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The combined use of the PLHC-1 cell line and the recombinant yeast assay to assess the environmental quality of estuarine and coastal sediments
2013
Schnell, Sabine | Olivares, Alba | Piña, Benjamin | Echavarri-Erasun, Beatriz | Lacorte, Silvia | Porte, Cinta
Sediment contamination poses a potential risk for both ecosystems and human health. Risk assessment is troublesome as sediments contain complex mixtures of toxicants, and traditional chemical analyses can neither provide information about potential hazards to organisms nor identify and measure all present contaminants. This work combines the use of the PLHC-1 cell line and the recombinant yeast assay (RYA) to assess the environmental quality of estuarine and coastal sediments. The application of multiple endpoints (cytotoxicity, generation of oxidative stress, presence of CYP1A inducing agents, micronucleus formation and estrogenicity) revealed that the organic extracts of those sediments affected by industrial activities or collected near harbours and untreated urban discharges showed significant cytotoxicity, micronuclei and CYP1A induction. The study highlights the usefulness of the applied bioassays to identify those sediments that could pose risk to aquatic organisms and that require further action to improve their environmental quality.
Afficher plus [+] Moins [-]Sample preparation methods for quantitative detection of DNA by molecular assays and marine biosensors
2013
Cox, Annie M. | Goodwin, Kelly D.
The need for quantitative molecular methods is growing in environmental, food, and medical fields but is hindered by low and variable DNA extraction and by co-extraction of PCR inhibitors. DNA extracts from Enterococcus faecium, seawater, and seawater spiked with E. faecium and Vibrio parahaemolyticus were tested by qPCR for target recovery and inhibition. Conventional and novel methods were tested, including Synchronous Coefficient of Drag Alteration (SCODA) and lysis and purification systems used on an automated genetic sensor (the Environmental Sample Processor, ESP). Variable qPCR target recovery and inhibition were measured, significantly affecting target quantification. An aggressive lysis method that utilized chemical, enzymatic, and mechanical disruption enhanced target recovery compared to commercial kit protocols. SCODA purification did not show marked improvement over commercial spin columns. Overall, data suggested a general need to improve sample preparation and to accurately assess and account for DNA recovery and inhibition in qPCR applications.
Afficher plus [+] Moins [-]Contaminants assessment in the coral reefs of Virgin Islands National Park and Virgin Islands Coral Reef National Monument
2013
Bargar, Timothy A. | Garrison, Virginia H. | Alvarez, David A. | Echols, Kathy R.
Coral, fish, plankton, and detritus samples were collected from coral reefs in Virgin Islands National Park (VIIS) and Virgin Islands Coral Reef National Monument (VICR) to assess existing contamination levels. Passive water sampling using polar organic chemical integrative samplers (POCIS) and semi-permeable membrane devices found a few emerging pollutants of concern (DEET and galaxolide) and polynuclear aromatic hydrocarbons. Very little persistent organic chemical contamination was detected in the tissue or detritus samples. Detected contaminants were at concentrations below those reported to be harmful to aquatic organisms. Extracts from the POCIS were subjected to the yeast estrogen screen (YES) to assess potential estrogenicity of the contaminant mixture. Results of the YES (estrogen equivalency of 0.17–0.31ng/L 17-β-estradiol) indicated a low estrogenicity likelihood for contaminants extracted from water. Findings point to low levels of polar and non-polar organic contaminants in the bays sampled within VICR and VIIS.
Afficher plus [+] Moins [-]Considerations on salts used for density separation in the extraction of microplastics from sediments
2021
Cutroneo, Laura | Reboa, Anna | Geneselli, Irene | Capello, Marco
Environmental contamination by plastics and microplastics is a recognised problem worldwide, and it is the focus of many research teams. In the quantification of microplastics in the environment (plastic items with dimensions between 1 μm and 5 mm), the search for shared and universally recognised protocols and methodologies is still ongoing. In this study, the use of a method for extracting microplastics from marine sediments based on density separation has been considered. Fifty studies were analysed to investigate the scenario of salts exploited during microplastic extraction. The most commonly used salts are NaCl (45.6%), ZnCl₂ (19.3%) and NaI (17.5%). Considerations related to cost, availability, hazards of the salts and thus the repeatability of the related extraction method are reported. In light of the findings, NaCl remains the most readily usable, economical and effective salt for the extraction of microplastics from marine sediments.
Afficher plus [+] Moins [-]Recent innovation in microbial source tracking using bacterial real-time PCR markers in shellfish
2013
Mauffret, A. | Mieszkin, S. | Morizur, M. | Alfiansah, Y. | Lozach, S. | Gourmelon, M.
We assessed the capacity of real-time PCR markers to identify the origin of contamination in shellfish. Oyster, cockles or clams were either contaminated with fecal materials and host-associated markers designed from Bacteroidales or Catellicoccus marimammalium 16S RNA genes were extracted from their intravalvular liquid, digestive tissues or shellfish flesh. Extraction of bacterial DNA from the oyster intravalvular liquid with FastDNA spin kit for soil enabled the selected markers to be quantified in 100% of artificially contaminated samples, and the source of contamination to be identified in 13 out of 38 naturally contaminated batches from European Class B and Class C areas. However, this protocol did not enable the origin of the contamination to be identified in cockle or clam samples. Although results are promising for extracts from intravalvular liquid in oyster, it is unlikely that a single protocol could be the best across all bacterial markers and types of shellfish.
Afficher plus [+] Moins [-]Heavy metal concentrations in consecutive saturation extracts of dredged sediment derived surface soils
1998
Tack, F.M.G. | Singh, S.P. | Verloo, M.G. (Laboratory for Analytical Chemistry and Applied Ecochemistry, University of Ghent, Coupure Links 653, B-9000 Gent (Belgium))
Combining Passive Sampling with a GC-MS-Database Screening Tool to Assess Trace Organic Contamination of Rivers: a Pilot Study in Melbourne, Australia
2015
This study assessed the suitability of passive sampler extracts for use with a GC-MS-database rapid screening technique for around 940 organic chemicals. Chemcatcher™ passive sampler systems containing either Empore™ SDB-XC or C18FF disks were deployed at 21 riverine sites in and near Melbourne, Victoria, Australia, for a period of 28 days during September–October 2008. Methanolic elution of the SDB-XC and C18FF disks produced an extract that, after evaporation and inversion into hexane, was compatible with the GC-MS-database method enabling over 30 chemicals to be observed. The sources of the non-agricultural chemicals are still unclear, but this study was conducted in a relatively dry season where total rainfall was approximately 40 % lower than the long-term mean for the catchment during the study period. Thus, the risks may be greater in wetter seasons, as greater quantities of chemicals are likely to reach waterways as the frequency, extent and intensity of surface run-off events increase. This study provides valuable information for policy and decision-makers, both in Australia and other regions of the world, in that passive sampling can be conveniently used prior to analysis by multi-residue techniques to produce data to assess the likely risks trace organic chemicals pose to aquatic ecosystems.
Afficher plus [+] Moins [-]Effects of metals on the microbial mineralization of organic acids
1997
BRYNHILDSEN, LENA | Rosswall, T. (Thomas)
The effects of chemical speciation on mineralization of organic compounds was studied using citric acid as a model substance. The degradation of ¹⁴C-labeled Al-, Co-, Cu-, and Zn-citrate was followed in chemically well-defined media inoculated with mixed cultures of microorganisms (soil extracts from two soils). The degradation of citrate was completely inhibited when the acid was bound to Zn, Cu, or Co and partly inhibited when bound to Al. The mineralization of citrate as well as histidine was also followed by incubation of the complexes (Cu, Zn, Al) in the two soils. No effect of metals on the degradation of histidine was seen. The degradation of citrate in soils was also unaffected when complexed to Cu and Zn, whereas Al exerted an inhibited decomposition in both soils.
Afficher plus [+] Moins [-]Insecticidal potency of Aspergillus terreus against larvae and pupae of three mosquito species Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti
2015
Ragavendran, Chinnasamy | Natarajan, Devarajan
Microbial control agents offer alternatives to chemical pest control, as they can be more selective than chemical insecticides. The present study evaluates the mosquito larvicidal and pupicidal potential of fungus mycelia using ethyl acetate and methanol solvent extracts produced by Aspergillus terreus against Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti. The A. terreus mycelia were extracted after 15 days from Sabouraud dextrose broth medium. The ethyl acetate extracts showed lethal concentration that kills 50 % of the exposed larvae (LC₅₀) and lethal concentration that kills 90 % of the exposed larvae (LC₉₀) values of the first, second, third, and fourth instar larvae of An. stephensi (LC₅₀ = 97.410, 102.551, 29.802, and 8.907; LC₉₀ = 767.957, 552.546, 535.474, and 195.677 μg/ml), Cx. quinquefasciatus (LC₅₀ = 89.584, 74.689, 68.265, and 67.40; LC₉₀ = 449.091, 337.355, 518.793, and 237.347 μg/ml), and Ae. aegypti (LC₅₀ = 83.541, 84.418, 80.407, and 95.926; LC₉₀ = 515.464, 443.167, 387.910, and 473.998 μg/ml). Pupicidal activity of mycelium extracts was tested against An. stephensi (LC₅₀ = 25.228, LC₉₀ = 140.487), Cx. quinquefasciatus (LC₅₀ = 54.525, LC₉₀ = 145.366), and Ae. aegypti (LC₅₀ = 10.536, LC₉₀ = 63.762 μg/ml). At higher concentration (500 μg/ml), mortality starts within the first 6 h of exposure. One hundred percent mortality occurs at 24-h exposure. The overall result observed that effective activity against selected mosquito larvae and pupae after 24 h was a dose and time-dependent activity. These ensure that the resultant mosquito population reduction is substantial even where the larvicidal and pupicidal potential is minimal. The FTIR spectra of ethyl acetate extract reflect prominent peaks (3448.32, 3000.36, 2914.59, 2118.73, 1668.21, 1436.87, 1409.02, 954.33, 901.13, and 704.67 cm⁻¹). The spectra showed a sharp absorption band at 1314.66 cm⁻¹ assigned to wagging vibration of the C–H group. The band at 1023.59 cm⁻¹ developed for C–O and C=N, respectively, and was commonly found in carboxylic acid and amine groups. GC–MS analysis of ethyl acetate extracts showed the presence of six compounds, of which the major compounds were identified as n-hexadecanoic acid (15.31 %) and methyl 12,15-octadecadienoate (31.989 %), based on their peak molecular weight. The HPLC analysis result highlights that the A. terreus ethyl acetate extract was compared with pure n-hexadecanoic acid which resulted in similar retention time of 19.52 and 19.38, respectively. Thus, the active compound produced by this species would be more useful against vectors responsible for diseases of public health importance. This is the first report on mosquito larvicidal and pupicidal activity of ethyl acetate extract produced by A. terreus species.
Afficher plus [+] Moins [-]Investigations on sediment toxicity of German rivers applying a standardized bioassay battery
2015
Hafner, Christoph | Gartiser, Stefan | Garcia-Käufer, Manuel | Schiwy, Sabrina | Hercher, Christoph | Meyer, Wiebke | Achten, Christine | Larsson, Maria | Engwall, Magnus | Keiter, Steffen | Hollert, Henner
River sediments may contain a huge variety of environmental contaminants and play a key role in the ecological status of aquatic ecosystems. Contaminants adsorbed to sediments and suspended solids may contribute directly or after remobilization to an adverse ecological and chemical status of surface water. In this subproject of the joint research project DanTox, acetonic Soxhlet extracts from three German river sediments from the River Rhine (Altrip and Ehrenbreitstein with moderate contamination) and River Elbe (Veringkanal Hamburg heavily contaminated) were prepared and redissolved in dimethyl sulfoxide (DMSO). These extracts were analyzed with a standard bioassay battery with organisms from different trophic levels (bacteria, algae, Daphnia, fish) as well as in the Ames test and the umuC test for bacterial mutagenicity and genotoxicity according to the respective OECD and ISO guidelines. In total, 0.01 % (standard) up to 0.25 % (only fish embryo test) of the DMSO sediment extract was dosed to the test systems resulting in maximum sediment equivalent concentrations (SEQ) of 2 up to 50 g l⁻¹. The sediment of Veringkanal near Hamburg harbor was significantly more toxic in most tests compared to the sediment extracts from Altrip and Ehrenbreitstein from the River Rhine. The most toxic effect found for Veringkanal was in the algae test with an EᵣC₅₀ (72 h) of 0.00226 g l⁻¹ SEQ. Ehrenbreitstein and Altrip samples were about factor 1,000 less toxic. In the Daphnia, Lemna, and acute fish toxicity tests, no toxicity at all was found at 2 g l⁻¹ SEQ. corresponding to 0.01 % DMSO. Only when increasing the DMSO concentration the fish embryo test showed a 22-fold higher toxicity for Veringkanal than for Ehrenbreitstein and Altrip samples, while the toxicity difference was less evident for the Daphnia test due to the overlaying solvent toxicity above 0.05 % dimethyl sulfoxide (DMSO). The higher toxicities observed with the Veringkanal sample are supported by the PAH and PCB concentrations analyzed in the sediments. The sediment extracts of Altrip and Veringkanal were mutagenic in the Ames tester strain TA98 with metabolic activation (S9-mix). The findings allow a better ecotoxicological characterization of the sediments extensively analyzed in all subprojects of the DanTox project (e.g., Garcia-Kaeufer et al. Environ Sci Pollut Res. doi: 10.1007/s11356-014-3894-4 , 2014; Schiwy et al. Environ Sci Pollut Res. doi: 10.1007/s11356-014-3185-0 , 2014; Hollert and Keiter 2015). In the absence of agreed limit values for sediment extracts in standard tests, further data with unpolluted reference sediments are required for a quantitative risk assessment of the investigated polluted sediments.
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