Native Water Samples Research Articles

Pesticide dynamics in three small agricultural creeks in Hesse, Germany.

Due to their high biodiversity, small water bodies play an important role for freshwater ecosystems. Nonetheless, systematic pesticide monitoring in small creeks with a catchment <30 km2 is rarely conducted. In this study, event-driven water samples were taken from May until November 2017 and March until July 2018 after 20 rain events at three sampling sites with catchment areas of <27 km2 in the Wetterau, a region with intensive agriculture in Southern Hesse, Germany. Additionally, enriched extracts of the native water samples from the campaign in 2018 were used for the Microtox assay to determine baseline toxicity to invertebrates over time and sum of toxic units (STU) were calculated to compare the potential toxicity of the samples. Overall, 37 pesticides and 17 transformation products were found, whereby the herbicide metamitron (79 µg/L) showed the highest concentration. Regularly, pesticide concentrations peaked at the time of the highest water level within each sampling event. Within each sampling event maximum pesticide concentration was mostly reached in water samples taken during the first two hours. The sum of the time-weighted mean concentration values of all pesticides was between 2.0 µg/L and 7.2 µg/L, whereby the measured concentrations exceeded their regulatory acceptable concentration (RAC) at 55% of all sampling events for at least one pesticide. The mean EC50 values varied between 28.6 ± 13.1 to 41.3 ± 12.1 REF (relative enrichment factor). The results indicated that several samples caused baseline toxicity, whereby the highest activity was measured at the time of highest water levels and pesticides concentrations, and then steadily decreased in parallel with the water level. Median STUs of invertebrates ranged from -2.10 to -3.91, of algae/aquatic plants from -0.79 to -1.84 and of fish from -2.47 to -4.24. For one of the three sampling sites, a significant linear correlation between baseline toxicity and STUinvertebratewas found (r2=0.48). The results of the present study suggest that (1) current pesticide monitoring programs underestimate risks posed by the exposure to pesticides for aquatic organisms and (2) pre-authorization regulatory risk assessment schemes are insufficient to protect aquatic environments.

Combined use of biomarkers to assess the impact of untreated wastewater from the Danube River, Serbia.

In this study a battery of bioassays, both in vivo (metals and metalloids concentrations, erythrocyte morphometry, comet assay, micronucleus assay, and histopathological analyses) on vimba bream Vimba vimba (L., 1758) and white bream Blicca bjoerkna (L., 1758), and in vitro (treatment of HepG2 cells with native water samples) was applied to assess the harmful potential of untreated wastewater. Faecal indicator bacteria were quantified to assess the microbiological water quality. Vimba bream had significantly higher Fe concentrations in both liver and muscle, while white bream had higher Ca and Cu concentrations in liver. Vimba bream had a significantly higher level of DNA damage in both liver and blood cells, in comparison to white bream. Low levels of micronucleus and nuclear abnormalities were observed in both species. Erythrocytes morphometry did not show significant interspecific differences. Histopathological analyses revealed a similar response of the studied species, with a significantly higher presence of ceroid pigments in the liver of vimba bream. Treatment of HepG2 cells revealed the high genotoxic potential of water downstream of the discharge point. The results of this study clearly demonstrate the importance of effect-based monitoring, in order to enforce more efficient management of natural resources and implementation of wastewater treatment systems.

High-performance thin-layer chromatography in combination with an acetylcholinesterase-inhibition bioassay with pre-oxidation of organothiophosphates to determine neurotoxic effects in storm, waste, and surface water

Pesticides such as organothiophosphates (OTPs) are neurotoxically active and enter the aquatic environment. Bioassays, using acetylcholinesterase (AChE), a suitable substrate and reactant, can be applied for the photometric detection of AChE-inhibiton (AChE-I) effects. The oxidized forms of OTPs, so-called oxons, have higher inhibition potentials for AChE. Therefore, a higher sensitivity is achieved for application of oxidized samples to the AChE assay. In this study, the oxidation of malathion, parathion, and chlorpyrifos by n-bromosuccinimide (NBS) was investigated in an approach combining high-performance thin-layer chromatography (HPTLC) with an AChE-I assay. Two AChE application approaches, immersion and spraying, were compared regarding sensitivity, precision, and general feasibility of the OTP effect detection. The oxidation by NBS led to an activation of the OTPs and a strong increase in sensitivity similar to the oxons tested. The sensitivity and precision of the two application techniques were similar, although the spray method was slightly more sensitive to the oxidized OTPs. The 10% inhibition concentrations (IC10) for the spray approach were 0.26, 0.75, and 0.35 ng/spot for activated malathion, parathion, and chlorpyrifos, respectively. AChE-I effect recoveries in samples from a stormwater retention basin and receiving stream were between 69 and 92% for malathion, parathion, and chlorpyrifos. The overall workflow, including sample enrichment by solid-phase extraction, HPTLC, oxidation of OTPs, and AChE-I assay, was demonstrated to be suitable for the detection of AChE-I effects in native water samples. An effect of unknown origin was found in a sample from a stormwater retention basin.

Chemical and Toxicological Analysis of Territories Affected by Radiation and Chemical Pollutants: III. Peroxide Oxidation of Lipids and Toxicity Effects of Natural Water (in vivo Studies)

The toxic properties of river water and groundwater samples taken in the Zhizdrinskii and Ul’yanovskii districts of the Kaluga Oblast, which were the areas most exposed to long-lived radioisotopes in consequence of the accident at the Chernobyl nuclear power plant, are studied using bioassay methods based on an analysis of the cumulative effect of toxic substances that are present in the analyzed water sample on the lipid peroxidation (LPO) processes. The concentrations of LPO products and extracellular DNA in the blood plasma of mice, the frequency of chromosomal aberrations in bone marrow cells of mice, the antioxidant status of mouse tissues, and the total number of alterations in the structure of DNA molecules of mouse blood lymphocytes, as estimated based on the value of the relative rate constant of alkaline elution, are determined in in vivo experiments with a single per os administration of hexane–ether extracts from drinking water to experimental animals. The concentration of LPO products in native water samples is determined in the in vitro experiments using LPO liposomes obtained from lecithin, i.e., egg yolk phospholipid, as a model test system. The total set of the obtained data indicates the presence of a relationship between the parameters of the LPO control system and DNA damage and makes it possible to propose a method for the initiated oxidation of liposomes produced from lecithin that can be used in the initial assessment of the intensity of LPO in an aqueous medium.

Chemical and Toxicological Analysis of Territories Affected by Radiation and Chemical Pollutants: II. Toxicity Effects of Natural Water (in vitro Studies)

The toxic properties of native water samples of surface (river) water and groundwater used to meet drinking water needs in settlements of the Zhizdrinskii and Ul’yanovskii districts of Kaluga Oblast, which were the areas most exposed to radiation after the accident at the Chernobyl nuclear power plant in 1986, are studied. The focus is on manifestations of genotoxic effects, as the most typical effects of radiation pollution of the territory. It is shown that water exhibits, in most of the analyzed samples, a reliable cytogenetic effect in relation to cells of transplanted cultures of the Chinese hamster (CHO and 237 clones). At the same time, the extractable ether-soluble organic substances contained in the analyzed water samples exhibit genotoxic activity only after they are concentrated by a factor of 100. In parallel, the mutagenic toxicity of water samples was analyzed using the Ames test. Without metabolic activation (in the absence of additions of the S9 fraction of the rat liver), most water samples from the drinking water supply sources show an acute toxicity to Salmonella strains (TA 97, TA 98, and TA 100). The results of biological tests are discussed in relation to previously obtained data on the radiation effects of the Chernobyl accident on the aquatic environment of the Dnieper reservoirs. It is concluded that the principal role in the manifestation of genotoxic properties may be played by radioactive isotopes that are hidden from traditional methods of radiation monitoring and are sources of alpha and beta radiation that lead to the formation of metal ions in the superoxidized state in the presence of environmental pollution by transition metals, in particular, to the formation of microcolloidal particles of mixed-valence manganese Mn(III, IV) that serve as carriers for the OH radical. The acute and chronic toxicity of the analyzed river and groundwater samples is established by bioassay methods using test organisms of various trophic levels (bacteria, ciliates, and Daphnia). No correlation with toxicity is detected in any of the traditionally monitored hydrochemical indices of water quality.

Role of Water-Soluble Forms of Toxicants in the Formation of Toxic Properties of Natural and Waste Water

Cytogenetic methods based on the analysis of chromosome changes in somatic mammalian cells are promising techniques for the early recognition of toxic effects arising from anthropogenic pollution of environmental objects. The genetic changes in somatic cells are an integral parameter of the mutagenic effect of the environment and the effectiveness of the immune system of organisms. In this work, the method of analysis of toxic properties based on determining the frequency and types of chromosomal aberrations during the short-term (within 2 h) exposure of mammalian cell cultures to a test water sample is used to study the toxic properties of natural and waste water with different levels of anthropogenic pollution. It is shown that the contribution to the genotoxicity of the analyzed native water samples from water-soluble substances that cannot be extracted with a mixture of hexane and ethyl ether can reach 50–75%, while the contribution of organic substances extracted with organic solvents can be only 2–3%. Water-soluble compounds of reduced sulfur play a key role in the formation of toxic properties of natural and waste waters. Using benzo[a]pyrene and 3,3',4,4'-tetrachlorobiphenyl (PCB 77) as an example, it is found that the genotoxic properties of even the most hydrophobic toxicants are determined by their dissolved form and can manifest themselves starting from a concentration of 10 ng/L. A comparative analysis with the Ames test, which is widely used in the study of the mutagenic activity of various toxicants, indicates that the cytogenetic method is a much more informative and sensitive method for monitoring the chemical safety of water bodies.

CONTENT OF URANIUM ISOTOPES IN THE GROUNDWATER SUPPLIES USED FOR POPULATION OF LENINGRAD REGION AND ST-PETERSBURG

The aim of the study was to obtain 234U and 238U specific activity in the water of groundwater supplies of Leningrad region and St-Petersburg and to estimate the possibility of their using from the point of view of radiation and chemical toxicity of the main dose-forming uranium isotopes usually containing in the underground waters. The study was focused on the water of functioning and exploratory groundwater supplies concerning the safety of water consumption taken into account radiation and chemical index. Data on the levels of natural uranium isotopes specific activity in the sources under investigation was obtained using radiochemical analysis with the subsequent alpha-spectrometric measurement of the counting samples. 58 samples of natural groundwater were analyzed during the period from February of 2011 till the May of 2017. The volume of the native water samples was from 10 l to 30 l, measuring time for each counting sample was 6 – 12 hours, which allowed measuring the radionuclides specific activity value at the level of 0.0002 Bq/kg. The values of 238U specific activity varied within 0.0002 Bq/kg – 0.06 Bq/kg, the average value was 0.007 Bq/kg. The values of 234U specific activity varied within 0.0006 Bq/kg – 0.111 Bq/kg, the average value was 0.013 Bq/kg. The average value of the specific activities ratio 234U/238U was 2.12 and the range of this value was 1.00 – 8.00. The results of carried out investigations demonstrates that the specific activity values of uranium isotopes could differ more then by an order of magnitude even in the borders of same aquifer of a small settlement. 238U specific activity value does not exceed 0.02 Bq/kg and 234U specific activity value does not exceed 0.04 Bq/kg for the vast majority of analyzed sample. The value of the ratio 234U/238U was 1.00 – 4.00 for 98.3% of analyzed sample. It is possible to conclude with a high probability that the water from the groundwater supplies of Leningrad region and St-Petersburg is safe for consumption by population concerning 234U and 238U content as from the point of view of radiological factor, so from the point of view of chemical toxicity.

Evaluation of genotoxic potential in the Velika Morava River Basin in vitro and in situ

The Velika Morava River is the greatest national Serbian river and the significant tributary of the Danube River. The major problems in the Velika Morava River Basin (VMRB) represent untreated industrial and municipal wastewaters. In this study, the level of genotoxic potential at the sites along the VMRB was evaluated by parallel in vitro and in situ approach. Within in vitro testing, genotoxicity of native water samples collected from the sites in VMRB was evaluated by SOS/umuC test on Salmonella typhimurium TA1535/pSK1002 and by the comet assay on HepG2 cells. DNA damage in situ was assessed in bleak (Alburnus alburnus) erythrocytes by the comet (alkaline and Fpg-modified comet) and micronucleus assays. Additionally, the concentration of heavy metals in fish tissue was measured and this data, compiled with the data of the physico-chemical parameters measured in water, was used as a measure of the pollution pressure at the sites.Results showed that applied in vitro tests with native water samples are less sensitive in comparison with in situ tests and should be taken with precaution when making predictions on the status of the ecosystem. Within applied battery of in situ assays differential sensitivity of assays was observed where alkaline comet assay showed the highest potential in differentiation of the sites based on genotoxic potential. Integrated biomarker response showed that usage of the battery of bioassays provides better insight in a genotoxic effects in animals, and consequently, that the holistic approach is more suitable for this type of study.

Enhancing Nitrification at Low Temperature with Zeolite in a Mining Operations Retention Pond

Ammonium nitrate explosives are used in mining operations at Diavik Diamond Mines Inc. in the Northwest Territories, Canada. Residual nitrogen is washed into the mine pit and piped to a nearby retention pond where its removal is accomplished by microbial activity prior to a final water treatment step and release into the sub-Arctic lake, Lac de Gras. Microbial removal of ammonium in the retention pond is rapid during the brief ice-free summer, but often slows under ice cover that persists up to 9 months of the year. The aluminosilicate mineral zeolite was tested as an additive to retention pond water to increase rates of ammonium removal at 4°C. Water samples were collected across the length of the retention pond monthly over a year. The structure of the microbial community (bacteria, archaea, and eukarya), as determined by denaturing gradient gel electrophoresis of PCR-amplified small subunit ribosomal RNA genes, was more stable during cold months than during July–September, when there was a marked phytoplankton bloom. Of the ammonia-oxidizing community, only bacterial amoA genes were consistently detected. Zeolite (10 g) was added to retention pond water (100 mL) amended with 5 mM ammonium and incubated at 12°C to encourage development of a nitrifying biofilm. The biofilm community was composed of different amoA phylotypes from those identified in gene clone libraries of native water samples. Zeolite biofilm was added to fresh water samples collected at different times of the year, resulting in a significant increase in laboratory measurements of potential nitrification activity at 4°C. A significant positive correlation between the amount of zeolite biofilm and potential nitrification activity was observed; rates were unaffected in incubations containing 1–20 mM ammonium. Addition of zeolite to retention ponds in cold environments could effectively increase nitrification rates year-round by concentrating active nitrifying biomass.

Direct injection of native aqueous matrices by achiral–chiral chromatography ion trap mass spectrometry for simultaneous quantification of pantoprazole and lansoprazole enantiomers fractions

A two-dimensional liquid chromatography system coupled to ion-trap tandem mass spectrometer (2DLC-IT–MS/MS) was employed for the simultaneous quantification of pantoprazole and lansoprazole enantiomers fractions. A restricted access media of bovine serum albumin octyl column (RAM-BSA C 8) was used in the first dimension for the exclusion of the humic substances, while a polysaccharide-based chiral column was used in the second dimension for the enantioseparation of both pharmaceuticals. The results described here show good selectivity, extraction efficiency, accuracy, and precision with detection limits of 0.200 and 0.150 μg L −1 for the enatiomers of pantoprazole and lansoprazole respectively, while employing a small amount (1.0 mL) of native water sample per injection. This work reports an innovative assay for monitoring work, studies of biotic and abiotic enantioselective degradation and temporal changes of enantiomeric fractions.

A column-switching method for quantification of the enantiomers of omeprazole in native matrices of waste and estuarine water samples

This work reports the use of a two-dimensional liquid chromatography (2D-LC) system for quantification of the enantiomers of omeprazole in distinct native aqueous matrices. An octyl restricted-access media bovine serum albumin column (RAM-BSA C 8) was used in the first dimension, while a polysaccharide-based chiral column was used in the second dimension with either ultraviolet (UV–vis) or ion-trap tandem mass spectrometry (IT-MS/MS) detection. An in-line configuration was employed to assess the exclusion capacity of the RAM-BSA columns to humic substances. The excluded macromolecules had a molecular mass in the order of 18 kDa. Good selectivity, extraction efficiency, accuracy, and precision were achieved employing a very small amount (500 μL or 1.00 mL) of native water sample per injection, with detection limits of 5.00 μg L −1, using UV–vis, and 0.0250 μg L −1, using IT-MS/MS. The total analysis time was only 35 min, with no time spent on sample preparation. The methods were successfully applied to analyze a series of waste and estuarine water samples. The enantiomers were detected in an estuarine water sample collected from the Douro River estuary (Portugal) and in an influent sample from the wastewater treatment plant (WWTP) of São Carlos (Brazil). As far as we are concerned, this is the first report of the occurrence of (+)-omeprazole and (−)-omeprazole in native aqueous matrices.

Transformation of the X-ray Contrast Medium Iopromide In Soil and Biological Wastewater Treatment

In water/soil systems, the iodinated contrast medium iopromide was quantitatively biotransformed into several transformation products (TPs). Twelve TPs were identified via HPLC-UV and LC tandem MS. The chemical structures of the TPs were elucidated via fragmentation in MS2 and MS3 of LC tandem MS with a linear ion trap and 1H and 13C NMR analyses. All TPs exhibited transformations at the side chains containing either carboxylic moieties and/or primary and secondary amide moieties, while the triiodoisophthalic acid structure remained unaltered. A transformation pathway was proposed based on the sequence of TP formation in aerobic batch experiments. Additionally, the occurrence of iopromide TPs was investigated in native water samples. All TPs identified were found in municipal WWTP effluents because of their formation during biological wastewater treatment with maximum concentrations of up to 3.7 +/- 0.9 microg/L (TP 819). Predominantly, those TPs were present at higher concentrations in WWTP effluents which were formed at the beginning of the transformation pathway. Furthermore, four TPs formed at the end of the transformation pathway (TP 759, 701A/B, and 643) were also found in bank filtrate up to 0.050 microg/L and in groundwater of an wastewater irrigation area up to 4.6 microg/L.

Integral assessment of estrogenic potentials of sediment-associated samples

Exogenic endocrine-active substances are also called 'Endocrine Disrupting Chemicals' (EDC). They imitate or hinder the function of natural endogenic hormones or disturb the synthesis or the metabolism of hormones or of hormone receptors. The Enzyme-Linked Receptor Assay (ELRA) can detect estrogenic and anti-estrogenic effects at the level of receptor binding and is a useful tool for the integrative detection of contaminant effects. Although the test system has been used repeatedly in sediment assessments, the questions have remained concerning how it responds to variations in the physico-chemical matrix. For some bioassays, the salinity of the sample is a critical factor. This is especially relevant when testing wastewater samples or when sediment-associated samples in the tidal reaches of rivers are tested. Sediments in the tidal reaches of rivers change their salinity several times a day. Against this background, it would be beneficial to have a test procedure of known salinity tolerance. On account of this, the salinity tolerance of the ELRA was tested, assessed with reference substances at several salinity levels, and compared with the E-Screen method and a Yeast Estrogen Screen (YES), which are also frequently applied in environmental testing. The aim of this paper was to explore when the salinity limits within these test procedures are applicable. The trials should reveal the working range to be expected, characterize the salinity-dependent variations in sensitivity of the test, and provide options for methodological adjustments to improve the stability against increased salinity. The ELRA was carried out with the human Estrogen Receptor alpha. (ER) using the same principle like a competitive immunoassay based on ligand-protein interaction. However, an essential difference is the use of a physiologically relevant receptor instead of an antibody as a linking protein. The ELRA measures the competition of sample estrogens and anti-estrogens against estradiol supplied as a BSA-coating conjugate for the binding site of dissolved ER. Estradiol or xeno-estrogen binding is quantified by a biotynilated anti-ER antibody and the subsequent measurement of peroxidase activity by a streptavidin-POD-biotin complex. The E-Screen was performed with the human breast cancer cell line MCF-7, which expresses the estrogen receptor constitutively. Cell proliferation depends on binding of estrogens or xeno-estrogens with the receptor. After incubation, estrogen-dependent cell growth was measured by sulforhodamin B staining. The YES was performed with a recombinant yeast strain, transfected with a receptor and a reporter plasmid bearing the estrogen receptor and a vitellogenin gene fused with the reporter gene lacZ. Estrogen or xeno-estrogen-dependent gene induction was measured indirectly by LacZ activity. The salinity levels were simulated in varying concentrations with NaCl from 0 to 40 per thousand or Artificial Sea Water (ASW) from 0 to 32 per thousand. The study characterized the factor 'salinity' for the prospective application fields of the ELRA. With reference substances such as 17-beta-estradiol, the ELRA showed classical sigmoidal concentration-effect relations in a range from 0.05 to 100 microg/l under physiological conditions. After a methodological adjustment to compensate decreasing receptor-binding affinity of estrogens and xeno-estrogens at higher salinity levels, the ELRA became applicable under salinity conditions up to concentrations of 20.5 per thousand. In tests, the ELRA reached under the influence of salinity a mean limit of detection of 0.062 microg/l 17-beta-estradiol. The mean relative inter-test error was around 11%. Above concentrations of 20.5 per thousand there is a risk of false negative assessment. Compared with the E-Screen method using the MCF7 cell line and the yeast estrogen test system (YES), the ELRA shows a lower sensitivity to 17-beta-estradiol. In the E-Screen, the cell proliferation was strongly reduced by sodium chloride induced cytotoxicity. In comparison with the E-Screen, the salinity tolerance of the YES and YAS methods is significantly higher. Despite adaption, total salinity tolerance could not be achieved with the ELRA. Freshwater samples were generally appraisable. Higher salinity levels above 20.5 per thousand would tend towards false negative results. The low inter-test error of 11% makes the ELRA suitable for the detection of estrogenic and anti-estrogenic potentials of single substances, substance mixtures, and of environmental samples. The ELRA is very fast and reproducible, it can be used for high-throughput screening in a microplate format at low cost, it is robust to microbial contamination, and is less susceptible to cytotoxic interferences than cell culture methods. In their established form, the YES and the E-Screen methods are not applicable for liquid phase testing at higher salinity conditions. The salinity-adapted test version of the ELRA described here shows a broader working range for samples. Native water samples of more or less brackish origin or high-salinity effluent samples are testable. Results of tests with sediment associated samples of different salinity will be subject of a forthcoming publication.

Comet assay with the fish cell line rainbow trout gonad‐2 for in vitro genotoxicity testing of xenobiotics and surface waters

The present study examines the potential of the comet assay using the rainbow trout gonad cell line-2 (RTG-2) as an in vitro indicator test for genotoxicity assessment of aquatic contaminants and native surface waters. Initially, the comet assay protocol was adapted to the RTG-2 cell line. An exposure period of 2 h was found to be optimal, because DNA damage decreased when exposure was prolonged. Then, the sensitivity of the comet assay with RTG-2 cells toward six genotoxic reference substances was evaluated. The lowest-observed-effect concentration values for the directly acting genotoxins, 4-nitroquinoline-N-oxide and N-methyl-N'-nitro-N-nitrosoguanidine, were in the low nanomolar range. The RTG-2 test system clearly was less sensitive for the indirectly acting genotoxins benzo[a]pyrene, nitrofurantoin, 2-acetylaminofluorene, and dimethylnitrosamine, despite the presence of xenobiotic metabolic capacities in RTG-2 cells. The two effect endpoints used, tail length (TL) and tail moment (TM), did not differ with respect to sensitivity, but the linearity of the concentration-response curve was better with TM than with TL. The overall reproducibility of the assay results was good. Finally, the applicability of the comet assay with RTG-2 cells for genotoxicity screening of native surface water samples was studied. The assay tolerated the use of nonsterile water samples and was able to detect genotoxic potentials in native water samples; that is, extraction and concentration of the samples were not needed. The results of the present study indicate the suitability of the comet assay with the fish cell line, RTG-2, as in vitro screen for detecting genotoxic potencies of xenobiotics and environmental samples.

Nitrite reduction and detection at a carbon paste electrode containing hemoglobin and colloidal gold.

A novel renewable reagentless nitrite biosensor based on the direct electron transfer of hemoglobin (Hb) and a new sensing mechanism was proposed by combining the advantageous features of colloidal gold nanoparticle and carbon paste technology. The direct electrochemistry of immobilized Hb displayed a pair of redox peaks with a formal potential of -42 mV (vs. NHE) in 0.2 mol dm(-3) NaAc-HAc buffer (pH 5.5). The immobilized Hb displayed an excellent response to the reduction of NO2(-) with one interfacial charge transfer followed by a chemical reaction (EC) mechanism. Under optimal conditions, the interfacial EC process could be used for the sensitive determination of NO2(-) with a linear range from 0.1 to 9.7 micromol dm(-3) and a detection limit of 0.06 [micro sign]mol dm(-3) at 3sigma. The amperometric determination of high concentrations of NO2(-) based on the irreversible reduction of NO could be performed at pH 4.0 with a linear range from 0.1 to 1.2 mmol dm(-3). The surface of biosensor could be renewed quickly and reproducibly by a simple polish step. The biosensor has been used satisfactorily for nitrite determination in native water samples.

A PCR‐BASED TEST TO ASSESS THE POTENTIAL FOR MICROCYSTIN OCCURRENCE IN CHANNEL CATFISH PRODUCTION PONDS1,2

Microcystis aeruginosa is a common form of cyanobacteria (blue‐green algae) capable of forming toxic heptapeptides (microcystins) that can cause illness or death. Occasionally, blooms of cyanobacteria have caused toxic fish‐kills in catfish production ponds. We have developed a PCR test that will detect the presence of microcystin‐producing cyanobacteria. Microcystin producers are detected by the presence of the microcystin peptide synthetase B gene (an obligate enzyme in the microcystin pathway), which appears to be present only in toxin‐producing cyanobacteria. These PCR amplifications can be performed in multiplex using purified DNA from pond waters or by two‐stage amplification from native water samples. A synoptic survey of 476 channel catfish production ponds from four states in the southeastern United States revealed that 31% of the ponds have the genetic potential to produce microcystins by toxic algae.

Tail Moment versus Tail Length—Application of an In Vitro Version of the Comet Assay in Biomonitoring for Genotoxicity in Native Surface Waters Using Primary Hepatocytes and Gill Cells from Zebrafish (Danio rerio)

In order to investigate the suitability of an in vitro version of the comet assay with primary hepatocytes and gill cells from zebrafish (Danio rerio), cells were isolated by immersion in trypsin/EDTA solution after whole-body perfusion with phosphate-buffered saline. Within the scope of an 18-month biomonitoring study, primary cells were used to identify the genotoxic potential of native water samples from different sites along the major German rivers, Rhine and Elbe, and to evaluate the sensitivity and practicability of the chosen assay. Depending on the endpoint measured, considerable differences were detected with respect to the number of genotoxic surface water samples: Whereas no differences could be recorded for tail moment and relative DNA contents of head and tail, the number of positively tested native surface water samples significantly increased with tail length as endpoint.