee wave production, and the response is in terms of internal tides. At high F _s, calculations with a narrow sill show that for a given F _s value, the lee wave response and internal mixing increase with increasing aspect ratio. Calculations using a narrow sill with constant U _s show that for small values of h _s, a near surface mixed layer can occur on the downstream side of the sill. For large values of h _s, a thick well-mixed bottom boundary layer occurs due to turbulence produced by the lee waves at the seabed. For intermediate values of h _s, “internal mixing” dominates the solution and controls across thermocline mixing.     

Gas Liquid Chromatographic Analysis of DDT Residues in the Kidney of Labeo rohita

Laboratory acclimated Labeo rohita, common fresh water edible fish, were exposed to a sublethal concentration (2.5 ppm) of DDT for 2, 4, 8, 16 and 32 days. Residues of three isomers of DDT were estimated in the kidney of Labeo rohita using the gas liquid chromatographic technique. The maximum residues of DDT, DDE and DDD were 1.4327, 0.4172 and 0.2913 ppm, respectively. The total DDT residues concentration was 2.1412 ppm on the final day of the experiment. The tabulated tcalculated t at all intervals.     

Behavioural response of blackfly larvae (Simuliidae, Diptera) to different current velocities

Video equipment was used to record and to compare the behaviour of different blackfly species at two flow velocities 0.25 m s⁻¹ and 1.35 m s⁻¹. Simulium noelleri Friederichs, Simulium ornatum (complex) Meigen and Simulium variegatum Meigen were studied. Effects of flow velocity on larval locomotive activties were analysed. Looping activity and the time needed to attach for filter feeding again were affectet to some extend, depending on species and flow velocity. While in S. ornatum no differences were recorded in locomotive activity, larvae of S. noelleri and S. variegatum were less active at high flow velocities. Larvae of S. noelleri also moved over shorter distances when exposed to high flow velocities. Consequences of the differences in behavioural response to flow velocities are discussed with respect to drift and silk pad adhesion.     

Accumulation of DDT Residues in the Testes of Channa punctatus (Bloch): A Gas Liquid Chromatographic Analysis

Residues of DDT isomers accumulated in the testes of Channa punctatus (BLOCH) were quantitatively estimated by exposing the fish to 2.5 ppm sublethal concentration for 32 days. The study revealed a high accumulation of DDT residues in the testes. The maximum concentration of residues was noted on the 32nd day of the experiment. The DDT isomer accumulated in maximum concentrations was followed by DDE and DDD isomers. The maximum residue concentrations were 0.3221 ppm (DDT), 0.1119 ppm (DDE) and 0.0695 ppm (DDD) on the 32nd day of the experiment. The total DDT residue concentration at the final interval was 0.563 ppm.     

Effect of atmospheric gases,surface albedo and cloud overlap on the absorbed solar radiation

Recent studies have provided new evidence that models may systematically underestimate cloud solar absorption compared to observations. This study extends previous work on this “absorption anomaly” by using observational data together with solar radiative transfer parameterisations to calculate f_s (the ratio of surface and top of the atmosphere net cloud forcings) and its latitudinal variation for a range of cloud types. Principally, it is found that (a) the zonal mean behaviour of f_s varies substantially with cloud type, with the highest values obtained for low clouds; (b) gaseous absorption and scattering can radically alter the pattern of the variation of f_s with latitude, but gaseous effects cannot in general raise f_s to the level of around 1.5 as recently determined; (c) the importance of the gaseous contribution to the atmospheric ASR is such that whilst f_s rises with surface albedo, the net cloud contribution to the atmospheric ASR falls; (d) the assumed form of the degree of cloud overlap in the model can substantially affect the cloud contribution to the atmospheric ASR whilst leaving the parameter f_s largely unaffected; (e) even large uncertainties in the observed optical depths alone cannot account for discrepancies apparent between modelled and newly observed cloud solar absorption. It is concluded that the main source of the anomaly may derive from the considerable uncertainties regarding impure droplet microphysics rather than, or together with, uncertainties in macroscopic quantities. Further, variable surface albedos and gaseous effects may limit the use of contemporaneous satellite and ground-based measurements to infer the cloud solar absorption from the parameter f_s.     

Short‐term stream water temperature observations permit rapid assessment of potential climate change impacts

Assessment of potential climate change impacts on stream water temperature (T_s) across large scales remains challenging for resource managers because energy exchange processes between the atmosphere and the stream environment are complex and uncertain, and few long‐term datasets are available to evaluate changes over time. In this study, we demonstrate how simple monthly linear regression models based on short‐term historical T_s observations and readily available interpolated air temperature (T_a) estimates can be used for rapid assessment of historical and future changes in T_s. Models were developed for 61 sites in the southeastern USA using ≥18 months of observations and were validated at sites with longer periods of record. The T_s models were then used to estimate temporal changes in T_s at each site using both historical estimates and future T_a projections. Results suggested that the linear regression models adequately explained the variability in T_s across sites, and the relationships between T_s and T_a remained consistent over 37 years. We estimated that most sites had increases in historical annual mean T_s between 1961 and 2010 (mean of +0.11 °C decade^?1). All 61 sites were projected to experience increases in T_s from 2011 to 2060 under the three climate projections evaluated (mean of +0.41 °C decade^?1). Several of the sites with the largest historical and future T_s changes were located in ecoregions home to temperature‐sensitive fish species. This methodology can be used by resource managers for rapid assessment of potential climate change impacts on stream water temperature. Copyright © 2014 John Wiley & Sons, Ltd.     

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.     

Beeinflussung der Enzymaktivität einer Protease durch Huminstoffe Influence of Humic Substances on the Activity of a Protease

This paper deals with the influence of different humic substances (HS) on the enzyme activity (EA) of the proteolytic enzyme pronase E. The EA was determined by analyzing the concentration of the amino acid valine hydrolyzed from caseine. For amino acid analysis, a flourescence detector was used after precolumn derivatization and HPLC-separation of the hydrolysis products. The data show that the presence of HS has no adverse effects on amino acid analysis. Based on this result, a decrease of EA was observed in the presence of HS acting as enzyme inhibitors. Clear inhibition effects could be seen for different fulvic acids (HO3 FA, FBR FA and FBR 1.0 FA) at concentrations of 150 mg/L. In case of FBI-2 FA, higher concentrations were necessary to decrease the EA significantly. At an inhibitor concentration of 1500 mg/L the EA was mostly found to be less than 50%. The standard deviations of the results don't allow a differentiation between the HS of different origin and treatment. Specifying HS according to their influence on EA has to remain uncertain. As shown in a kinetic study of the hydrolysis of L-Leucine-2-naphthylamide-hydrochloride, the inhibitor HO3 FA decreases the maximum velocity of the reaction but has no effect on the Michaelis constant. These results indicate that this HS acts as a noncompetitive inhibitor.     

Runoff modelling at two field slopes: use of in situ measurements of air permeability to characterize spatial variability of saturated hydraulic conductivity

The time required at a field site to obtain a few measurements of saturated hydraulic conductivity (K_s) will allow for many measurements of soil air permeability (k_a). This study investigates if k_a measured in situ (k_{a, in situ}) can be a substitute for measurement of K_s in relation to infiltration and surface runoff modelling. Measurements of k_{a, in situ} were carried out in two small agricultural catchments. A spatial correlation of the log‐transformed values existed having a range of approximately 100 m. A predictive relationship between K_s and k_a measured on 100‐cm³ soil samples in the laboratory was derived for one of the field slopes and showed good agreement with an earlier suggested predictive K_s–k_a relationship. In situ measurements of K_s and k_a suggested that the predictive relationships also could be used at larger scale. The K_s–k_a relationships together with the k_{a, in situ} data were applied in a distributed surface runoff (DSR) model, simulating a high‐intensity rainfall event. The DSR simulation results were highly dependent on whether the geometric average of k_{a, in situ} or kriged values of k_{a, in situ} was used as model input. When increasing the resolution of K_s in the DSR model, a limit of 30–40 m was found for both field slopes. Below this limit, the simulated runoff and hydrograph peaks were independent of resolution scale. If only a few randomly chosen values of K_s were used to represent the spatial variation within the field slope, very large deviations in repeated DSR simulation results were obtained, both with respect to peak height and hydrograph shape. In contrast, when using many predicted K_s values based on a K_s–k_a relationship and measured k_{a, in situ} data, the DSR model generally captured the correct hydrograph shape although simulations were sensitive to the chosen K_s–k_a relationship. As massive measurement efforts normally will be required to obtain a satisfactory representation of the spatial variability in K_s, the use of k_{a, in situ} to assess spatial variability in K_s appears a promising alternative. Copyright © 2004 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.     

The ‘missing’ member of Hough's functions in diurnal tides

Summary It is shown that for azimuthal wave numbers and period 0.5 (s+1) days, Hough's functions for tidal oscillations are the same as the associated Legendre polynomialsP _s+1 ^s and Hough's functions are shown to form a complete set of orthogonal functions. The implication of this on the vertical variation of tides is discussed.     

Origin of Dissolved Organic Carbon Studied by UV‐vis Spectroscopy

Dissolved organic carbon (DOC) distributions in water from Lake Ipê, MS, Brazil, were investigated. The samplings were performed monthly (surface, 1 m depth, and bottom) from June 1999 to June 2000. Absorbance at 285 nm and DOC concentrations in mg dm^—3, p(DOC), were highly correlated for the three depths. 77% of the surface, 85% for 1 m and bottom samples presented a variation between 20 dm³ g^—1 cm^—1 and 50 dm³ g^—1 cm^—1 of A(285 nm)/p(DOC), that characterizes the dissolved organic matter in lake water as essentially fulvic. The ratio A(254 nm)/p(DOC) was also sensitive for fulvic matter, and an A(250 nm)/A(365 nm) = 4 ratio was characteristic of strongly colored waters. The ratios A(436 nm)/p(DOC) for the three depths also showed a significant correlation. The predominance of fulvic acid is explained by environmental characteristics such as the tropical climate, temperatures above 18 °C, and the lake environment. It was demonstrated that the variation in the water carbon content due to different compartments in the lake can be monitored by UV‐vis spectroscopy ratios.     

Atrazine fate and transport within the coastal zone in southeastern Puerto Rico

Agrichemical transport to coastal waters may have adverse ecological impact. This work examined atrazine fate and transport in a field adjacent to Puerto Rico’s Jobos Bay National Estuarine Research Reserve. The herbicide’s use was linked to residue detection in shallow groundwater and movement toward the estuary; however, data indicated that transport via this pathway was small. In contrast, surface runoff as tropical storm systems moved through the area appeared to have high potential for atrazine transport. In this case, transport to the estuary was limited by runoff event timing relative to atrazine application and very rapid atrazine dissipation (DT₅₀ = 1–3 days) in field soil. Soil incubation studies showed that accelerated degradation conditions had developed in the field due to repeated atrazine treatment. To improve weed management, atrazine replacement with other herbicide(s) is recommended. Use of products that have greater soil persistence may increase runoff risk.     

Significance of an In Vivo Genotoxicity Test for Surface Waters Is Increased when the Physiological Characteristics of the Test Organism (Bivalvia: Mollusca) Are Taken into Account?

The alkaline filter elution assay using the gills of the freshwater clam species Corbicula fluminea detects breaks in single‐stranded DNA and is thus a good method for determining the genotoxic potential of surface waters. In attempting to standardize the procedure, a wide range of factors which could have an influence on the uptake of genotoxic substances by the exposed clams were studied. The most important parameters of the static exposure in relation to the rate of filtration by the animals turned out to be the temperature, the volume of the water, and the exposure time. Differences in body size and in the amount of suspended particles in surface waters did not play a significant role. The results demonstrate that the in vivo test system can be quite sensitive and its results reproducible when the relevant species‐specific characteristics of the test organisms are brought into consideration, even if the test organism belongs to a biologically more advanced group. A clear dose‐response relationship to the reference substances 4‐nitroquinoline‐1‐oxide (NQO) and N‐methyl‐N‐nitro‐N‐nitrosoguanidine (MNNG) can be observed even after a short exposure time of between four and twenty hours. No genotoxic effects were observed when using this method on surface waters from the area of Cologne (including water from the Rhine River and within the protection zone 2 of the Cologne waterworks).     

Atrazine effects on meiobenthic assemblages of a modular estuarine mesocosm

Atrazine is a widely used herbicide in the US found at levels ranging from <10 ng/L to 62.5 microg/L in estuaries throughout the southeast. Effects of atrazine on estuarine meiobenthic assemblages chronically exposed to environmentally relevant concentrations are unknown. The purpose of our research was to assess effects of atrazine on meiobenthos at concentrations near the proposed USEPA SWQC (26 microg/L) using modular estuarine salt marsh mesocosms as a field surrogate. Indigenous copepod and nematode densities were assessed after 28 days of exposure in transplanted colonization chambers. Cluster analysis showed a group characterized by low copepod densities, mostly atrazine exposed chambers, and a group containing all but one control chamber. The later group included chambers with high densities of the copepods Paronychocamptus wilsoni and Enhydrosoma baruchi. Compared to controls, copepod densities was approximately 70% lower in atrazine chambers, with three of the most common copepod species (E. baruchi, Onychocamptus sp. and P. wilsoni) showing an average of 50-70% reduction in population densities (p<0.05). Although nematode density did not differ between atrazine and control chambers, the nematode-to-copepod ratio was significantly higher in atrazine (9.95+/-7.61; p=0.011) than in control chambers (0.61+/-0.35). Our findings suggest that chronic exposures over multiple generations to atrazine at concentrations near the proposed USEPA SWQC could have significant effects on the abundance and composition of estuarine meiobenthic copepod assemblages.