High throughput and miniaturised systems for biodegradability assessments

The society demands safer products with a better ecological profile. Regulatory criteria have been developed to prevent risks for human health and the environment, for example, within the framework of the European regulation REACH (Regulation (EC) No 1907, 2006). This has driven industry to consider the development of high throughput screening methodologies for assessing chemical biodegradability. These new screening methodologies must be scalable for miniaturisation, reproducible and as reliable as existing procedures for enhanced biodegradability assessment. Here, we evaluate two alternative systems that can be scaled for high throughput screening and conveniently miniaturised to limit costs in comparison with traditional testing. These systems are based on two dyes as follows: an invasive fluorescent dyes that serves as a cellular activity marker (a resazurin-like dye reagent) and a noninvasive fluorescent oxygen optosensor dye (an optical sensor). The advantages and limitations of these platforms for biodegradability assessment are presented. Our results confirm the feasibility of these systems for evaluating and screening chemicals for ready biodegradability. The optosensor is a miniaturised version of a component already used in traditional ready biodegradability testing, whereas the resazurin dye offers an interesting new screening mechanism for chemical concentrations greater than 10 mg/l that are not amenable to traditional closed bottle tests. The use of these approaches allows generalisation of high throughput screening methodologies to meet the need of developing new compounds with a favourable ecological profile and also assessment for regulatory purpose.     

Biochemical and standard toxic effects of acetaminophen on the macrophyte species Lemna minor and Lemna gibba

Acetaminophen is globally one of the most prescribed drugs due to its antipyretic and analgesic properties. However, it is highly toxic when the dosage surpasses the detoxification capability of an exposed organism, with involvement of an already described oxidative stress pathway. To address the issue of the ecotoxicity of acetaminophen, we performed acute exposures of two aquatic plant species, Lemna gibba and Lemna minor, to this compound. The selected biomarkers were number of fronds, biomass, chlorophyll content, lipid peroxidation (TBARS assay), and proline content. Our results showed marked differences between the two species. Acetaminophen caused a significant decrease in the number of fronds (EC₅₀?=?446.6 mg/L), and the establishment of a dose-dependent peroxidative damage in L. minor, but not in L. gibba. No effects were reported in both species for the indicative parameters chlorophyll content and total biomass. However, the proline content in L. gibba was substantially reduced. The overall conclusions point to the occurrence of an oxidative stress scenario more prominent for L. minor. However, the mechanisms that allowed L. gibba to cope with acetaminophen exposure were distinct from those reported for L. minor, with the likely involvement of proline as antioxidant.     

Monitoring the quality of the marine environment

As part of the monitoring program from Instituto Hidrográfico, since 1981 sediment and water samples have been collected from four different estuarine areas located along the continental coast of Portugal. The concentrations of different parameters were measured in the water and sediment samples. After normalization, the concentrations of chromium, mercury, lead and zinc in the sediments from the different areas were compared.     

Heavy metal pollution in surface sediment and mussel samples in the Gulf of Gemlik

Different sources of pollution in Karasu Creek were investigated to obtain the water quality and ratio of contamination in this region. To achieve the main objectives of the present study, water samples were collected from Karasu Creek, starting from flow pattern at the upstream site of Akkaya Dam to the end of the dam, crossing the place where the Creek drains into. Dissolved oxygen, electrical conductivity, temperature and maximum/minimum pH were measured systematically for 12 months in the stations, where the water samples were collected. Chemical analyses of the water samples were carried out by using Cadas 50 S brand UV spectrometer to find out the Pb, Fe, Cu, Zn, Ni, Cr, Cd, S, F and Cn concentrations. These concentration were determined in μg/lt as 80–850; 180–4,920; 10–6,100; 440–25,530; 130–2,400; 120–280; 20–150; 214,250–1,113,580; 1,560–4,270 and 40–690, respectively. To determine metal levels of the water samples, multivariate analyses (element coefficient correlation, coefficient correlation dendrogram, hierarchical cluster analysis dendrogram, model summary and ANOVA) were used. The analyses yielded highly accurate results. There were positive correlations between some elements and their possible sources were the same. The stations which resembled each other along the creek were divided into three groups. The water quality of the creek was low and had toxic qualities. Eutrophication developed in Akkaya Dam along the creek. The source of pollution was thought to be industrial and residential wastes. Absolute (0–100 m), short distance (100–500 m) and medium distance (500–2,000 m) conservation areas should be determined in pollution areas along Karasu Creek and they should be improved.     

Comparison of periphytic biofilm and filter-feeding bivalve metal bioaccumulation (Cd and Zn) to monitor hydrosystem restoration after industrial remediation: a year of biomonitoring

Despite a significant decrease in the metallic waste emissions from an industrial site and a remediation process initiated in 2007, the Riou-Mort watershed (southwest France) still exhibits high Cd and Zn concentrations. Metal wastes have long been proven to significantly disturb aquatic communities. In this study, bioaccumulation capacities and responses to the chemical improvement of the hydrosystem were assessed for a year along the contamination gradient through the comparison of two biological models: Corbicula fluminea and periphytic biofilms, both considered as good bioindicators. Bioaccumulation results confirmed the persistence of water contamination in Corbicula fluminea and biofilms with, respectively, maximum Cd concentrations reaching 80.6 and 861.2 μg gDW(-1), and Zn concentrations 2.0 and 21.3 mg gDW(-1). Biofilms exhibited bioaccumulation in close correlation with water contamination, while Corbicula fluminea presented Cd bioaccumulation clearly regulated by water temperature and metal concentrations, affecting the ventilatory activity, as revealed by condition indices measurements. Also, a linear regression using Cd bioaccumulation and temperature () showed that below approximately 6 °C Corbicula fluminea did not appear to accumulate metals. Bioconcentration factors (BCFs) were higher in biofilms in comparison with Corbicula fluminea and showed the great accumulation capacity of suspended particulate matter in biofilms. However, bioaccumulation capacities are known to be influenced by many factors other than metal concentrations, such as temperature, water oxygenation or plankton and nutrient concentrations. Thus, this study demonstrates the power of a combined assessment using both Corbicula fluminea and biofilms as bioindicators to give a more integrated view of water quality assessment. Finally, when comparing our results with previous studies, the start of hydrosystem restoration could be shown by decreasing bioaccumulation in organisms.     

A one year investigation of the occurrence of illicit drugs in wastewater from Brussels, Belgium

Daily 24 hour composite influent wastewater samples were collected from the wastewater treatment plant of Brussels-North (Belgium) for eight months to study variations in the concentrations and mass loads of nine illicit drugs and metabolites: cocaine (COC) and its metabolites benzoylecgonine (BE) and ecgonine methyl ester (EME), amphetamine (AMP), methylenedioxymethamphetamine (MDMA), methamphetamine (METH), methadone (MTD) and its metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and the specific metabolite of heroin, 6-monoacetylmorphine (6-MAM). Samples were analyzed using a validated analytical method based on solid-phase extraction and hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry. The selected compounds could be detected and quantified in all samples. The measured concentrations were in agreement with earlier published concentrations, when available. All illicit drugs showed significant differences in mass loads between months. Daily variations were observed for MDMA, AMP and COC and metabolites, with higher mass loads observed during the weekend. This is probably related to their recreational consumption pattern and was further confirmed when holiday periods (New Year's Eve, Belgian national holiday) were in detail investigated. For METH, 6-MAM, MTD and EDDP, stable and consistent amounts were observed during the week. In all samples, the ratio of the concentration of the parent compound to its metabolite(s) was calculated to evaluate whether measured concentrations reflect human excretion of the illicit drugs. For MTD, the ratio of parent compound/metabolite was in agreement with the excretion pattern, while for COC some deviations were observed, resulting from excretion pattern uncertainties, and stability and discharge issues.     

Microbial activity and water-soluble trace element species in the rhizosphere of spring wheat (Triticum aestivum cv. USU-Perigee)

The influence of microbial activity on the concentration and speciation of trace elements (TEs) was assessed in a study on the bioavailability of TEs for edible plants. A growth chamber experiment with spring wheat (Triticum aestivum cv. USU-Perigee) was conducted and the bulk (Bk) and the rhizosphere (Rz) soil components were collected at maturity. A characterization of the microbial activity and population was made by measuring the microbial biomass, enzymes (acid phosphatase, arylsulfatase, dehydrogenase and urease) and 16S rDNA DGGE profiles. In soil water extracts, major solutes (H(+), Ca, Mg, Na, NH(4), K, Cl, NO(3), SO(4), total N, DON and DOC) and trace elements (Al, As, Cd, Ce, Cr, Cu, Fe, Pb, Tl, and Zn) including monomeric Al species, free Cu(2+) and labile Zn were determined. The partition of the variation indicated that 12.1% of the distribution of TEs in the Bk soil was significantly and exclusively explained by chemical properties while this value was less than 0.1% for the Rz soil. To the contrary, microbial properties contributed significantly to 12.3% of the distribution of TEs in the Rz soil whereas it explained less than 0.1% for the Bk soil. Detailed redundancy analyses identified several potential mechanisms (e.g. weathering of primary mineral, solubilisation of sesquioxides, bacterial effect on the redox status) explaining the fate of TEs in the Bk and Rz soils. This study revealed that microbial activity is strongly associated to the speciation of trace elements in the Rz of edible plants and points to some microbial processes influencing TE speciation.     

Evaluation of diffusive gradients in thin films technique (DGT) for measuring Al, Cd, Co, Cu, Mn, Ni, and Zn in Amazonian rivers

Studies concerning the lability and bioavailability of trace metals have played a prominent role in the search for contamination of water resources. This work describes the first application yet of the diffusive gradients in thin films technique (DGT) to the determination of the fraction of free plus labile metals in waters from the Amazon Basin. Due to the complexity of the use of DGT for samples with low ionic strength and high organic matter content (characteristic of Amazonian rivers), a new analytical procedure was developed. The method is based on the determinations of apparent diffusion coefficients (D_ap) in the laboratory, by performing deployments in samples collected in the corresponding sites of study. The D_ap thereby determined is then used for in situ measurements. The suitability of the proposed approach for determination of labile Al, Cd, Co, Cu, Mn, Ni, and Zn in the Amazon River and Rio Negro (English: Black River) was evaluated. Except for Co, Mn (in a deployment at Rio Negro), Ni and Zn (in a deployment at Amazon River), labile in situ measurements were lower or similar to dissolved concentrations, indicating suitability of the proposed approach.     

Investigation of the bacterial load and antibiotic susceptibility of dental units

The aim of this study was to evaluate the bacterial contamination level and to determine the antibiotic susceptibility of the isolated bacteria from dental unit waterlines (DUWLs) in Istanbul. Bacterial quality of DUWLs is very important, as patients and dental staff are regularly exposed to water and aerosols generated by the unit. If opportunistic pathogens such as Staphylococcus aureus, Pseudomonas aeruginosa, and Legionella pneumophila are present in DUWLs, patient and dental staff can be infected. One hundred water samples were collected from high-speed drills and input waters from 50 dental units. Aerobic heterotrophic bacteria counts and the presence of Legionella, Pseudomonas, oral streptococci, and Staphylococcus were investigated in dental unit waters and aerosol samples. In addition, the antibiotic susceptibility of the isolated and identified bacteria from DUWLs was examined. This research found that 37 out of 50 dental unit water samples exceeded the American Dental Association’s limit of 200 colony-forming units (CFU)/mL^?1. Legionella, oral streptococci, and S. aureus were not detected in any water or aerosol samples, but P. aeruginosa was isolated in three DUWLs. Also, Pseudomonas and Staphylococcus were found in water and aerosol samples. Cefoperazone, ofloxacin, gentamicin, ciprofloxacin, and piperacillin were the most effective antibiotics against the isolated bacteria from DUWLs.     

Monitoring The Turbidity and Surface Temperature Changes and Effects of the 17 August 1999 Earthquake in the İzmİt Gulf,Turkey by the Landsat TM/ETM Data

The temporal turbidity and surface temperature changes and effects of the 17 August 1999 earthquake in the İzmit Gulf, Turkey have been investigated using Landsat TM/ETM data. The gulf is in the Mediterranean–Black Sea transition climatic zone and is partially surrounded by green vegetation cover and degraded and densely urbanized-industrialized areas. Landsat TM/ETM data acquired in 1990–1999 confirms increase in turbidity. Turbidity is always low in the southern part and high in the northern part of the gulf, because the more urbanized and industrialized areas are located in the northern part. The Landsat-7 ETM data acquired in the same year (1999) shows seasonal changes in turbidity. Moreover, the two high turbidity and surface temperature anomalies, one of which is parallel to the 17 August 1999 earthquake surface rupture (east–west) and the other which is in the northwest–southeast direction were mapped from Landsat-5 TM data acquired the day (18.08.1999) following the earthquake in the east end of the gulf. On the basis of turbidity implying the sea bottom movement, it is possible to state that a second rupture in the northwest and southeast direction could have occurred at the sea bottom during the earthquake. The distribution of the seismicity centers and the orientation of the lineaments in the area support this finding.