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Sample records for embryo toxicity test

  1. Use of blue crab (Callinectes sapidus) embryos for toxicity testing

    SciTech Connect

    Lee, R.; O`Malley, K.

    1995-12-31

    After fertilization, blue crab embryos develop in egg sacs attached to the female pleopods, often referred to as the sponge. Lipovitellin and lipid droplets in the egg sacs provide energy and nutrition for the developing embryos. Embryos were removed from the sponge and transferred to 24 well culture plates containing sea water with or without toxicants, Each well contained 10 embryos. After 7 to 10 days, embryos hatched to swimming zoea. The effects of toxicants at various concentrations on hatching were determined and the EC{sub 50} calculated. For example, the EC{sub 50} for tributyltin, fenvalerate and mercuric chloride were 50, 30 and 90 ng/liter, respectively. The hatching success of control embryos ranged from 95 to 98%. Formation of the heart, eyespot formation, appendage formation and utilization rate of lipovitellin were also effected by exposure to toxicants. At a low concentration of mercuric ion (30ng/liter) the heart formed, but there was no heart beat. Eyespot formation was abnormal in the presence of high concentrations of cadmium (2 {micro}g/liter) and zinc (5 {micro}g/liter), Crab embryos offer many advantages for toxicity testing of pure compounds or mixtures in water, including toxicity testing of sediment pore water. The crab embryos may also serve as models to understand the effect of specific toxicants on the heart and eye spots of crustaceans.

  2. IMPROVED SEA URCHIN DNA-BASED EMBRYO GROWTH TOXICITY TEST

    EPA Science Inventory

    The article presents an improved DNA-based embryo toxicity test in which growth is measured as the increase in DNA during a 5-h postfertilization period. The test, using direct fluorometric determination of DNA rather than thymidine incorporation, is fast, sensitive, easy to cond...

  3. Toxicity testing of human assisted reproduction devices using the mouse embryo assay.

    PubMed

    Punt-van der Zalm, J P E M; Hendriks, J C M; Westphal, J R; Kremer, J A M; Teerenstra, S; Wetzels, A M M

    2009-04-01

    Systems to assess the toxicity of materials used in human assisted reproduction currently lack efficiency and/or sufficient discriminatory power. The development of 1-cell CBA/B6 F1 hybrid mouse embryos to blastocysts, expressed as blastocyst rate (BR), is used to measure toxicity. The embryos were divided into control and test groups, and were exposed to either control medium or to a potentially toxic test medium. Inferences on toxicity were based on differences in BR between the two groups. The mouse embryo assay followed a stratified (mouse), randomized (embryo), and balanced (equal number of embryos per group and per mouse) design. The number of embryos needed was calculated using power analysis. The basal BR of the hybrid strain was determined in a historical population. Sixty-nine mouse embryos per group were required to detect toxic materials with sufficient sensitivity and to account for the considerable inter-mouse variation in blastocyst development. Fifty-two samples, divided over batches of seven different products were tested before use in the study IVF centre and five of these were found to be toxic. This test system, presented as the Nijmegen mouse embryo assay (NMEA), can be used to detect embryo-toxic materials in daily IVF practice, and this report may provide a starting point for standardization. PMID:19400995

  4. COMPARATIVE EVALUATION OF THREE RAPID MARINE TOXICITY TESTS: SEA URCHIN EARLY EMBRYO GROWTH TEST, SEA URCHIN SPERM CELL TOXICITY TEST AND MICROTOX

    EPA Science Inventory

    Three rapid marine toxicity tests were evaluated to determine their potential usefulness in a toxicity testing program: early embryo growth test and sperm cell toxicity test, both using the sea urchin Arbacia punctulata, and Microtox. Toxicity values (EC50s) were derived for eigh...

  5. Fish embryo toxicity test: identification of compounds with weak toxicity and analysis of behavioral effects to improve prediction of acute toxicity for neurotoxic compounds.

    PubMed

    Klüver, Nils; König, Maria; Ortmann, Julia; Massei, Riccardo; Paschke, Albrecht; Kühne, Ralph; Scholz, Stefan

    2015-06-01

    The fish embryo toxicity test has been proposed as an alternative for the acute fish toxicity test, but concerns have been raised for its predictivity given that a few compounds have been shown to exhibit a weak acute toxicity in the fish embryo. In order to better define the applicability domain and improve the predictive capacity of the fish embryo test, we performed a systematic analysis of existing fish embryo and acute fish toxicity data. A correlation analysis of a total of 153 compounds identified 28 compounds with a weaker or no toxicity in the fish embryo test. Eleven of these compounds exhibited a neurotoxic mode of action. We selected a subset of eight compounds with weaker or no embryo toxicity (cyanazine, picloram, aldicarb, azinphos-methyl, dieldrin, diquat dibromide, endosulfan, and esfenvalerate) to study toxicokinetics and a neurotoxic mode of action as potential reasons for the deviating fish embryo toxicity. Published fish embryo LC50 values were confirmed by experimental analysis of zebrafish embryo LC50 according to OECD guideline 236. Except for diquat dibromide, internal concentration analysis did not indicate a potential relation of the low sensitivity of fish embryos to a limited uptake of the compounds. Analysis of locomotor activity of diquat dibromide and the neurotoxic compounds in 98 hpf embryos (exposed for 96 h) indicated a specific effect on behavior (embryonic movement) for the neurotoxic compounds. The EC50s of behavior for neurotoxic compounds were close to the acute fish toxicity LC50. Our data provided the first evidence that the applicability domain of the fish embryo test (LC50s determination) may exclude neurotoxic compounds. However, neurotoxic compounds could be identified by changes in embryonic locomotion. Although a quantitative prediction of acute fish toxicity LC50 using behavioral assays in fish embryos may not yet be possible, the identification of neurotoxicity could trigger the conduction of a conventional fish acute toxicity test or application of assessment factors while considering the very good fish embryo-acute fish toxicity correlation for other compounds. PMID:25939044

  6. Use of fish embryo toxicity tests for the prediction of acute fish toxicity to chemicals.

    PubMed

    Belanger, Scott E; Rawlings, Jane M; Carr, Gregory J

    2013-08-01

    The fish embryo test (FET) is a potential animal alternative for the acute fish toxicity (AFT) test. A comprehensive validation program assessed 20 different chemicals to understand intra- and interlaboratory variability for the FET. The FET had sufficient reproducibility across a range of potencies and modes of action. In the present study, the suitability of the FET as an alternative model is reviewed by relating FET and AFT. In total, 985 FET studies and 1531 AFT studies were summarized. The authors performed FET-AFT regressions to understand potential relationships based on physical-chemical properties, species choices, duration of exposure, chemical classes, chemical functional uses, and modes of action. The FET-AFT relationships are very robust (slopes near 1.0, intercepts near 0) across 9 orders of magnitude in potency. A recommendation for the predictive regression relationship is based on 96-h FET and AFT data: log FET median lethal concentration (LC50) = (0.989 × log fish LC50) - 0.195; n = 72 chemicals, r = 0.95, p < 0.001, LC50 in mg/L. A similar, not statistically different regression was developed for the entire data set (n = 144 chemicals, unreliable studies deleted). The FET-AFT regressions were robust for major chemical classes with suitably large data sets. Furthermore, regressions were similar to those for large groups of functional chemical categories such as pesticides, surfactants, and industrial organics. Pharmaceutical regressions (n = 8 studies only) were directionally correct. The FET-AFT relationships were not quantitatively different from acute fish-acute fish toxicity relationships with the following species: fathead minnow, rainbow trout, bluegill sunfish, Japanese medaka, and zebrafish. The FET is scientifically supportable as a rational animal alternative model for ecotoxicological testing of acute toxicity of chemicals to fish. PMID:23606235

  7. Optimization of high-throughput nanomaterial developmental toxicity testing in zebrafish embryos

    EPA Science Inventory

    Nanomaterial (NM) developmental toxicities are largely unknown. With an extensive variety of NMs available, high-throughput screening methods may be of value for initial characterization of potential hazard. We optimized a zebrafish embryo test as an in vivo high-throughput assay...

  8. Progress Towards the Development of a Fathead Minnow Embryo Test and Comparison to the Zebrafish Embryo Test for Assessing Acute Fish Toxicity

    EPA Science Inventory

    The Zebrafish Embryo Test (ZFET) for acute fish toxicity is a well developed method nearing adoption as an OECD Test Guideline. Early drafts of the test guideline (TG) envisioned a suite of potential test species to be covered including zebrafish, fathead minnow, Japanese Medaka...

  9. Development of a new integrative toxicity index based on an improvement of the sea urchin embryo toxicity test.

    PubMed

    Morroni, L; Pinsino, A; Pellegrini, D; Regoli, F; Matranga, V

    2016-01-01

    The sea urchin embryo toxicity test is classically used to assess the noxious effects of contaminated marine waters and sediments. In Italian guidelines on quality of dredged sediments, the standard toxicity criteria used for this assay are based on a single endpoint at 48 hours of development, corresponding to the pluteus stage. Different typologies of abnormalities, including those which occur at earlier stages, are not categorized, thus preventing the evaluation of the actual teratogenic hazards. A new integrative toxicity index has been developed in this study based on the analysis of two developmental stages, at 24 and 48h post-fertilization, and the differentiation between development delays and germ layers impairments: the new toxicity index is calculated by integrating the frequency of abnormal embryos with the severity of such abnormalities. When tested on dredged sediments, the evaluation of increasing levels of toxicity affecting embryonic outcomes enhanced the capability to discriminate different samples, appearing particularly relevant to validate the sea urchin embryo toxicity assay, and supporting its utility in practical applications such as the sediments classification in harbor areas. PMID:26477574

  10. PAH toxicity at aqueous solubility in the fish embryo test with Danio rerio using passive dosing.

    PubMed

    Seiler, Thomas-Benjamin; Best, Nina; Fernqvist, Margit Mller; Hercht, Hendrik; Smith, Kilian E C; Braunbeck, Thomas; Mayer, Philipp; Hollert, Henner

    2014-10-01

    As part of the risk assessment process within REACh, prior to manufacturing and distribution of chemical substances their (eco)toxicological impacts have to be investigated. The fish embryo toxicity test (FET) with the zebrafish Danio rerio has gained a high significance as an in vitro alternative to animal testing in (eco)toxicology. However, for hydrophobic organic chemicals it remains a technical challenge to ensure constant freely dissolved concentration at the maximum exposure level during such biotests. Passive dosing with PDMS silicone was thus applied to control the freely dissolved concentration of ten PAHs at their saturation level in the FET. The experiments gave repeatable results, with the toxicity of the PAHs generally increasing with the maximum chemical activities of the PAHs. HPLC analysis confirmed constant exposure at the saturation level. In additional experiments, fish embryos without direct contact to the silicone surface showed similar mortalities as those exposed with direct contact to the silicone. Silicone oil overlaying the water phase as a novel passive dosing phase had no observable effects on the development of the fish embryos until hatching. This study provides further data to support the close relationship between the chemical activity and the toxicity of hydrophobic organic compounds. Passive dosing from PDMS silicone enabled reliable toxicity testing of (highly) hydrophobic substances at aqueous solubility, providing a practical way to control toxicity exactly at the maximum exposure level. This approach is therefore expected to be useful as a cost-effective initial screening of hydrophobic chemicals for potential adverse effects to freshwater vertebrates. PMID:25048891

  11. OECD validation study to assess intra- and inter-laboratory reproducibility of the zebrafish embryo toxicity test for acute aquatic toxicity testing.

    PubMed

    Busquet, François; Strecker, Ruben; Rawlings, Jane M; Belanger, Scott E; Braunbeck, Thomas; Carr, Gregory J; Cenijn, Peter; Fochtman, Przemyslaw; Gourmelon, Anne; Hübler, Nicole; Kleensang, André; Knöbel, Melanie; Kussatz, Carola; Legler, Juliette; Lillicrap, Adam; Martínez-Jerónimo, Fernando; Polleichtner, Christian; Rzodeczko, Helena; Salinas, Edward; Schneider, Katharina E; Scholz, Stefan; van den Brandhof, Evert-Jan; van der Ven, Leo T M; Walter-Rohde, Susanne; Weigt, Stefan; Witters, Hilda; Halder, Marlies

    2014-08-01

    The OECD validation study of the zebrafish embryo acute toxicity test (ZFET) for acute aquatic toxicity testing evaluated the ZFET reproducibility by testing 20 chemicals at 5 different concentrations in 3 independent runs in at least 3 laboratories. Stock solutions and test concentrations were analytically confirmed for 11 chemicals. Newly fertilised zebrafish eggs (20/concentration and control) were exposed for 96h to chemicals. Four apical endpoints were recorded daily as indicators of acute lethality: coagulation of the embryo, lack of somite formation, non-detachment of the tail bud from the yolk sac and lack of heartbeat. Results (LC50 values for 48/96h exposure) show that the ZFET is a robust method with a good intra- and inter-laboratory reproducibility (CV<30%) for most chemicals and laboratories. The reproducibility was lower (CV>30%) for some very toxic or volatile chemicals, and chemicals tested close to their limit of solubility. The ZFET is now available as OECD Test Guideline 236. Considering the high predictive capacity of the ZFET demonstrated by Belanger et al. (2013) in their retrospective analysis of acute fish toxicity and fish embryo acute toxicity data, the ZFET is ready to be considered for acute fish toxicity for regulatory purposes. PMID:24874798

  12. SIMPLE TEST FOR TOXICITY OF NUMBER 2 FUEL OIL AND OIL DISPERSANTS TO EMBRYOS OF GRASS SHRIMP, PALAEMONETES PUGIO

    EPA Science Inventory

    A simple test, using embryos of the grass shrimp Palaemonetes pugio, was employed to determine the toxicity of two commercial oil dispersants (Corexit 7664 and Corexit 9527) and toxicity of the water soluble fraction of Number 2 fuel oil (WSF oil) prepared with and without the ad...

  13. Automated Lab-on-a-Chip Technology for Fish Embryo Toxicity Tests Performed under Continuous Microperfusion (μFET).

    PubMed

    Zhu, Feng; Wigh, Adriana; Friedrich, Timo; Devaux, Alain; Bony, Sylvie; Nugegoda, Dayanthi; Kaslin, Jan; Wlodkowic, Donald

    2015-12-15

    The fish embryo toxicity (FET) biotest has gained popularity as one of the alternative approaches to acute fish toxicity tests in chemical hazard and risk assessment. Despite the importance and common acceptance of FET, it is still performed in multiwell plates and requires laborious and time-consuming manual manipulation of specimens and solutions. This work describes the design and validation of a microfluidic Lab-on-a-Chip technology for automation of the zebrafish embryo toxicity test common in aquatic ecotoxicology. The innovative device supports rapid loading and immobilization of large numbers of zebrafish embryos suspended in a continuous microfluidic perfusion as a means of toxicant delivery. Furthermore, we also present development of a customized mechatronic automation interface that includes a high-resolution USB microscope, LED cold light illumination, and miniaturized 3D printed pumping manifolds that were integrated to enable time-resolved in situ analysis of developing fish embryos. To investigate the applicability of the microfluidic FET (μFET) in toxicity testing, copper sulfate, phenol, ethanol, caffeine, nicotine, and dimethyl sulfoxide were tested as model chemical stressors. Results obtained on a chip-based system were compared with static protocols performed in microtiter plates. This work provides evidence that FET analysis performed under microperfusion opens a brand new alternative for inexpensive automation in aquatic ecotoxicology. PMID:26506399

  14. Simple test for toxicity of number 2 fuel oil and oil dispersants to embryos of grass shrimp, palaemonetes pugio

    SciTech Connect

    Fisher, W.S.; Foss, S.S.

    1993-01-01

    A simple test, using embryos of the grass shrimp Palaemonetes pugio, was employed to determine the toxicity of two commercial oil dispersants (Corexit 7664 and Corexit 9527) and toxicity of the water soluble fraction of Number 2 fuel oil (WSF oil) prepared with and without the addition of the dispersants. Tests revealed P. pugio embryos were similar to previously measured life stages in their sensitivity to WSF oil prepared without dispersants. They were approximately ten times more sensitive to water soluble fractions of dispersed oil, which may have been due to the approximately ten-fold increases in total hydrocarbons measured analytically. Both temperatures and salinity of the sea water affected toxicity of WSF prepared with dispersants, the most obvious effect being earlier onset of mortalities at higher temperatures. (Copyright (c) 1993 Pergamon Press Ltd.)

  15. Large-Scale Assessment of the Zebrafish Embryo as a Possible Predictive Model in Toxicity Testing

    PubMed Central

    Ali, Shaukat; van Mil, Harald G. J.; Richardson, Michael K.

    2011-01-01

    Background In the drug discovery pipeline, safety pharmacology is a major issue. The zebrafish has been proposed as a model that can bridge the gap in this field between cell assays (which are cost-effective, but low in data content) and rodent assays (which are high in data content, but less cost-efficient). However, zebrafish assays are only likely to be useful if they can be shown to have high predictive power. We examined this issue by assaying 60 water-soluble compounds representing a range of chemical classes and toxicological mechanisms. Methodology/Principal Findings Over 20,000 wild-type zebrafish embryos (including controls) were cultured individually in defined buffer in 96-well plates. Embryos were exposed for a 96 hour period starting at 24 hours post fertilization. A logarithmic concentration series was used for range-finding, followed by a narrower geometric series for LC50 determination. Zebrafish embryo LC50 (log mmol/L), and published data on rodent LD50 (log mmol/kg), were found to be strongly correlated (using Kendall's rank correlation tau and Pearson's product-moment correlation). The slope of the regression line for the full set of compounds was 0.73403. However, we found that the slope was strongly influenced by compound class. Thus, while most compounds had a similar toxicity level in both species, some compounds were markedly more toxic in zebrafish than in rodents, or vice versa. Conclusions For the substances examined here, in aggregate, the zebrafish embryo model has good predictivity for toxicity in rodents. However, the correlation between zebrafish and rodent toxicity varies considerably between individual compounds and compound class. We discuss the strengths and limitations of the zebrafish model in light of these findings. PMID:21738604

  16. ADAPTING THE MEDAKA EMBRYO ASSAY TO A HIGH-THROUGHPUT APPROACH FOR DEVELOPMENTAL TOXICITY TESTING.

    EPA Science Inventory

    Chemical exposure during embryonic development may cause persistent effects, yet developmental toxicity data exist for very few chemicals. Current testing procedures are time consuming and costly, underlining the need for rapid and low cost screening strategies. While in vitro ...

  17. Extensive review of fish embryo acute toxicities for the prediction of GHS acute systemic toxicity categories.

    PubMed

    Scholz, Stefan; Ortmann, Julia; Klüver, Nils; Léonard, Marc

    2014-08-01

    Distribution and marketing of chemicals require appropriate labelling of health, physical and environmental hazards according to the United Nations global harmonisation system (GHS). Labelling for (human) acute toxicity categories is based on experimental findings usually obtained by oral, dermal or inhalative exposure of rodents. There is a strong societal demand for replacing animal experiments conducted for safety assessment of chemicals. Fish embryos are considered as alternative to animal testing and are proposed as predictive model both for environmental and human health effects. Therefore, we tested whether LC50s of the fish embryo acute toxicity test would allow effectively predicting of acute mammalian toxicity categories. A database of published fish embryo LC50 containing 641 compounds was established. For these compounds corresponding rat oral LD50 were identified resulting in 364 compounds for which both fish embryo LC50 and rat LD50 was available. Only a weak correlation of fish embryo LC50 and rat oral LD50 was obtained. Fish embryos were also not able to effectively predict GHS oral acute toxicity categories. We concluded that due to fundamental exposure protocol differences (single oral dose versus water-borne exposure) a reverse dosimetry approach is needed to explore the predictive capacity of fish embryos. PMID:24929227

  18. Comparison of embryo toxicity using two classes of aquatic vertebrates.

    PubMed

    Carlsson, Gunnar; Norrgren, Leif

    2014-01-01

    Toxicity tests of musk ketone (MK) and tetrabromobisphenol-A (TBBPA) on embryos were conducted in two amphibian species, Xenopus (Silurana) tropicalis and the Swedish native species Rana arvalis. TBBPA was also tested on fish embryos of Danio rerio. All species were tested in similar experimental setup. Musk ketone caused decreased heart rates at concentrations from 10 and 100 μg/L in R. arvalis and X. tropicalis, respectively. TBBPA caused effects at 1000 μg/L in all three species. The responses were comparable between all three species which supports the relevance for using data from non-native species in national risk assessment. PMID:24291369

  19. Comparison of the mouse Embryonic Stem cell Test, the rat Whole Embryo Culture and the Zebrafish Embryotoxicity Test as alternative methods for developmental toxicity testing of six 1,2,4-triazoles.

    PubMed

    de Jong, Esther; Barenys, Marta; Hermsen, Sanne A B; Verhoef, Aart; Ossendorp, Bernadette C; Bessems, Jos G M; Piersma, Aldert H

    2011-06-01

    The relatively high experimental animal use in developmental toxicity testing has stimulated the search for alternatives that are less animal intensive. Three widely studied alternative assays are the mouse Embryonic Stem cell Test (EST), the Zebrafish Embryotoxicity Test (ZET) and the rat postimplantation Whole Embryo Culture (WEC). The goal of this study was to determine their efficacy in assessing the relative developmental toxicity of six 1,2,4-triazole compounds,(1) flusilazole, hexaconazole, cyproconazole, triadimefon, myclobutanil and triticonazole. For this purpose, we analyzed effects and relative potencies of the compounds in and among the alternative assays and compared the findings to their known in vivo developmental toxicity. Triazoles are antifungal agents used in agriculture and medicine, some of which are known to induce craniofacial and limb abnormalities in rodents. The WEC showed a general pattern of teratogenic effects, typical of exposure to triazoles, mainly consisting of reduction and fusion of the first and second branchial arches, which are in accordance with the craniofacial malformations reported after in vivo exposure. In the EST all triazole compounds inhibited cardiomyocyte differentiation concentration-dependently. Overall, the ZET gave the best correlation with the relative in vivo developmental toxicities of the tested compounds, closely followed by the EST. The relative potencies observed in the WEC showed the lowest correlation with the in vivo developmental toxicity data. These differences in the efficacy between the test systems might be due to differences in compound kinetics, in developmental stages represented and in the relative complexity of the alternative assays. PMID:21443896

  20. Comparison of the mouse Embryonic Stem cell Test, the rat Whole Embryo Culture and the Zebrafish Embryotoxicity Test as alternative methods for developmental toxicity testing of six 1,2,4-triazoles

    SciTech Connect

    Jong, Esther de; Barenys, Marta; Hermsen, Sanne A.B.; Verhoef, Aart; Ossendorp, Bernadette C.; Bessems, Jos G.M.; Piersma, Aldert H.

    2011-06-01

    The relatively high experimental animal use in developmental toxicity testing has stimulated the search for alternatives that are less animal intensive. Three widely studied alternative assays are the mouse Embryonic Stem cell Test (EST), the Zebrafish Embryotoxicity Test (ZET) and the rat postimplantation Whole Embryo Culture (WEC). The goal of this study was to determine their efficacy in assessing the relative developmental toxicity of six 1,2,4-triazole compounds, flusilazole, hexaconazole, cyproconazole, triadimefon, myclobutanil and triticonazole. For this purpose, we analyzed effects and relative potencies of the compounds in and among the alternative assays and compared the findings to their known in vivo developmental toxicity. Triazoles are antifungal agents used in agriculture and medicine, some of which are known to induce craniofacial and limb abnormalities in rodents. The WEC showed a general pattern of teratogenic effects, typical of exposure to triazoles, mainly consisting of reduction and fusion of the first and second branchial arches, which are in accordance with the craniofacial malformations reported after in vivo exposure. In the EST all triazole compounds inhibited cardiomyocyte differentiation concentration-dependently. Overall, the ZET gave the best correlation with the relative in vivo developmental toxicities of the tested compounds, closely followed by the EST. The relative potencies observed in the WEC showed the lowest correlation with the in vivo developmental toxicity data. These differences in the efficacy between the test systems might be due to differences in compound kinetics, in developmental stages represented and in the relative complexity of the alternative assays.

  1. Toxicity of trihalomethanes to common carp embryos

    SciTech Connect

    Mattice, J.S.; Tsai, S.C.; Burch, M.B.; Beauchamp, J.J.

    1981-03-01

    Trihalomethanes recently have been identified in real and simulated effluents from power plants where chlorine is used for biofouling control. Toxicity of the four chlorine- or bromine-containing trihalomethanes (chloroform, bromodichloromethane, dibromochloromethane, and bromoform) to developing common carp (Cyprinus carpio) embryos was determined under conditions of intermittent (8-hour) toxicant renewal, based on percent hatch as the end point. Nominal median lethal concentrations (LC50) ranged from 161 mg/liter for chloroform to 53 mg/liter for dibromochloromethane. Decay studies conducted under conditions similar to those used for the toxicity studies, but in distilled water, indicated that (1) half-lives of the trihalomethanes ranged from 4.4 to 6.9 hours; (2) decay was due primarily to volatilization; (3) higher relative toxicity of dibromochloromethane probably was due to formation of a degradation product (likely Br/sub 2/). Correction of the nominal LC50 values to time-weighted mean concentrations over the period between toxicant changes gave weighted LC50 values of 97.2, 67.4, 33.5, and 52.3 mg/liter for chloroform, bromodichloromethane, dibromochloromethane, and bromoform, respectively. In addition, the period of water-hardening of fertilized eggs was not critical for expression of toxicity of dibromochloromethane. Comparison of these and other published data on effluent and toxic concentrations, persistence, and bioaccumulation of water-chlorination products suggests that trihalomethanes are not as environmentally critical as other chlorinated organic compounds or residual chlorine.

  2. The fish embryo toxicity test as a replacement for the larval growth and survival test: A comparison of test sensitivity and identification of alternative endpoints in zebrafish and fathead minnows.

    PubMed

    Jeffries, Marlo K Sellin; Stultz, Amy E; Smith, Austin W; Stephens, Dane A; Rawlings, Jane M; Belanger, Scott E; Oris, James T

    2015-06-01

    The fish embryo toxicity (FET) test has been proposed as an alternative to the larval growth and survival (LGS) test. The objectives of the present study were to evaluate the sensitivity of the FET and LGS tests in fathead minnows (Pimephales promelas) and zebrafish (Danio rerio) and to determine if the inclusion of sublethal metrics as test endpoints could enhance test utility. In both species, LGS and FET tests were conducted using 2 simulated effluents. A comparison of median lethal concentrations determined via each test revealed significant differences between test types; however, it could not be determined which test was the least and/or most sensitive. At the conclusion of each test, developmental abnormalities and the expression of genes related to growth and toxicity were evaluated. Fathead minnows and zebrafish exposed to mock municipal wastewater-treatment plant effluent in a FET test experienced an increased incidence of pericardial edema and significant alterations in the expression of genes including insulin-like growth factors 1 and 2, heat shock protein 70, and cytochrome P4501A, suggesting that the inclusion of these endpoints could enhance test utility. The results not only show the utility of the fathead minnow FET test as a replacement for the LGS test but also provide evidence that inclusion of additional endpoints could improve the predictive power of the FET test. PMID:25929752

  3. Toxic effects of brominated indoles and phenols on zebrafish embryos.

    PubMed

    Kammann, U; Vobach, M; Wosniok, W

    2006-07-01

    Organobromine compounds in the marine environment have been the focus of growing attention in past years. In contrast to anthropogenic brominated flame retardants, other brominated compounds are produced naturally, e.g., by common polychaete worms and algae. Brominated phenols and indoles assumed to be of biogenic origin have been detected in water and sediment extracts from the German Bight. These substances as well as some of their isomers have been tested with the zebrafish embryo test and were found to cause lethal as well as nonlethal malformations. The zebrafish test was able to detect a log K(OW)-related toxicity for bromophenols, suggesting nonpolar narcosis as a major mode of action. Different effect patterns could be observed for brominated indoles and bromophenols. The comparison of effective concentrations in the zebrafish embryo test with the concentrations determined in water samples suggests the possibility that brominated indoles may affect early life stages of marine fish species in the North Sea. PMID:16418895

  4. Evaluation of MWNT toxic effects on daphnia and zebrafish embryos

    NASA Astrophysics Data System (ADS)

    Olasagasti, Maider; Alvarez, Noelia; Vera, Carolina; Rainieri, Sandra

    2009-05-01

    Organisms of daphnia (Daphnia magna) and zebrafish (Danio rerio) embryos were exposed to a range of different concentrations of COOH-functionalized MWCNT suspended in an aqueous solution of Tween 20. Immobilization of daphnia and growth retardation, inhibition and malformation of zebrafish embryos were the endpoints tested after 24 and 48 hours. Immobilization of daphnia could be observed from 3 to 16 ppm and an increasing mortality of zebrafish embryo was detected at all the concentration tested. To identify more subtle toxic effects, we took advantage of the extensive information available on the zebrafish genome and monitored by RT-PCR the expression patterns of different zebrafish genes that could act as toxicity bio-markers. At some of the concentrations tested, changes in the expression profiles of the genes examined were detected. Our results suggest that MWCNT could potentially represent a risk to human health and environment, therefore a wider range of concentrations and further testing of this molecules should be carried out to define possible limitations in their use.

  5. Embryo toxicity and teratogenicity of formaldehyde.

    PubMed

    Thrasher, J D; Kilburn, K H

    2001-01-01

    C-14 formaldehyde crosses the placenta and enters fetal tissues. The incorporated radioactivity is higher in fetal organs (i.e., brain and liver) than in maternal tissues. The incorporation mechanism has not been studied fully, but formaldehyde enters the single-carbon cycle and is incorporated as a methyl group into nucleic acids and proteins. Also, formaldehyde reacts chemically with organic compounds (e.g., deoxyribonucleic acid, nucleosides, nucleotides, proteins, amino acids) by addition and condensation reactions, thus forming adducts and deoxyribonucleic acid-protein crosslinks. The following questions must be addressed: What adducts (e.g., N-methyl amino acids) are formed in the blood following formaldehyde inhalation? What role do N-methyl-amino adducts play in alkylation of nuclear and mitochondrial deoxyribonucleic acid, as well as mitochondrial peroxidation? The fact that the free formaldehyde pool in blood is not affected following exposure to the chemical does not mean that formaldehyde is not involved in altering cell and deoxyribonucleic acid characteristics beyond the nasal cavity. The teratogenic effect of formaldehyde in the English literature has been sought, beginning on the 6th day of pregnancy (i.e., rodents) (Saillenfait AM, et al. Food Chem Toxicol 1989, pp 545-48; Martin WJ. Reprod Toxicol 1990, pp 237-39; Ulsamer AG, et al. Hazard Assessment of Chemicals; Academic Press, 1984, pp 337-400; and U.S. Department of Health and Human Services. Toxicological Profile of Formaldehyde; ATSDR, 1999 [references 1-4, respectively, herein]). The exposure regimen is critical and may account for the differences in outcomes. Pregnant rats were exposed (a) prior to mating, (b) during mating, (c) or during the entire gestation period. These regimens (a) increased embryo mortality; (b) increased fetal anomalies (i.e., cryptochordism and aberrant ossification centers); (c) decreased concentrations of ascorbic acid; and (d) caused abnormalities in enzymes of mitochondria, lysosomes, and the endoplasmic reticulum. The alterations in enzymatic activity persisted 4 mo following birth. In addition, formaldehyde caused metabolic acidosis, which was augmented by iron deficiency. Furthermore, newborns exposed to formaldehyde in utero had abnormal performances in open-field tests. Disparities in teratogenic effects of toxic chemicals are not unusual. For example, chlorpyrifos has not produced teratogenic effects in rats when mothers are exposed on days 6-15 (Katakura Y, et al. Br J Ind Med 1993, pp 176-82 [reference 5 herein]) of gestation (Breslin WJ, et al. Fund Appl Toxicol 1996, pp 119-30; and Hanley TR, et al. Toxicol Sci 2000, pp 100-08 [references 6 and 7, respectively, herein]). However, either changing the endpoints for measurement or exposing neonates during periods of neurogenesis (days 1-14 following birth) and during subsequent developmental periods produced adverse effects. These effects included neuroapoptosis, decreased deoxyribonucleic acid and ribonucleic acid synthesis, abnormalities in adenylyl cyclase cascade, and neurobehavioral effects (Johnson DE, et al. Brain Res Bull 1998, pp 143-47; Lassiter TL, et al. Toxicol Sci 1999, pp 92-100; Chakraborti TK, et al. Pharmacol Biochem Behav 1993, pp 219-24; Whitney KD, et al. Toxicol Appl Pharm 1995, pp 53-62; Chanda SM, et al. Pharmacol Biochem Behav 1996, pp 771-76; Dam K, et al. Devel Brain Res 1998, pp 39-45; Campbell CG, et al. Brain Res Bull 1997, pp 179-89; and Xong X, et al. Toxicol Appl Pharm 1997, pp 158-74 [references 8-15, respectively, herein]). Furthermore, the terata caused by thalidomide is a graphic human example in which the animal model and timing of exposure were key factors (Parman T, et al. Natl Med 1999, pp 582-85; and Brenner CA, et al. Mol Human Repro 1998, pp 887-92 [references 16 and 17, respectively, herein]). Thus, it appears that more sensitive endpoints (e.g., enzyme activity, generation of reactive oxygen species, timing of exposure) for the measurement of toxic effects of environmental agents on embryos, fetuses, and neonates are more coherent than are gross terata observations. The perinatal period from the end of organogenesis to the end of the neonatal period in humans approximates the 28th day of gestation to 4 wk postpartum. Therefore, researchers must investigate similar stages of development (e.g., neurogenesis occurs in the 3rd trimester in humans and neonatal days occur during days 1-14 in rats and mice, whereas guinea pigs behave more like humans). Finally, screening for teratogenic events should also include exposure of females before mating or shortly following mating. Such a regimen is fruitful inasmuch as environmental agents cause adverse effec PMID:11572272

  6. Toxic effects of several phthalate esters on the embryos and larvae of abalone Haliotis diversicolor supertexta

    NASA Astrophysics Data System (ADS)

    Yang, Zhihui; Zhang, Xiangjing; Cai, Zhonghua

    2009-05-01

    As the most widely used plasticizers in the world, phthalate esters (PAEs) are potential endocrine disruption compounds (EDCs). In the present study, the toxicity of dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP), di (2-ethylhexyl) phthalate (DEHP) on embryogenesis and larvae development of the marine univalve Haliotis diversicolor supertexta was examined in laboratory. The results show that the malformation of embryos appeared during the experiment, such as embryos died or lysed, small transparent flocculent rings studded on the periphery of the embryo, and the larvae could failed to hatch. In embryo toxic test, embryos incubated at the highest concentration of DMP, DEP and DBP solutions showed significantly high abnormal rate compared with the control, while DEHP solutions displayed no significant difference. In larval toxic test, in all concentrations of DMP, DEP and DBP solutions, larval settlement rates were low significantly than that of the control. Similarly, DEHP solutions show nearly no effect on the larval settlement. The order of toxicity on embryos and larvae is DBP>DEP>DMP>DEHP. Being a simple and easy stimulation to indoor spawn, sensitive to environmental factors, and short culture time, the embryos of H. diversicolor supertexta can be used to indicate toxicity of the PAEs.

  7. Toxicity of cryoprotectants agents in freshwater prawn embryos of Macrobrachium amazonicum.

    PubMed

    Ferreira, Arthur Vinícius Lourenço; Castro, Elias José Teles; Barbosa, Mariana Silva Alves; de Sousa, Míriam Luzia Nogueira Martins; de Araújo Neto, Manoel Paiva; Filho, Aldeney Andrade Soares; de Souza Sampaio, Celia Maria

    2015-12-01

    The process of cooling and cryopreservation of prawn embryos is a viable alternative for a continuous supply of larvae for freshwater prawn farming ponds. However, studies involving the application of those techniques as well as on toxicity of cryoprotectants in freshwater prawn embryos are scarce. Thus, this study aims to test the toxicity of methylic alcohol (MET), dimethyl sulfoxide (DMSO) and ethylene glycol (EG) on Macrobrachium amazonicum embryos. For the present experiment, pools of embryos were taken from 15 M. amazonicum females and were divided into three groups and tested in duplicate at concentrations of 10, 5, 3; 1, 0.5 or 0.1%. Toxicity tests were conducted for 24 h in Falcon® pipes to obtain the lethal concentration for 50% of the larvae (LC50). After the set period for testing, random samples of embryos were removed for morphological analysis under stereoscopic microscopes. Results were analysed using analysis of variance (ANOVA) and Tukey's test at a 5% significance level and Trimmed Spearman-Karber Analysis to determine LC50-24 h. DMSO toxicity tests revealed that 5% and 10% concentrations showed the highest toxicity and differed from the control (P ≤ 0.05), 24h-LC50 was 437.4 ± 14.4 µL. MET was less toxic among the tested cryoprotectants and concentrations did not allow the determination of its LC50-24h. For tests with EG, concentrations of 3, 5 or 10% solutions resulted in a 100% mortality to tested embryos; EG was the tested cryoprotectant with the highest toxicity, with an LC50-24h average of 81.91 ± 35.3 µl. PMID:25255785

  8. Rapid aquatic toxicity assay utilizing labeled thymidine incorporation in sea urchin embryos

    SciTech Connect

    Jackim, E.; Nacci, D.

    1984-01-01

    Aquatic toxicity was evaluated in the sea urchin embryo (Arbacea punctulata) by the inhibition of tritiated thymidine incorporation after a brief exposure to toxic chemicals. Arbacia is a useful surrogate species for assay: well-studied, easily cultured and fertile virtually year round. The simplicity and speed of this test system lends itself to screening large numbers of compounds, mixtures or water samples.

  9. DEVELOPMENT OF ISOLATED MAMMALIAN EMBRYO TECHNIQUES FOR TOXIC SUBSTANCES SCREENING

    EPA Science Inventory

    A potential screen for assessing teratogenic potential of compounds in mammals was tested. The technique involves testing isolated mammalian embryos in culture by direct exposure to agents. Embryos from three species of mammals were used; the mouse, rabbit and swine. Four differe...

  10. DEVELOPMENTAL TOXICITY OF COPPER SULFATE AND METHYLENE CHLORIDE TO SHRIMP EMBRYOS

    EPA Science Inventory

    The embryos of the grass shrimp (Palaemonetes pugio) have shown sensitivity to the water-soluble fraction of Number 2 fuel oil which indicates they may be a useful test species in estuarine developmental toxicity tests. Detailed concentration-response curves for copper sulfate an...

  11. A novel contact assay for testing aryl hydrocarbon receptor (AhR)-mediated toxicity of chemicals and whole sediments in zebrafish (Danio rerio) embryos.

    PubMed

    Schiwy, Sabrina; Bräunig, Jennifer; Alert, Henriette; Hollert, Henner; Keiter, Steffen H

    2015-11-01

    The European Water Framework Directive aims to achieve a good ecological and chemical status in surface waters until 2015. Sediment toxicology plays a major role in this intention as sediments can act as a secondary source of pollution. In order to fulfill this legal obligation, there is an urgent need to develop whole-sediment exposure protocols, since sediment contact assays represent the most realistic scenario to simulate in situ exposure conditions. Therefore, in the present study, a vertebrate sediment contact assay to determine aryl hydrocarbon receptor (AhR)-mediated activity of particle-bound pollutants was developed. Furthermore, the activity and the expression of the CYP1 family in early life stages of zebrafish after exposure to freeze-dried sediment samples were investigated. In order to validate the developed protocol, effects of β-naphthoflavone and three selected sediment on zebrafish embryos were investigated. Results documented clearly AhR-mediated toxicity after exposure to β-naphthoflavone (β-NF) and to the sediment from the Vering canal. Upregulation of mRNA levels was observed for all investigated sediment samples. The highest levels of all investigated cyp genes (cyp1a, cyp1b1, cyp1c1, and cyp1c2) were recorded after exposure to the sediment sample of the Vering canal. In conclusion, the newly developed sediment contact assay can be recommended for the investigation of dioxin-like activities of single substances and the bioavailable fraction of complex environmental samples. Moreover, the exposure of whole zebrafish embryos to native (freeze-dried) sediment samples represents a highly realistic and ecologically relevant exposure scenario. PMID:24958532

  12. Developmental toxicity assay using high content screening of zebrafish embryos.

    PubMed

    Lantz-McPeak, Susan; Guo, Xiaoqing; Cuevas, Elvis; Dumas, Melanie; Newport, Glenn D; Ali, Syed F; Paule, Merle G; Kanungo, Jyotshna

    2015-03-01

    Typically, time-consuming standard toxicological assays using the zebrafish (Danio rerio) embryo model evaluate mortality and teratogenicity after exposure during the first 2 days post-fertilization. Here we describe an automated image-based high content screening (HCS) assay to identify the teratogenic/embryotoxic potential of compounds in zebrafish embryos in vivo. Automated image acquisition was performed using a high content microscope system. Further automated analysis of embryo length, as a statistically quantifiable endpoint of toxicity, was performed on images post-acquisition. The biological effects of ethanol, nicotine, ketamine, caffeine, dimethyl sulfoxide and temperature on zebrafish embryos were assessed. This automated developmental toxicity assay, based on a growth-retardation endpoint should be suitable for evaluating the effects of potential teratogens and developmental toxicants in a high throughput manner. This approach can significantly expedite the screening of potential teratogens and developmental toxicants, thereby improving the current risk assessment process by decreasing analysis time and required resources. PMID:24871937

  13. Estradiol uptake, toxicity, metabolism, and adverse effects on cadmium-treated amphibian embryos.

    PubMed Central

    Fridman, Osvaldo; Corró, Lucrecia; Herkovits, Jorge

    2004-01-01

    The exposure of Bufo arenarum embryos to 25 micromol/L 17beta-estradiol (E2) resulted in 100% lethality within 48 hr, whereas 10 micromol//L E2 was the no observed effect concentration value for short-term chronic (7 days) exposure. The toxicity profile curves show that lethal effects were proportional to the E2 concentration and the time of exposure. The E2 uptake resulted in 20.1 ng E2/mg embryo at 8 hr posttreatment, but 67.3% of this value was achieved during the first 30 min of incubation with this estrogen. Regarding metabolism, the embryos synthesize estrone (E1) from E2 by means of 17beta-hydroxysteroid dehydrogenase. Simultaneous treatments of Bufo arenarum embryos with 1 mg/L Cd2+ and 0.1, 1, or 10 micromol/L E2 enhanced the lethality exerted by cadmium in 76.7, 80, and 83.3% of embryos, respectively. The results indicate that estrogenic endocrine disruptors could have an adverse effect on amphibian embryos and enhance the toxic effect of Cd on amphibian embryos. This study points to the possibility of using the AMPHITOX test as a screening method for potential endocrine disruption as well as the combined effects of chemical mixtures. PMID:15175173

  14. Virtual Embryo: Systems Modeling in Developmental Toxicity

    EPA Science Inventory

    High-throughput and high-content screening (HTS-HCS) studies are providing a rich source of data that can be applied to in vitro profiling of chemical compounds for biological activity and potential toxicity. EPA’s ToxCast™ project, and the broader Tox21 consortium, in addition t...

  15. Virtual Embryo: Systems Modeling in Developmental Toxicity

    EPA Science Inventory

    High-throughput and high-content screening (HTS-HCS) studies are providing a rich source of data that can be applied to in vitro profiling of chemical compounds for biological activity and potential toxicity. EPAs ToxCast project, and the broader Tox21 consortium, in addition t...

  16. Nanomaterial Toxicity Screening in Developing Zebrafish Embryos

    EPA Science Inventory

    To assess nanomaterial vertebrate toxicity, a high-content screening assay was created using developing zebrafish, Danio rerio. This included a diverse group of nanomaterials (n=42 total) ranging from metallic (Ag, Au) and metal oxide (CeO2, CuO, TiO2, ZnO) nanoparticles, to non...

  17. Combined endosulfan and cypermethrin-induced toxicity to embryo-larval development of Rhinella arenarum.

    PubMed

    Svartz, Gabriela V; Aronzon, Carolina M; Pérez Coll, Cristina S

    2016-01-01

    The combined effects of two widely used pesticides, endosulfan and cypermethrin, on survival of embryo-larval development of the South American toad (Rhinella arenarum) were examined. The toxicity bioassays were performed according to the AMPHITOX test. Embryos and larvae were exposed to mixtures of these pesticides at equitoxic ratios from acute or chronic exposure to evaluate interaction effects. The results were analyzed using both Marking's additive index and combination index (CI)-isobologram methods. Acute (96-h) and intermediate (168-h) toxicity of endosulfan-cypermethrin mixtures remained almost constant for larvae and embryos, but when exposure duration was increased, there was a significant elevation in toxicity, obtaining chronic (240-h) no-observed-effect concentrations (NOEC) values of 0.045 and 0.16 mg/L for embryos and larvae, respectively. These are environmentally relevant concentrations that reflect a realistic risk of this pesticide mixture to this native amphibian species. The toxicity increment with the exposure duration was coincident with the central nervous system development on embryos reaching the larval period, the main target organ of these pesticides. The interactions of the pesticide mixtures at acute and chronic exposure were antagonistic for embryo development (CI > 1), and additive (CI = 1) for larvae, while chronic exposure interactions were synergistic (CI < 1) for both developmental periods. Data indicated that endosulfan-cypermethrin mixtures resulted in different interaction types depending on duration and developmental stage exposed. As a general pattern and considering conditions of overall developmental period and chronic exposure, this pesticide mixture usually applied in Argentine crop fields is synergistic with respect to toxicity for this native amphibian species. PMID:26914601

  18. In Vivo Nanotoxicity Testing using the Zebrafish Embryo Assay

    PubMed Central

    Rizzo, Larissa Y.; Golombek, Susanne K.; Mertens, Marianne E.; Pan, Yu; Laaf, Dominic; Broda, Janine; Jayapaul, Jabadurai; Möckel, Diana; Subr, Vladimir; Hennink, Wim E.; Storm, Gert; Simon, Ulrich; Jahnen-Dechent, Willi; Kiessling, Fabian; Lammers, Twan

    2013-01-01

    Nanoparticles are increasingly used for biomedical purposes. Many different diagnostic and therapeutic applications are envisioned for nanoparticles, but there are often also serious concerns regarding their safety. Given the fact that numerous new nanomaterials are being developed every day, and that not much is known about the long-term toxicological impact of exposure to nanoparticles, there is an urgent need to establish efficient methods for nanotoxicity testing. The zebrafish (Danio rerio) embryo assay has recently emerged as an interesting ‘intermediate’ method for in vivo nanotoxicity screening, enabling (semi-) high-throughput analyses in a system significantly more complex than cultured cells, but at the same time also less ‘invasive’ and less expensive than large-scale biocompatibility studies in mice or rats. The zebrafish embryo assay is relatively well-established in the environmental sciences, but it has not yet gained wide notice in the nanomedicine field. Using prototypic polymeric drug carriers, gold-based nanodiagnostics and nanotherapeutics, and iron oxide-based nanodiagnostics, we here show that toxicity testing using zebrafish embryos is easy, efficient and informative, and faithfully reflects, yet significantly extends, cell-based toxicity testing. We therefore expect that the zebrafish embryo assay will become a popular future tool for in vivo nanotoxicity screening. PMID:24179674

  19. Tissue-specific direct microtransfer of nanomaterials into Drosophila embryos as a versatile in vivo test bed for nanomaterial toxicity assessment.

    PubMed

    Vega-Alvarez, Sasha; Herrera, Adriana; Rinaldi, Carlos; Carrero-Martínez, Franklin A

    2014-01-01

    Nanomaterials are the subject of intense research, focused on their synthesis, modification, and biomedical applications. Increased nanomaterial production and their wide range of applications imply a higher risk of human and environmental exposure. Unfortunately, neither environmental effects nor toxicity of nanomaterials to organisms are fully understood. Cost-effective, rapid toxicity assays requiring minimal amounts of materials are needed to establish both their biomedical potential and environmental safety standards. Drosophila exemplifies an efficient and cost-effective model organism with a vast repertoire of in vivo tools and techniques, all with high-throughput scalability and screening feasibility throughout its life cycle. Here we report tissue specific nanomaterial assessment through direct microtransfer into target tissues. We tested several nanomaterials with potential biomedical applications such as single-wall carbon nanotubes, multiwall carbon nanotubes, silver, gold, titanium dioxide, and iron oxide nanoparticles. Assessment of nanomaterial toxicity was conducted by evaluating progression through developmental morphological milestones in Drosophila. This cost-effective assessment method is amenable to high-throughput screening. PMID:24790441

  20. Tissue-specific direct microtransfer of nanomaterials into Drosophila embryos as a versatile in vivo test bed for nanomaterial toxicity assessment

    PubMed Central

    Vega-Alvarez, Sasha; Herrera, Adriana; Rinaldi, Carlos; Carrero-Martínez, Franklin A

    2014-01-01

    Nanomaterials are the subject of intense research, focused on their synthesis, modification, and biomedical applications. Increased nanomaterial production and their wide range of applications imply a higher risk of human and environmental exposure. Unfortunately, neither environmental effects nor toxicity of nanomaterials to organisms are fully understood. Cost-effective, rapid toxicity assays requiring minimal amounts of materials are needed to establish both their biomedical potential and environmental safety standards. Drosophila exemplifies an efficient and cost-effective model organism with a vast repertoire of in vivo tools and techniques, all with high-throughput scalability and screening feasibility throughout its life cycle. Here we report tissue specific nanomaterial assessment through direct microtransfer into target tissues. We tested several nanomaterials with potential biomedical applications such as single-wall carbon nanotubes, multiwall carbon nanotubes, silver, gold, titanium dioxide, and iron oxide nanoparticles. Assessment of nanomaterial toxicity was conducted by evaluating progression through developmental morphological milestones in Drosophila. This cost-effective assessment method is amenable to high-throughput screening. PMID:24790441

  1. Virtual Embryo: Systems Modeling in Developmental Toxicity

    EPA Science Inventory

    High-throughput screening (HTS) studies are providing a rich source of data that can be applied to chemical profiling to address sensitivity and specificity of molecular targets, biological pathways, cellular and developmental processes. EPA’s ToxCast project is testing 960 uniq...

  2. Virtual Embryo: Systems Modeling in Developmental Toxicity

    EPA Science Inventory

    High-throughput screening (HTS) studies are providing a rich source of data that can be applied to chemical profiling to address sensitivity and specificity of molecular targets, biological pathways, cellular and developmental processes. EPAs ToxCast project is testing 960 uniq...

  3. Use of Medaka in Toxicity Testing

    PubMed Central

    Cowden, John; Hinton, David E.; Johnson, Rodney; Flynn, Kevin; Hardman, Ronald C.; Yuen, Bonny; Law, Sheran; Kullman, Seth W.; Au, Doris W.T.

    2015-01-01

    Small aquarium fishes are increasingly used as animal models, and one of these, Japanese Medaka (Oryzias latipes), is frequently utilized for toxicity testing. While these vertebrates have many similarities with their terrestrial counterparts, there are differences that must be considered if these organisms are to be used to their highest potential. Testing commonly may employ either the developing embryo or adults; both are easy to use and to work with. We present here three main protocols to illustrate the utility and breadth of toxicity testing possible using medaka fish. The first protocol assesses neurotoxicity in developing embryos. The second protocol describes the sexual genotyping of medaka to evaluate toxicant effects on sexual phenotype after treatment with endocrine disrupting chemicals. The third protocol assesses hepatotoxicity in adult fish after treatment with a model hepatotoxicant. The methods run the gamut from immunohistology through PCR to basic histological techniques. PMID:20922755

  4. Ultraviolet Radiation Enhances the Toxicity of Deepwater Horizon Oil to Mahi-mahi (Coryphaena hippurus) Embryos.

    PubMed

    Alloy, Matthew; Baxter, David; Stieglitz, John; Mager, Edward; Hoenig, Ronald; Benetti, Daniel; Grosell, Martin; Oris, James; Roberts, Aaron

    2016-02-16

    The 2010 Deepwater Horizon oil spill resulted in the accidental release of millions barrels of crude oil into the Gulf of Mexico. Photoinduced toxicity following coexposure to ultraviolet (UV) radiation is one mechanism by which polycyclic aromatic hydrocarbons (PAHs) from oil spills may exert toxicity. Mahi-mahi (Coryphaena hippurus), an important fishery resource, have positively buoyant, transparent eggs. These characteristics may result in mahi-mahi embryos being at particular risk from photoinduced toxicity. The goal of this study was to determine whether exposure to ultraviolet radiation as natural sunlight enhances the toxicity of crude oil to embryonic mahi-mahi. Mahi-mahi embryos were exposed to several dilutions of water accommodated fractions (WAF) from slick oil collected during the 2010 spill and gradations of natural sunlight in a fully factorial design. Here, we report that coexposure to natural sunlight and WAF significantly reduced percent hatch in mahi-mahi embryos. Effect concentrations of PAH in WAF were within the range of surface PAH concentrations reported in the Gulf of Mexico during the Deepwater Horizon spill. These data suggest that laboratory toxicity tests that do not include UV may underestimate the toxicity of oil spills to early lifestage fish species. PMID:26784438

  5. Rapid aquatic toxicity assay using incorporation of tritiated-thymidine into sea urchin, Arbacia punctulata, embryo: evaluation of toxicant exposure procedures

    SciTech Connect

    Nacci, D.E.; Jackim, E.

    1985-01-01

    Toxicity of substances in seawater was measured using growth inhibition of embryonic sea urchins during a short period after fertilization. Growth of Arbacia punctulata embryos was monitored by incorporation of tritium-labeled thymidine. The paper presents a comparison of toxicant exposure procedures using the Arbacia embryo thymidine incorporation test. Toxicant exposure began before, at the time of, or after fertilization and continued for 4 h following fertilization. In addition to the eight organic chemicals tested for comparison to acute toxicity values for other species, several chemicals with embryotoxic potentials (tumor promoters and teratogens) were tested to determine differential sensitivities of exposed life-stages: unfertilized egg, fertilization, and early embryo. EC50 values for any one substance were not significantly changed by exposure modification. Toxicity values for exposures that included fertilization as well as early embryo growth were at least as sensitive as post-fertilization exposure values for all compounds tested except one. Because of technical ease and potential sensitivity, toxicant exposure that includes fertilization as well as early embryo growth (but not unfertilized egg exposure) is recommended for future testing.

  6. DEVELOPMENTAL TOXICITY TESTING

    EPA Science Inventory

    Contemporary developmental toxicity testing focuses on the evaluation of a variety of adverse developmental effects which include structural malformations, intrauterine death, growth retardation, and deficits in postnatal function. n the extrapolation of information from animal s...

  7. ENGINEERING BULLETIN: BIOLOGICAL TOXICITY TESTING

    EPA Science Inventory

    This Engineering Bulletin is intended to provide site managers with information on ecological assessment and biological toxicity testing, applicability of biological toxicity testing, planning effective biological toxicity assessments, descriptions of test methods, limitations, c...

  8. Toxicity of urban highway runoff in Shanghai to Zebrafish (Danio rerio ) embryos and luminous bacteria (Vibrio qinghaiensis.Q67).

    PubMed

    Wu, Lingling; Jiang, Yue; Zhang, Lili; Chen, Ling; Zhang, Haiping

    2014-02-01

    Pollution from urban highway runoff has been identified as one of the major causes of the deterioration of receiving water quality. The purpose of this study is to assess the toxicity of urban storm water samples in Shanghai using the zebrafish (Danio rerio ) embryo test and the bacterial luminescence (Vibrio qinghaiensis ) assay. The toxicity of highway runoff from seventeen storm events was investigated in both grab and composite samples. Zebrafish embryos were exposed to the runoff samples and development parameters including lethality, spontaneous movements in 20 s, heart beat rate, hatching rate, and abnormality of zebrafish embryos were observed after 24, 48, 72, and 96 h of exposure. Inhibition rates of luminescence intensity were also recorded. The results showed that in the zebrafish embryo toxicity tests, both grab and composite samples increased the lethality, reduced the percentage with spontaneous movements and heart beats, inhibited the hatching of embryos, and induced morphological abnormalities. In the Vibrio qinghaiensis toxicity test, all the grab samples inhibited the luminescence, while some of the composite samples promoted it, which indicated that different types of toxicants might have been affecting the species. The multivariate statistics analysis indicated that heavy metal (zinc, manganese, and copper) and PAHs might mainly contribute to the toxicity of runoff samples. PMID:24122161

  9. Status and applications of echinoid (phylum echinodermata) toxicity test methods

    SciTech Connect

    Bay, S.; Burgess, R.; Nacci, D.

    1993-01-01

    The use of echinoderms for toxicity testing has focused primarily on sea urchins and sand dollars (Strongylocentrotus purpuratus, Arbacia punctulata, Lytechinus pictus, and Dendraster excentricus, for example). The status and relative sensitivity of various test methods are described. The most frequently used test methods consist of short-term exposures of sea urchin sperm or embryos; these tests can be easily conducted at all times of the year by using species with complementary spawning cycles or laboratory conditioned populations of a single species. Data from reference toxicant and effluent toxicity tests are summarized. Information on the precision and sensitivity of echinoid test methods are limited and preclude rigorous comparisons with other test methods. The available data indicate that the sensitivity and precision of these methods are comparable to short-term chronic methods for other marine invertebrates and fish. Recent application of the sperm test in toxicity identification evaluations (TIEs) and studies of effluent toxicity decay and sediment toxicity illustrate the versatility of this rapid (10 to 60 min exposure) test method. Embryo tests typically use a 48 to 96 h exposure period and measure the occurrence of embryo malformations. Most recent applications of the embryo test have been for the assessment of sediment elutriate toxicity. Adult echinoderms are not frequently used to assess effluent or receiving water toxicity. Recent studies have had success in using the adult life stage of urchins and sand dollars to assess the effects of contaminated sediment on growth, behavior, and bioaccumulation.

  10. Developmental toxicity and alteration of gene expression in zebrafish embryos exposed to PFOS

    SciTech Connect

    Shi Xiongjie; Du Yongbing; Lam, Paul K.S.; Wu, Rudolf S.S.; Zhou Bingsheng

    2008-07-01

    Perfluorooctanesulfonate (PFOS) is a persistent organic pollutant, the potential toxicity of which is causing great concern. In the present study, we employed zebrafish embryos to investigate the developmental toxicity of this compound. Four-hour post-fertilization (hpf) zebrafish embryos were exposed to 0.1, 0.5, 1, 3 and 5 mg/L PFOS. Hatching was delayed and hatching rates as well as larval survivorship were significantly reduced after the embryos were exposed to 1, 3 and 5 mg/L PFOS until 132 hpf. The fry displayed gross developmental malformations, including epiboly deformities, hypopigmentation, yolk sac edema, tail and heart malformations and spinal curvature upon exposure to PFOS concentrations of 1 mg/L or greater. Growth (body length) was significantly reduced in the 3 and 5 mg/L PFOS-treated groups. To test whether developmental malformation was mediated via apoptosis, flow cytometry analysis of DNA content, acridine orange staining and TUNEL assay was used. These techniques indicated that more apoptotic cells were present in the PFOS-treated embryos than in the control embryos. Certain genes related to cell apoptosis, p53 and Bax, were both significantly up-regulated upon exposure to all the concentrations tested. In addition, we investigated the effects of PFOS on marker genes related to early thyroid development (hhex and pax8) and genes regulating the balance of androgens and estrogens (cyp19a and cyp19b). For thyroid development, the expression of hhex was significantly up-regulated at all concentrations tested, whereas pax8 expression was significantly up-regulated only upon exposure to lower concentrations of PFOS (0.1, 0.5, 1 mg/L). The expression of cyp19a and of cyp19b was significantly down-regulated at all exposure concentrations. The overall results indicated that zebrafish embryos constitute a reliable model for testing the developmental toxicity of PFOS, and the gene expression patterns in the embryos were able to reveal some potential mechanisms of developmental toxicity.

  11. Effects-driven chemical fractionation of heavy fuel oil to isolate compounds toxic to trout embryos.

    PubMed

    Bornstein, Jason M; Adams, Julie; Hollebone, Bruce; King, Thomas; Hodson, Peter V; Brown, R Stephen

    2014-04-01

    Heavy fuel oil (HFO) spills account for approximately 60% of ship-source oil spills and are up to 50 times more toxic than medium and light crude oils. Heavy fuel oils contain elevated concentrations of polycyclic aromatic hydrocarbons (PAHs) and alkyl-PAHs, known to be toxic to fish; however, little direct characterization of HFO toxicity has been reported. An effects-driven chemical fractionation was conducted on HFO 7102 to separate compounds with similar chemical and physical properties, including toxicity, to isolate the groups of compounds most toxic to trout embryos. After each separation, toxicity tests directed the next phase of fractionation, and gas chromatography-mass spectrometry analysis correlated composition with toxicity, with a focus on PAHs. Low-temperature vacuum distillation permitted the separation of HFO into 3 fractions based on boiling point ranges. The most toxic of these fractions underwent wax precipitation to remove long-chain n-alkanes. The remaining PAH-rich extract was further separated using open column chromatography, which provided distinct fractions that were grouped according to increasing aromatic ring count. The most toxic of these fractions was richest in PAHs and alkyl-PAHs. The results of the present study were consistent with previous crude oil studies that identified PAH-rich fractions as the most toxic. PMID:24375845

  12. Acute toxicity of agricultural pesticides to embryo-larval and juvenile African catfish Clarias gariepinus.

    PubMed

    Agbohessi, P T; Imorou Toko, I; Houndji, A; Gillardin, V; Mandiki, S N M; Kestemont, P

    2013-05-01

    Acute toxicities of Tihan 175 O-TEQ, as well as its active ingredients flubendiamide and spirotetramat, and of Thionex 350 EC (active compound endosulfan) were measured for embryo-larval and juvenile stages of the African catfish Clarias gariepinus to assess risks of pesticide use in the cotton basin in Benin (West Africa). For embryo-larval stages, Tihan was more toxic (LC5048h 20 ppm) than Thionex (LC5048h 56 ppm), and flubendiamide was more toxic (LC5048h 2.0 ppm) than spirotetramat (LC5048h 8.44 ppm). All decreased hatching rates. Tihan and spirotetramat disturbed larval swimming coordination; flubendiamide induced tail cleavage. For juvenile fish, Thionex was more toxic (LC5096h 0.22 ppm) than Tihan (LC5096h 8.8 ppm), and flubendiamide (LC5096h 4.7 ppm) was more toxic than spirotetramat (LC5096h 6.0 ppm). Eggs were more resistant than juvenile fish to all tested pesticides except flubendiamide. Although Thionex was more toxic to juvenile fish, replacing Thionex with Tihan may be undesirable for survival of eggs and larvae. PMID:23334459

  13. Relative embryotoxic potency of p-substituted phenols in the embryonic stem cell test (EST) and comparison to their toxic potency in vivo and in the whole embryo culture (WEC) assay.

    PubMed

    Strikwold, Marije; Woutersen, Ruud A; Spenkelink, Bert; Punt, Ans; Rietjens, Ivonne M C M

    2012-09-01

    The applicability of the embryonic stem cell test (EST) as an alternative for in vivo embryotoxicity testing was evaluated for a series of five p-substituted phenols. To this purpose, the potency ranking for this class of compounds derived from the inhibition of cardiomyocyte differentiation in the EST was compared to in vivo embryotoxic potency data obtained from literature and to the potency ranking defined in the in vitro whole embryo culture (WEC) assay. From the results obtained it appears that the EST was able to identify the embryotoxic potential for p-substituted phenols, providing an identical potency ranking compared to the WEC assay. However, the EST was not able to predict an accurate ranking for the phenols compared to their potency observed in vivo. Only phenol, the least potent compound within this series, was correctly ranked. Furthermore, p-mercaptophenol was correctly identified as a relative potent congener of the phenols tested, but its ranking was distorted by p-heptyloxyphenol, of which the toxicity was overestimated in the EST. It is concluded that when attempting to explain the observed disparity in potency rankings between in vitro and in vivo embryotoxicity, the in vitro models should be combined with a kinetic model describing in vivo absorption, distribution, metabolism and excretion processes of the compounds. PMID:22820428

  14. Unexpected interaction with dispersed crude oil droplets drives severe toxicity in Atlantic haddock embryos.

    PubMed

    Sørhus, Elin; Edvardsen, Rolf B; Karlsen, Ørjan; Nordtug, Trond; van der Meeren, Terje; Thorsen, Anders; Harman, Christopher; Jentoft, Sissel; Meier, Sonnich

    2015-01-01

    The toxicity resulting from exposure to oil droplets in marine fish embryos and larvae is still subject for debate. The most detailed studies have investigated the effects of water-dissolved components of crude oil in water accommodated fractions (WAFs) that lack bulk oil droplets. Although exposure to dissolved petroleum compounds alone is sufficient to cause the characteristic developmental toxicity of crude oil, few studies have addressed whether physical interaction with oil micro-droplets are a relevant exposure pathway for open water marine speices. Here we used controlled delivery of mechanically dispersed crude oil to expose pelagic embryos and larvae of a marine teleost, the Atlantic haddock (Melanogrammus aeglefinus). Haddock embryos were exposed continuously to two different concentrations of dispersed crude oil, high and low, or in pulses. By 24 hours of exposure, micro-droplets of oil were observed adhering and accumulating on the chorion, accompanied by highly elevated levels of cyp1a, a biomarker for exposure to aromatic hydrocarbons. Embryos from all treatment groups showed abnormalities representative of crude oil cardiotoxicity at hatch (5 days of exposure), such as pericardial and yolk sac edema. Compared to other species, the frequency and severity of toxic effects was higher than expected for the waterborne PAH concentrations (e.g., 100% of larvae had edema at the low treatment). These findings suggest an enhanced tissue uptake of PAHs and/or other petroleum compounds from attached oil droplets. These studies highlight a novel property of haddock embryos that leads to greater than expected impact from dispersed crude oil. Given the very limited number of marine species tested in similar exposures, the likelihood of other species with similar properties could be high. This unanticipated result therefore has implications for assessing the ecological impacts of oil spills and the use of methods for dispersing oil in the open sea. PMID:25923774

  15. Unexpected Interaction with Dispersed Crude Oil Droplets Drives Severe Toxicity in Atlantic Haddock Embryos

    PubMed Central

    Sørhus, Elin; Edvardsen, Rolf B.; Karlsen, Ørjan; Nordtug, Trond; van der Meeren, Terje; Thorsen, Anders; Harman, Christopher; Jentoft, Sissel; Meier, Sonnich

    2015-01-01

    The toxicity resulting from exposure to oil droplets in marine fish embryos and larvae is still subject for debate. The most detailed studies have investigated the effects of water-dissolved components of crude oil in water accommodated fractions (WAFs) that lack bulk oil droplets. Although exposure to dissolved petroleum compounds alone is sufficient to cause the characteristic developmental toxicity of crude oil, few studies have addressed whether physical interaction with oil micro-droplets are a relevant exposure pathway for open water marine speices. Here we used controlled delivery of mechanically dispersed crude oil to expose pelagic embryos and larvae of a marine teleost, the Atlantic haddock (Melanogrammus aeglefinus). Haddock embryos were exposed continuously to two different concentrations of dispersed crude oil, high and low, or in pulses. By 24 hours of exposure, micro-droplets of oil were observed adhering and accumulating on the chorion, accompanied by highly elevated levels of cyp1a, a biomarker for exposure to aromatic hydrocarbons. Embryos from all treatment groups showed abnormalities representative of crude oil cardiotoxicity at hatch (5 days of exposure), such as pericardial and yolk sac edema. Compared to other species, the frequency and severity of toxic effects was higher than expected for the waterborne PAH concentrations (e.g., 100% of larvae had edema at the low treatment). These findings suggest an enhanced tissue uptake of PAHs and/or other petroleum compounds from attached oil droplets. These studies highlight a novel property of haddock embryos that leads to greater than expected impact from dispersed crude oil. Given the very limited number of marine species tested in similar exposures, the likelihood of other species with similar properties could be high. This unanticipated result therefore has implications for assessing the ecological impacts of oil spills and the use of methods for dispersing oil in the open sea. PMID:25923774

  16. Formation of copper complexes in landfill leachate and their toxicity to zebrafish embryos

    SciTech Connect

    Fraser, J.K.; Butler, C.A.; Timperley, M.H.; Evans, C.W.

    2000-05-01

    Toxic metal organic complexes have not been found in natural waters, although some organic acids form bioavailable lipophilic and metabolite-type metal complexes. Landfill leachates usually contain organic acids and in the urban environment these leachates, when mixed with storm waters containing Cu, could be a source of toxic Cu organic complexes in streams and estuaries. The authors investigated the formation of Cu complexes in the leachate from an active urban landfill and found that some of the complexes formed were toxic to zebrafish embryos. High and low nominal molecular weight (NMWT) fractions; >5,000 Da and <700 Da, of leachate both formed Cu complexes with almost identical Cu complexing characteristics but the toxicity was due solely to the low NMWT complexes formed in the <700 Da fraction. Chemical equilibrium modeling with MINTEQA2 and H and Cu complex conditional association constants and ligand concentrations obtained from pH and Cu titrations with a Cu ion-selective electrode and van den Berg-Ruzic analyses of the titration data was used to calculate the copper speciation in the embryo test solutions. This calculated speciation, which was confirmed by measurements of Cu{sup 2+} in the test solutions, enabled the toxicity due to the free Cu ion and to the Cu complexes to be distinguished.

  17. Embryo Toxic Effects of Depleted Uranium on the Morphology of the Mouse Fetus

    PubMed Central

    Mirderikvand, Nina; Mohammadzadeh Asl, Baharak; Naserzadeh, Parvaneh; Shaki, Fatemeh; Shokrzadeh, Mohammad; Pourahmad, Jalal

    2014-01-01

    Although the biokinetics, metabolism, and chemical toxicity of uranium are well known, until recently little attention was paid to the potential toxic effects of uranium on reproduction and development in mammals. In recent years, it has been shown that uranium is a developmental toxicant when given orally or subcutaneously (SC) to mice. Decreased fertility, embryo/fetal toxicity including teratogenicity, and reduced growth of the offspring have been observed following uranium exposure at different gestation periods. For investigating the effects of DU on pregnant animals, three groups (control, sham and test) of NMRI mice were chosen. In test group 4 mg/Kg of DU were administered intraperitonealy at 11 day of gestation, in sham group only normal saline injected to interior peritoneum as indicated in the test group and in Control group which was considered as the comparison base line of our research, no injection was made. Caesarean sections were performed at 15 day of the gestation; and their placentas were examined externally. Base on our results DU caused significant external anomalies, and caused a significant decrease (p < 0.05) in the weight and diameter of placentas, the number of the embryos, their body weight and crown-rump length of fetuses. PMID:24734072

  18. Benomyl induction of brain aromatase and toxic effects in the zebrafish embryo.

    PubMed

    Kim, Dong-Jae; Seok, Seung-Hyeok; Baek, Min-Won; Lee, Hui-Young; Na, Yi-Rang; Park, Sung-Hoon; Lee, Hyun-Kyoung; Dutta, Noton Kumar; Kawakami, Koichi; Park, Jae-Hak

    2009-05-01

    Benomyl is a benzimidazole fungicide that has been widely used on a variety of food crops and ornamental plants. It is known to cause adverse effects on reproductive systems, including decreased testicular and epididymal weights and reduced epididymal sperm counts and fertility. The brain aromatase gene is up-regulated by estrogens and estrogen mimics and considered a target gene to screen estrogen mimics. This study was designed to test the estrogenic potential and toxic effects of benomyl in the zebrafish system, and validated this system as a model that may correspond to the effect of benomyl in rodents. Concentrations of 20 x 10(-6), 40 x 10(-6) and 80 x 10(-6) M of benomyl-treated embryos showed decreased survival, hatching and heart rates, and increased incidence of malformations, such as pericardial edema, spinal lordosis, elongated heart, head edema, eye lens protrusion and caudal fin disappearance. Benomyl induced enhanced green fluorescent protein (EGFP) expression in the mediobasal hypothalamus (MBH) in transient zebrafish embryos with a brain aromatase-based reporter gene. In this study, we determined that benomyl has estrogenic potential based on zebrafish brain aromatase gene induction, and that benomyl is toxic at 20 x 10(-6) M concentration and higher. These results demonstrate the usefulness of zebrafish embryos as an in vivo system to examine the estrogenic and developmental toxic potential of unknown compounds. PMID:19058295

  19. Toxicity of crude oil chemically dispersed in a wave tank to embryos of Atlantic herring (Clupea harengus).

    PubMed

    Greer, Colleen D; Hodson, Peter V; Li, Zhengkai; King, Thomas; Lee, Kenneth

    2012-06-01

    Tests of crude oil toxicity to fish are often chronic, exposing embryos from fertilization to hatch to oil solutions prepared using standard mixing procedures. However, during oil spills, fish are not often exposed for long periods and the dynamic nature of the ocean is not easily replicated in the lab. Our objective was to determine if brief exposures of Atlantic herring (Clupea harengus) embryos to dispersed oil prepared by standard mixing procedures was as toxic as oil dispersed in a more realistic model system. Embryos were first exposed to chemically dispersed Alaska North Slope crude and Arabian light crude oil for 2.4 h to 14 d from fertilization to determine if exposure time affected toxicity. Toxicity increased with exposure time, but 2.4-h exposures at realistic concentrations of oil induced blue-sac disease and reduced the percentage of normal embryos at hatch; there was little difference in toxicity between the two oils. Secondly, oil was chemically dispersed in a wave tank to determine if the resultant oil solutions were as toxic to herring embryos as laboratory-derived dispersed oil using a single exposure period of 24 h. Samples taken 15 min postdispersion were more toxic than laboratory-prepared solutions, but samples taken at 5, 30, and 60 min postdispersion were less toxic. Overall, the laboratory- and wave tank-derived solutions of dispersed oil provided similar estimates of toxicity despite differences in the methods for preparing test solutions, suggesting that laboratory and wave tank data are a reliable basis for ecological risk assessments of spilled oil. PMID:22488782

  20. Optimisation of the bovine whole in vitro embryo system as a sentinel for toxicity screening: a cadmium challenge.

    PubMed

    Jorssen, Ellen P A; Vergauwen, Lucia; Goossens, Karen; Hagenaars, An; Van Poucke, Mario; Petro, Evi; Peelman, Luc; Knapen, Dries; Leroy, Jo L M R; Bols, Peter E J

    2015-05-01

    Developmental toxicity testing could greatly benefit from the availability of an in vitro alternative model based on the use of animal embryos that have better human-like physiology than the currently-used alternative models. These current models are insufficient, as extrapolation of the results can be challenging. Therefore, an in vitro bovine embryo culture system was used to expose individual morulae to test substances, and to study developmental characteristics up to the blastocyst stage. Cadmium was chosen as the reference toxicant to investigate the sensitivity of the bovine morulae to various concentrations and exposure times. Oocytes from slaughterhouse-obtained bovine ovaries, were maturated, fertilised and cultured up until the morula stage. Morulae were exposed to different cadmium concentrations for 18 or 70 hours, and developmental competence, embryo quality and the expression of cadmium exposure-related genes were evaluated. Cadmium exposure hampered embryonic developmental competence and quality. Compared with the 18-hour exposure, the 70-hour exposure induced a 20-fold higher toxic response with regard to developmental competence and a more 'cadmium-typical' transcript expression. The bovine morula might be a promising tool for toxicity testing as, following exposure, the embryos reacted in a sensitive and 'cadmium-typical' manner to our reference toxicant. PMID:25995012

  1. Differential developmental toxicity of naphthoic acid isomers in medaka (Oryzias latipes) embryos.

    PubMed

    Carney, Michael W; Erwin, Kyle; Hardman, Ron; Yuen, Bonny; Volz, David C; Hinton, David E; Kullman, Seth W

    2008-01-01

    Polycyclic aromatic hydrocarbons (PAHs) are widespread persistent pollutants that readily undergo biotic and abiotic conversion to numerous transformation products in rivers, lakes and estuarine sediments. Here we characterize the developmental toxicity of four PAH transformation products each structural isomers of hydroxynaphthoic acid: 1H2NA, 2H1NA, 2H3NA, and 6H2NA. Medaka fish (Oryzias latipes) embryos and eleutheroembryos were used to determine toxicity. A 96-well micro-plate format was used to establish a robust, statistically significant platform for assessment of early life stages. Individual naphthoic acid isomers demonstrated a rank order of toxicity with 1H2NA>2H1NA>2H3NA>6H2NA being more toxic. Abnormalities of circulatory system were most pronounced including pericardial edema and tube heart. To determine if HNA isomers were AhR ligands, spatial-temporal expression and activity of CYP1A was measured via in vivo EROD assessments. qPCR measurement of CYP1A induction proved different between isomers dosed at respective concentrations affecting 50% of exposed individuals (EC50s). In vitro, all ANH isomers transactivated mouse AhR using a medaka CYP1A promoter specific reporter assay. Circulatory abnormalities followed P450 induction and response was consistent with PAH toxicity. A 96-well micro-plates proved suitable as exposure chambers and provided statistically sound evaluations as well as efficient toxicity screens. Our results demonstrate the use of medaka embryos for toxicity analysis thereby achieving REACH objectives for the reduction of adult animal testing in toxicity evaluations. PMID:18433798

  2. Toxicity of seabird guano to sea urchin embryos and interaction with Cu and Pb.

    PubMed

    Rial, Diego; Santos-Echeandía, Juan; Álvarez-Salgado, Xosé Antón; Jordi, Antoni; Tovar-Sánchez, Antonio; Bellas, Juan

    2016-02-01

    Guano is an important source of marine-derived nutrients to seabird nesting areas. Seabirds usually present high levels of metals and other contaminants because the bioaccumulation processes and biotic depositions can increase the concentration of pollutants in the receiving environments. The objectives of this study were to investigate: the toxicity of seabird guano and the joint toxicity of guano, Cu and Pb by using the sea urchin embryo-larval bioassay. In a first experiment, aqueous extracts of guano were prepared at two loading rates (0.462 and 1.952 g L(-1)) and toxicity to sea-urchin embryos was tested. Toxicity was low and not dependent of the load of guano used (EC50 0.42 ± 0.03 g L(-1)). Trace metal concentrations were also low either in guano or in aqueous extracts of guano and the toxicity of extracts were apparently related to dissolved organic matter. In a second experiment, the toxicity of Cu-Pb mixtures in artificial seawater and in extracts of guano (at two loadings: 0.015 and 0.073 g L(-1)), was tested. According to individual fittings, Cu added to extracts of guano showed less toxicity than when dissolved in artificial seawater. The response surfaces obtained for mixtures of Cu and Pb in artificial seawater, and in 0.015 g L(-1) and 0.073 g L(-1) of guano, were better described by Independent Action model adapted to describe antagonism, than by the other proposed models. This implied accepting that EC50 for Cu and Pb increased with the load of guano and with a greater interaction for Cu than for Pb. PMID:26692516

  3. Marine toxicity tests development with a New Zealand echinoid

    SciTech Connect

    Nipper, M.G.; Roper, D.S.; Martin, M.L.; Williams, E.K.

    1995-12-31

    The generally low levels of contamination around New Zealand lead to the search for a sensitive toxicity test, which could be used to screen effluent and to detect contaminant effects in coastal waters and sediments. Echinoid early life stage tests were considered ideal candidates. However, the adaptation of international toxicity test methods to indigenous species has not been straightforward or troublefree! The echinoid Fellaster zelandiae was selected because it is abundant around New Zealand and is fertile year round. Fertilization tests showed that gamete density, rather than sperm/egg ratio, was a crucial factor for successful control fertilization rates. This method, however, presented several problems related to (1) temporal variability in the quality of sperm batches, (2) rapid reduction of egg quality and viability after spawning, and (3) highly variable sensitivity in reference toxicant tests. Embryo tests were more reliable, with good control results (> 80% normal embryos) and consistent sensitivity to a reference toxicant, zinc sulfate. EC{sub 50} values averaged 0.06 mg Zn/L, comparable to the sensitivity of echinoid species used elsewhere. Brine prepared by freezing seawater was suitable for adjusting the salinity of effluents, with more than 90% normal embryos developing in brine diluted with UV-treated deionized water as a test-control. The assessment of the embryo development test as a tool for screening sediment toxicity (using sediment pore water), is presently underway, concurrently with growth and behavioral endpoint tests using indigenous amphipods and bivalves.

  4. Zebrafish Embryo Toxicity Microscale Model for Ichthyotoxicity Evaluation of Marine Natural Products.

    PubMed

    Bai, Hong; Kong, Wen-Wen; Shao, Chang-Lun; Li, Yun; Liu, Yun-Zhang; Liu, Min; Guan, Fei-Fei; Wang, Chang-Yun

    2016-04-01

    Marine organisms often protect themselves against their predators by chemical defensive strategy. The second metabolites isolated from marine organisms and their symbiotic microbes have been proven to play a vital role in marine chemical ecology, such as ichthyotoxicity, allelopathy, and antifouling. It is well known that the microscale models for marine chemoecology assessment are urgently needed for trace quantity of marine natural products. Zebrafish model has been widely used as a microscale model in the fields of environment ecological evaluation and drug safety evaluation, but seldom reported for marine chemoecology assessment. In this work, zebrafish embryo toxicity microscale model was established for ichthyotoxicity evaluation of marine natural products by using 24-well microplate based on zebrafish embryo. Ichthyotoxicity was evaluated by observation of multiple toxicological endpoints, including coagulation egg, death, abnormal heartbeat, no spontaneous movement, delayed hatch, and malformation of the different organs during zebrafish embryogenesis periods at 24, 48, and 72 h post-fertilization (hpf). 3,4-Dichloroaniline was used as the positive control for method validation. Subsequently, the established model was applied to test the ichthyotoxic activity of the compounds isolated from corals and their symbiotic microbes and to isolate the bioactive secondary metabolites from the gorgonian Subergorgia mollis under bioassay guidance. It was suggested that zebrafish embryo toxicity microscale model is suitable for bioassay-guided isolation and preliminary bioactivity screening of marine natural products. PMID:26838966

  5. Assessment of biocompatibility of 3D printed photopolymers using zebrafish embryo toxicity assays.

    PubMed

    Macdonald, N P; Zhu, F; Hall, C J; Reboud, J; Crosier, P S; Patton, E E; Wlodkowic, D; Cooper, J M

    2016-01-01

    3D printing has emerged as a rapid and cost-efficient manufacturing technique to enable the fabrication of bespoke, complex prototypes. If the technology is to have a significant impact in biomedical applications, such as drug discovery and molecular diagnostics, the devices produced must be biologically compatible to enable their use with established reference assays and protocols. In this work we demonstrate that we can adapt the Fish Embryo Test (FET) as a new method to quantify the toxicity of 3D printed microfluidic devices. We assessed the biocompatibility of four commercially available 3D printing polymers (VisiJetCrystal EX200, Watershed 11122XC, Fototec SLA 7150 Clear and ABSplus P-430), through the observation of key developmental markers in the developing zebrafish embryos. Results show all of the photopolymers to be highly toxic to the embryos, resulting in fatality, although we do demonstrate that post-printing treatment of Fototec 7150 makes it suitable for zebrafish culture within the FET. PMID:26646354

  6. Drosophila Embryos as Model to Assess Cellular and Developmental Toxicity of Multi-Walled Carbon Nanotubes (MWCNT) in Living Organisms

    PubMed Central

    Liu, Boyin; Campo, Eva M.; Bossing, Torsten

    2014-01-01

    Different toxicity tests for carbon nanotubes (CNT) have been developed to assess their impact on human health and on aquatic and terrestrial animal and plant life. We present a new model, the fruit fly Drosophila embryo offering the opportunity for rapid, inexpensive and detailed analysis of CNTs toxicity during embryonic development. We show that injected DiI labelled multi-walled carbon nanotubes (MWCNTs) become incorporated into cells in early Drosophila embryos, allowing the study of the consequences of cellular uptake of CNTs on cell communication, tissue and organ formation in living embryos. Fluorescently labelled subcellular structures showed that MWCNTs remained cytoplasmic and were excluded from the nucleus. Analysis of developing ectodermal and neural stem cells in MWCNTs injected embryos revealed normal division patterns and differentiation capacity. However, an increase in cell death of ectodermal but not of neural stem cells was observed, indicating stem cell-specific vulnerability to MWCNT exposure. The ease of CNT embryo injections, the possibility of detailed morphological and genomic analysis and the low costs make Drosophila embryos a system of choice to assess potential developmental and cellular effects of CNTs and test their use in future CNT based new therapies including drug delivery. PMID:24558411

  7. Developmental toxicity of three hexabromocyclododecane diastereoisomers in embryos of the marine medaka Oryzias melastigma.

    PubMed

    Hong, Haizheng; Shen, Rong; Liu, Wanxin; Li, Dongmei; Huang, Lingming; Shi, Dalin

    2015-12-15

    The composition of major hexabromocyclododecane (HBCD) diastereoisomers, i.e. ?-, ?-, and ?-HBCDs, in marine biota is different from that of the commercially available form (technical HBCD), which is used extensively for toxicological studies. To properly evaluate the impact of HBCDs, the embryos of Oryzias melastigma were used to examine the developmental toxicity of the individual diastereoisomers. Results showed that HBCD diastereoisomers at the environmentally realistic concentrations in the embryos induced malformation rate and heartbeat, and caused the appearance of apoptotic heart. In addition, ?-, ?-, and ?-HBCDs had similar potency to stimulate the generation of reactive oxygen species, consequently leading to apoptosis in O. melastigma embryos. The order of the developmental toxicity of ?-, ?-, and ?-HBCDs in O. melastigma embryos was different from that in zebrafish embryos studied previously, which highlighted the importance of using species from both fresh and salt water for toxicity assessment. PMID:26563546

  8. VIRTUAL EMBRYO: SYSTEMS MODELING IN DEVELOPMENTAL TOXICITY - Symposium: SOT 2012

    EPA Science Inventory

    High-throughput screening (HTS) studies are providing a rich source of data that can be applied to in vitro profiling of chemical compounds for biological activity and potential toxicity. Chemical profiling in ToxCast covered 965 drugs-chemicals in over 500 diverse assays testing...

  9. Toxicity of binary mixtures of oil fractions to sea urchin embryos.

    PubMed

    Rial, Diego; Vázquez, José A; Menduiña, Araceli; García, Ana M; González, M Pilar; Mirón, Jesús; Murado, Miguel A

    2013-12-15

    The assumption of additive toxicity for oil compounds is related to a narcotic mode of action. However, the joint toxicity of oil fractions has not been fully investigated. A fractionation of Maya crude oil into aliphatics, aromatics and polars was performed, fractions were dissolved in dimethyl sulfoxide (DMSO) and subsequently toxicity of single fractions and binary mixtures was assessed using the sea urchin embryo test. The descriptive ability of Concentration Addition (CA), Independent Action (IA) and modifications of both models for describing the joint toxicity of mixtures has also been evaluated. The hydrocarbon content extractable with dichloromethane of the fractions dissolved in DMSO was: 12.0 ± 1.8 mg mL(-1), 39.0 ± 0.5 mg mL(-1) and 20.5 ± 2.5 mg mL(-1) for aliphatics, aromatics and polars, respectively. The toxicity of the extracts in DMSO of the fractions as EC50 (μLL(-1)) was: aliphatics (165.8-242.3)toxicity of oil fractions. In addition, synergistic or antagonistic effects were not observed. PMID:24231335

  10. Effects of photoinduced toxicity of fluoranthene on amphibian embryos and larvae

    SciTech Connect

    Hatch, A.C.; Burton, G.A. Jr.

    1998-09-01

    Embryos and newly hatched larvae of three amphibian species, the spotted salamander (Ambystoma maculatum), the northern leopard frog (Rana pipiens), and the African clawed frog (Xenopus laevis), were exposed to fluoranthene and ultraviolet (UV) light in two scenarios. Embryos were exposed in a laboratory setting from an early developmental stage through hatching under artificial UV light, and newly hatched larvae were exposed outdoors in varying sunlight intensity levels. Outdoor exposures indicated greater sensitivity in the toxic response than did laboratory exposures. In the laboratory, mortality and malformation of X. laevis were the most sensitive indicators of exposure. Xenopus laevis was also the most sensitive species tested to the effects of UV light alone. Hatching success of R. pipiens was monitored outdoors and was not a useful predictive endpoint in the determination of photoinduced toxicity; however, newly hatched larvae were sensitive to the effects of photoinduced toxicity. Amybstoma maculatum and X. laevis larvae were affected by low ({micro}g/L) concentrations of fluoranthene in sunlight. These findings suggest that low levels of polycyclic aromatic hydrocarbons could be acting synergistically with environmental factors such as UV light to place young amphibians at risk.

  11. Effects of environmental toxicants on development of a teleost embryo

    SciTech Connect

    Crawford, R.B.; Guarino, A.M.

    1985-11-01

    Embryos of the teleost Fundulus heteroclitus are shown to be useful model systems for monitoring the effects of xenobiotic compounds on development. Fourteen different substances were tested: malathion, aroclor, aldrin, diquat, parathion, pentachlorophenol, sevin, toxaphene, lindane, 2,4-D, DDT, paraquat, 2,4,5-T, and aminotriazole. Concentrations used for each of these was from 0.01 to 10.0 ppm in the incubation dishes. The variety of effects on development observed depended on the compound and its concentration. These effects included inhibition of gastrulation, abnormal axis formation, diminished pigmentation, slowed rate of development, reduced frequency of hatching, loss of neuromuscular control, and reduction or inhibition of heart beat. Possible modes of action of some of these compounds are discussed. It is also shown that embryogenesis is not always the most susceptible part of the organism's life cycle.

  12. Effects of storage method and duration on the toxicity of marine sediments to embryos of Crassostrea gigas oysters.

    PubMed

    Geffard, O; His, E; Budzinski, H; Chiffoleau, J F; Coynel, A; Etcheber, H

    2004-06-01

    The objective of laboratory sediment bioassays is to estimate in situ toxicity. This goal is difficult to achieve, as one of the main limitations of sediment toxicity tests is disruption of sediment geochemistry during sampling, handling and preservation. The effects of storage on the estimation of marine sediment toxicity to Crassostrea gigas embryos and larvae were investigated. Three storage methods and four storage periods were compared with three different sediment types contaminated by heavy metals, polycyclic aromatic hydrocarbons (PAHs) and both contaminants. Freezing and freeze-drying considerably increased the toxicity of decanted sediments and their elutriates as compared to the toxicity obtained with fresh sediments. Concerning the elutriates, the toxicity found with frozen and freeze-dried sediments was correlated with DOC, ammonia and PAH contents. However, the toxicity of fresh sediments kept at 4 degrees C increased with increasing duration of storage and was also correlated with the amount of ammonia in the elutriates. PMID:15016466

  13. DEVELOPMENTAL TOXICITY OF DI- AND TETRACHLOROETHANE AND DICHLOROPROPANE IN EMBRYO CULTURE

    EPA Science Inventory

    DEVELOPMENTAL TOXICITY OF DI- AND TETRACHLOROETHANE AND DICHLOROPROPANE IN EMBRYO CULTURE. JE Andrews, H Nichols, and ES Hunter. Reproductive Toxicology Division, NHEERL, USEPA, RTP, NC.

    Disinfection of drinking water with chlorine results in numerous chlorinated byprodu...

  14. Mixture toxicity of water contaminants-effect analysis using the zebrafish embryo assay (Danio rerio).

    PubMed

    Schmidt, Susanne; Busch, Wibke; Altenburger, Rolf; Küster, Eberhard

    2016-06-01

    Three water contaminants were selected to be tested in the zebrafish embryo toxicity test (DarT) in order to investigate the sensitivity of the zebrafish embryo toxicity test with respect to mixture effect detection. The concentration-response curves for the observed effects lethality and hypo-pigmentation were calculated after an exposure of the embryos for 96 h with a fungicide (carbendazim), a plasticizer or propellent precursor (2,4-DNT: 2,4- dinitrotoluene) and an aromatic compound (AαC: 2-amino-9H-pyrido[2,3-b]indol), respectively. Follow-up mixture tests were based on the calculated LC50 or EC50 of the single compounds and combined effects were predicted according to the mixture concepts of concentration addition (CA) and independent action (IA). The order of toxicity for the single substances was carbendazim (LC50 = 1.25 μM) < AαC (LC50 = 8.16 μM) < 2,4-DNT (LC50 = 177.05 μM). For AαC and 2,4 DNT hypo-pigmentation was observed in addition (AαC EC50 = 1.81 μM; 2,4-DNT EC50 = 8.81 μM). Two binary and one ternary mixture were studied on lethality and one on hypo-pigmentation: 2,4-DNT/AαC (LC50 = 119.21 μM, EC50 = 5.37 μM), carbendazim/AαC (LC50 = 4.49 μM) and AαC/Carbendazim/2,4 DNT (LC50 = 108.62 μM). Results showed that the effects were in agreement with the CA model when substances were tested in mixtures. Therefore, in a reasonable worst case scenario substance combination effects in fish embryos were at maximum only prone to overestimation when using CA as the mixture concept. PMID:27011319

  15. Toxicity of buprofezin on the survival of embryo and larvae of African catfish, Clarias gariepinus (Bloch).

    PubMed

    Marimuthu, Kasi; Muthu, Narmataa; Xavier, Rathinam; Arockiaraj, Jesu; Rahman, M Aminur; Subramaniam, Sreeramanan

    2013-01-01

    Buprofezin is an insect growth regulator and widely used insecticide in Malaysia. The present study evaluated the toxic effects of buprofezin on the embryo and larvae of African catfish (Clarias gariepinus) as a model organism. The embryos and larvae were exposed to 7 different concentrations (0, 0.05, 0.5, 5, 25, 50 and 100 mg/L) of buprofezin. Each concentration was assessed in five replicates. Eggs were artificially fertilized and 200 eggs and larvae were subjected to a static bath treatment for all the concentrations. The mortality of embryos was significantly increased with increasing buprofezin concentrations from 5 to 100 mg/L (p< 0.05). However, the mortality was not significantly different (p<0.05) among the following concentrations: 0 (control), 0.05, 0.5 and 5 mg/L. Data obtained from the buprofezin acute toxicity tests were evaluated using probit analysis. The 24 h LC50 value (with 95% confidence limits) of buprofezin for embryos was estimated to be 6.725 (3.167-15.017) mg/L. The hatching of fish embryos was recorded as 68.8, 68.9, 66.9, 66.4, 26.9, 25.1 and 0.12% in response to 7 different concentrations of buprofezin, respectively. The mortality rate of larvae significantly (p<0.05) increased with increasing buprofezin concentrations exposed to 24-48 h. The 24 and 48 h LC50 values (with 95% confidence limits) of buprofezin for the larvae was estimated to be 5.702 (3.198-8.898) and 4.642 (3.264-6.287) mg/L respectively. There were no significant differences (p>0.05) in the LC50 values obtained at 24 and 48 h exposure times. Malformations were observed when the embryos and larvae exposed to more than 5 mg/L. The results emerged from the study suggest that even the low concentration (5 mg/L) of buprofezin in the aquatic environment may have adverse effect on the early embryonic and larval development of African catfish. PMID:24098390

  16. Rethinking guideline toxicity testing.

    PubMed

    Saghir, Shakil Ahmed

    2015-07-01

    The guidelines for risk assessment of plant protection products (PPPs) and other non-pharmaceuticals were developed over three decades ago and have generally not been updated to incorporate advancements in toxicology and exposure sciences. These guidelines recommend using maximum-tolerated-dose (MTD) even when human relevance of such high doses is mostly limited due to orders of magnitude margin-of-exposure. Conducting animal studies at such high doses often requires further mode-of-action (MoA) studies elucidating human relevance. In order to improve data, ILSI/HESI-ACSA technical committee proposed a tiered approach with emphasis on determining systemic dose of parent and/or metabolite(s) in test animals as biological effects are reflective of systemic rather than administered dose. Any deviation from linearity in systemic dose (saturation of absorption or elimination) in animal studies may have profound toxic effect(s) not expected to occur in likely human exposure scenarios and should be avoided. Toxicity studies should ideally be conducted at kinetically linear doses or slightly above the point of departure from linearity or kinetically-derived maximum dose (KMD) as the systemic dose nonlinearity is a more sensitive parameter occurring much earlier than the MTD endpoints. Therefore, determining systemic dose, especially KMD, in study animals is an improvement to hazard assessment of PPPs and other non-pharmaceuticals allowing toxicologists to better understand findings in animals at systemically linear as well as nonlinear doses to likely human exposures which can easily be accomplished using core study animals as outlined below. Determining systemic dose in studies will also increase the understanding of initial potential MoA of a PPPs and other non-pharmaceuticals and reduce the use of animals by avoiding unnecessary additional MoA studies. PMID:25980640

  17. Evaluation of the toxicity and teratogenity of six commercial textile dyes using the frog embryo teratogenesis assay-Xenopus.

    PubMed

    Birhanli, Ayse; Ozmen, Murat

    2005-01-01

    Potential developmental toxicities of six different textile dyes were evaluated using the frog embryo teratogenesis assay-Xenopus (FETAX). Xenopus laevis embryos were exposed to astrazon red FBL, astrazon blue FGRL, remazol red RR, remazol turquoise blue G-A, cibacron red FN-3G, and cibacron blue FN-R from stage 8 to 11 for a 96-h exposure period in static renewal test conditions. A minimum of 17 concentration-response tests were performed with tested dyes, excluding a control group for each dye. Median lethal concentration (LC50), malformation (EC50), non observed adverse effect concentration (NOAEC), and lowest observed adverse effect concentration (LOAEC) were calculated. Also, teratogenic index (TI), minimum concentration to inhibit growth (MCIG), and MCIG/LC50 values were determined for each of the tested dyes. Characteristic abnormalities induced by a given test material were determined by the relationship between concentration and dye in the study. Results from these studies suggested that each tested dye is teratogenic for X. laevis embryos. The lowest LC50 was determined for astrazon red exposure corresponding to a value of 4.73 mg/L. The LC50 value was similar for this dye and astrazon blue; the highest TI was calculated for astrazon blue exposure. Tests with X. laevis indicated that each of the tested compounds possessed teratogenic potential with varying degrees of potency: astrazon blue FGRL > remazol turquoise blue G-A > astrazon red FBL > cibacron blue FN-R > cibacron red FN-3G > remazol red RR. Different types of malformations occurred in the embryos, depending on concentration and dye. From these results, we can suggest that astrazon blue is the most toxic compound, but that the others are also highly toxic and teratogenic substances for X. laevis embryos. Results of the study confirmed that the FETAX assay can be useful in an integrated biological hazard assesment for the preliminary screening of textile dye stuff. PMID:15720035

  18. The VIRTUAL EMBRYO. A Computational Framework for Developmental Toxicity

    EPA Science Inventory

    EPAs Virtual Embryo Project (v-Embryo) is focused on the predictive toxicology of childrens health and developmental defects following prenatal exposure to environmental chemicals. The research is motivated by scientific principles in systems biology as a framework for the g...

  19. Gene expression profiling in zebrafish embryos exposed to diclofenac, an environmental toxicant.

    PubMed

    De Felice, Bruna; Copia, Luisa; Guida, Marco

    2012-03-01

    Pharmaceuticals are continually released in the environment and therefore pollution from drugs is a pressing problem in the environment. Diclofenac, 2-[(2,6-dichlorophenyl)amino]phenylacetic acid is a FDA approved non-steroidal anti-inflammatory drug (NSAID) for the treatment of inflammation. This pharmaceutical has been found as pollutant in superficial waters. Danio rerio (zebrafish) embryo has been used as a model organism for acute pollutant toxicity tests in order to identify morphological alterations in development and death rate. Through the combination of mRNA differential display and quantitative Real Time experiments, we analyzed the alterations of gene expression in zebrafish embryos left to develop in the presence of diclofenac and thereby assess the molecular mechanism involved in ecotoxicity of diclofenac polluted waters. This approach, in embryos exposed to 1.25 mg/l drug for 48 h, allowed identifying 36 different genes, with both known and unknown functions, whose transcription is differentially regulated. The identity and ontological classification of these genes is presented. The wide variety of functional classes of transcripts isolated in this screen reflects the diverse spectrum of influences operating across diclofenac exposure. Of these 36 genes, several have been selected for detailed quantitative Real Time analysis to validate the screen. Our results, for the first time, provide an insight into some of the varied and novel molecular networks following zebrafish exposure to diclofenac polluted waters. PMID:21643954

  20. Developmental toxic effects of monocrotophos, an organophosphorous pesticide, on zebrafish (Danio rerio) embryos.

    PubMed

    Pamanji, Rajesh; Bethu, M S; Yashwanth, B; Leelavathi, S; Venkateswara Rao, J

    2015-05-01

    The present study examined the response of zebrafish embryos exposed to different concentrations (10, 20, 30, 40, 50, and 60 mg/L) of monocrotophos under static conditions for 96 h. We found that mortality had occurred within 48 h at all test concentrations, later insignificant mortality was observed. Monocrotophos (MCP) can be rated as moderately toxic to the Zebrafish embryos with a 96-h median lethal concentration (LC50) of 37.44??3.32 mg/L. In contrast, it greatly affected the development of zebrafish embryos by inducing several developmental abnormalities like pericardial edema, altered heart development, spinal and vertebral anomalies in a concentration-dependent manner. A significant percent reduction in length by 9-48% and heart beats by 18-51% was observed in hatchlings exposed to LC10 and LC50 concentrations at 96 h when compared to controls. The process of looping formation of heart at embryonic stage was greatly affected by the LC50 concentration of MCP. The neurotoxic potentiality of MCP was assessed by using a marker enzyme, acetylcholinesterase in both in vitro and in vivo experiments. MCP was found to be the most potent inhibitor of AChE in vitro with an IC50 value of 4.3??10(-4) M. The whole-body AChE enzyme activity in vivo was significantly inhibited during the exposure tenure with the maximum inhibition of 62% at 24 h. PMID:25604565

  1. Toxicity test method development in southeast Asia

    SciTech Connect

    McPherson, C.A.

    1995-12-31

    Use of aquatic toxicity tests is relatively new in southeast Asia. As part of the ASEAN-Canada Cooperative Programme on Marine Science -- Phase 2, which includes development of marine environmental criteria, a need for tropical toxicity data was identified. A step-wise approach was used for test method development (simple, acute tests and easily measured endpoints first, then more complex short-term chronic methods), for test specific selection (using species found throughout the region first, and then considering species with narrower geographic distribution), and for integration of quality assurance/quality control (QA/QC) practices into all laboratory activities. Development of test protocols specifically for tropical species included acute and chronic toxicity tests with marine fish, invertebrates and algae. Criteria for test species selection will be reviewed. Method development was based on procedures and endpoints already widely used in North America and Europe (e.g., 96-h LC50 with fish), but adapted for use with tropical species. For example, a bivalve larval development test can use the same endpoints but the duration is only 24 hours. Test method development included research on culture and holding procedures, determination of test conditions (e.g., duration, test containers), and identification of appropriate endpoints. Acute tests with fish and invertebrates were developed first. The next step was development of short-term chronic tests to measure phytoplankton growth, bivalve and echinoderm embryo or larval development, and larval fish growth. The number of species and types of tests was increased in a staged approach, as laboratories became better equipped and personnel gained practical experience. In most cases, method development coincided with training workshops to introduce the principles of toxicity testing.

  2. Artifacts in ambient toxicity testing

    SciTech Connect

    Kszos, L.A.; Stewart, A.J.

    1992-01-01

    Short-term toxicity tests with fathead minnow (Pimephales promelas) larvae and Ceriodaphnia dubia can be used to estimate the acute or chronic toxicity of effluents or receiving water. The results of effluent toxicity tests may need to be interpreted differently from the results of ambient toxicity tests. In this paper we provide examples of common artifacts, which can cause either false positives or false negatives, that we have encountered when these tests are used in ambient assessments. The examples we provide are drawn from diverse effluent and ambient water toxicity tests conducted at the Oak Ridge National Laboratory from March, 1985 through November, 1991. Three types of artifacts which have been encountered when using these tests in ambient applications are explored here. One type involves unusual replicate-specific variance in survival of fathead minnow larvae. The second and third types of artifacts affect the C. dubia test and appear to be related to food availability.

  3. Artifacts in ambient toxicity testing

    SciTech Connect

    Kszos, L.A.; Stewart, A.J.

    1992-10-01

    Short-term toxicity tests with fathead minnow (Pimephales promelas) larvae and Ceriodaphnia dubia can be used to estimate the acute or chronic toxicity of effluents or receiving water. The results of effluent toxicity tests may need to be interpreted differently from the results of ambient toxicity tests. In this paper we provide examples of common artifacts, which can cause either false positives or false negatives, that we have encountered when these tests are used in ambient assessments. The examples we provide are drawn from diverse effluent and ambient water toxicity tests conducted at the Oak Ridge National Laboratory from March, 1985 through November, 1991. Three types of artifacts which have been encountered when using these tests in ambient applications are explored here. One type involves unusual replicate-specific variance in survival of fathead minnow larvae. The second and third types of artifacts affect the C. dubia test and appear to be related to food availability.

  4. Differential toxicity of three PCB congeners in developing sea urchin embryos and implication of TEQ approach

    SciTech Connect

    Schweitzer, L.; Suffet, I.; Hose, J.; Bay, S.

    1995-12-31

    The relationship between body burden and toxicity of three individual PCB congeners in developing sea urchin embryos was investigated to evaluate the validity of current predictive models of PCB toxicity in an invertebrate system. The uptake and accumulation of radiolabeled PCB congeners from sea water was measured in the sea urchin embryo tissues and the relative toxicity determined. According to the toxic equivalents (TEQ) approach of assessing risk to mammals, congener 77, a nonortho-substituted congener, is predicted to be more toxic than the diortho-substituted congeners 47 and 153. Using a 72 hour embryo development assay, congener 47 was found to be at least four times as toxic as congener 77, with EC50s of 15.7 and > 72.5 mmol/kg, respectively. Congener 153, a hexachlorobiphenyl, was virtually nontoxic even at the highest dose used. Cytologic and cytogenetic anomalies were studied to find a possibly more sensitive endpoint and to suggest a mechanism of toxicity. The cytogenetic analysis revealed that the PCBs inhibited mitosis. At the highest doses, complete mitotic arrest was observed. Congener 77 was found to be at least two times more toxic than congener 153 but not as toxic as congener 47 using mitotic activity as the endpoint. Thus, the two endpoints of toxicity did not change the order in which the congeners are toxic, but established different EC50s. The relative toxicities of these congeners in this study contradict the structure-activity prediction of the mammalian-based TEQ approach.

  5. Bivalve embryo bioassay to assess the potential toxicity of dredged material before dumping

    SciTech Connect

    Quiniou, F.

    1995-12-31

    Dredged harbor sediments frequently contain a wide spectrum of contaminants in addition to a significant percentage of organic matter. Also, dredging and dumping activities into sea water, of these highly contaminated soil may induce a harmful effect on the environment. In France, in accordance with Oslo convention guidelines, a working group on dredging activities and environment (GEODE) created since 1991 decided to set up a pilot research program to assess the intrinsic toxicity of four harbor sludges. Intrinsic toxicity of harbor muds were tested by solid phase (whole sediment) and aqueous extract bioassays (sea water elutriates) using the sublethal toxicity test bivalve embryo bioassay (Crassostrea gigas). Elutriates enable them to detect the toxicity of contaminants which may be released in the soluble form into the water column during dredging operations. While, whole sediment integrate the synergistic effects of all the contaminants (hydrophilic and hydrophobic) including pore water. Bioassays results, correlated to chemical analysis, are compared to contaminant levels determined by French working group GEODE and Canadian sediment quality criteria.

  6. Effects of storage temperature and duration on toxicity of sediments assessed by Crassostrea gigas oyster embryo bioassay

    SciTech Connect

    Beiras, R.; His, E.; Seaman, M.N.L.

    1998-10-01

    The effects of temperature and duration of storage on the toxicity of estuarine sediments were investigated with the Crassostrea gigas oyster embryo bioassay. Sediments ranging from unpolluted (controls) to extremely polluted with heavy metals (>100 ppm Hg, Cu, Zn, and Pb) and total hydrocarbons (>1,000 ppm) were collected from sites in southwest France and northern Spain, Control sediments were toxic only at the highest concentrations tested and after freezing in liquid nitrogen ({minus}196 C). Polluted sediments significantly reduced the success of oyster embryogenesis. Analysis of variance showed that the effect of storage temperature on toxicity increased with the prolongation of storage. Prolonged storage of fresh (4 C) sediments resulted in a loss of toxicity, which was more rapid in the less-polluted sediments. Deep-frozen sediments ({minus}196 C) were highly toxic regardless of origin and storage time, and because deep-freezing causes spurious toxicity in the control samples, it cannot be recommended for toxicological studies. In the context of the assessment of sediment toxicity by embryo-larval bioassays, fresh (4 C) storage is recommended when sediments need to be stored for no longer than a few days. The advisable duration of fresh storage to avoid false-negative results is directly related to the degree of toxicity. Should the sediments require prolonged storage, freezing at {minus}20 C appears to be the best choice.

  7. Nickel and Copper Toxicity to Embryos of the Long-Spined Sea Urchin, Diadema savignyi.

    PubMed

    Rosen, G; Rivera-Duarte, I; Colvin, M A; Dolecal, R E; Raymundo, L J; Earley, P J

    2015-07-01

    The sensitivity of long-spined sea urchins (Diadema savignyi) collected from Guam (Northern Marianas Islands), USA, to nickel and copper in seawater was explored using 48-h embryo-larval development toxicity tests. The median effective concentrations (EC50) averaged 94 µg L(-1) for nickel, and 19 µg L(-1) from a single exposure to copper, and suggest relatively high sensitivity of this species to nickel compared with other sea urchin genera, but similar sensitivity to copper. Ambient nickel and copper concentrations concurrently sampled from 16 near-shore locations around Guam were one to two orders of magnitude lower than those that would be expected to result in adverse effects to D. savignyi embryos. Although nationally recommended chronic ambient water quality criteria, currently 8.2 and 3.1 µg L(-1) for nickel and copper, respectively, were not exceeded, recently derived qualifying toxicity data should be considered for updating these criteria to ensure protectiveness of sensitive tropical species. PMID:25573279

  8. vEmbryo In Silico Models: Predicting Vascular Developmental Toxicity

    EPA Science Inventory

    The cardiovascular system is the first to function in the vertebrate embryo, reflecting the critical need for nutrient delivery and waste removal during organogenesis. Blood vessel development occurs by complex interacting signaling networks, including extra-cellular matrix remod...

  9. RAPID AQUATIC TOXICITY ASSAY UTILIZING LABELED THYMIDINE INCORPORATION IN SEA URCHIN EMBRYOS

    EPA Science Inventory

    Aquatic toxicity was evaluated in the sea urchin embryo (Arbacea punctulata) by the inhibition of tritiated thymidine incorporation after a brief exposure to toxic chemicals. Arbacia is a useful surrogate species for assay: well-studied, easily cultured and fertile virtually year...

  10. Comparative toxicity of lead (Pb2+), copper (Cu2+), and mixtures of lead and copper to zebrafish embryos on a microfluidic chip

    PubMed Central

    Li, Yinbao; Yang, Xiujuan; Zhang, Beibei; Pan, Jianbin; Li, Xinchun; Yang, Fan; Sun, Duanping

    2015-01-01

    Investigations were conducted to determine acute effects of Pb2+ and Cu2+ presented individually and collectively on zebrafish embryos. Aquatic safety testing requires a cheap, fast, and highly efficient platform for real-time evaluation of single and mixture of metal toxicity. In this study, we have developed a microfluidic system for phenotype-based evaluation of toxic effects of Pb2+ and Cu2+ using zebrafish (Danio rerio) embryos. The microfluidic chip is composed of a disc-shaped concentration gradient generator and 24 culture chambers, which can generate one blank solution, seven mixture concentrations, and eight single concentrations for each metal solution, thus enabling the assessment of zebrafish embryos. To test the accuracy of this new chip platform, we have examined the toxicity and teratogenicity of Pb2+ and Cu2+ on embryos. The individual and combined impact of Pb2+ and Cu2+ on zebrafish embryonic development was quantitatively assessed by recording a series of physiological indicators, such as spontaneous motion at 22 hours post fertilization (hpf), mortality at 24 hpf, heartbeat and body length at 96 hpf, etc. It was found that Pb2+ or Cu2+ could induce deformity and cardiovascular toxicity in zebrafish embryos and the mixture could induce more severe toxicity. This chip is a multiplexed testing apparatus that allows for the examination of toxicity and teratogenicity for substances and it also can be used as a potentially cost-effective and rapid aquatic safety assessment tool. PMID:25825620

  11. Potential toxicity of environmentally relevant perfluorooctane sulfonate (PFOS) concentrations to yellow-legged gull Larus michahellis embryos.

    PubMed

    Parolini, Marco; Colombo, Graziano; Valsecchi, Sara; Mazzoni, Michela; Possenti, Cristina Daniela; Caprioli, Manuela; Dalle-Donne, Isabella; Milzani, Aldo; Saino, Nicola; Rubolini, Diego

    2016-01-01

    Perfluooctane sulfonate (PFOS) is considered an emerging pollutant because of its wide distribution in both aquatic and terrestrial ecosystems, as well as its potential toxicity to living organisms. Although PFOS environmental levels and the adverse effects on classical model organisms in toxicological studies are well known, including developmental alterations and alteration of oxidative status, its toxicity to free-living species has been seldom investigated. The aim of this study was to assess the potential toxicity of environmental levels of PFOS to yellow-legged gull (Larus michahellis) embryos under field experimental conditions. In a within-clutch experimental design, we injected two PFOS concentrations (100 ng PFOS/g egg weight and 200 ng PFOS/g egg weight) in ovo soon after laying. Eggs were collected when they reached the cracking stage. We investigated the effects of PFOS treatment, laying order and sex on both morphological and biochemical endpoints of embryos. Specifically, we assessed changes in embryo body mass and tarsus length, as well as in liver and brain mass. Moreover, the imbalance of oxidative status was evaluated in both liver and brain from embryos by measuring total antioxidant capacity (TAC) and total oxidant status (TOS), while the levels of protein carbonyl content (PCO) and DNA fragmentation were measured as oxidative and genetic damage endpoints, respectively. The concentrations of PFOS we tested did not significantly alter the morphological endpoints, independently of laying order and sex. Similarly, embryo oxidative status and oxidative and genetic damage were not significantly affected by PFOS in ovo exposure. These findings suggest that current environmental PFOS levels do not affect early development of yellow-legged gull embryos. PMID:26310703

  12. Influence of sediment composition on PAH toxicity using zebrafish (Danio rerio) and Japanese medaka (Oryzias latipes) embryo-larval assays.

    PubMed

    Perrichon, Prescilla; Le Bihanic, Florane; Bustamante, Paco; Le Menach, Karyn; Budzinski, Hélène; Cachot, Jérôme; Cousin, Xavier

    2014-12-01

    Due to hydrophobic and persistent properties, polycyclic aromatic hydrocarbons (PAHs) have a high capacity to accumulate in sediment. Sediment quality criteria, for the assessment of habitat quality and risk for aquatic life, include understanding the fate and effects of PAHs. In the context of European regulation (REACH and Water Framework Directive), the first objective was to assess the influence of sediment composition on the toxicity of two model PAHs, benzo[a]pyrene and fluoranthene using 10-day zebrafish embryo-larval assay. This procedure was undertaken with an artificial sediment in order to limit natural sediment variability. A suitable sediment composition might be then validated for zebrafish and proposed in a new OECD guideline for chemicals testing. Second, a comparative study of toxicity responses from this exposure protocol was then performed using another OECD species, the Japanese medaka. The potential toxicity of both PAHs was assessed through lethal (e.g., survival, hatching success) and sublethal endpoints (e.g., abnormalities, PMR, and EROD) measured at different developmental stages, adapted to the embryonic development time of both species. Regarding effects observed for both species, a suitable artificial sediment composition for PAH toxicity testing was set at 92.5 % dry weight (dw) silica of 0.2-0.5-mm grain size, 5 % dw kaolin clay without organic matter for zebrafish, and 2.5 % dw blond peat in more only for Japanese medaka. PAH bioavailability and toxicity were highly dependent on the fraction of organic matter in sediment and of the K ow coefficients of the tested compounds. The biological responses observed were also dependent of the species under consideration. Japanese medaka embryos appeared more robust than zebrafish embryos for understanding the toxicity of PAHs following a sediment contact test, due to the longer exposure duration and lower sensitivity of sediment physical properties. PMID:25175355

  13. USING THE MEDAKA EMBRYO ASSAY TO INVESTIGATE DEVELOPMENTAL ETHANOL TOXICITY.

    EPA Science Inventory

    Ethanol (EtOH) is a well-known developmental toxicant that produces a range of abnormal phenotypes. While the toxic potential of developmental EtOH exposure is well characterized, the effect of the timing of exposure on the extent of toxicity remains unknown. Fish models such as ...

  14. The VIRTUAL EMBRYO. A Computational Framework for Developmental Toxicity

    EPA Science Inventory

    EPA’s ‘Virtual Embryo Project’ (v-Embryo™) is focused on the predictive toxicology of children’s health and developmental defects following prenatal exposure to environmental chemicals. The research is motivated by scientific principles in systems biology as a framework for the g...

  15. Toxic effects of phencyclidine on developing chick embryo brain.

    PubMed

    Ahmad, G; Zamenhof, S

    1987-09-14

    We studied the effects of Phencyclidine (PCP, Angel Dust) on the developing chick embryo brain. In Group-1, the eggs were injected with PCP on the 7th day of incubation and the embryo brains were studied on the 10th day. In Group-2, eggs were injected twice; first on the 7th day and then on the 10th day of incubation. Group-2 brains were then studied on the 16th day of incubation. PCP significantly depressed the development of embryo brains. Cerebral hemisphere weight, total protein and total DNA were significantly lower on day 10 of incubation in Group-1. Similar results were observed in Group-2. Concomitantly, the concentration of brain serotonin at day 10 was also significantly reduced when PCP was injected into the eggs on the 7th day of incubation. Since serotonin has been reported to influence development of the chick embryo brain, the present finding of the effect of PCP on brain development might be a secondary phenomenon. The possible implications of the effects of PCP on human brain development are also discussed. PMID:3626765

  16. USE OF WHOLE EMBRYO CULTURE FOR EVALUATING TOXICITY AND TERATOGENICITY

    EPA Science Inventory

    The use of rodent embryos cultured in vitro for the evaluation of the teratogenicity of compounds is discussed in detail. Time and concentration of exposure are delineated as factors of importance, as well as metabolism and embryonic stage. The economic feasibility of using such ...

  17. Aquatic toxicity assessment of single-walled carbon nanotubes using zebrafish embryos

    NASA Astrophysics Data System (ADS)

    Pan, Huichin; Lin, Yu-Jun; Li, Meng-Wei; Chuang, Han-Ni; Chou, Cheng-Chung

    2011-07-01

    Zebrafish embryos selected at the 64-cell stage were exposed to various concentrations of amide functionalized single-walled carbon nanotubes (SWCNTs) ranging from 1 to 10 μg/ml dissolved in 1% Pluronic F-68 (a cell culture grade surfactant), and the development of embryos was examined from 24 to 120 hours post fertilization (hpf). Incubation of embryos in 1% F-68 did not induce overt abnormal phenotype as compared to the wild-type; neither did it cause significant mortality during the exposure period. Generally, there was a slight developmental delay in larvae treated with SWCNTs of 5 μg/ml or above. Only larvae exposed to >= 5 μg/ml SWCNTs showed significantly reduced survival rates. About 50% of the embryos exposed to 5 μg/ml showed abnormal phenotypes at 24 hpf as compared to the control group. As development proceeds to 120 hpf, more embryos displayed defective morphology. A slight hatching delay was observed in embryos exposed to concentrations above 5 μg/ml. There was a general reduction of body axes, including narrowed somite and shortened yolk stalk. In addition, pigmentation in the ventral trunk area was less than that observed in control group. The body lengths of the exposed embryos were decreased significantly at 48 hpf (3.11 mm in control vs. 3.00 mm in SWCNTs-exposed embryos). However, exposure to SWCNTs did not affect the number of somites. Other features that were noticed in the SWCNTs-exposed embryos included edema and shrinkage and blebbling of the epidermal lining. Most of these observed phenotypes persisted from 48 hpf through 120 hpf. Overall, the aforementioned results indicate that soluble amide-functionalized SWCNTs are toxic to zebrafish embryos at a minimum concentration of 5 μg/ml.

  18. METAL TOXICITY TO EMBRYOS AND LARVAE OF EIGHT SPECIES OF FRESHWATER FISH--II: COPPER

    EPA Science Inventory

    Fish larvae and early juveniles of all species tested (brook trout, rainbow trout, brown trout, lake trout, northern pike, white sucker, herring and smallmouth bass) were more sensitive to copper than the embryos. Embryo survival was affected only at the higher concentrations tes...

  19. [Toxic effects of CdSe/ZnS QDs to zebrafish embryos].

    PubMed

    Chen, Mu-Fei; Huang, Cheng-Zhi; Pu, De-Yong; Zheng, Chao-Yi; Yuan, Kai-Mi; Jin, Xing-Xing; Zhang, Yao-Guang; Jin, Li

    2015-02-01

    The toxic effects of CdSe/ZnS QDs on zebrafish (Danio rerio) embryos at different developmental stages were investigated in this study. The voluntary movement frequency, body length, hatching rate, mortality and malformation rate, SOD activities, MDA contents, mRNA expression of metallothionein (MT) and heat stress protein 70 (Hsp70) were used as indicators. The results showed that the EC50 was 316.994 nmol x L(-1) for zebrafish embryos (72 hpf) when exposed to CdSe/ZnS QDs. After the CdSe/ZnS QDs exposure, the embryos showed a significant increase in mortality and malformation rate, a decrease in hatching rate and body length, an advance in hatching time, and a changing in the spontaneous movement frequency, and many other toxic effects, such as the condensation of embryonic eggs, the formation of pericardial cysts and curvature of the spine. Moreover, it was found that the MDA contents in the embryos in CdSe/ZnS QDs groups were significantly increased, and the SOD activities were changed. In addition, the mRNA expression level of MT and Hsp70 were up-regulated. All the information suggests that exposure of CdSe/ZnS QDs can cause toxic effects on zebrafish embryos, and the effects may be related to the releasing of Cd2+, particle size and oxidative stress. PMID:26031104

  20. Phytochemicals reduce aflatoxin-induced toxicity in chicken embryos

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins (AF) are toxic metabolites produced by molds, Aspergillus flavus and Aspergillus parasicitus, which frequently contaminate chicken feed ingredients. Ingestion of AF-contaminated feed by chickens leads to deleterious effects, including decreased chicken performance and reduced egg producti...

  1. GLUCOCORTICOID RECEPTOR REGULATION IN THE RAT EMBRYO: A POTENTIAL SITE FOR DEVELOPMENTAL TOXICITY?

    EPA Science Inventory

    Glucocorticoid receptor regulation in the rat embryo: a potential site for developmental toxicity?

    Ghosh B, Wood CR, Held GA, Abbott BD, Lau C.

    National Research Council, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711, USA.

  2. Sediment spiking for toxicity testing

    SciTech Connect

    Murdoch, M.H.; Norman, D.M.; Chapman, P.M.; Norman, D.M.; Quintino, V.M.

    1994-12-31

    Sediment toxicity testing integrates responses to sediment variables and hence does not directly indicate cause-and-effect. One tool for determining cause-and-effect is sediment spiking in which relatively uncontaminated sediment is amended with known amounts of contaminants, then tested for toxicity. Based on the concentration-response relationship(s), the relative toxicity of the spiked contaminants and their significance in sediment mixtures can be assessed. However, sediment spiking methods vary considerably. The present study details an appropriate methodology for amending sediments with a range of organic contaminant concentrations including different solvent schemes and an equilibration period. This methodology is described as appropriate because predicted and actual concentrations were similar, and responses in an acute 10-d amphipod test matched predictions and other data.

  3. The future of toxicity testing.

    PubMed

    Andersen, Melvin E; Al-Zoughool, Mustafa; Croteau, Maxine; Westphal, Margit; Krewski, Daniel

    2010-02-01

    In 2007, the U.S. National Research Council (NRC) released a report, "Toxicity Testing in the 21st Century: A Vision and a Strategy," that proposes a paradigm shift for toxicity testing of environmental agents. The vision is based on the notion that exposure to environmental agents leads to adverse health outcomes through the perturbation of toxicity pathways that are operative in humans. Implementation of the NRC vision will involve a fundamental change in the assessment of toxicity of environmental agents, moving away from adverse health outcomes observed in experimental animals to the identification of critical perturbations of toxicity pathways. Pathway perturbations will be identified using in vitro assays and quantified for dose response using methods in computational toxicology and other recent scientific advances in basic biology. Implementation of the NRC vision will require a major research effort, not unlike that required to successfully map the human genome, extending over 10 to 20 years, involving the broad scientific community to map important toxicity pathways operative in humans. This article provides an overview of the scientific tools and technologies that will form the core of the NRC vision for toxicity testing. Of particular importance will be the development of rapidly performed in vitro screening assays using human cells and cell lines or human tissue surrogates to efficiently identify environmental agents producing critical pathway perturbations. In addition to the overview of the NRC vision, this study documents the reaction by a number of stakeholder groups since 2007, including the scientific, risk assessment, regulatory, and animal welfare communities. PMID:20574896

  4. Cadmium: toxic effects on the reproductive system and the embryo.

    PubMed

    Thompson, Jennifer; Bannigan, John

    2008-04-01

    The heavy metal cadmium (Cd) is a pollutant associated with several modern industrial processes. Cd is absorbed in significant quantities from cigarette smoke, and is known to have numerous undesirable effects on health in both experimental animals and humans, targeting the kidneys, liver and vascular systems in particular. However, a wide spectrum of deleterious effects on the reproductive tissues and the developing embryo has also been described. In the testis, changes due to disruption of the blood-testis barrier and oxidative stress have been noted, with onset of widespread necrosis at higher dosage exposures. Incorporation of Cd into the chromatin of the developing gamete has also been demonstrated. Ovarian Cd concentration increases with age, and has been associated with failure of progression of oocyte development from primary to secondary stage, and failure to ovulate. A further mechanism by which ovulation could be rendered ineffective is by failure of pick-up of the oocyte by the tubal cilia due to suboptimal expansion of the oocyte-cumulus complex and mis-expression of cell adhesion molecules. Retardation of trophoblastic outgrowth and development, placental necrosis and suppression of steroid biosynthesis, and altered handling of nutrient metals by the placenta all contribute to implantation delay and possible early pregnancy loss. Cd has been shown to accumulate in embryos from the four-cell stage onwards, and higher dosage exposure inhibits progression to the blastocyst stage, and can cause degeneration and decompaction in blastocysts following formation, with apoptosis and breakdown in cell adhesion. Following implantation, exposure of experimental animals to oral or parenteral Cd causes a wide range of abnormalities in the embryo, depending on the stage of exposure and dose given. Craniofacial, neurological, cardiovascular, gastrointestinal, genitourinary, and limb anomalies have all been described in placentates, with axial abnormalities and defects in somite structure noted in fish and ventral body wall defect and vertebral malformation occurring in the chick. In this paper, we examine the mechanisms by which Cd can affect reproductive health, and consider the use of micronutrients in prevention of these problems. PMID:18367374

  5. Toxicity testing using Caenorhabditis elegans

    SciTech Connect

    Middendorf, P.J.; Dusenbery, D.B.; Williams, P.L.

    1995-12-31

    Caenorhabditis elegans is a small free-living nematode that is representative of what may be the most abundant animal group. It has been promoted as a possible model organism for toxicity testing in the laboratory and in field evaluations in part because more is known about its biology than any other animal, Toxicity tests using C. elegans have been developed with lethality, reproduction, and behavior as end points. The tests have also been developed to varying degrees using standard laboratory media, water, and soil. The results of the tests when exposing C. elegans to a variety of metals, inorganic, and organic compounds indicate it is typically at least as sensitive as other species currently used, such as Daphnia and earthworms, and is generally much easier to maintain in the laboratory. The advantages and disadvantages of C. elegans and the state of development of the tests will be discussed.

  6. Differential toxicity of three polychlorinated biphenyl congeners in developing sea urchin embryos

    SciTech Connect

    Schweitzer, L.E.; Suffet, I.H.; Hose, J.E.; Bay, S.M.

    1997-07-01

    The relationship between body burden and toxicity of three individual polychlorinated biphenyl (PCB) congeners in developing sea urchin embryos was investigated to evaluate the validity of current predictive models of PCB toxicity in an invertebrate system. Body burdens of radiolabeled PCB congeners (IUPAC-47, 77, and 153) accumulated from a seawater were used to determine median effective concentrations (EC50s) for developmental and cytogenetic effects following a 72-h exposure. Congener 47, a di-ortho-substituted tetrachlorobiphenyl, was found to be at least four times more toxic than congener 77, a non-ortho-substituted (coplanar) tetrachlorobiphenyl, with EC50s of 47 and >218 mmol/kg, respectively, using an embryo development assay. This result contradicts the structure-activity prediction of the mammalian-based toxic equivalents (TEQs) approach, demonstrating the need for an ecotoxicologic model. Congener 153, a di-ortho-substituted hexachlorobiphenyl, was virtually nontoxic in terms of developmental effects at the highest dose achievable at its limit of water solubility. Cytogenetic analysis was a more sensitive method for assessing toxicity than the embryo development assay. Dose-response relationships were established with mitotic activity being the most sensitive endpoint because the PCBs appeared to inhibit mitosis. At the highest doses, complete mitotic arrest was observed. Congener 77 was found to be at least two times more toxic than congener 153 but not as toxic as congener 47 using mitotic activity as the endpoint for toxicity. Thus, the developmental and cytogenetic endpoints ranked the toxicity of the congeners similarly, but established different EC50s.

  7. Using zebrafish in systems toxicology for developmental toxicity testing.

    PubMed

    Nishimura, Yuhei; Inoue, Atsuto; Sasagawa, Shota; Koiwa, Junko; Kawaguchi, Koki; Kawase, Reiko; Maruyama, Toru; Kim, Soonih; Tanaka, Toshio

    2016-01-01

    With the high cost and the long-term assessment of developmental toxicity testing in mammals, the vertebrate zebrafish has become a useful alternative model organism for high-throughput developmental toxicity testing. Zebrafish is also very favorable for the 3R perspective in toxicology; however, the methodologies used by research groups vary greatly, posing considerable challenges to integrative analysis. In this review, we discuss zebrafish developmental toxicity testing, focusing on the methods of chemical exposure, the assessment of morphological abnormalities, housing conditions and their effects on the production of healthy embryos, and future directions. Zebrafish as a systems toxicology model has the potential to elucidate developmental toxicity pathways, and to provide a sound basis for human health risk assessments. PMID:26537640

  8. Linking embryo toxicity with genotoxic responses in the freshwater snail Physa acuta: single exposure to benzo(a)pyrene, fluoxetine, bisphenol A, vinclozolin and exposure to binary mixtures with benzo(a)pyrene.

    PubMed

    Sánchez-Argüello, Paloma; Aparicio, Natalia; Fernández, Carlos

    2012-06-01

    Genotoxic effects on fauna after waterborne pollutant exposure have been demonstrated by numerous research programmes. Less effort has been focused on establishing relationship between genotoxicity and long-term responses at higher levels of biological organization. Taking into account that embryos may be more sensitive indicators of reproductive impairment than alterations in fertility, we have developed two assays in multiwell plates to address correlations between embryo toxicity and genotoxicity. The potential teratogenicity was assessed by analyzing abnormal development and mortality of Physa acuta at embryonic stage. Genotoxicity was measured by the micronucleus (MN) test using embryonic cells. Our results showed that linkage between genotoxicity and embryo toxicity depends on mechanisms of action of compounds under study. Embryo toxic responses showed a clear dose-related tendency whereas no clear dose-dependent effect was observed in micronucleus induction. The higher embryo toxicity was produced by benzo(a)pyrene exposure followed by fluoxetine and bisphenol A. Vinclozolin was the lower embryo toxic compound. Binary mixtures with BaP always resulted in higher embryo toxicity than single exposures but antagonistic effects were observed for MN induction. Benzo(a)pyrene produced the higher MN induction at 0.04 mg/L, which also produced clear embryo toxic effects. Fluoxetine did not induce cytogenetic effects but 0.25mg/L altered embryonic development. Bisphenol A significantly reduced hatchability at 0.5mg/L while MN induction appeared with higher treatments than those that start causing teratogenicity. Much higher concentration of vinclozolin (5mg/L) reduced hatchability and induced maximum MN formation. In conclusion, while validating one biomarker of genotoxicity and employing one ecologically relevant effect, we have evaluated the relative sensitivity of a freshwater mollusc for a range of chemicals. The embryo toxicity test is a starting point for the development of a life cycle test with freshwater snails even for undertaking multigeneration studies focused on transgenerational effects. PMID:22417675

  9. Further development of rodent whole embryo culture: solvent toxicity and water insoluble compound delivery system.

    PubMed

    Kitchin, K T; Ebron, M T

    1984-02-14

    In order to study the in vitro embryotoxicity and dysmorphogenesis of water insoluble compounds, solvents or chemical delivery systems of low toxicity and teratogenicity to the developing embryo must be found. Therefore, day 10.5 rat embryos were cultured for 2 days in whole rat serum containing 0.1, 0.5 and 2.5 vol.% of ethyl alcohol, dimethylsulfoxide, acetone, Tween 80, corn oil and 10% acetone/90% corn oil. No adverse effects occurred with any of the solvents at the 0.1% concentration level. At 0.5% ethyl alcohol and Tween 80 significantly reduced embryonic growth and increased the incidence of embryonic abnormalities. With the exception of corn oil and acetone/corn oil, embryos cultured in media containing 2.5% of various solvents failed to grow, did not differentiate, and died during the culture period. Corn oil suspended in rat serum by use of ultrasound was non-toxic even at concentrations of 2.5% and 10%. Growth parameters of embryos cultured in serum containing corn oil were indistinguishable from controls and overall morphogenesis was good (particularly at 2.5%). The order of increasing embryotoxicity and dysmorphogenesis of the studied liquids was corn oil less than acetone/corn oil less than dimethylsulfoxide less than ethyl alcohol, acetone less than Tween 80. Any of the 4 water miscible solvents (at 0.1%) or a sonicated suspension of corn oil in serum (up to 2.5%) met the criteria of a non-toxic and non-teratogenic water insoluble compound delivery system for in vitro embryo culture. PMID:6701904

  10. Opportunities for embryo transfer in the age of DNA testing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Embryo transfer (ET) has contributed to increasing selection intensity in cattle breeding for many years. Preimplantation DNA testing offers the opportunity to increase selection response further through increasing within-family selection intensity. Further increases in between-family selection inte...

  11. Developmental toxicity testing of vaccines.

    PubMed

    Barrow, Paul C; Allais, Linda

    2013-01-01

    Preventative and therapeutic vaccines are increasingly used during pregnancy and present special considerations for developmental toxicity testing. The various components of the vaccine formulation (i.e., protein or polysaccharide antigen, adjuvants, and excipients) need to be assessed for direct effects on the developing conceptus. In addition, possible adverse influences of the induced antibodies on fetal and/or postnatal development need to be evaluated. A guidance document on the preclinical testing of preventative and therapeutic vaccines for developmental toxicity was issued by the FDA in 2006. Preclinical studies are designed to assess possible influences of vaccines on pre- and postnatal development. The choice of model animal for these experiments is influenced by species differences in the timing and extent of the transfer of the induced maternal antibodies to the fetus. The cross-placental transport of maternal immunoglobulins generally only occurs in late gestation and tends to be greater in humans and monkeys than in non-primate species. For many vaccines, the rabbit shows a greater rate of prenatal transfer of the induced antibodies than rodents. For biotechnology-derived vaccines that are not immunogenic in lower species, nonhuman primates may be the only appropriate models. It may be advisable to test new adjuvants using the ICH study designs for conventional pharmaceuticals in addition to the developmental toxicity study with the final vaccine formulation. PMID:23138897

  12. Embryo-fetal development toxicity of honokiol microemulsion intravenously administered to pregnant rats.

    PubMed

    Zhang, Qianqian; Ye, Xiangfeng; Wang, Lingzhi; Peng, Bangjie; Zhang, Yingxue; Bao, Jie; Li, Wanfang; Wei, Jinfeng; Wang, Aiping; Jin, Hongtao; Chen, Shizhong

    2016-02-01

    The aim of this study was to evaluate the embryo-fetal development toxicity of honokiol microemulsion. The drug was intravenously injected to pregnant SD rats at dose levels of 0, 200, 600 and 2000 μg/kg/day from day 6-15 of gestation. All the pregnant animals were observed for body weights and any abnormal changes and subjected to caesarean-section on gestation day (GD) 20; all fetuses obtained from caesarean-section were assessed by external inspection, visceral and skeletal examinations. No treatment-related external alterations as well as visceral and skeletal malformations were observed in honokiol microemulsion groups. There was no significant difference in the body weight gain of the pregnant rats, average number of corpora lutea, and the gravid uterus weight in the honokiol microemulsion groups compared with the vehicle control group. However, at a dose level of 2000 μg/kg/day, there was embryo-fetal developmental toxicity observed, including a decrease in the body length and tail length of fetuses. In conclusion, the no-observed-adverse-effect level (NOAEL) of honokiol microemulsion is 600 μg/kg/day, 75 times above the therapeutic dosage and it has embryo-fetal toxicity at a dose level of 2000 μg/kg/day, which is approximately 250 times above the therapeutic dosage. PMID:26619782

  13. 670 nanometer light treatment attenuates dioxin toxicity in the developing chick embryo.

    PubMed

    Yeager, Ronnie L; Lim, Jinhwan; Millsap, Deborah S; Jasevicius, Ashley V; Sanders, Ruth A; Whelan, Harry T; Watkins, John B; Eells, Janis T; Henshel, Diane S

    2006-01-01

    2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is an acutely toxic anthropogenic chemical. Treatment with a red to near-infrared (630-1000 nm) light-emitting diode (LED) attenuates the toxicant-induced oxidative stress and energy deficit in neuronal cell culture. For this study, fertile chicken (Gallus gallus) eggs were injected once at the start of incubation with sunflower oil vehicle or 200 pg TCDD/g egg (200 parts per trillion), an environmentally relevant dose. Daily LED treatment after TCDD exposure reduced embryonic mortality by 47%. LED treatment of TCDD-exposed eggs also decreased the hepatic oxidized-to-reduced glutathione ratio by 88%. Activities of other hepatic indicators of oxidative stress, such as glutathione reductase and catalase, were increased after LED treatment of TCDD-exposed eggs. Our study demonstrates that 670 nm phototherapy can mitigate the oxidative stress and energy deficit resulting from developmental exposure to TCDD while reducing TCDD-induced embryo mortality. Moreover, LED treatment restores hepatic enzyme activities to control levels in TCDD-exposed embryos. The effective attenuation of TCDD-induced embryo toxicity by LED treatment could extend to mitigating the effects of other teratogens that induce oxidative and energy stress. PMID:17163486

  14. Developmental and reproductive toxicity testing of vaccines.

    PubMed

    Barrow, Paul

    2012-03-01

    The majority of new preventative and therapeutic vaccines are now assessed for developmental toxicity according to guidelines issued by the FDA in 2006. Despite the absence of confirmed effects in humans, vaccines are frequently suspected of having adverse side-effects on the development of children. Such suspicions are perhaps unavoidable considering the extremely widespread use of vaccines. The preclinical developmental toxicology studies are designed to assess possible influences of each component of the vaccine formulation-and the induced antibodies-on the development of the conceptus, neonate and suckling organism. Immune modulation by a vaccine or an adjuvant could, for instance, affect the outcome of pregnancy by interfering with the natural shift in immune balance of the mother during gestation. Maternal immunoglobulins are transferred from the mother to the offspring in order to confer passive immunity during early life. This maternal antibody transport is prenatal in humans and monkeys, but tends to be delayed until after birth in other species. Therefore, a suitable model species needs to be chosen for preclinical studies in order to ensure exposure of the foetus to the induced maternal antibodies following vaccination. Rabbits are the best laboratory model for prenatal immunoglobulin transfer, but rodents are more practical for the necessary postnatal investigations. Non-human primates are the only appropriate models for the testing of vaccines that are not immunogenic in lower species. It is advisable to test new adjuvants separately according to the ICH S5(R2) guidelines. Preclinical paediatric investigations are not currently required for vaccines, even though most vaccines are given to children. Other areas of regulatory concern include developmental immunotoxicity and effects on the preimplantation embryo. Because of the limitations of the available animal models for developmental toxicity testing, pharmacovigilance is essential. PMID:22233769

  15. Toxicity to embryo and adult zebrafish of copper complexes with two malonic acids as models for dissolved organic matter

    SciTech Connect

    Palmer, F.B.; Evans, C.W.; Butler, C.A.; Timperley, M.H.

    1998-08-01

    The toxicity to embryo and adult zebrafish (Danio rerio) of Cu complexes with two substituted malonic acids, benzyl- and n-hexadecyl-, chosen as models for low-molecular-weight natural dissolved organic matter, were investigated. Toxicity test solutions at pH 6.5 {+-} 0.1 with the required Cu ion-specific electrode. In the absence of malonic acids, concentrations of Cu{sup 2+} up to 1.13 {mu}mol/L increased the embryo hatching time from approx. 2 d in control solutions (no Cu or malonic acid) and solutions containing malonic acids without Cu to approx. 8 d. The Cu-benzylmalonic acid complex in the presence of inorganic Cu species did not delay hatching beyond that attributable to Cu{sup 2+}. In contrast, 0.60 {mu}mol/L Cu-n-hexadecylmalonic complexes delayed hatching by 5.5 d in excess of that attributable to 1.13 {mu}mol/L Cu{sup 2+}, assuming that the hatching delays caused by the different Cu species were additive, possibly because of Cu entry into the embryo as the lipophilic Cu-n-hexadecylmalonic complex. None of the Cu-malonic acid complexes was acutely toxic to adult zebrafish at concentrations up to 1.4 {mu}mol/L, possibly because Cu was removed from the Cu-malonic acid complexes by stronger chelating groups at the gill surface. Substituted malonic acids with similar proton and Cu association constants can be readily prepared with a variety of simple substituents, radiolabeled if required. Their results show that these acids could be useful ligands for investigating intracellular transport and metabolism of metal-organic complexes.

  16. Protective effects of glucose-6-phosphate and NADP against alpha-chaconine-induced developmental toxicity in Xenopus embryos.

    PubMed

    Rayburn, J R; Bantle, J A; Qualls, C W; Friedman, M

    1995-12-01

    In previous studies a metabolic activation system (MAS) composed of Aroclor 1254-induced rat liver microsomes led to an apparent reduction of potato glycoalkaloid developmental toxicity in the frog embryo teratogenesis assay-Xenopus (FETAX). The reasons for this reduction were investigated in this study. The effect of the exogenous MAS on glycoalkaloid developmental toxicity was examined in two experiments in which a concentration series of alpha-chaconine was tested with a MAS with and without a reduced nicotinamide adenine dinucleotide (NADPH) generator system consisting of NADPH, oxidized nicotinamide adenine dinucleotide (NADP), glucose-6-phosphate (G6P) and glucose-6-phosphate dehydrogenase. The NADPH generator system and each of its individual components were tested at a single high concentration of alpha-chaconine to evaluate their potential effects on toxicity. The findings indicated that the protective effect of the MAS was not the result of detoxification by microsomal enzyme systems, but was caused by two components of the NADPH generator system, namely NADP and G6P. G6P was more protective of alpha-chaconine-induced toxicity than NADP at the concentrations tested. Thus, FETAX with a MAS must be performed with appropriate controls that take into account the possible interactions with individual components of the system. PMID:8846997

  17. Embryo toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin to the wood duck (Aix sponsa)

    USGS Publications Warehouse

    Augspurger, T.P.; Tillitt, D.E.; Bursian, S.J.; Fitzgerald, S.D.; Hinton, D.E.; Di Giulio, R.T.

    2008-01-01

    We examined the sensitivity of the wood duck (Aix sponsa) embryo to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) by injecting the toxicant into their eggs. Six groups of wood duck eggs (n = 35 to 211 per trial) were injected with 0 to 4600 pg TCDD/g egg between 2003 and 2005. Injections were made into yolk prior to incubation, and eggs were subsequently incubated and assessed weekly for mortality. Significant TCDD-induced mortality was not observed through day 25 (90% of incubation). Liver, heart, eye, and brain histology were generally unremarkable. Hepatic ethoxyresorufin-O-deethylase activity, a biomarker of dioxin-like compound exposure, was induced by 12-fold in the 4600 pg/g treatment relative to controls. The median lethal dose for chicken (Gallus domesticus) eggs we dosed identically to wood duck eggs was about 100 pg/g, similar to other assessments of chickens. Among dioxin-like compound embryo lethality data for 15 avian genera, the wood duck 4600 pg/g no-observed-effect level ranks near the middle. Because no higher doses were tested, wood ducks may be like other waterfowl (order Anseriformes), which are comparatively tolerant to embryo mortality from polychlorinated dibenzo-p-dioxins and dibenzofurans when exposed by egg injection. ?? 2008 US Government.

  18. Stage-dependent toxicity of bisphenol a on Rhinella arenarum (anura, bufonidae) embryos and larvae.

    PubMed

    Wolkowicz, Ianina R Hutler; Herkovits, Jorge; Pérez Coll, Cristina S

    2014-02-01

    The acute and chronic toxicity of bisphenol A (BPA) was evaluated on the common South American toad Rhinella arenarum embryos and larvae by means of continuous and pulse exposure treatments. Embryos were treated continuously from early blastula (S.4) up to complete operculum (S.25), during early larval stages and by means of 24 h pulse exposures of BPA in concentrations ranging between 1.25 and 40 mg L(-1) , in order to evaluate the susceptibility to this compound in different developmental stages. For lethal effects, S.25 was the most sensitive and gastrula was the most resistant to BPA. The Teratogenic Index for neurula, the most sensitive embryonic stage for sublethal effects was 4.7. The main morphological alterations during early stages were: delayed or arrested development, reduced body size, persistent yolk plug, microcephaly, axial/tail flexures, edemas, blisters, waving fin, underdeveloped gills, mouth malformations, and cellular dissociation. BPA caused a remarkable narcotic effect from gill circulation stage (S.20) onwards in all the organisms exposed after 3 h of treatment with 10 mg L(-1) BPA. After recovering, the embryos exhibited scarce response to stimuli, erratic or circular swimming, and spasmodic contractions from 5 mg L(-1) onwards. Our results highlight the lethal and sublethal effectsof BPA on R. arenarum embryos and larvae, in the last case both at structural and functional levels. PMID:22052622

  19. Aryl hydrocarbon receptor 2 mediates the toxicity of Paclobutrazol on the digestive system of zebrafish embryos.

    PubMed

    Wang, Wen-Der; Chen, Guan-Ting; Hsu, Hwei-Jan; Wu, Chang-Yi

    2015-02-01

    Paclobutrazol (PBZ), a trazole-containing fungicide and plant growth retardant, has been widely used for over 30 years to regulate plant growth and promote early fruit setting. Long-term usage of PBZ in agriculture and natural environments has resulted in residual PBZ in the soil and water. Chronic exposure to waterborne PBZ can cause various physiological effects in fish, including hepatic steatosis, antioxidant activity, and disruption of spermatogenesis. We have previously shown that PBZ also affects the rates of zebrafish embryonic survival and hatching, and causes developmental failure of the head skeleton and eyes; here, we further show that PBZ has embryonic toxic effects on digestive organs of zebrafish, and describe the underlying mechanisms. PBZ treatment of embryos resulted in dose-dependent morphological and functional abnormalities of the digestive organs. Real-time RT-PCR and in situ hybridization were used to show that PBZ strongly induces cyp1a1 expression in the digestive system, and slightly induces ahr2 expression in zebrafish embryos. Knockdown of ahr2 with morpholino oligonucleotides prevents PBZ toxicity. Thus, the toxic effect of PBZ on digestive organs is mediated by AhR2, as was previously reported for retene and TCDD. These findings have implications for understanding the potential toxicity of PBZ during embryogenesis, and thus the potential impact of fungicides on public health and the environment. PMID:25500619

  20. Chronic toxicity of heavy fuel oils to fish embryos using multiple exposure scenarios.

    PubMed

    Martin, Jonathan D; Adams, Julie; Hollebone, Bruce; King, Thomas; Brown, R Stephen; Hodson, Peter V

    2014-03-01

    The chronic toxicity to rainbow trout (Oncorhynchus mykiss) embryos of heavy fuel oil (HFO) 6303, weathered HFO 6303, HFO 7102, and medium South American (MESA) crude oil was assessed by different exposure regimes. These included water accommodated fractions (WAF; water in contact with floating oil), chemically enhanced WAF (CEWAF; oil dispersed with Corexit 9500), and effluent from columns of gravel coated with stranded oil. Heavy fuel oil WAF was nontoxic and did not contain detectable concentrations of hydrocarbons, likely because the high density and viscosity of HFO prevented droplet formation. In contrast, chemically dispersed HFO and effluent from columns of stranded HFO contained measurable concentrations of alkyl polycyclic aromatic hydrocarbons (PAH), coincident with embryo toxicity. These exposure regimes enhanced the surface area of oil in contact with water, facilitating oil-water partitioning of hydrocarbons. Heavy fuel oil was consistently more toxic to fish than crude oil and the rank order of alkyl PAH concentrations in whole oil were sufficient to explain the rank order of toxicity, regardless of exposure method. Thus, the propensity of HFO to sink and strand in spawning shoals creates a long-term risk to developing fish because of the sustained release of PAHs from HFO to interstitial waters. Further, PAH monitoring is key to accurate risk assessment. PMID:24464524

  1. Nanosilver-coated socks and their toxicity to zebrafish (Danio rerio) embryos.

    PubMed

    Gao, Jiejun; Sepúlveda, Maria S; Klinkhamer, Christopher; Wei, Alexander; Gao, Yu; Mahapatra, Cecon T

    2015-01-01

    Silver nanoparticles (AgNPs) are being incorporated and are known to be released from various consumer products such as textiles. However, no data are available on the toxicity of AgNPs released from any of these commercial products. In this study, we quantified total silver released from socks into wash water by inductively coupled plasma mass spectrometry (ICP-MS) and determined the presence of AgNPs using transmission electron microscopy (TEM). We then exposed zebrafish (Danio rerio) embryos for 72 h to either this leachate ("sock-AgNP") or to the centrifugate ("spun-AgNP") free of AgNPs and compared their toxicity to that of ionic silver (Ag(+)). Our data suggest that AgNPs do get released into the wash water, and centrifugation eliminated AgNPs but did not decrease total silver concentrations, indicating that most of the silver in the sock-AgNP solution was in the ionic form. All embryos died during the first 24 h when exposed to undiluted sock-AgNP and spun-AgNP solutions resulting in significantly lower LC50 values (0.14 and 0.26 mg L(-1)) compared to AgNO3 (0.80 mg L(-1)). Similarly, at 72 hpf, both sock-derived solutions were more potent at affecting hatching and inducing abnormal development. These results suggest that both sock-AgNP and spun-AgNP solutions were more toxic than AgNO3. Previous studies have consistently shown the opposite, i.e., AgNPs are about 10 times less toxic that Ag(+). All together our results show that the high toxicity induced by the leachate of these socks is likely not caused by AgNPs or Ag(+). More studies are needed to evaluate the toxicity of the myriad of AgNP-coated commercial products that are now estimated to be close to 500. PMID:25303653

  2. Toxicity assessment and bioaccumulation in zebrafish embryos exposed to carbon nanotubes suspended in Pluronic® F-108.

    PubMed

    Wang, Ruhung; N Meredith, Alicea; Lee, Michael; Deutsch, Dakota; Miadzvedskaya, Lizaveta; Braun, Elizabeth; Pantano, Paul; Harper, Stacey; Draper, Rockford

    2016-08-01

    Carbon nanotubes (CNTs) are often suspended in Pluronic® surfactants by sonication, which may confound toxicity studies because sonication of surfactants can create degradation products that are toxic to mammalian cells. Here, we present a toxicity assessment of Pluronic® F-108 with and without suspended CNTs using embryonic zebrafish as an in vivo model. Pluronic® sonolytic degradation products were toxic to zebrafish embryos just as they were to mammalian cells. When the toxic Pluronic® fragments were removed, there was little effect of pristine multi-walled CNTs (pMWNTs), carboxylated MWNTs (cMWNTs) or pristine single-walled carbon nanotubes (pSWNTs) on embryo viability and development, even at high concentrations. A gel electrophoretic method coupled with Raman imaging was developed to measure the bioaccumulation of CNTs by zebrafish embryos, and dose-dependent uptake of CNTs was observed. These data indicate that embryos accumulate pMWNTs, cMWNTs and pSWNTs yet there is very little embryo toxicity. PMID:26559437

  3. Toxicity of spill-treating agents and oil to sea urchin embryos.

    PubMed

    Rial, Diego; Vázquez, José A; Murado, Miguel A

    2014-02-15

    The aim of this study was to assess the joint toxicity of a Maya crude oil and four spill-treating agents (STAs) (CytoSol, Finasol OSR51, Agma OSD569 and OD4000). The acute toxicity of the binary mixtures of the water accommodated fractions (WAFs) obtained independently for the oil and each STA was assessed. The toxicity of the chemically enhanced WAF (CEWAF) of oil and Finasol OSR51 at several dispersant to oil ratios (1:2, 1:10 and 1:100) was also evaluated. The toxicity (EC50) obtained for the WAFs of the STAs was: CytoSol (15.1 mL/L)toxicity of binary mixtures was obtained by the following models: Concentration Addition (Agma OSD569 and CytoSol), Independent Action (Finasol OSR51) and extended Concentration Addition model to describe antagonistic effects (OD4000). The CEWAFs showed an intermediate position between the more moderate effects of the WAF of oil and the higher toxicity of the WAFs of Finasol OSR51, which suggested the high sensitivity of the sea urchin embryo toward the dispersant. PMID:24295747

  4. Assessment of trifloxystrobin uptake kinetics, developmental toxicity and mRNA expression in rare minnow embryos.

    PubMed

    Zhu, Bin; Liu, Guang-Lu; Liu, Lei; Ling, Fei; Wang, Gao-Xue

    2015-02-01

    Trifloxystrobin (TFS) is the widely used strobilurin fungicide. However, little information is so far available regarding the uptake kinetics and developmental toxicity of TFS to fish. The present study was conducted to investigate the uptake kinetics, potential environment risk and toxicity of TFS on Gobiocypris rarus embryos. Results revealed that increased malformation, decreased body length and heart rate, affected spontaneous movement and swimming speed provide a gradual concentration-dependent manner; values of 144 h LC50 (median lethal concentration) and EC50 (median effective concentration) were 1.11 and 0.86 μg L(-1). Continuous exposure to TFS resulted in a steady accumulation with no evidence of elimination. Enzyme activities were significantly changed; reactive oxygen species and DNA damage were significantly induced after TFS treatment. Certain genes related to cell apoptosis (p53), metabolism (cyp1a), stress response (hsp70) and blood vessels (vezf1) development were all significantly up-regulated. This is the first study to define uptake kinetics and to focus on behavioral consequences, physiological changes and mRNA expression following TFS exposure in the early life stages of fish. Our results suggest that TFS is highly toxic to fish embryos. PMID:25240160

  5. Effects of ionization on the toxicity of silver nanoparticles to Japanese medaka (Oryzias latipes) embryos.

    PubMed

    Lee, Byoung-Cheun; Kim, Jungkon; Cho, Jae-Gu; Lee, Jae-Woo; Duong, Cuong N; Bae, Eunjoo; Yi, Jongheop; Eom, Ig-Chun; Choi, Kyunghee; Kim, Pilje; Yoon, Junheon

    2014-01-01

    Increase in the use of manufactured nanomaterials (NMs) has led to concerns about the environmental impacts. Especially, hazard of metal-based NMs is more severe due to ions released from surface by water quality parameters and physicochemical properties after entering into the water environment. However, little is known about the effects of ionization on the toxicity of metal-based NMs in the water environment. To address this question, we prepared the suspensions of silver nanoparticles (AgNP) at 25?g L(-1) containing different concentrations of Ag(+) (5, 10, 20, 45, and 75% Ag(+) to total Ag), and evaluated their toxicity to Japanese medaka (Oryzias latipes) embryos. Higher Ag(+) ratios in the AgNP suspension, suggesting the lower number of particles, led to the higher adverse effects on embryos and sac-fries. In addition, histopathology analysis revealed that AgNPs penetrated through chorion of eggs and skin membrane, and were distributed into the tissues. The results imply that the ionization could decrease the toxicity of metal-based NMs in the water environment. PMID:24279620

  6. Potential protective effect of L-cysteine against the toxicity of acrylamide and furan in exposed Xenopus laevis embryos: an interaction study

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The embryo toxicities of two food-processing-induced toxic compounds, acrylamide and furan, with and without added L-cysteine were examined individually and in mixtures using the frog embryo teratogenesis assay-Xenopus (FETAX). The following measures of developmental toxicity were used (a) 96-h LC5...

  7. Amphiphilic nanoparticles of resveratrol-norcantharidin to enhance the toxicity in zebrafish embryo.

    PubMed

    Yan, Deyue; Ni, Lin-Kai; Chen, Ho-Lun; Chen, Li-Chou; Chen, Yau-Hung; Cheng, Chien-Chung

    2016-02-01

    Direct coupling of a hydrophobic drug and a hydrophilic natural product via an ester bond produced an amphiphilic adduct that formed liposomes. Liposomes of resveratrol-norcantharidin adduct are capable of forming a tadpole-like nanoparticle and exhibited high toxicity in zebrafish embryos to give the better transportation and the effective concentration into cells. Using fluorescent chromophore showed the liposome in the stomach and intestinal villi rather than in the skin and muscle. This result may provide an insight into the mechanism of action of traditional Chinese medicines, which often contain a significant amount of flavonoids and polyphenol analogs. PMID:26764188

  8. Joint toxicity of permethrin and cypermethrin at sublethal concentrations to the embryo-larval zebrafish.

    PubMed

    Yang, Ye; Ma, Huihui; Zhou, Jinghua; Liu, Jing; Liu, Weiping

    2014-02-01

    Pyrethroids, the widely used pesticides, are highly toxic to aquatic organisms. However, little information is so far available regarding the joint toxicity of type I and type II pyrethroids to fish. Zebrafish is a well-accepted aquatic vertebrate model for toxicity assessment due to small size, easy husbandry, high fecundity and transparent embryos. In this study, we utilized embryo-larval zebrafish to elucidate the combined effects of sublethal concentrations of permethrin (PM) and cypermethrin (CP), which are the most frequently used type I and type II pyrethroids, respectively. Fish were exposed from 3h postfertilization (hpf) to 144 hpf to binary mixtures of nominal concentrations of 100, 200, 300μgL(-1) PM (PM100, PM200, PM300) and 10, 20, 30μgL(-1) CP (CP10, CP20, CP30). Analytical data of the real concentrations of the chemicals showed a significant degradation of the pyrethroids but an obvious recovery after the renewal of the exposure solution. Defect rates of embryos exposed to these low concentrations of single PM or CP exhibited no statistically significant difference from the control,while the application of combination of PM and CP resulted in deleterious effects on zebrafish embryonic development. In all PM200 and PM300 exposure groups, increasing CP concentrations acted additively to the action of PM in terms of all sublethal endpoints. Co-treatment of embryos with the specific sodium channel blocker MS-222 and pyrethroids (individuals or the mixture) caused a decline in the incidences of body axis curvature and spasms compared to treatment of animals with pyrethroids alone, suggesting that the developmental toxicity of PM and CP to zebrafish was related to disruption of ion channels. We further revealed that mixture of the two pyrethroids caused greater down-regulation in the mRNA levels of proneural genes. The individual pesticides had no effect on the activity of superoxide dismutase (SOD), while the mixture exposure caused significant induction. Treatment with CP or the mixture increased the activity of catalase (CAT). Taken together, our data indicated that the mixture of PM and CP caused higher incidence of morphological defects, greater inhibition in proneural gene expression and more oxidative stress, compared to the single chemical at the corresponding doses. Our findings suggest that the combination of type I and type II pyrethroids poses a greater risk to fish in the water column. PMID:24184047

  9. Low-dose agrochemicals and lawn-care pesticides induce developmental toxicity in murine preimplantation embryos.

    PubMed Central

    Greenlee, Anne R; Ellis, Tammy M; Berg, Richard L

    2004-01-01

    Occupational exposures to pesticides may increase parental risk of infertility and adverse pregnancy outcomes such as spontaneous abortion, preterm delivery, and congenital anomalies. Less is known about residential use of pesticides and the risks they pose to reproduction and development. In the present study we evaluate environmentally relevant, low-dose exposures to agrochemicals and lawn-care pesticides for their direct effects on mouse preimplantation embryo development, a period corresponding to the first 5-7 days after human conception. Agents tested were those commonly used in the upper midwestern United States, including six herbicides [atrazine, dicamba, metolachlor, 2,4-dichlorophenoxyacetic acid (2,4-D)], pendimethalin, and mecoprop), three insecticides (chlorpyrifos, terbufos, and permethrin), two fungicides (chlorothalonil and mancozeb), a desiccant (diquat), and a fertilizer (ammonium nitrate). Groups of 20-25 embryos were incubated 96 hr in vitro with either individual chemicals or mixtures of chemicals simulating exposures encountered by handling pesticides, inhaling drift, or ingesting contaminated groundwater. Incubating embryos with individual pesticides increased the percentage of apoptosis (cell death) for 11 of 13 chemicals (p embryo for 3 of 13 agents (p embryos (p embryo (p

  10. Toxicity of cylindrospermopsin, and other apparent metabolites from Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum, to the zebrafish (Danio rerio) embryo

    PubMed Central

    Berry, John P.; Gibbs, Patrick D.L.; Schmale, Michael C.; Saker, Martin L.

    2012-01-01

    Cyanobacteria produce a diverse array of toxic or otherwise bioactive compounds that pose growing threats to human and environmental health. We utilized the zebrafish (Danio rerio) embryo, as a model of vertebrate development, to investigate the inhibition of development pathways (i.e. developmental toxicity) by the cyanobacterial toxin, cylindrospermopsin (CYN), as well as extracts from various isolates of Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum. CYN was toxic only when injected directly into embryos, but not by direct immersion at doses up to 50 μg/ml. Despite the dose dependency of toxicity observed following injection of CYN, no consistent patterns of developmental defects were observed, suggesting that toxic effects of CYN may not target specific developmental pathways. In contrast, direct immersion of embryos in all of the extracts resulted in both increased mortality and reproducible, consistent, developmental dysfunctions. Interestingly, there was no correlation of developmental toxicity observed for these extracts with the presence of CYN or with previously reported toxicity for these strains. These results suggest that CYN is lethal to zebrafish embryos, but apparently inhibits no specific developmental pathways, whereas other apparent metabolites from C. raciborskii and A. ovalisporum seem to reproducibly inhibit development in the zebrafish model. Continued investigation of these apparent, unknown metabolites is needed. PMID:19087885

  11. Toxicity, uptake kinetics and behavior assessment in zebrafish embryos following exposure to perfluorooctanesulphonicacid (PFOS)

    PubMed Central

    Huang, Haihua; Huang, Changjiang; Wang, Lijun; Ye, Xiaowei; Bai, Chenglian; Simonich, Michael T.; Tanguay, Robert L.; Dong, Qiaoxiang

    2014-01-01

    Perfluorooctanesulphonicacid (PFOS), a persistent organic contaminant, has been widely detected in the environment, wildlife and humans, but few studies have assessed its effect on aquatic organisms. The present study evaluated the effect of PFOS on zebrafish embryos. Zebrafish embryos exhibited bent spine and developmental toxicity after exposure to various PFOS concentrations (0.01-16.0 μM) from 6 to 120 hour post-fertilization (hpf). The LC50 at 120 hpf was 4.39 μM and the EC50 at 120 hpf was 2.23 μM. PFOS induced apoptosis at 24 hpf was consistently located in the brain, eye, and tail region of embryos. PFOS elevated the basal rate of swimming after 4 days of exposure, and larvae exposed to PFOS (0.5-8.0μM) for only 1 h at 6 dpf swam faster with increasing PFOS concentration. Larvae exposed to 16.0 μM PFOS for 24 h periods from 1 to 121 hpf showed the highest incidence of malformations in the 97-121 hpf window. Continuous exposure to PFOS from 1 to 121 hpf resulted in a steady accumulation with no evidence of elimination. Our results further the understanding of the health risks of PFOS to aquatic organisms and identify additional research needed on PFOS toxicology. PMID:20171748

  12. Toxicity screening of produced water extracts in a zebrafish embryo assay.

    PubMed

    Carlsson, G; Norrgren, L; Hylland, K; Tollefsen, K E

    2014-01-01

    Produced water is the largest effluent discharge from oil and gas/condensate production facilities in the North Sea. There is concern that contaminants originating from the reservoir and chemicals used in the production process may affect marine organisms. Developmental toxicity of extractable organic compounds in produced water effluents from oil and gas/condensate production platforms in the Norwegian sector of the North Sea was assessed in a temporal and spatial manner using zebrafish (Danio rerio) embryos. Large-scale solid-phase extraction (SPE) and on-column fractionation of water-soluble fraction (WSF) and an oil/particulate fraction was used in a rapid screening bioassay for embryotoxicity. Exposure to produced water extracts increased rate of mortality and reduced pigmentation and heart rate, as well as delaying time to hatch. The oil/particulate fraction was 10-fold less toxic than WSF, indicating that toxicity was predominantly produced by moderately polar and bioavailable compounds. Large spatial and temporal variation in produced water toxicity was observed, displaying considerable variability in the reservoir, oil well, and effluent composition over time. The noted toxicity did not correlate well with either reported produced water composition or parameters such as total hydrocarbons, thus challenging chemical measurements as a reliable source of information for predicting complex effects. Although embryotoxicity was observed following exposure to the extracts, dilution and transformation of produced water in the recipient are expected to rapidly reduce the concentrations of compounds in the effluents to levels below the thresholds of observed effects. PMID:24754395

  13. 7 day chronic ceriodaphnia toxicity test -- reproductive

    SciTech Connect

    Not Available

    1989-01-01

    This toxicity test was conducted to determine if the effluent causes death (acute toxicity) or reduction in the reproduction of the test organisms (chronic toxicity) during a seven day period. A series of dilutions of the effluent are set to determine how much the effluent must be diluted before toxic effects are no longer noted. Acute toxicity is checked by statistically analyzing whether significantly more organisms die in the effluent dilutions than in the control treatment, and, if significantly more die, how much the effluent must be diluted so as to kill only 50% of the test organisms (the LC50). Chronic toxicity is checked by statistically analyzing whether significantly fewer young are produced by test organisms exposed to the effluent dilutions. Results indicate the lowest effluent concentration which shows a toxic effect (the LOEC) and the highest effluent concentration which does not demonstrate an effect (NOEC).

  14. Evaluation of the detoxication efficiencies for acrylonitrile wastewater treated by a combined anaerobic oxic-aerobic biological fluidized tank (A/O-ABFT) process: Acute toxicity and zebrafish embryo toxicity.

    PubMed

    Na, Chunhong; Zhang, Ying; Deng, Minjie; Quan, Xie; Chen, Shuo; Zhang, Yaobin

    2016-07-01

    Acrylonitrile (ACN) wastewater generated during ACN production has been reported to be toxic to many aquatic organisms. However, few studies have evaluated toxicity removal of ACN wastewater during and after the treatment process. In this study, the detoxication ability of an ACN wastewater treatment plant (WWTP) was evaluated using Daphnia magna, Danio rerio and zebrafish embryo. This ACN WWTP has a combined anaerobic oxic-aerobic biological fluidized tank (A/O-ABFT) process upgraded from the traditional anaerobic oxic (A/O) process. Moreover, the potential toxicants of the ACN wastewaters were identified by gas chromatography-mass spectrometry (GC-MS). The raw ACN wastewater showed high acute and embryo toxicity. 3-Cyanopyridine, succinonitrile and a series of nitriles were detected as the toxic contributors of ACN wastewater. The A/O process was effective for the acute and embryo toxicity removal, as well as the organic toxicants. However, the A/O effluent still showed acute and embryo toxicity which was attributed by the undegraded and the newly generated toxicants during the A/O process. The residual acute and embryo toxicity as well as the organic toxicants in the A/O effluent were further reduced after going through the downstream ABFT process system. The final effluent displayed no significant acute and embryo toxicity, and less organic toxicants were detected in the final effluent. The upgrade of this ACN WWTP results in the improved removal efficiencies for acute and embryo toxicity, as well as the organic toxicants. PMID:27037768

  15. 9 CFR 113.37 - Detection of pathogens by the chicken embryo inoculation test.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... embryo inoculation test. 113.37 Section 113.37 Animals and Animal Products ANIMAL AND PLANT HEALTH... VECTORS STANDARD REQUIREMENTS Standard Procedures § 113.37 Detection of pathogens by the chicken embryo...-serum mixture shall be inoculated into each of at least 20 fully susceptible chicken embryos. (1)...

  16. 9 CFR 113.37 - Detection of pathogens by the chicken embryo inoculation test.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... embryo inoculation test. 113.37 Section 113.37 Animals and Animal Products ANIMAL AND PLANT HEALTH... VECTORS STANDARD REQUIREMENTS Standard Procedures § 113.37 Detection of pathogens by the chicken embryo...-serum mixture shall be inoculated into each of at least 20 fully susceptible chicken embryos. (1)...

  17. 9 CFR 113.37 - Detection of pathogens by the chicken embryo inoculation test.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... embryo inoculation test. 113.37 Section 113.37 Animals and Animal Products ANIMAL AND PLANT HEALTH... VECTORS STANDARD REQUIREMENTS Standard Procedures § 113.37 Detection of pathogens by the chicken embryo...-serum mixture shall be inoculated into each of at least 20 fully susceptible chicken embryos. (1)...

  18. 9 CFR 113.37 - Detection of pathogens by the chicken embryo inoculation test.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... embryo inoculation test. 113.37 Section 113.37 Animals and Animal Products ANIMAL AND PLANT HEALTH... VECTORS STANDARD REQUIREMENTS Standard Procedures § 113.37 Detection of pathogens by the chicken embryo...-serum mixture shall be inoculated into each of at least 20 fully susceptible chicken embryos. (1)...

  19. 9 CFR 113.37 - Detection of pathogens by the chicken embryo inoculation test.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... embryo inoculation test. 113.37 Section 113.37 Animals and Animal Products ANIMAL AND PLANT HEALTH... VECTORS STANDARD REQUIREMENTS Standard Procedures § 113.37 Detection of pathogens by the chicken embryo...-serum mixture shall be inoculated into each of at least 20 fully susceptible chicken embryos. (1)...

  20. [Embryo-fetus development toxicity of a novel PPAR-δ agonist in rat].

    PubMed

    Gong, Hua-Yun; Zhu, Yong; Li, Zong-He; Fan, Xiao-Yan; Fan, Rong; Wang, Fang-Tong

    2014-11-01

    The study aims to investigate the embryo-fetus development toxicity of the novel PPAR-δ agonist HS060098 on SD rats. The pregnant rats that were randomly divided into the solvent control group (1% hydroxypropyl methyl cellulose water solution) and HS060098 suspension groups (10, 30 and 100 mg x kg(-1) xd(-1)) were orally administered with HS060098 suspension or vehicle during the gestation of 6 -15 days (GD6-15). At termination (GD20), female rats were sacrificed. The pregnant females were evaluated by corpora lutea count, implantation sites, existence and death of embryos. Fetal sex, weight, externals, variations and malformations of viscus and skeleton were observed. The results show that there were no significant abnormality in maternal general conditions and fetal appearance as well as viscera, but in the 100 mg x kg(-1) x d(-1) group, the maternal weight gain decreased greatly (P < 0.01) and the skeletal ossification delayed remarkably (P < 0.01); in the 30 mg x kg(-1) xd(-1) group, the fatal and litter number of incompletely ossified sternebrae II was higher than those of the control group (P < 0.05); the skeletal malformations occurred in all dose groups, which indicate that the novel PPAR-δ agonist HS060098 had maternal toxicity and adversely effected fetal skeletal development under the experimental conditions. PMID:25757278

  1. Toxicity and cardiac effects of carbaryl in early developing zebrafish (Danio rerio) embryos

    SciTech Connect

    Lin, C.C.; Hui, Michelle N.Y.; Cheng, S.H. E-mail: bhcheng@cityu.edu.hk

    2007-07-15

    Carbaryl, an acetylcholinesterase inhibitor, is known to be moderately toxic to adult zebrafish and has been reported to cause heart malformations and irregular heartbeat in medaka. We performed experiments to study the toxicity of carbaryl, specifically its effects on the heart, in early developing zebrafish embryos. LC50 and EC50 values for carbaryl at 28 h post-fertilization were 44.66 {mu}g/ml and 7.52 {mu}g/ml, respectively, and 10 {mu}g/ml carbaryl was used in subsequent experiments. After confirming acetylcholinesterase inhibition by carbaryl using an enzymatic method, we observed red blood cell accumulation, delayed hatching and pericardial edema, but not heart malformation as described in some previous reports. Our chronic exposure data also demonstrated carbaryl-induced bradycardia, which is a common effect of acetylcholinesterase inhibitors due to the accumulation of acetylcholine, in embryos from 1 day post-fertilization (dpf) to 5 dpf. The distance between the sinus venosus, the point where blood enters the atrium, and the bulbus arteriosus, the point where blood leaves the ventricle, indicated normal looping of the heart tube. Immunostaining of myosin heavy chains with the ventricle-specific antibody MF20 and the atrium-specific antibody S46 showed normal development of heart chambers. At the same time, acute exposure resulted in carbaryl-induced bradycardia. Heart rate dropped significantly after a 10-min exposure to 100 {mu}g/ml carbaryl but recovered when carbaryl was removed. The novel observation of carbaryl-induced bradycardia in 1- and 2-dpf embryos suggested that carbaryl affected cardiac function possibly through an alternative mechanism other than acetylcholinesterase inhibition such as inhibition of calcium ion channels, since acetylcholine receptors in zebrafish are not functional until 3 dpf. However, the exact nature of this mechanism is currently unknown, and thus further studies are required.

  2. BIOEQUIVALENCE APPROACH FOR WHOLE EFFLUENT TOXICITY TESTING

    EPA Science Inventory

    Increased use of whole effluent toxicity (WET) tests in the regulatory arena has brought increased concern over the statistical analysis of WET test data and the determination of toxicity. One concern is the issue of statistical power. A number of WET tests may pass the current...

  3. The status of toxicity tests with sediment in Brazil

    SciTech Connect

    Araujo, R.P.A.

    1995-12-31

    The earliest studies in Brazil aiming to evaluate sediment quality through toxicity tests started in the beginning of the 80`s. These were developed by the Environmental Sanitation Agency of Sao Paulo State (CETESB) in Cubatao River and Billings Reservoir, which are located in industrialized and populous regions. Elutriate phase sediment toxicity tests were run with Daphnia similis. In the Cubatao River Basin the combination of toxicity, chemistry data and benthic community structure provided clear indications of sites with different levels of pollution. At this time there was a consensus that the study of a complex compartment such as sediment needed improvements in sampling and analysis procedures. Only in the 90`s the investigations involving sediment toxicity assessment were resumed by CETESB, and it was clear that integrative studies were needed in order to make environmental quality assessment. This kind of studies were conducted by CETESB in some highly polluted areas of Sao Paulo State, Ceriodaphnia dubia and Photobacterium phosphoreum interstitial water tests and Hyalella sp whole sediment tests were run, and the results correlated with several sediment organic and inorganic contaminants. The Sediment Quality Triad proposed by Chapman was applied in one of these studies. This approach was extremely useful in interpreting the data. At the same time marine sediment toxicity tests were developed by CETESB in collaboration with Sao Paulo University, and tests were run with the amphipods Tiburonella viscana, Battyporeiapus bisetosus; tanaidacean Kalliapseudes shubarti and the echinoderm Lytechinus variegatus. The embryo test with L. variegatus was the most adequate in these studies. Nowadays there are other groups in some universities developing sediment toxicity tests with Hyalella and Chironomus in response to a growing concern in Brazil to establish adequate sediment quality assessment guidelines.

  4. POREWATER TOXICITY TESTING: AN OVERVIEW

    EPA Science Inventory

    Sediments act as sinks for contaminants, where they may build up to toxic levels. Sediments containing toxic levels of contaminants pose a risk to aquatic life, human health, and wildlife. There is an overwhelming amount of evidence that demonstrates chemicals in sediments are re...

  5. Development of toxicant identification procedures for whole sediment toxicity tests

    SciTech Connect

    Mount, D.R.; Henke, C.E.; Ingersoll, C.G.; Besser, J.M.; Ankley, G.T.; Norberg-King, T.J.; West, C.W.

    1995-12-31

    To effectively assess and manage contaminated sediments, identifying the specific contaminants responsible for sediment toxicity is highly desirable. Though effective toxicity identification evaluation (TIE) methods are well established for water column toxicity, new TIE methodologies are needed that address the special characteristics of whole sediment toxicity tests. Much of the effort to date has focused on the assessment of ammonia toxicity. Whereas pH manipulation is a key tool used to characterize ammonia toxicity in water column TIE, control of pH in interstitial water is much more challenging. Direct addition of hard acid has shown undesirable side effects (e.g., liberation and oxidation of iron), while CO{sub 2}-enrichment is limited in penetration of fine-grained sediments. Biological buffers (MES and POPSO) incorporated into the sediment are effective at altering interstitial pH without causing direct toxicity to Chironomus tentans, Lumbriculus variegatus, and to a lesser extent Hyalella azteca, but the range of pH control achieved has been small ({+-} 0.5 units). Introduction of aquatic plants reduces ammonia concentrations in the water column, but may not provide sufficient control of interstitial water. To date, the most promising results have been achieved using zeolite; adding zeolite to sediment produces moderate reductions in interstitial ammonia concentrations and is non-toxic to the organisms referenced above. Attempts to induce microbial removal of ammonia have been unsuccessful thus far. This presentation will review these and other sediment TIE methods currently under development in laboratories.

  6. Synthesis, physicochemical studies, embryos toxicity and DNA interaction of some new Iron(II) Schiff base amino acid complexes

    NASA Astrophysics Data System (ADS)

    Abdel-Rahman, Laila H.; El-Khatib, Rafat M.; Nassr, Lobna A. E.; Abu-Dief, Ahmed M.

    2013-05-01

    New Fe(II) Schiff base amino acid complexes derived from the condensation of o-hydroxynaphthaldehyde with L-alanine, L-phenylalanine, L-aspartic acid, L-histidine and L-arginine were synthesized and characterized by elemental analysis, IR, electronic spectra, and conductance measurements. The stoichiometry and the stability constants of the complexes were determined spectrophotometrically. The investigated Schiff bases exhibited tridentate coordination mode with the general formulae [Fe(HL)2]nH2O for all amino acids except L-histidine. But in case of L-histidine, the ligand acts as tetradentate ([FeL(H2O)2]2H2O), where HL = mono anion and L = dianion of the ligand. The structure of the prepared complexes is suggested to be octahedral. The prepared complexes were tested for their toxicity on chick embryos and found to be safe until a concentration of 100 ?g/egg with full embryos formation. The interaction between CT-DNA and the investigated complexes were followed by spectrophotometry and viscosity measurements. It was found that, the prepared complexes bind to DNA via classical intercalative mode and showed a different DNA cleavage activity with the sequence: nhi > nari > nali > nasi > nphali. The thermodynamic Profile of the binding of nphali complex and CT-DNA was constructed by analyzing the experimental data of absorption titration and UV melting studies with the McGhee equation, van't Hoff's equation, and the Gibbs-Helmholtz equation.

  7. In Vivo Quantitative Study of Sized-Dependent Transport and Toxicity of Single Silver Nanoparticles Using Zebrafish Embryos

    PubMed Central

    Lee, Kerry J.; Browning, Lauren M.; Nallathamby, Prakash D.; Desai, Tanvi; Cherukui, Pavan K.; Xu, Xiao-Hong Nancy

    2012-01-01

    Nanomaterials possess distinctive physicochemical properties (e.g., small sizes, high surface area-to-volume ratios) and promise a wide variety of applications, ranging from design of high quality consumer products to effective disease diagnosis and therapy. These properties can lead to toxic effects, potentially hindering advance in nanotechnology. In this study, we have synthesized and characterized purified and stable (non-aggregation) silver nanoparticles (Ag NPs, 41.6±9.1 nm in average diameters), and utilized early-developing (cleavage-stage) zebrafish embryos (critical aquatic and eco- species) as in vivo model organisms to probe diffusion and toxicity of Ag NPs. We found that single Ag NPs (30–72 nm diameters) passively diffused into the embryos through chorionic pores via random Brownian motion and stayed inside the embryos throughout their entire development (120 hours-post-fertilization, hpf). Dose and size dependent toxic effects of the NPs on embryonic development were observed, showing the possibility of tuning biocompatibility and toxicity of the NPs. At lower concentrations of the NPs (≤ 0.02 nM), 75–91% of embryos developed to normal zebrafish. At the higher concentrations of NPs (≥ 0.20 nM), 100% of embryos became dead. At the concentrations in between (0.02–0.2 nM), embryos developed to various deformed zebrafish. Number and sizes of individual Ag NPs embedded in tissues of normal and deformed zebrafish at 120 hpf were quantitatively analyzed, showing deformed zebrafish with higher number of larger NPs than normal zebrafish, and size-dependent nanotoxicity. By comparing with our previous studies of smaller Ag NPs (11.6±3.5 nm), the results further demonstrate striking size-dependent nanotoxicity that, at the same molar concentration, the larger Ag NPs (41.6±9.1 nm) are more toxic than the smaller Ag NPs (11.6±3.5 nm). PMID:22486336

  8. GUIDELINES FOR CONDUCTING EARLY LIFE STAGE TOXICITY TESTS WITH JAPANESE MEDAKA (ORYZIAS LATIPES)

    EPA Science Inventory

    This manual represents a procedural guide for conducting embryo-larval early life stage (ELS) toxicity tests with Japanese medaka (Oryzias latipes). hese procedures are based upon evaluation of published papers and recent methods development work conducted at our laboratory in Du...

  9. Toxicity of endosulfan on embryo-larval development of the South American toad Rhinella arenarum.

    PubMed

    Svartz, Gabriela V; Wolkowicz, Ianina R Hutler; Coll, Cristina S Pérez

    2014-04-01

    Endosulfan is a widely used pesticide despite its extreme toxicity to a variety of taxa and its worldwide ban. The aim of the present study was to evaluate the acute and chronic toxicity of endosulfan on the embryonic-larval development of the common South American toad Rhinella arenarum. The results showed that lethal and sublethal effects increased with concentration and exposure time. The sensitivity to endosulfan increased during the larval period, the complete operculum stage (S.25) being the most sensitive (504-h median lethal concentration [LC50] = 0.01 mg endosulfan/L; 10% lethal concentration [LC10] = 0.004 mg endosulfan/L). Endosulfan exposure caused morphological abnormalities such as general underdevelopment, edema, gill malformations, and cellular dissociation as well as neurotoxicity. Our results also showed that larvae exposed to concentrations of 0.005 mg endosulfan/L and 0.01 mg endosulfan/L completed metamorphosis earlier than controls, but with underdevelopment. The 240-h teratogenic index was 6.13, implying a high risk for embryos to be malformed in the absence of significant embryonic lethality. Because the hazard quotients for chronic exposure were over 1, the level of concern value and toxicity endpoints obtained in the present study for R. arenarum occurred at concentrations lower than the levels of endosulfan reported in the environment, this pesticide should be considered a potential risk for this species. PMID:24375551

  10. Toxicity of crude oil and pyrene to the embryos of beach spawning capelin (Mallotus villosus).

    PubMed

    Frantzen, Marianne; Falk-Petersen, Inger-Britt; Nahrgang, Jasmine; Smith, Timothy J; Olsen, Gro H; Hangstad, Thor Arne; Camus, Lionel

    2012-02-01

    Due to a northward shift in oil and gas activities, there is an increasing need to understand the potential anthropogenic impacts of oil-related compounds on sub-Arctic and Arctic organisms, particularly those in coastal habitats. Capelin (Mallotus villosus), a key fish species in the Barents Sea ecosystem, undertakes aggregated spawning at both intertidal and subtidal coastal localities in northern Norway. To investigate the sensitivity of capelin embryos to oil compounds, newly fertilized capelin eggs were collected from a spawning beach and exposed until hatch (32 days) to either the water soluble fraction of crude oil or the single PAH compound, pyrene. Threshold levels for egg mortality, development and hatching success were determined. Concentrations of 40 μg/L crude oil (∑26 PAHs) and 55 μg/L pyrene significantly increased embryonic mortality rates and decreased hatching success, compared with controls, indicating that a potential oil spill in the vicinity of capelin spawning grounds may cause significant impacts. No significant incidence of adverse effects such as yolk sac oedema, pericardia oedema, haemorrhages, craniofacial abnormalities, premature hatch or inhibited growth was observed. Histological studies of hatched larvae did not reveal specific sublethal effects in tissues and organs. Developmental delays and subsequent embryo death were noticed at the period of eye pigmentation in affected groups. Early life-history stages of capelin are sensitive indicators of PAH impacts, but the mechanisms responsible for the toxic effects require further investigation. PMID:22037118

  11. Acute toxicity and sublethal effects of the mixture glyphosate (Roundup Active) and Cosmo-Flux 411F to anuran embryos and tadpoles of four Colombian species.

    PubMed

    Henao Muñoz, Liliana Marcela; Montes Rojas, Claudia Marsela; Bernal Bautista, Manuel Hernando

    2015-03-01

    Glyphosate is the most widely used herbicide in the world with application in agriculture, forestry, industrial weed control, garden and aquatic environments. However, its use is highly controversial for the possible impact on not-target organisms, such as amphibians, which are vanishing at an alarming and rapid rate. Due to the high solubility in water and ionic nature, the glyphosate requires of surfactants to increase activity. In addition, for the control of coca (Erythroxylum coca) and agricultural weeds in Colombia, formulated glyphosate is mixed and sprayed with the adjuvant Cosmo-Flux 411F to increase the penetration and activity of the herbicide. This study evaluates the acute toxic and sublethal effects (embryonic development, tadpole body size, tadpole swimming performance) of the mixture of the formulated glyphosate Roundup Active and Cosmo-Flux 411F to anuran embryos and tadpoles of four Colombian species under 96h laboratory standard tests and microcosms, which are more similar to field conditions as they include soil, sand and macrophytes. In the laboratory, embryos and tadpoles of Engystomops pustulosus were the most tolerant (LC50 = 3904 microg a.e./L; LC50=2 799 pg a.e./L, respectively), while embryos and tadpoles of Hypsiboas crepitans (LC50=2 203 microg a.e./L; LC50=1424 microgg a.e./L, respectively) were the most sensitive. R. humboldti and R. marina presented an intermediate toxicity. Embryos were significantly more tolerant to the mixture than tadpoles, which could be likely attributed to the exclusion of chemicals by the embryonic membranes and the lack of organs, such as gills, which are sensitive to surfactants. Sublethal effects were observed for the tadpole body size, but not for the embryonic development and tadpole swimming performance. In microcosms, no toxicity (LC50 could not be estimated), or sublethal responses were observed at concentrations up to fourfold (14.76 kg glyphosate a.e./ha) the highest field application rate of 3.69 kg glyphosate a.e./ha. Thus, toxicity was less in the microcosms than in laboratory tests, which may be attributed to the presence of sediments and organic matter which rapidly adsorb glyphosate and surfactants such as POEA. It is concluded that the mixture of glyphosate (Roundup Active) and Cosmo-Flux*411F, as used in the field, has a negligible toxic effect to embryos and tadpoles of the species tested in this study. PMID:26299127

  12. RAPID AQUATIC TOXICITY ASSAY USING INCORPORATION OF TRITIATED-THYMIDINE INTO SEA URCHIN, 'ARBACIA PUNCTULATA', EMBRYO: EVALUATION OF TOXICANT EXPOSURE PROCEDURES

    EPA Science Inventory

    Toxicity of substances in seawater was measured using growth inhibition of embryonic sea urchins during a short period after fertilization. Growth of Arbacia punctulata embryos was monitored by incorporation of tritium-labeled thymidine. The paper presents a comparison of toxican...

  13. Sulfide tolerance of four marine bioassay test species used to evaluate sediment toxicity

    SciTech Connect

    Knezovich, J.P.; Jelinski, J.A.; Anderson, S.L.; Steichen, D.J.

    1994-12-31

    Hydrogen sulfide is a potential positive interference associated with marine sediment toxicity tests. It is necessary, therefore, to evaluate the possible contribution of sulfides to sediment toxicity before toxicity attributed to anthropogenic contaminants can be determined. The sulfide tolerances of several marine bioassay test species (R. abronius, E. estuaris, M. edulis and S. purpuratus) commonly used in bulk sediment and porewater toxicity tests have not been reported. The authors used a flow-through exposure system to expose these organisms to constant concentrations of sulfide during 48-h exposure periods. Significant abnormalities occurred at 3 to 5 micromoles total sulfide per liter ({mu}M/L) in M. edulis embryos, and at 6{mu}M/L in S. purpuratus embryos. Survivorship of amphipods was significantly reduced at total sulfide concentrations of 45 and 126 {mu}M/L for R. abronius and E estuaris, respectively. These tolerances provide guidelines for evaluating the potential for sulfide toxicity prior to the initiation of toxicity tests. Because sulfide concentrations in bulk sediment and pore-water may exceed these values, care must be taken to ensure that sulfide toxicity is not contributing to toxicity observed when using these test organisms.

  14. Searching for biomarkers of developmental toxicity with microarrays: normal eye morphogenesis in rodent embryos

    SciTech Connect

    Nemeth, Kimberly A.; Singh, Amar V.; Knudsen, Thomas B. . E-mail: thomas.knudsen@louisville.edu

    2005-08-07

    Gene expression arrays reveal the potential linkage of altered gene expression with specific adverse effects leading to disease phenotypes. But how closely do microarray data reflect early physiological or pharmacological measures that predict toxic event(s)? To explore this issue, we have undertaken experiments in early mouse embryos exposed to various teratogens during neurulation stages with the aim of correlating large-scale changes in gene expression across the critical period during exposure. This study reports some of the large-scale changes in gene expression that can be detected in the optic rudiment of the developing mouse and rat embryo across the window of development during which the eye is exceedingly sensitive to teratogen-induced micro-/anophthalmia. Microarray analysis was performed on RNA from the headfold or ocular region at the optic vesicle and optic cup stages when the ocular primordium is enriched for Pax-6, a master control gene for eye morphogenesis. Statistical selection of differentially regulated genes and various clustering techniques identified groups of genes in upward or downward trajectories in the normal optic primordium during early eye development in mouse and rat species. We identified 165 genes with significant differential expression during eye development, and a smaller subset of 58 genes that showed a tight correlation between mouse-rat development. Significantly over-represented functional categories included fatty acid metabolism (up-regulated) and glycolysis (down-regulated). From studies such as these that benchmark large-scale gene expression during normal embryonic development, we may be able to identify the panel of biomarkers that best correlate with species differences and the risks for developmental toxicity.

  15. Toxicity assessment of TiO₂ nanoparticles in zebrafish embryos under different exposure conditions.

    PubMed

    Clemente, Z; Castro, V L S S; Moura, M A M; Jonsson, C M; Fraceto, L F

    2014-02-01

    The popularity of TiO2 nanoparticles (nano-TiO2) lies in their wide range of nanotechnological applications, together with low toxicity. Meanwhile, recent studies have shown that the photocatalytic properties of this material can result in alterations in their behavior in the environment, causing effects that have not yet been fully elucidated. The objective of this study was to evaluate the toxicity of two formulations of nano-TiO2 under different illumination conditions, using an experimental model coherent with the principle of the three Rs of alternative animal experimentation (reduction, refinement, and replacement). Embryos of the fish Danio rerio were exposed for 96h to different concentrations of nano-TiO2 in the form of anatase (TA) or an anatase/rutile mixture (TM), under either visible light or a combination of visible and ultraviolet light (UV). The acute toxicity and sublethal parameters evaluated included survival rates, malformation, hatching, equilibrium, and overall length of the larvae, together with biochemical biomarkers (specific activities of catalase (CAT), glutathione S-transferase (GST), and acid phosphatase (AP)). Both TA and TM caused accelerated hatching of the larvae. Under UV irradiation, there was greater mortality of the larvae of the groups exposed to TM, compared to those exposed to TA. Exposure to TM under UV irradiation altered the equilibrium of the larvae. Alterations in the activities of CAT and GST were indicative of oxidative stress, although no clear dose-response relationship was observed. The effects of nano-TiO2 appeared to depend on both the type of formulation and the illumination condition. The findings contribute to elucidation of the factors involved in the toxicity of these nanoparticles, as well as to the establishment of protocols for risk assessments of nanotechnology. PMID:24418748

  16. Literature review on duckweed toxicity testing

    SciTech Connect

    Wang, W. )

    1990-06-01

    Duckweed commonly refers to a group of floating, flowering plants of the family Lemnaceae. Duckweed plants are fast growing and widely distributed. They are easy to culture and to test. Some reports suggest that duckweed plants are tolerant to environmental toxicity. Other studies, however, indicate that duckweed plants are as sensitive to toxicity as other aquatic species. Duckweed plants are especially suitable for use in complex effluent bioassays, and for testing herbicide pollution in the aquatic environment, lake and river pollution, sediment toxicity, and the like. Duckweed and algae represent different levels of complexity in the plant kingdom. They complement each other as phytotoxicity test organisms, instead of mutually excluding each other. Many duckweed species have been studied, primarily of the Lemna and Spirodela genera. Lemna minor and L. gibba have been recommended as standard test species. Differences in duckweed test methodology occur with regard to test types, test vessels, control tests, nutrient media, end points, and applications. 76 references.

  17. Folic acid protects against arsenic-mediated embryo toxicity by up-regulating the expression of Dvr1

    PubMed Central

    Ma, Yan; Zhang, Chen; Gao, Xiao-Bo; Luo, Hai-Yan; Chen, Yang; Li, Hui-hua; Ma, Xu; Lu, Cai-Ling

    2015-01-01

    As a nutritional factor, folic acid can prevent cardiac and neural defects during embryo development. Our previous study showed that arsenic impairs embryo development by down-regulating Dvr1/GDF1 expression in zebrafish. Here, we investigated whether folic acid could protect against arsenic-mediated embryo toxicity. We found that folic acid supplementation increases hatching and survival rates, decreases malformation rate and ameliorates abnormal cardiac and neural development of zebrafish embryos exposed to arsenite. Both real-time PCR analysis and whole in-mount hybridization showed that folic acid significantly rescued the decrease in Dvr1 expression caused by arsenite. Subsequently, our data demonstrated that arsenite significantly decreased cell viability and GDF1 mRNA and protein levels in HEK293ET cells, while folic acid reversed these effects. Folic acid attenuated the increase in subcellular reactive oxygen species (ROS) levels and oxidative adaptor p66Shc protein expression in parallel with the changes in GDF1 expression and cell viability. P66Shc knockdown significantly inhibited the production of ROS and the down-regulation of GDF1 induced by arsenite. Our data demonstrated that folic acid supplementation protected against arsenic-mediated embryo toxicity by up-regulating the expression of Dvr1/GDF1, and folic acid enhanced the expression of GDF1 by decreasing p66Shc expression and subcellular ROS levels. PMID:26537450

  18. DEVELOPMENTAL TOXICITY OF COPPER CHLORIDE, METHYLENE CHLORIDE,AND 6-AMINONICOTINAMIDE TO EMBRYOS OF THE GRASS SHRIMPPALAEMONETES PUGIO

    EPA Science Inventory

    Embryos of estuarine grass shrimp Palaemonetes pugio have demonstrated sensitivity to various solvents and petroleum products, indicating utility for evaluating estuarine contamination. Testing was performed to establish concentration-response curves for methylene chloride, cop...

  19. A comparison of standard acute toxicity tests with rapid-screening toxicity tests

    SciTech Connect

    Toussaint, M.W.; Shedd, T.R.; Schalie, W.H. van der; Leather, G.R.

    1995-05-01

    This study compared the relative sensitivity of five inexpensive, rapid toxicity tests to the sensitivity of five standard aquatic acute toxicity tests through literature review and testing. The rapid toxicity tests utilized organisms that require little culturing or handling prior to testing: a freshwater rotifer (Branchionus calyciflorus); brine shrimp (Artemia salina); lettuce (Lactuca sativa); and two microbial tests (Photobacterium phosphoreum--Microtox{reg_sign} test, and a mixture of bacterial species--the Polytox{reg_sign} test). Standard acute toxicity test species included water fleas (Daphnia magna and Ceriodaphnia dubia), green algae (Selenastrum capricornutum), fathead minnows (Pimephales promelas), and mysid shrimp (Mysidopsis bahia). Sensitivity comparisons between rapid and standard acute toxicity tests were based on LC50/EC50 data from 11 test chemicals. Individually, the lettuce and rotifer tests ranked most similar in sensitivity to the standard tests, while Microtox fell just outside the range of sensitivities represented by the group of standard acute toxicity tests. The brine shrimp and Polytox tests were one or more orders of magnitude different from the standard acute toxicity tests for most compounds. The lettuce, rotifer, and Microtox tests could be used as a battery for preliminary toxicity screening of chemicals. Further evaluation of complex real-world environmental samples is recommended.

  20. Comparison of standard acute toxicity tests with rapid-screening toxicity tests

    SciTech Connect

    Toussaint, M.W.; Shedd, T.R.; VanDerSchal, W.H.; Leather, G.R.

    1995-10-01

    This study compared the relative sensitivity of five inexpensive, rapid toxicity tests to the sensitivity of five standard aquatic acute toxicity tests through literature review and testing. The rapid toxicity tests utilized organisms that require little culturing or handling prior to testing: a freshwater rotifer (Branchionus ccalyciflorus); brine shrimp (Artemia salina); lettuce (Lactuca sativa); and two microbial tests (Photo bacterium phosphoreum - Microtox test, and a mixture of bacterial species - the polytox test). Standard acute toxicity test species included water fleas (Daphnia magna and Ceriadaphnta dubia), green algae (Setenastrum capricarnutum), fathead minnows (Pimephalespromelas), and mysid shrimp (Mysidopsis bahia). Sensitivity comparisons between rapid and standard acute toxicity tests were based on LC5O/EC50 data from 11 test chemicals. Individually, the lettuce and rotifer tests ranked most similar in sensitivity to the standard tests, while Microtox fell just outside the range of sensitivities represented by the group of standard acute toxicity tests. The brine shrimp and Polytox tests were one or more orders of magnitude different from the standard acute toxicity tests for most compounds. The lettuce, rotifer, and Microtox tests could be used as a battery for preliminary toxicity screening of chemicals. Further evaluation of complex real-world environmental samples is recommended.

  1. REGULATORY APPLICATIONS OF POREWATER TOXICITY TESTING

    EPA Science Inventory

    The purpose of this chapter is to evaluate the use of porewater toxicity tests in regulatory applications, including their potential use in the development of sediment quality guideline (SQG) values. Specifically, the following discussion focuses on the appropriateness and readin...

  2. Automated Test Systems for Toxic Vapor Detectors

    NASA Technical Reports Server (NTRS)

    Mattson, C. B.; Hammond, T. A.; Schwindt, C. J.

    1997-01-01

    The NASA Toxic Vapor Detection Laboratory (TVDL) at the Kennedy Space Center (KSC), Florida, has been using Personal Computer based Data Acquisition and Control Systems (PCDAS) for about nine years. These systems control the generation of toxic vapors of known concentrations under controlled conditions of temperature and humidity. The PCDAS also logs the test conditions and the test article responses in data files for analysis by standard spreadsheets or custom programs. The PCDAS was originally developed to perform standardized qualification and acceptance tests in a search for a commercial off-the-shelf (COTS) toxic vapor detector to replace the hydrazine detectors for the Space Shuttle launch pad. It has since become standard test equipment for the TVDL and is indispensable in producing calibration standards for the new hydrazine monitors at the 10 part per billion (ppb) level. The standard TVDL PCDAS can control two toxic vapor generators (TVG's) with three channels each and two flow/ temperature / humidity (FTH) controllers and it can record data from up to six toxic vapor detectors (TVD's) under test and can deliver flows from 5 to 50 liters per minute (L/m) at temperatures from near zero to 50 degrees Celsius (C) using an environmental chamber to maintain the sample temperature. The concentration range for toxic vapors depends on the permeation source installed in the TVG. The PCDAS can provide closed loop control of temperature and humidity to two sample vessels, typically one for zero gas and one for the standard gas. This is required at very low toxic vapor concentrations to minimize the time required to passivate the sample delivery system. Recently, there have been several requests for information about the PCDAS by other laboratories with similar needs, both on and off KSC. The purpose of this paper is to inform the toxic vapor detection community of the current status and planned upgrades to the automated testing of toxic vapor detectors at the Kennedy Space Center.

  3. Automated Test Systems for Toxic Vapor Detectors

    NASA Technical Reports Server (NTRS)

    Mattson, C. B.; Hammond, T. A.; Schwindt, C. J.

    1997-01-01

    The NASA Toxic Vapor Detection Laboratory (TVDL) at the Kennedy Space Center (KSC), Florida, has been using Personal Computer based Data Acquisition and Control Systems (PCDAS) for about nine years. These systems control the generation of toxic vapors of known concentrations under controlled conditions of temperature and humidity. The PCDAS also logs the test conditions and the test article responses in data files for analysis by standard spreadsheets or custom programs. The PCDAS was originally developed to perform standardized qualification and acceptance tests in a search for a commercial off-the-shelf (COTS) toxic vapor detector to replace the hydrazine detectors for the Space Shuttle launch pad. It has since become standard test equipment for the TVDL and is indispensable in producing calibration standards for the new hydrazine monitors at the 10 part per billion (ppb) level. The standard TVDL PCDAS can control two toxic vapor generators (TVG's) with three channels each and two flow/temperature/humidity (FIFH) controllers and it can record data from up to six toxic vapor detectors (TVD's) under test and can deliver flows from 5 to 50 liters per minute (L/m) at temperatures from near zero to 50 degrees Celsius (C) using an environmental chamber to maintain the sample temperature. The concentration range for toxic vapors depends on the permeation source installed in the TVG. The PCDAS can provide closed loop control of temperature and humidity to two sample vessels, typically one for zero gas and one for the standard gas. This is required at very low toxic vapor concentrations to minimize the time required to passivate the sample delivery system. Recently, there have been several requests for information about the PCDAS by other laboratories with similar needs, both on and off KSC. The purpose of this paper is to inform the toxic vapor detection community of the current status and planned upgrades to the automated testing of toxic vapor detectors at the Kennedy Space Center.

  4. Automated Zebrafish Chorion Removal and Single Embryo Placement: Optimizing Throughput of Zebrafish Developmental Toxicity Screens

    PubMed Central

    Mandrell, David; Truong, Lisa; Jephson, Caleb; Sarker, Mushfiqur R.; Moore, Aaron; Lang, Christopher; Simonich, Michael T.; Tanguay, Robert L.

    2012-01-01

    The potential of the developing zebrafish model for toxicology and drug discovery is limited by inefficient approaches to manipulating and chemically exposing zebrafish embryos—namely, manual placement of embryos into 96- or 384-well plates and exposure of embryos while still in the chorion, a barrier of poorly characterized permeability enclosing the developing embryo. We report the automated dechorionation of 1600 embryos at once at 4 h postfertilization (hpf) and placement of the dechorionated embryos into 96-well plates for exposure by 6 hpf. The process removed ≥95% of the embryos from their chorions with 2% embryo mortality by 24 hpf, and 2% of the embryos malformed at 120 hpf. The robotic embryo placement allocated 6-hpf embryos to 94.7% ± 4.2% of the wells in multiple 96-well trials. The rate of embryo mortality was 2.8% (43 of 1536) from robotic handling, the rate of missed wells was 1.2% (18 of 1536), and the frequency of multipicks was <0.1%. Embryo malformations observed at 24 hpf occurred nearly twice as frequently from robotic handling (16 of 864; 1.9%) as from manual pipetting (9 of 864; 1%). There was no statistical difference between the success of performing the embryo placement robotically or manually. PMID:22357610

  5. ECVAM and new technologies for toxicity testing.

    PubMed

    Bouvier d'Yvoire, Michel; Bremer, Susanne; Casati, Silvia; Ceridono, Mara; Coecke, Sandra; Corvi, Raffaella; Eskes, Chantra; Gribaldo, Laura; Griesinger, Claudius; Knaut, Holger; Linge, Jens P; Roi, Annett; Zuang, Valérie

    2012-01-01

    The development of alternative empirical (testing) and non-empirical (non-testing) methods to traditional toxicological tests for complex human health effects is a tremendous task. Toxicants may potentially interfere with a vast number of physiological mechanisms thereby causing disturbances on various levels of complexity of human physiology. Only a limited number of mechanisms relevant for toxicity ('pathways' of toxicity) have been identified with certainty so far and, presumably, many more mechanisms by which toxicants cause adverse effects remain to be identified. Recapitulating in empirical model systems (i.e., in vitro test systems) all those relevant physiological mechanisms prone to be disturbed by toxicants and relevant for causing the toxicity effect in question poses an enormous challenge. First, the mechanism(s) of action of toxicants in relation to the most relevant adverse effects of a specific human health endpoint need to be identified. Subsequently, these mechanisms need to be modeled in reductionist test systems that allow assessing whether an unknown substance may operate via a specific (array of) mechanism(s). Ideally, such test systems should be relevant for the species of interest, i.e., based on human cells or modeling mechanisms present in humans. Since much of our understanding about toxicity mechanisms is based on studies using animal model systems (i.e., experimental animals or animal-derived cells), designing test systems that model mechanisms relevant for the human situation may be limited by the lack of relevant information from basic research. New technologies from molecular biology and cell biology, as well as progress in tissue engineering, imaging techniques and automated testing platforms hold the promise to alleviate some of the traditional difficulties associated with improving toxicity testing for complex endpoints. Such new technologies are expected (1) to accelerate the identification of toxicity pathways with human relevance that need to be modeled in test methods for toxicity testing (2) to enable the reconstruction of reductionist test systems modeling at a reduced level of complexity the target system/organ of interest (e.g., through tissue engineering, use of human-derived cell lines and stem cells etc.), (3) to allow the measurement of specific mechanisms relevant for a given health endpoint in such test methods (e.g., through gene and protein expression, changes in metabolites, receptor activation, changes in neural activity etc.), (4) to allow to measure toxicity mechanisms at higher throughput rates through the use of automated testing. In this chapter, we discuss the potential impact of new technologies on the development, optimization and use of empirical testing methods, grouped according to important toxicological endpoints. We highlight, from an ECVAM perspective, the areas of topical toxicity, skin absorption, reproductive and developmental toxicity, carcinogenicity/genotoxicity, sensitization, hematopoeisis and toxicokinetics and discuss strategic developments including ECVAM's database service on alternative methods. Neither the areas of toxicity discussed nor the highlighted new technologies represent comprehensive listings which would be an impossible endeavor in the context of a book chapter. However, we feel that these areas are of utmost importance and we predict that new technologies are likely to contribute significantly to test development in these fields. We summarize which new technologies are expected to contribute to the development of new alternative testing methods over the next few years and point out current and planned ECVAM projects for each of these areas. PMID:22437818

  6. Individual and joint toxic effects of cadmium sulfate and α-naphthoflavone on the development of zebrafish embryo*

    PubMed Central

    Yin, Jian; Yang, Jian-ming; Zhang, Feng; Miao, Peng; Lin, Ying; Chen, Ming-li

    2014-01-01

    This paper aims to evaluate the individual and joint toxicities of cadmium sulfate (CdSO4) and α-naphthoflavone (ANF) in zebrafish embryos. As a result, CdSO4 caused both lethal and sub-lethal effects, such as 24 h post-fertilization (hpf) death and 72 hpf delayed hatching. However, ANF only caused sub-lethal effects, including 48 hpf cardiac edema and 72 hpf delayed hatching. Taking 24 hpf death and 48 hpf cardiac edema as endpoints, the toxicities of CdSO4 and ANF were significantly enhanced by each other. Consistently, both CdSO4 and ANF caused significant oxidative stress, including decreases in the reduced glutathione (GSH) level, inhibition of superoxide dismutase (SOD) activity, as well as increases in malondialdehyde (MDA) content in zebrafish embryos, but these mixtures produced much more significant alterations on the biomarkers. Co-treatment of CdSO4 and ANF significantly down-regulated the mRNA level of multidrug resistance-associated protein (mrp) 1 and cytochrome P450 (cyp) 1a, which constituted the protective mechanisms for zebrafish embryos to chemical toxins. In conclusion, co-treatment of CdSO4 and ANF exhibited a much more severe damage in zebrafish embryos than individual treatment. Meanwhile, production of oxidative stress and altered expression of mrp1 and cyp1a could be important components of such joint toxicity. PMID:25183031

  7. A high-throughput lab-on-a-chip interface for zebrafish embryo tests in drug discovery and ecotoxicology

    NASA Astrophysics Data System (ADS)

    Zhu, Feng; Akagi, Jin; Hall, Chris J.; Crosier, Kathryn E.; Crosier, Philip S.; Delaage, Pierre; Wlodkowic, Donald

    2013-12-01

    Drug discovery screenings performed on zebrafish embryos mirror with a high level of accuracy. The tests usually performed on mammalian animal models, and the fish embryo toxicity assay (FET) is one of the most promising alternative approaches to acute ecotoxicity testing with adult fish. Notwithstanding this, conventional methods utilising 96-well microtiter plates and manual dispensing of fish embryos are very time-consuming. They rely on laborious and iterative manual pipetting that is a main source of analytical errors and low throughput. In this work, we present development of a miniaturised and high-throughput Lab-on-a-Chip (LOC) platform for automation of FET assays. The 3D high-density LOC array was fabricated in poly-methyl methacrylate (PMMA) transparent thermoplastic using infrared laser micromachining while the off-chip interfaces were fabricated using additive manufacturing processes (FDM and SLA). The system's design facilitates rapid loading and immobilization of a large number of embryos in predefined clusters of traps during continuous microperfusion of drugs/toxins. It has been conceptually designed to seamlessly interface with both upright and inverted fluorescent imaging systems and also to directly interface with conventional microtiter plate readers that accept 96-well plates. We also present proof-of-concept interfacing with a high-speed imaging cytometer Plate RUNNER HD® capable of multispectral image acquisition with resolution of up to 8192 x 8192 pixels and depth of field of about 40 μm. Furthermore, we developed a miniaturized and self-contained analytical device interfaced with a miniaturized USB microscope. This system modification is capable of performing rapid imaging of multiple embryos at a low resolution for drug toxicity analysis.

  8. Low-level pyrene exposure causes cardiac toxicity in zebrafish (Danio rerio) embryos.

    PubMed

    Zhang, Youyu; Wang, Chonggang; Huang, Lixing; Chen, Rong; Chen, Yixin; Zuo, Zhenghong

    2012-06-15

    It is widely accepted that the most abundant polycyclic aromatic hydrocarbons (PAHs) in weathered crude oils is cardiotoxic. Although PAHs toxic endpoints show strong correlation with the aryl hydrocarbon receptor (AhR), a ligand activated transcription factor, and is thought to be a potent inducer of cytochrome P4501A, the action mechanism of PAHs on vertebrate cardiovascular development and disease is unclear. Herein, we address the cardiac developmental effects of exposure to the weak AhR agonist pyrene on the early life-stages of zebrafish. Embryos were exposed to 0, 0.05, 0.5, 5, and 50 nmol/L pyrene up to 72h post-fertilization (hpf). Pyrene-treated embryos showed dose-dependent heart abnormalities, such as pericardial edema and cardiac looping defects. Changes in AhR1a, AhR1b, AhR2, and Cyp1A expression were assessed by real-time RT-PCR. The results showed that low-level pyrene failed to alter these genes expression. However, the homeodomain transcription factor Nkx2.5, which plays an essential role in the development of the cardiovascular system, was down-regulated in a dose-dependent manner by pyrene exposure. The bone morphogenetic protein 2b (Bmp2b), which has been identified as the upstream gene of Nkx2.5, also was inhibited in a dose-dependent manner after treatment with pyrene. Taken together, these data indicated that embryonic exposure of zebrafish to low-level environmental pyrene disrupt normal cardiac development and alter expression of defective cardiac differentiation related genes. PMID:22446823

  9. Estimating the gasoline components and formulations toxicity to microalgae (Tetraselmis chuii) and oyster (Crassostrea rhizophorae) embryos: An approach to minimize environmental pollution risk

    SciTech Connect

    Paixao, J.F.; Nascimento, I.A. . E-mail: iracema@ftc.br; Pereira, S.A.; Leite, M.B.L.; Carvalho, G.C.; Silveira, J.S.C.; Reboucas, M.; Matias, G.R.A.; Rodrigues, I.L.P.

    2007-03-15

    Even though petrochemical contamination frequently occurs in the form of oil spills, it is thought that a greater danger to coastal habitats is posed by chronic petrochemical toxicity associated with urban run-off, in which gasoline water-soluble-fraction (WSF) plays an important role. The hypothesis of the entrepreneurs, who were associated to the scientists uncharged of this research, was that recycled petrochemical waste may provide different gasoline formulations, having different toxic properties; the correlation between the gasoline formulations and their components' toxicological effects might contribute to the reformulation of the products, in such a way that the gasoline generated could be less toxic and less harmful to the environment. The aim of this research was to determine the toxic effects of 14 different types of gasoline (formulated, in accordance with National Petroleum Agency standards, from petrochemical waste), on Tetraselmis chuii (microalgae culture) and Crassostrea rhizophorae (embryos). Microalgae and oyster embryos were exposed to different gasoline formulations water-soluble fractions (WSF) at a range of concentrations (0%, 4.6%, 10.0%, 22.0%, 46.0%, and 100%), for 96 and 24 h, respectively. The tests were carried out under controlled conditions. End-points have been CI50-96h (concentration causing 50% growth inhibition in microalgae cultures) and EC50-24h (concentration causing abnormalities on 50% of the exposed embryos). Through these procedures, gasoline formulations, which represent the lowest environmental risk, were selected. Bioassays carried out on the 8 different gasoline components aimed to correlate gasoline toxicity with the toxic potential of its components. The analysis of principal components showed that the C9DI, a mixture of aromatic hydrocarbons of 9 carbon atoms, had the highest level of toxic potential, followed by C9S (a mixture of aromatics with 9-11 carbon atoms) and heavy naphtha. The results showed gasoline formulations 1-4 (monoaromatic hydrocarbons being the most conspicuous components) to be the least toxic, whilst formulations 12-14 (having higher content of C9DI, C9S and naphtha) were found to be the most harmful to organisms. This study led to the identification of the most toxic WSF gasoline components (C9DI and C9S), and to the possibility of developing more eco-compatible gasoline formulations.

  10. Developmental toxicity, acute toxicity and mutagenicity testing in freshwater snails Biomphalaria glabrata (Mollusca: Gastropoda) exposed to chromium and water samples.

    PubMed

    Tallarico, Lenita de Freitas; Borrely, Sueli Ivone; Hamada, Natália; Grazeffe, Vanessa Siqueira; Ohlweiler, Fernanda Pires; Okazaki, Kayo; Granatelli, Amanda Tosatte; Pereira, Ivana Wuo; Pereira, Carlos Alberto de Bragança; Nakano, Eliana

    2014-12-01

    A protocol combining acute toxicity, developmental toxicity and mutagenicity analysis in freshwater snail Biomphalaria glabrata for application in ecotoxicological studies is described. For acute toxicity testing, LC50 and EC50 values were determined; dominant lethal mutations induction was the endpoint for mutagenicity analysis. Reference toxicant potassium dichromate (K2Cr2O7) was used to characterize B. glabrata sensitivity for toxicity and cyclophosphamide to mutagenicity testing purposes. Compared to other relevant freshwater species, B. glabrata showed high sensitivity: the lowest EC50 value was obtained with embryos at veliger stage (5.76mg/L). To assess the model applicability for environmental studies, influent and effluent water samples from a wastewater treatment plant were evaluated. Gastropod sensitivity was assessed in comparison to the standardized bioassay with Daphnia similis exposed to the same water samples. Sampling sites identified as toxic to daphnids were also detected by snails, showing a qualitatively similar sensitivity suggesting that B. glabrata is a suitable test species for freshwater monitoring. Holding procedures and protocols implemented for toxicity and developmental bioassays showed to be in compliance with international standards for intra-laboratory precision. Thereby, we are proposing this system for application in ecotoxicological studies. PMID:25259848

  11. Bioequivalence approach for whole effluent toxicity testing

    SciTech Connect

    Shukla, R.; Wang, Q.; Fulk, F.; Deng, C.; Denton, D.

    2000-01-01

    Increased use of whole effluent toxicity (WET) tests in the regulatory arena has brought increased concern over the statistical analysis of WET test data and the determination of toxicity. One concern is the issue of statistical power. A number of WET tests may pass the current hypothesis test approach because they lack statistical power to detect relevant toxic effects because of large within-test variability. Additionally, a number of WET tests may fail the current approach because they possess excessive statistical power, as a result of small within-test variability, and detect small differences that may not be biologically relevant. The strengths and limitations of both the traditional hypothesis test approach and the bioequivalence approach for use in the National Pollutant Discharge Elimination System program were evaluated. Data from 5,213 single-concentration, short-term chronic WET tests with Ceriodaphnia dubia provided the database for analysis. Comparison of results between the current approach and the bioequivalence approach indicates that the current approach to WET testing is generally sound but that adopting the proposed bioequivalence approach resolves concerns of statistical power. Specifically, within this data set, applying the bioequivalence approach resulted in failure for tests with relatively large test variability and a pass for tests with relatively small within-test variability.

  12. Avian models for toxicity testing

    USGS Publications Warehouse

    Hill, E.F.; Hoffman, D.J.

    1984-01-01

    The use of birds as test models in experimental and environmental toxicology as related to health effects is reviewed, and an overview of descriptive tests routinely used in wildlife toxicology is provided. Toxicologic research on birds may be applicable to human health both directly by their use as models for mechanistic and descriptive studies and indirectly as monitors of environmental quality. Topics include the use of birds as models for study of teratogenesis and embryotoxicity, neurotoxicity, behavior, trends of environmental pollution, and for use in predictive wildlife toxicology. Uses of domestic and wild-captured birds are discussed.

  13. MINIATURIZED SEDIMENT PROCEDURES FOR ASESSING TOXICITY USING MARINE AND FRESHWATER AMPHIPODS AND EMBRYO/LARVAL FISH

    EPA Science Inventory

    Sediment toxicity tests are needed that can be conducted with less sediment volume and fewer organisms. Bench scale remediation techniques often produce less sediment than is required to perform the standardized sediment methods and the excess sediments that are generated present...

  14. TOXIC SUBSTANCES CONTROL ACT TEST SUBMISSIONS (TSCATS)

    EPA Science Inventory

    Toxic Substances Control Act Test Submissions (TSCATS) is an online index to unpublished, nonconfidential studies covering chemical testing results and adverse effects of chemicals on health and ecological systems. The studies are submitted by U.S. industry to EPA under several s...

  15. Toxicity of petroleum crude oils and their effect on xenobiotic metabolizing enzyme activities in the chicken embryo in ovo.

    PubMed

    Lee, Y Z; O'Brien, P J; Payne, J F; Rahimtula, A D

    1986-02-01

    Microliter quantities of a Prudhoe Bay crude oil (PBCO) applied to the shell of fertile chick eggs during various stages of development induced cytochrome P-450 levels and mixed-function oxidase activities within the liver of the embryo. PBCO (5 microliter) applied on Day 11 of incubation was found to maximally induce within 24 hr embryo hepatic cytochrome P-450 levels (fourfold), naphthalene hydroxylase (sixfold), benzo[a]pyrene 3-hydroxylase (14-fold), and 7-ethoxyresorufin O-deethylase (24-fold). Glutathione S-transferase was not induced. Crude oils are known to be highly toxic to avian embryos, especially during the early stages of development. The LD50 of PBCO and Hibernia crude oil applied to the egg shell on Day 8 of incubation was found to be 1.3 and 2.2 microliter, respectively. Mixed-function oxidase-dependent metabolism of crude oil components may be required for toxicity since administration of 20 micrograms of disulfiram in dioxane 1 hr prior to application of 1.3 microliter of PBCO reduced embryo mortality from 60 to 20%. PMID:3943505

  16. Comparative toxicities of benzene, chlorobenzene, and dichlorobenzenes to sea urchin embryos and sperm

    SciTech Connect

    Pagano, G.; Cipollaro, M.; Corsale, G.; Esposito, A.; Giordano, G.G.; Ragucci, E.; Trieff, N.M.

    1988-04-01

    The leukemogenicity and myelotoxicity of benzene are well-known and the major cause of benzene's banning from most industrial applications. Various benzene derivatives such as alkylbenzenes and chlorobenzenes, however, continue to be used as chemical intermediates, solvents, pesticides, etc. in spite of incomplete knowledge of their chronic toxicity. This study was designed to obtain comparative data on developmental, genetic and reproductive toxicities of benzene (B), chlorobenzene (CB) and dichlorobenzenes (o-, m-, and p-DCB) in the sea urchin bioassay. This test system, permits observation of a number of biological endpoints on mitotic activity, embryogenesis and fertilization, and thus provides multi-parametric toxicological data on xenobiotics.

  17. New Zealand sediment toxicity testing methodologies

    SciTech Connect

    Hickey, C.W.; Roper, D.S.; Nipper, M.; Martin, M.L.

    1995-12-31

    Sediment toxicity testing in New Zealand is developing against a background of an increasing public desire for environmental protection and strict legislative requirements that contaminant discharges should not have any significant adverse effects on aquatic life. The importance of sediment contamination and its potential immediate and long term adverse effects on aquatic biota in general is becoming widely recognized, This has lead to an effort to develop acute and chronic sediment toxicity tests with organisms representative of the New Zealand indigenous biota. An amphipod species occurring in both freshwater and estuarine environments, Chaetocorophium cf lucasi, and the marine bivalve Macomona liliana, a common inhabitant of intertidal sandflats, have been evaluated for their sensitivity to natural sediment characteristics. The amphipod and bivalve are presently being used for testing sediment acute (10d) and chronic toxicity (20--30d), with survival and growth as test endpoints, and the bivalve has shown to be a useful organism for behavioral tests with burial and sediment avoidance by movement and drifting as endpoints. The estuarine bivalve Arthritica bifurca, abundant in muddy sediments, is a self-fertilizing hermaphroditic species and its suitability for sediment tests with a reproductive endpoint is underway. Freshwater sphaeriid bivalves, Sphaerium novazelandiae, are also being used for survival, growth, reproduction and behavioral endpoints. Sensitivity to reference toxicants and results for contaminated sediments will be presented and discussed in relation to sediment quality criteria developed elsewhere.

  18. Intra- and intertreatment variability in reference toxicant tests: Implications for whole effluent toxicity testing programs

    SciTech Connect

    Moore, D.R.J.; Warren-Hicks, W.; Parkhurst, B.R.; Teed, R.S.; Baird, R.B.; Berger, R.; Denton, D.L.; Pletl, J.J.

    2000-01-01

    Whole effluent toxicity tests are used in permitting programs across the US to determine whether effluents are potentially toxic to aquatic biota in receiving environments. In cases where whole effluent toxicity tests indicate unacceptable toxicity, corrective measures or further testing may be required. To be consistent and fair to permit holders, whole effluent toxicity test outcomes should not be strongly influenced by intra- and interlaboratory variability. In this study, the authors quantified intra- and interlaboratory variability for four species-data type combinations using the results of reference toxicant tests compiled from many laboratories in recent years. For each set of test results, they conducted a regression analysis using the generalized linear models framework. The results indicated that the coefficient of variation for intralaboratory 25% effective concentration values varied from 15.7% for number of young of Ceriodaphnia dubia in laboratory CD4 to 122% for mortality of Menidia beryllina in laboratory MB3. Interlaboratory variability was small for both mortality and number of young of C. dubia. Interlaboratory variability for mortality and biomass of M. beryllina, however, was very high. Their study shows that permit toxicity limits can be exceeded because of factors other than effluent toxicity, particularly when the limits are based on testing of M. beryllina.

  19. Toxicity of polychlorinated diphenyl ethers in hydra attenuata and in rat whole-embryo culture. Master's thesis

    SciTech Connect

    Becker, M.C.

    1991-05-01

    Polychlorinated diphenyl ethers (PCDEs) are a class of biaryl compounds that have little commercial application, but appear to be widespread in the environment. They have been found in wood preservative waste dumpsites and in fly ash from municipal waste incinerators. They have been detected in bird eggs and tissues, fish, and other edible marine organisms in the United States, Canada, and Europe. There are limited reports in the extant literature on the toxicity of PCDEs. This study was designed to evaluate the toxicity of selected PCDEs in cultures of Hydra attenuata and post-implantation rat whole embryos. The toxicity of several closely related polychlorinated biphenyls (PCBs) was evaluated in both cultures and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was evaluated in whole embryo culture. Embryonic growth and development parameters (yolk sac diameter, crown-rump length, somite count, and DNA and protein content) and gross morphology were determined. Findings indicated that these chemicals were neither embryotoxic nor teratogenic. Thus, the PCDEs, which elicit other diverse toxic and biochemical responses in rodents, are relatively inactive in these bioassays for developmental toxicity.

  20. Validation of alternative methods for toxicity testing.

    PubMed Central

    Bruner, L H; Carr, G J; Curren, R D; Chamberlain, M

    1998-01-01

    Before nonanimal toxicity tests may be officially accepted by regulatory agencies, it is generally agreed that the validity of the new methods must be demonstrated in an independent, scientifically sound validation program. Validation has been defined as the demonstration of the reliability and relevance of a test method for a particular purpose. This paper provides a brief review of the development of the theoretical aspects of the validation process and updates current thinking about objectively testing the performance of an alternative method in a validation study. Validation of alternative methods for eye irritation testing is a specific example illustrating important concepts. Although discussion focuses on the validation of alternative methods intended to replace current in vivo toxicity tests, the procedures can be used to assess the performance of alternative methods intended for other uses. Images Figure 1 PMID:9599695

  1. Cardiac toxicity by sublethal 2,3,7,8-tetrachlorodibenzo-p-dioxin correlates with its anti-proliferation effect on cardiomyocytes in zebrafish embryos.

    PubMed

    Chen, Jing

    2015-02-01

    The cardiac toxicity of zebrafish embryos in response to the lethal dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been well characterized. Dioxin contamination levels in nature are usually lower, however, and sublethal TCDD toxicity is less investigated. The present study found that the nonlethal doses of TCDD for 72-h-postfertilization (hpf) zebrafish embryos were 25 pg/mL and lower. For the present study, sublethal TCDD concentrations of 10 pg/mL and 25 pg/mL were selected, and their toxicity was then characterized. The results showed that embryos still exhibited acute and subchronic cardiac toxicity at these 2 dosages. The stroke volume and cardiac output of these embryos significantly declined early until 8 d postexposure. Embryos' heart size became smaller, and the hearts contained fewer cardiomyocytes per heart, with decreased cardiomyocyte proliferation. Apoptosis was not detected either in the TCDD-treated or the control hearts. Real-time polymerase chain reaction (PCR) revealed that the transcription of a battery of cell-cycle-related genes was suppressed within the sublethal TCDD-treated heart. In contrast, embryonic jaw development seemed not to be affected. The present study suggests that dioxin contamination, even at lower levels, might lead to cardiac toxicity in fish embryos. Such cardiac toxicity presents as disrupted normal heart function, originating from the anti-proliferative effect of sublethal TCDD on cardiomyocytes. PMID:25477153

  2. Toxicity test of a dental commercial composite

    PubMed Central

    Ponce-Bravo, Santa; Martínez-Rivera, José-Luis; Garcés-Ortíz, Maricela

    2015-01-01

    Background International rules must be followed for testing biosecurity in dental materials. A new brand of restorative material appeared in the market and regulations indicated that it should be tested for toxicity. Objectives The aim of this study was to determine the 90-day sub chronic toxicity of one triethylene glycol dimethacrylate containing composite (MEDENTAL Light-Cure Composite™) orally administered to rats according to Organization for Economic Co-Operation and Development no. 48 guidelines and the requirements specified in the ISO 10993-11. Material and Methods Wistar rats ate the polymerized composite during 90 days and were observed to determine changes in their behavior, eye and skin signs and other attitudes such as aggressiveness, posture, walking and response to handling. After 90 days were sacrificed to ascertain blood alterations, we did special hematological tests and assessed microscopic slides from 33 different organs. Results We recorded no significant changes in clinical behavior of the animals. Microscopic review of the H&E stained slides obtained from the analyzed organs showed no abnormal inflammatory or cytological changes and all hematological special tests were within normal limits. Conclusions Results of this study show that under our experimental conditions the MEDENTAL Light-Cure Composite™ does not produce inflammatory or cytological changes suggestive of toxicity. Key words:Dental materials, composite resin, toxicity, inflammation, TEGDMA. PMID:26155348

  3. Assessment of the bioavailability and toxicity of sediment-associated polycyclic aromatic hydrocarbons and heavy metals applied to Crassostrea gigas embryos and larvae.

    PubMed

    Geffard, O; Geffard, A; His, E; Budzinski, H

    2003-04-01

    Sediments represent a vast sink for contaminants in aquatic systems, and may pose a threat to pelagic and benthic organisms. The objective of this research was to determine the bioavailability and toxicity of sediment-associated PAHs and heavy metals, using embryos and larvae of the oyster Crassostrea gigas, exposed to two sediment fractions: the whole sediment and the elutriate. The percentages of abnormal larvae, the contaminant accumulation and, (in the case of metal contamination), the induction of metallothionein in the larvae, were investigated. Sediment-associated PAHs and heavy metals were available for exposure, as indicated by their accumulation in C. gigas larvae and by the abnormalities induced during larval development. The critical body burden of PAHs (Fluo, Pyr, BaA, Triph, Chrys, BbF, BkF, BjK, BeP, BaP, Per, IP, BPer and the DahA) in the larvae was 0.3 micro g g(-1), above which abnormalities were observed. This value corresponds to concentrations observed for most vertebrate and invertebrate species. The bioavailability of PAHs is determined by their solubility; only the soluble fraction of PAHs is accumulated by the embryos. The bioavailability of metals for the larvae is substantiated by MT induction, correlated with cytosolic metal concentrations. MT induction provided a better early-warning response than the embryotoxicity test currently used for evaluating environmental contamination by metals. This study recommends choosing oyster embryos as a particularly sensitive tool for evaluating sediment quality. PMID:12705922

  4. Neurobehavioral aspects of developmental toxicity testing.

    PubMed Central

    Ulbrich, B; Palmer, A K

    1996-01-01

    Tests for detection of neurobehavioral changes in the offspring have been a regulatory requirement in developmental toxicity testing of drugs for almost 20 years. Keeping their purpose of hazard identification and risk assessment for humans in mind, investigators and agency reviewers have become deeply ingrained with some stereotyped behaviors with respect to such relevant issues as choice of animal species and data evaluation. Other problematic areas of study design and conduct, selection of litter representatives for testing, what methods to combine in a testing battery, and statistical treatment of results and their interpretation, will need more research and discussion in the future. PMID:9182049

  5. Mechanisms of Nickel Toxicity in the Highly Sensitive Embryos of the Sea Urchin Evechinus chloroticus, and the Modifying Effects of Natural Organic Matter.

    PubMed

    Blewett, Tamzin A; Smith, D Scott; Wood, Chris M; Glover, Chris N

    2016-02-01

    A 96 h toxicity test showed that the embryos of the New Zealand sea urchin (Evechinus chloroticus) are the most sensitive of all studied marine species to waterborne nickel (Ni), with the EC50 for the development of fully formed pluteus larvae found to be 14 μg L(-1). Failure to develop a standard larval shape suggested skeletal impairment. Whole body ions (Na, Mg) increased with Ni exposure and calcium influx was depressed. The effects of natural organic matter (NOM) on Ni accumulation and toxicity were also examined in three different seawater sources (nearshore, offshore, and near the outlet of a "brown water" stream). At low dissolved organic carbon (DOC) concentrations the brown water NOM was protective against Ni toxicity, however at higher DOC concentrations it exacerbated developmental toxicity in the presence of Ni. These results show that sea urchin development is highly sensitive to Ni via a mechanism that involves ionoregulatory disturbance, and that Ni toxicity is influenced by environmental factors such as NOM. These data will be critical for the development of water quality guidelines for Ni in the marine environment. PMID:26730609

  6. Contact tests for pentachlorophenol toxicity to earthworms

    SciTech Connect

    Spontak, D.A.

    1994-12-31

    The standardized contact filter paper test (EEC and OECD) provides an effective screening test for toxicity to earthworms in a laboratory setting. A need exists for a reliable and inexpensive technique for non-laboratory settings where screening is desired, but facilities cannot provide for the acquisition and maintenance of the glass vials required by the standardized test. This study evaluated two modifications of the standardized test using clear polyethylene bags, with and without filter paper, with Eisenia fetida and domesticated surface-feeding earthworms. The tests were conducted according to EEC and OECD guidelines. Results of the modified tests corresponded in dose and effect to the standardized contact filter paper test indicating the usefulness of the modified tests.

  7. Estimation of toxicity using the Toxicity Estimation Software Tool (TEST)

    EPA Science Inventory

    Tens of thousands of chemicals are currently in commerce, and hundreds more are introduced every year. Since experimental measurements of toxicity are extremely time consuming and expensive, it is imperative that alternative methods to estimate toxicity are developed.

  8. Teratogenic and toxic effects of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.:Fr.) P. Karst. (higher Basidiomycetes), on zebrafish embryo as model.

    PubMed

    Dulay, Rich Milton R; Kalaw, Sofronio P; Reyes, Renato G; Alfonso, Noel F; Eguchi, Fumio

    2012-01-01

    This paper highlights the teratogenic and toxic effects of Ganoderma lucidum (Lingzhi or Reishi mushroom) extract on zebrafish embryos. Hatchability, malformations, and lethality rate of zebrafish embryos were assessed to provide valuable information regarding the potential teratogenic activity of G. lucidum. Hatching was completed 48 h post treatment application (hpta) at 1% or lower concentrations of extract and embryo water. The hatching rate of embryos treated with 5% or higher concentrations was significantly lower (p> 0.05) than the control. Tail malformation was the most marked morphological abnormality in embryos at 72 hpta, which was obviously caused by 1% extract (55.56% tail malformation) and was observed in all embryos exposed to 5% of extract. Growth retardation was evident in embryos exposed to 5%, 10%, and 20%. However, lethal effect of extract of G. lucidum was dependent on dose and time of exposure. Mortality rates of embryos treated with 5% (44.44%) or higher concentrations of the extract was significantly higher (p > 0.05) than that of the control embryos at 72 hpta. These results suggest that G. lucidum extract has lethal and sub-lethal effects on zebrafish embryos. PMID:23510220

  9. Toxicity assessment of sediments from three European river basins using a sediment contact test battery.

    PubMed

    Tuikka, A I; Schmitt, C; Höss, S; Bandow, N; von der Ohe, P C; de Zwart, D; de Deckere, E; Streck, G; Mothes, S; van Hattum, B; Kocan, A; Brix, R; Brack, W; Barceló, D; Sormunen, A J; Kukkonen, J V K

    2011-01-01

    The toxicity of four polluted sediments and their corresponding reference sediments from three European river basins were investigated using a battery of six sediment contact tests representing three different trophic levels. The tests included were chronic tests with the oligochaete Lumbriculus variegatus, the nematode Caenorhabditis elegans and the mudsnail Potamopyrgus antipodarum, a sub-chronic test with the midge Chironomus riparius, an early life stage test with the zebra fish Danio rerio, and an acute test with the luminescent bacterium Vibrio fischeri. The endpoints, namely survival, growth, reproduction, embryo development and light inhibition, differed between tests. The measured effects were compared to sediment contamination translated into toxic units (TU) on the basis of acute toxicity to Daphnia magna and Pimephales promelas, and multi-substance Potentially Affected Fractions of species (msPAF) as an estimate for expected community effects. The test battery could clearly detect toxicity of the polluted sediments with test-specific responses to the different sediments. The msPAF and TU-based toxicity estimations confirmed the results of the biotests by predicting a higher toxic risk for the polluted sediments compared to the corresponding reference sediments, but partly having a different emphasis from the biotests. The results demonstrate differences in the sensitivities of species and emphasize the need for data on multiple species, when estimating the effects of sediment pollution on the benthic community. PMID:20833427

  10. Assessment of silver nanoparticle toxicity for common carp (Cyprinus carpio) fish embryos using a novel method controlling the agglomeration in the aquatic media.

    PubMed

    Oprsal, Jakub; Blaha, Ludek; Pouzar, Miloslav; Knotek, Petr; Vlcek, Milan; Hrda, Katerina

    2015-12-01

    Formation of agglomerates and their rapid sedimentation during aquatic ecotoxicity testing of nanoparticles is a major issue with a crucial influence on the risk assessment of nanomaterials. The present work is aimed at developing and testing a new approach based on the periodic replacement of liquid media during an ecotoxicological experiment which enabled the efficient monitoring of exposure conditions. A verified mathematical model predicted the frequencies of media exchanges which checked for formation of agglomerates from silver nanoparticles AgNP with 50 nm average size of the original colloid. In the model experiments, embryos of common carp Cyprinus carpio were exposed repeatedly for 6 h to AgNPs (5-50 μm Ag L(-1)) either under semistatic conditions (exchange of media after 6 h) or in variants with frequent media exchanges (varying from 20 to 300 min depending on the AgNP colloid concentration and the desired maximum agglomerate size of 200 or 400 nm). In contrast to other studies, where dissolved free metals are usually responsible for toxic effects, our 144-h experiments demonstrated the importance of AgNP agglomerates in the adverse effects of nanosilver. Direct adsorption of agglomerates on fish embryos locally increased Ag concentrations which resulted in pronounced toxicity particularly in variants with larger 400 nm agglomerates. The present study demonstrates the suitability of the novel methodology in controlling the conditions during aquatic nanomaterial toxicity testing. It further provided insights into the mechanisms underlying the effects of AgNP, which rank on a global scale among the most widely used nanomaterials. PMID:26233755

  11. Embryo-Fetal Developmental Toxicity Studies with Pregabalin in Mice and Rabbits.

    PubMed

    Morse, Dennis C

    2016-04-01

    Pregabalin was evaluated for potential developmental toxicity in mice and rabbits. Pregabalin was administered once daily by oral gavage to female albino mice (500, 1250, or 2500 mg/kg) and New Zealand White rabbits (250, 500, or 1250 mg/kg) during organogenesis (gestation day 6 through 15 [mice] or 6 through 20 [rabbits]). Fetuses were evaluated for viability, growth, and morphological development. Pregabalin administration to mice did not induce maternal or developmental toxicity at doses up to 2500 mg/kg, which was associated with a maternal plasma exposure (AUC0-24 ) of 3790 μg•hr/ml, ≥30 times the expected human exposure at the maximum recommended daily dose (MRD; 600 mg/day). In rabbits, treatment-related clinical signs occurred at all doses (AUC0-24 of 1397, 2023, and 4803 μg•hr/ml at 250, 500, and 1250 mg/kg, respectively). Maternal toxicity was evident at all doses and included ataxia, hypoactivity, and cool to touch. In addition, abortion and females euthanized moribund with total resorption occurred at 1250 mg/kg. There were no treatment-related malformations at any dose. At 1250 mg/kg, compared with study and historical controls, the percentage of fetuses with retarded ossification was significantly increased and the mean number of ossification sites was decreased, which correlated with decreased fetal and placental weights, consistent with in utero growth retardation. Therefore, the no-effect dose for developmental toxicity in rabbits was 500 mg/kg, which produced systemic exposure approximately 16-times human exposure at the MRD. These findings indicate that pregabalin, at the highest dose tested, was not teratogenic in mice or rabbits. PMID:27044003

  12. Enantioselectivity in Developmental Toxicity of rac-metalaxyl and R-metalaxyl in Zebrafish (Danio rerio) Embryo.

    PubMed

    Zhang, Yinjun; Zhang, Yi; Chen, An; Zhang, Wei; Chen, Hao; Zhang, Quan

    2016-06-01

    Enantioselectivity of chiral pesticides in environmental safety has attracted more and more attention. In this study, we evaluated the enantioselective toxicity of rac-metalaxyl and R-metalaxyl to zebrafish (Danio rerio) embryos through various malformations including pericardial edema, yolk sac edema, crooked body, and short tails. The results showed that there were significant differences in toxicity to zebrafish embryos caused by rac-metalaxyl and R-metalaxyl, and the LC50 s at 96 h are 416.41 (353.91, 499.29) mg · L(-1) and 320.650 (279.80, 363.46) mg · L(-1) , respectively. In order to explore the possible mechanism of the development defects, the genes involved in the hypothalamic-pituitary-gonadal axis (vtg1, vtg2, cyp17, cyp19a, cyp19b) and hypothalamic-pituitary-thyroid axis (dio1, dio2, nis, tg, tpo) were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). The results revealed that there were no significant differences in the expression of vtg1, vtg2, cyp17, cyp19a, and cyp19b after exposure to rac-metalaxyl. However, the expression of vtg1, cyp19a, and cyp19b decreased significantly after exposure to R-metalaxyl. And likewise, rac-metalaxyl only caused the upregulation of dio2, while R-metalaxyl suppressed the expression of dio1 and tpo and induced the expression of dio2 and nis. The change of gene expression may cause the enantioselectivity in developmental toxicity in zebrafish embryo. The data provided here will be helpful for us to comprehensively understand the potential ecological risks of the currently used chiral fungicides. Chirality 28:489-494, 2016. © 2016 Wiley Periodicals, Inc. PMID:27103609

  13. Ocular Toxicity Testing of Lunar Dust

    NASA Technical Reports Server (NTRS)

    Meyers, Valerie E.

    2010-01-01

    This slide presentation reviews the use of ocular testing to determine the toxicity of lunar dust. The OECD recommendations are reviewed. With these recommendations in mind the test methodology was to use EpiOcular, tissues derived from normal human epidermal keratinocytes, the cells of which have been differentiated on cell culture inserts to form a multi-layered structure, which closely parallels the corneal epithelium and to dose the tissue with 100 mg dust from various sources. The in-vitro study provides evidence that lunar dust is not severely corrosive or irritating, however, in vitro tests have limitations, and in vivo tests provides a more complete scenario, and information, it is recommended that in vivo tests be performed.

  14. MAMMALIAN WILDLIFE (MINK AND FERRET) TOXICITY TEST PROTOCOLS (LC50, REPRODUCTION, AND SECONDARY TOXICITY)

    EPA Science Inventory

    Protocols describing guidelines for conducting dietary LC50 and reproduction toxicity tests and for assessing the primary versus secondary toxicity of a test substance using carnivorous mammalian wildlife, specifically mink (Mustela vison) or European ferrets (Mustela putorius fu...

  15. CHIRONOMIDAE TOXICITY TESTS--BIOLOGICAL BACKGROUND AND PROCEDURES

    EPA Science Inventory

    Toxicity tests must be based on an understanding of the test animal's life cycle. The first section of this report describes the biological information needed to develop toxicity test procedures. The second section describes three categories of toxicity test systems - short-expos...

  16. 16 CFR 1500.40 - Method of testing toxic substances.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 16 Commercial Practices 2 2012-01-01 2012-01-01 false Method of testing toxic substances. 1500.40... testing toxic substances. The method of testing the toxic substances referred to in § 1500.3(c) (1)(ii)(C... may vary in size to accommodate smaller or larger subjects. In the testing of unctuous materials...

  17. 16 CFR 1500.40 - Method of testing toxic substances.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 16 Commercial Practices 2 2011-01-01 2011-01-01 false Method of testing toxic substances. 1500.40... testing toxic substances. The method of testing the toxic substances referred to in § 1500.3(c) (1)(ii)(C... may vary in size to accommodate smaller or larger subjects. In the testing of unctuous materials...

  18. 16 CFR 1500.40 - Method of testing toxic substances.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 16 Commercial Practices 2 2010-01-01 2010-01-01 false Method of testing toxic substances. 1500.40... testing toxic substances. The method of testing the toxic substances referred to in § 1500.3(c) (1)(ii)(C... may vary in size to accommodate smaller or larger subjects. In the testing of unctuous materials...

  19. A novel one-pot green synthesis of selenium nanoparticles and evaluation of its toxicity in zebrafish embryos.

    PubMed

    Kalishwaralal, Kalimuthu; Jeyabharathi, Subhaschandrabose; Sundar, Krishnan; Muthukumaran, Azhaguchamy

    2016-03-01

    Over the last 50 years, compelling evidence has accumulated on the beneficial role of selenium in human health. In the present study, different proteins were evaluated as reducing agents for the eco-friendly synthesis of selenium nanoparticles from an aqueous solution of sodium selenite. This method is a simple, low cost green synthesis alternative to chemical synthesis. The high conversion of selenium ions to selenium nanoparticles (SeNPs) was achieved by a reaction mixture of 0.1 g bovine serum albumin and 0.1 g sodium selenite at a reaction temperature of 121°C for 20 min duration. The selenium nanoparticles were characterized by fourier transform infrared (FTIR), scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy. The FTIR spectral bands were sharp with strong absorption peaks at 1649 and 1551 cm(- 1). SEM analysis of the synthesized selenium nanoparticles clearly showed the spherical shape with an average size ranging from 500 to 600 nm. The toxicity of SeNPs was evaluated using zebrafish embryos as a model system. SeNPs induced malformations in zebrafish embryos in a concentration-dependent manner. Selenium nanoparticles at 15-25 μg/ml concentration caused pericardial edema, tail malformation and decrease in heart rate in zebrafish embryos. Treatments with lower concentrations did not alter the heart rate or display any heart abnormalities. This study underlines the importance of identifying optimal SeNP concentration that could have potential therapeutic applications. PMID:25287880

  20. Acute toxic effects of the herbicide formulation and the active ingredient used in cycloxydim-tolerant maize cultivation on embryos and larvae of the African clawed frog, Xenopus laevis.

    PubMed

    Wagner, Norman; Lötters, Stefan; Veith, Michael; Viertel, Bruno

    2015-04-01

    Most genetically engineered herbicide-tolerant crops are still awaiting approval in Europe. There is, however, a recent trend for the cultivation of cycloxydim-tolerant maize hybrids for use in maize production. We studied the acute toxic effects of the complementary herbicide Focus(®) Ultra and its active ingredient cycloxydim on embryos and early-stage larvae of the African clawed frog (Xenopus laevis). The results indicate that the herbicide formulation is significantly more toxic than the active ingredient alone. Therefore, it is suggested that the added substances either solely or in a synergistic action with the active ingredient are responsible for adverse effects. The formulation was found to be moderately toxic to embryos but highly toxic to early larvae. Based on calculated teratogenic indices, both cycloxydim and Focus(®) Ultra seem to be non-teratogenic and also the minimum Focus(®) Ultra concentration to inhibit growth in embryos and larvae was close to the LC50 values. The data suggest that tests with the rainbow trout are not in all cases appropriate to assess the risk in aquatically developing anurans. This is demonstrated by 96-h LC50 values, which are for rainbow trout more than 50- to 20-fold higher than for early X. laevis larvae. However, based on worst-case predicted environmental concentrations for surface waters, there is apparently a large safety margin in field use of Focus(®) Ultra if buffer strips between the farm land and the amphibian habitats are regarded. PMID:25634323

  1. Toxicity of Uranium Adsorbent Materials using the Microtox Toxicity Test

    SciTech Connect

    Park, Jiyeon; Jeters, Robert T.; Gill, Gary A.; Kuo, Li-Jung; Bonheyo, George T.

    2015-10-01

    The Marine Sciences Laboratory at the Pacific Northwest National Laboratory evaluated the toxicity of a diverse range of natural and synthetic materials used to extract uranium from seawater. The uranium adsorbent materials are being developed as part of the U. S. Department of Energy, Office of Nuclear Energy, Fuel Resources Program. The goal of this effort was to identify whether deployment of a farm of these materials into the marine environment would have any toxic effects on marine organisms.

  2. 40 CFR 797.1050 - Algal acute toxicity test.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Algal acute toxicity test. 797.1050 Section 797.1050 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) ENVIRONMENTAL EFFECTS TESTING GUIDELINES Aquatic Guidelines § 797.1050 Algal acute toxicity test. (a) Purpose. The...

  3. 40 CFR 797.1300 - Daphnid acute toxicity test.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Daphnid acute toxicity test. 797.1300 Section 797.1300 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) ENVIRONMENTAL EFFECTS TESTING GUIDELINES Aquatic Guidelines § 797.1300 Daphnid acute toxicity test. (a) Purpose....

  4. 40 CFR 797.1300 - Daphnid acute toxicity test.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 32 2011-07-01 2011-07-01 false Daphnid acute toxicity test. 797.1300 Section 797.1300 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) ENVIRONMENTAL EFFECTS TESTING GUIDELINES Aquatic Guidelines § 797.1300 Daphnid acute toxicity test. (a) Purpose....

  5. 40 CFR 797.1300 - Daphnid acute toxicity test.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 32 2014-07-01 2014-07-01 false Daphnid acute toxicity test. 797.1300 Section 797.1300 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) ENVIRONMENTAL EFFECTS TESTING GUIDELINES Aquatic Guidelines § 797.1300 Daphnid acute toxicity test. (a) Purpose....

  6. THE USE OF EMBRYOS OF PALAEMONETES PUGIO IN EVALUATING THE DEVELOPMENTAL TOXICITY OF COMPOUNDS

    EPA Science Inventory

    The grass shrimp, Palaemonetes pugio, is a common, well distributed species along the eastern coast of America that is an important part of the food web. The embryos of P. pugio have shown sensitivity to the water-soluble fraction of Number 2 fuel oil, which indicates they may be...

  7. TOXICITY TESTS FOR SEDIMENT QUALITY ASSESSMENTS

    EPA Science Inventory

    Toxic sediments have contributed to a wide-variety of environmental problems around the world. The observed effects include direct toxic effects to aquatic life, bio-magnification of toxicants in the food chain, and economic impacts. This chapter discusses the use of toxicity...

  8. SAND SPIKED WITH COPPER AS A REFERENCE TOXICANT MATERIAL FOR SEDIMENT TOXICITY TESTING: A PRELIMINARY EVALUATION

    EPA Science Inventory

    Routine use of solid-phase sediment toxicity tests for scientific and regulatory purposes necessitates the development of solid-phase reference toxicant materials. n order to evaluate an approach for developing such materials, 12 solid-phase 96-h reference toxicant tests were con...

  9. Role of oxidative stress and antioxidant defense in 3,3',4,4',5-pentachlorobiphenyl-induced toxicity and species-differential sensitivity in chicken and duck embryos.

    PubMed

    Jin, X; Kennedy, S W; Di Muccio, T; Moon, T W

    2001-05-01

    The role of oxidative stress and antioxidant defense in 3,3',4,4',5-pentachlorobiphenyl (PCB 126)-induced toxicity and species-specific sensitivity was examined in White Leghorn chicken (Gallus domesticus) and Pekin duck (Anas platyrhynchos) embryos. Eggs were injected into the air cell with 0.4-1.6 microgram PCB 126/kg egg in corn oil prior to incubation. Lipid peroxidation measured by thiobarbituric acid reactive substances (TBARS), the GSSG:GSH ratio, and glutathione peroxidase (GPox) activities were determined in liver and adipose tissue of day 19 chicken and day 26 duck embryos. In chicken embryos, PCB 126 increased mortality and the incidence of edema and liver lesions, decreased embryo size, increased eye and head malformations, and markedly reduced fat storage. In contrast, no effects on the endpoints were observed in duck embryos even at the highest dose used in chicken embryos. PCB 126 increased hepatic 7-ethoxyresorufin-O-deethylase (EROD) activity in a dose-dependent manner in chicken but not duck embryos. PCB 126 significantly increased TBARS levels in liver and to a greater degree in adipose tissue of chicken embryos, indicating that adipose tissue is a sensitive target for this compound. Increases in lipid peroxidation by PCB 126 were associated with significant decreases in GPox activity in these tissues. These biochemical changes support oxidative stress playing a role in PCB 126-induced embryo toxicity while antioxidant defenses provided protection against oxidative damage induced by this compound. Ducks, the less-sensitive species, showed higher basal levels of hepatic GPox than chickens, suggesting that this antioxidant enzyme may contribute to the differences in sensitivity to this compound between the two species. PMID:11312653

  10. Effect of pH on pentachlorophenol toxicity to embryos and larvae of zebrafish (Brachydanio rerio)

    SciTech Connect

    Dave, G.

    1984-11-01

    The hydrogen ion is an important determinant of toxicity in the aquatic environment, especially for weak acids or bases. Pentachlorophenol is a weak acid and is consequently more toxic at low pH because of increasing proportions of free phenol. For some heavy metals the toxic interaction with ambient pH is complicated by complexation with organic and inorganic compounds. These complexes are affected by pH, and bioassays may be the easiest and most reliable method of determining the influence of pH in such cases. The objective of this study was to find a simple laboratory procedure for determination of the effect of pH on the toxicity of various chemicals added to or already present in various samples of ground and surface waters. Pentachlorophenol was selected as a reference toxicant and positive control because of its established pH-toxicity interaction in fish.

  11. EVALUATING THE EFFECTS OF FLY ASH EXPOSURE ON FISH EARLY LIFE STAGES: FATHEAD MINNOW EMBRYO-LARVAL TESTS

    SciTech Connect

    Greeley Jr, Mark Stephen; Elmore, Logan R; McCracken, Kitty

    2012-05-01

    On December 22, 2008, a dike containing fly ash and bottom ash in an 84-acre complex of the Tennessee Valley Authority's (TVA) Kingston Steam Plant in East Tennessee failed and released a large quantity of ash into the adjacent Emory River. Ash deposits extended as far as 4 miles upstream (Emory River mile 6) of the Plant, and some ash was carried as far downstream as Tennessee River mile 564 ({approx}4 miles downstream of the Tennessee River confluence with the Clinch River). A byproduct of coal burning power plants, fly ash contains a variety of metals and other elements which, at sufficient concentrations and in specific forms, can be toxic to biological systems. The effects of fly ash contamination on exposed fish populations depend on the magnitude and duration of exposure, with the most significant risk considered to be the effects of specific ash constituents, especially selenium, on fish early life stages. Uptake by adult female fish of fly ash constituents through the food chain and subsequent maternal transfer of contaminants to the developing eggs is thought to be the primary route of selenium exposure to larval fish (Woock and others 1987, Coyle and others 1993, Lemly 1999, Moscatello and others 2006), but direct contact of the fertilized eggs and developing embryos to ash constituents in river water and sediments is also a potential risk factor (Woock and others 1987, Coyle and others 1993, Jezierska and others 2009). To address the risk of fly ash from the Kingston spill to the reproductive health of downstream fish populations, ORNL has undertaken a series of studies in collaboration with TVA including: (1) a field study of the bioaccumulation of fly ash constituents in fish ovaries and the reproductive condition of sentinel fish species in reaches of the Emory and Clinch Rivers affected by the fly ash spill; (2) laboratory tests of the potential toxicity of fly ash from the spill area on fish embryonic and larval development (reported in the current technical manuscript); (3) additional laboratory experimentation focused on the potential effects of long-term exposures to fly ash on fish survival and reproductive competence; and (4) a combined field and laboratory study examining the in vitro developmental success of embryos and larvae obtained from fish exposed in vivo for over two years to fly ash in the Emory and Clinch Rivers. These fish reproduction and early life-stage studies are being conducted in conjunction with a broader biological monitoring program administered by TVA that includes a field study of the condition of larval fish in the Emory and Clinch Rivers along with assessments of water quality, sediment composition, ecotoxicological studies, terrestrial wildlife studies, and human and ecological risk assessment. Information and data generated from these studies will provide direct input into risk assessment efforts and will also complement and help support other phases of the overall biomonitoring program. Fish eggs, in general, are known to be capable of concentrating heavy metals and other environmental contaminants from water-borne exposures during embryonic development (Jezierska and others 2009), and fathead minnow embryos in particular have been shown to concentrate methylmercury (Devlin 2006) as well as other chemical toxicants. This technical report focuses on the responses of fathead minnow embryos to simple contact exposures to fly ash in laboratory toxicity tests adapted from a standard fathead minnow (Pimephales promelas) 7-d embryo-larval survival and teratogenicity test (method 1001.0 in EPA 2002) with mortality, hatching success, and the incidences of developmental abnormalities as measured endpoints.

  12. Exposure Science for Chemical Prioritization and Toxicity Testing

    EPA Science Inventory

    Currently, a significant research effort is underway to apply new technologies to screen and prioritize chemicals for toxicity testing as well as to improve understanding of toxicity pathways (Dix et al. 2007, Toxicol Sci; NRC, 2007, Toxicity Testing in the 21st Century; Collins ...

  13. Comparison of bulk sediment and sediment elutriate toxicity testing methods

    EPA Science Inventory

    Elutriate bioassays are among numerous methods that exist for assessing the potential toxicity of sediments in aquatic systems. In this study, interlaboratory results were compared from 96-hour Ceriodaphnia dubia and Pimephales promelas static-renewal acute toxicity tests conduct...

  14. Effect of water hardness on peracetic toxicity to zebrafish, Danio rerio, embryos

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The use of peracetic acid (PAA) in aquaculture has been suggested as an alternative therapeutic agent as use of previous therapeutants becomes restricted. Few data are available concerning fish toxicity by PAA or factors that affect this toxicity. The aim of this study was to investigate the influ...

  15. Rabbit whole embryo culture.

    PubMed

    Marshall, Valerie A; Carney, Edward W

    2012-01-01

    Although the rabbit is used extensively in developmental toxicity testing, relatively little is known about the fundamental developmental biology of this species let alone mechanisms underlying developmental toxicity. This paucity of information about the rabbit is partly due to the historic lack of whole embryo culture (WEC) methods for the rabbit, which have only been made available fairly recently. In rabbit WEC, early somite stage embryos (gestation day 9) enclosed within an intact amnion and attached to the visceral yolk sac are dissected from maternal tissues and placed in culture for up to 48 h at approximately 37C and are continuously exposed to an humidified gas atmosphere mixture in a rotating culture system. During this 48 h culture period, major phases of organogenesis can be studied including cardiac looping and segmentation, neural tube closure, and development of anlagen of the otic system, eyes and craniofacial structures, somites and early phases of limb development (up to bud stage), as well as expansion and closure of the visceral yolk sac around the embryo. Following completion of the culture period, embryos are evaluated based on several growth and development parameters and also are assessed for morphological abnormalities. The ability to sustain embryo development independent of the maternal system allows for exposure at precise development stages providing the opportunity study the direct action of a teratogen or one of its metabolites on the developing embryo. Rabbit WEC is perhaps most useful when used in conjunction with rodent WEC methods to investigate species-specific mechanisms of developmental toxicity. PMID:22669668

  16. Toxicity of glyphosate as Glypro and LI700 to red-eared slider (trachemys scripta elegans) embryos and early hatchlings.

    PubMed

    Sparling, Donald W; Matson, Cole; Bickham, John; Doelling-Brown, Paige

    2006-10-01

    More than 8.2 billion ha of cropland, gardens, and forests are treated with the herbicide glyphosate each year. Whereas the toxicity of glyphosate and associated adjuvants has been measured in other vertebrates, few, if any, studies have looked at their effects in reptiles. In some instances, management of turtle habitat requires control of successional stages through application of herbicides. Adults and juvenile turtles may be exposed directly, whereas embryos may contact the chemicals through the soil. In the present study, we exposed eggs of red-eared sliders (Trachemys scripta elegans) to single applications of herbicide ranging from 0 to 11,206 ppm wet weight of glyphosate in Glypro and 0 to 678 ppm of the surfactant, LI700. Hatching success at the highest concentration was significantly lower (73%) than in other treatments (80-100%). At hatch, turtles at the highest concentration weighed less than those at other concentrations. During a 14-d holding period, we observed dose-response relationships in the ability of hatchlings to right themselves when turned on their backs. At the end of the holding period, hatchlings at the highest dose level were still lighter, and somatic indices were lower, than those in other treatments. Genetic damage, as measured by flow cytometry, increased with treatment concentration except for the highest dose. We conclude that because of the high concentrations needed to produce effects and the protection offered by several centimeters of soil or sediment, glyphosate with LI700 poses low levels of risk to red-eared slider embryos under normal field operations with regards to the endpoints measured in the present study. Carelessness in handling glyphosate or failure to follow label directions may produce adverse effects. There also is a risk that the health of turtle embryos may be affected in ways not measured in the present study. PMID:17022419

  17. Selenoneine, a novel selenium-containing compound, mediates detoxification mechanisms against methylmercury accumulation and toxicity in zebrafish embryo.

    PubMed

    Yamashita, Michiaki; Yamashita, Yumiko; Suzuki, Tamami; Kani, Yoko; Mizusawa, Nanami; Imamura, Shintaro; Takemoto, Kenji; Hara, Tatsuro; Hossain, Md Anwar; Yabu, Takeshi; Touhata, Ken

    2013-10-01

    The selenium (Se)-containing antioxidant selenoneine (2-selenyl-N α,N α,N α-trimethyl-L-histidine) has recently been discovered to be the predominant form of organic Se in tuna blood. Although dietary intake of fish Se has been suggested to reduce methylmercury (MeHg) toxicity, the molecular mechanism of MeHg detoxification by Se has not yet been determined. Here, we report evidence that selenoneine accelerates the excretion and demethylation of MeHg, mediated by a selenoneine-specific transporter, organic cations/carnitine transporter-1 (OCTN1). Selenoneine was incorporated into human embryonic kidney HEK293 cells transiently overexpressing OCTN1 and zebrafish blood cells by OCTN1. The K m for selenoneine uptake was 13.0 μM in OCTN1-overexpressing HEK293 cells and 9.5 μM in zebrafish blood cells, indicating high affinity of OCTN1 for selenoneine in human and zebrafish cells. When such OCTN1-expressing cells and embryos were exposed to MeHg-cysteine (MeHgCys), MeHg accumulation was decreased and the excretion and demethylation of MeHg were enhanced by selenoneine. In addition, exosomal secretion vesicles were detected in the culture water of embryos that had been microinjected with MeHgCys, suggesting that these may be responsible for MeHg excretion and demethylation. In contrast, OCTN1-deficient embryos accumulated MeHg, and MeHg excretion and demethylation were decreased. Furthermore, Hg accumulation was decreased in OCTN1-overexpressing HEK293 cells, but not in mock vector-transfected cells, indicating that selenoneine and OCTN1 can regulate MeHg detoxification in human cells. Thus, the selenoneine-mediated OCTN1 system regulates secretory lysosomal vesicle formation and MeHg demethylation. PMID:23709046

  18. Developmental toxicity of metaldehyde in the embryos of Lymnaea stagnalis (Gastropoda: Pulmonata) co-exposed to the synergist piperonyl butoxide.

    PubMed

    Hallett, Katrina C; Atfield, Andrew; Comber, Sean; Hutchinson, Thomas H

    2016-02-01

    Metaldehyde is a widely used molluscicide in countries where damage to crops from slugs and snails is a major problem associated with warm and wet winters. In the UK it is estimated that over 8% of the area covered by arable crops is treated with formulated granular bait pellets containing metaldehyde as the main active ingredient. Metaldehyde is hydrophilic (log Kow=0.12), water soluble (200 mg·L(-1) at 17 °C) and has been detected in UK surface waters in the concentration range of typically 0.2-0.6 μg·L(-1) (maximum 2.7 μg·L(-1)) during 2008-2011. In the absence of chronic data on potential hazards to non-target freshwater molluscs, a laboratory study was conducted to investigate the impact of metaldehyde on embryo development in the gastropod Lymnaea stagnalis (RENILYS strain) and using zinc as a positive control. L. stagnalis embryos were exposed to metaldehyde under semi-static conditions at 20±1 °C and hatching success and growth (measured as shell height and intraocular distance) examined after 21 d. Exposure concentrations were verified using HPLC and gave 21 d (hatching)NOEC and (hatching)LOEC mean measured values of 36 and 116 mg MET·L(-1), respectively (equal to the 21 d (shell height)NOEC and (shell height)LOEC values). For basic research purposes, a second group of L. stagnalis embryos was co-exposed to metaldehyde and the pesticide synergist piperonyl butoxide (PBO). Co-exposure to the PBO (measured concentrations between 0.47-0.56 mg·L(-1)) reduced hatching success from 100% to 47% and resulted in a 30% reduction in embryo growth (shell height) in snail embryos co-exposed to metaldehyde at 34-36 mg·L(-1) over 21 d. In conclusion, these data suggest mollusc embryos may have some metabolic detoxication capacity for metaldehyde and further work is warranted to explore this aspect in order to support the recent initiative to include molluscs in the OECD test guideline programme. PMID:26575636

  19. 16 CFR 1500.40 - Method of testing toxic substances.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... not require animals, are presented in the CPSC's animal testing policy set forth in 16 CFR 1500.232. A... test animals. The method of testing the toxic substances referred to in § 1500.3(c)(1)(ii)(C) and (c)(2... 16 Commercial Practices 2 2014-01-01 2014-01-01 false Method of testing toxic substances....

  20. 16 CFR 1500.40 - Method of testing toxic substances.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... that does not require animals, are presented in the CPSC's animal testing policy set forth in 16 CFR... number of test animals. The method of testing the toxic substances referred to in § 1500.3(c)(1)(ii)(C... 16 Commercial Practices 2 2013-01-01 2013-01-01 false Method of testing toxic substances....

  1. Assessing differences in toxicity and teratogenicity of three phthalates, Diethyl phthalate, Di-n-propyl phthalate, and Di-n-butyl phthalate, using Xenopus laevis embryos.

    PubMed

    Gardner, Steven T; Wood, Andrew T; Lester, Rachel; Onkst, Paitra E; Burnham, Nathaniel; Perygin, Donna H; Rayburn, James

    2016-01-01

    Phthalates, compounds used to add flexibility to plastics, are ubiquitous in the environment. In particular, the diethyl (DEP), di-n-propyl (DnPP), and di-n-butyl (DBP) phthalates were found to exert detrimental effects in both mammalian and non-mammalian studies, with toxic effects varying according to alkyl chain length. Embryos of Xenopus laevis, the African clawed frog, have been used to assess toxicity and teratogenicity of several compounds and serves as a model for assessing adverse and teratogenic effects of ortho-phthalate esters. The purpose of this study was to develop a model for comparison of developmentally toxic effects of ortho-phthalate esters using Xenopus embryos. In this study developing Xenopus laevis embryos were exposed to increasing concentrations of DEP, DnPP, and DBP using the 96-h Frog Embryo Teratogenesis Assay-Xenopus (FETAX), with 96-h lethal concentrations, effective concentrations to induce malformations, teratogenic indices, and concentrations to inhibit growth determined. DEP, DnPP, and DBP showed enhanced toxicity with increasing ester length. Developing Xenopus laevis exposed to DEP, DnPP, and DBP showed similar malformations that also occurred at lower concentrations with increasing alkyl chain length. Teratogenic risk did not change markedly with alkyl chain length, with data showing only DBP to be teratogenic. PMID:26730679

  2. A Simple "in Vitro" Culture of Freshwater Prawn Embryos for Laboratory Investigations

    ERIC Educational Resources Information Center

    Porntrai, Supaporn; Damrongphol, Praneet

    2008-01-01

    Giant freshwater prawn ("Macrobrachium rosenbergii" De Man) embryos can be cultured "in vitro" to hatching in 15% (v/v) artificial seawater (ASW). This technique can be applied as a bioassay for testing toxicity or for the effects of various substances on embryo development and can be used as a simple and low-cost model for studying embryo

  3. Bacterial and enzymatic bioassays for toxicity testing in the environment.

    PubMed

    Bitton, G; Koopman, B

    1992-01-01

    Microbioassays using bacteria or enzymes are increasingly applied to measure chemical toxicity in the environment. Attractive features of these assays may include low cost, rapid response to toxicants, high sample throughput, modest laboratory equipment and space requirements, low sample volume, portability, and reproducible responses. Enzymatic tests rely on measurement of either enzyme activity or enzyme biosynthesis. Dehydrogenases are the enzymes most used in toxicity testing. Assay of dehydrogenase activity is conveniently carried out using oxidoreduction dyes such as tetrazolium salts. Other enzyme activity tests utilize ATPases, esterases, phosphatases, urease, luciferase, beta-galactosidase, protease, amylase, or beta-glucosidase. Recently, the inhibition of enzyme (beta-galactosidase, tryptophanase, alpha-glucosidase) biosynthesis has been explored as a basis for toxicity testing. Enzyme biosynthesis was found to be generally more sensitive to organic chemicals than enzyme activity. Bacterial toxicity tests are based on bioluminescence, motility, growth, viability, ATP, oxygen uptake, nitrification, or heat production. An important aspect of bacterial tests is the permeability of cells to environmental toxicants, particularly organic chemicals of hydrophobic nature. Physical, chemical, and genetic alterations of the outer membrane of E. coli have been found to affect test sensitivity to organic toxicants. Several microbioassays are now commercially available. The names of the assays and their basis are: Microtox (bioluminescence), Polytox (respiration), ECHA Biocide Monitor (dehydrogenase activity), Toxi-Chromotest (enzyme biosynthesis), and MetPAD (enzyme activity). An important feature common to these tests is the provision of standardized cultures of bacteria in freeze-dried form. Two of the more recent applications of microbioassays are in sediment toxicity testing and toxicity reduction evaluation. Sediment pore water may be assayed directly or solvents may be used to extract the toxicants. Some of the solvents used for extraction of organic chemicals are themselves toxic to bacteria (e.g., dichloromethane), requiring exchange with a less toxic solvent (e.g., ethanol, methanol, DMSO). A modification of the Microtox test allows direct assay of solid-phase samples such as sediments. The toxicity reduction evaluation (TRE) must be carried out at wastewater treatment plants whose effluents fail toxicity standards. The TREs require numerous and repeated toxicity assays, thus favoring application of microbioassays. Presently, no single microbioassay can detect all categories of environmental toxicants with equal sensitivity. Therefore, a battery of tests approach is recommended. The differential sensitivity of alternative tests may, in fact, be exploited. Further research is needed to construct strains of genetically engineered microorganisms or isolate microorganisms or enzymes that respond to specific classes of toxicants. These can be combined into batteries appropriate for different environments or test objectives. PMID:1509175

  4. Toxicity of pristine graphene in experiments in a chicken embryo model

    PubMed Central

    Sawosz, Ewa; Jaworski, Slawomir; Kutwin, Marta; Hotowy, Anna; Wierzbicki, Mateusz; Grodzik, Marta; Kurantowicz, Natalia; Strojny, Barbara; Lipińska, Ludwika; Chwalibog, André

    2014-01-01

    Evaluation of the potential cytotoxicity of graphene is a key factor for medical applications, where flakes or a surface of graphene may be used as bioactive molecules, drug carriers, or biosensors. In the present work, effects of pristine graphene (pG) on the development of a living organism, with an emphasis on morphological and molecular states of the brain, were investigated using a chicken embryo model. Fertilized chicken eggs were divided into the control group and groups administered with pG suspended in milli-Q water at concentrations of 50 μg/L, 100 μg/L, 500 μg/L, 1,000 μg/L, 5,000 μg/L, and 10,000 μg/L (n=30 per group). The experimental solutions were injected in ovo into the albumin and then the eggs were incubated. After 19 days of incubation, the survival, weight of the body and organs, and blood serum biochemical indices were measured. The brain samples were collected for microscopic examination of brain ultrastructure and measurements of gene and protein expression. Survival of embryos was significantly decreased after treatment with pG, but the body and organ weights as well as biochemical indices were not affected. In all treatment groups, some atypical ultrastructures of the brain were observed, but they were not enhanced by the increasing concentrations of pG. Expression of proliferating cell nuclear antigen at the messenger ribonucleic acid level was downregulated, and the number of proliferating cell nuclear antigen-positive nuclei was significantly reduced in the 500–10,000 μg/L groups compared with the control group, indicating a decreased rate of deoxyribonucleic acid synthesis in the brain. The present results demonstrate some harmful effects of the applied pG flakes on the developing organism, including brain tissue, which ought to be considered prior to any medical applications. PMID:25152621

  5. Nickel toxicity in embryos and larvae of the South American toad: effects on cell differentiation, morphogenesis, and oxygen consumption.

    PubMed

    Sztrum, Abelardo Andrés; D'Eramo, José Luis; Herkovits, Jorge

    2011-05-01

    Nickel, a widely distributed heavy metal in the biosphere, produces systemic, carcinogenic, and teratogenic effects. The objectives of the present study are to report the acute, short-term chronic, and chronic toxicity of Ni in Rhinella arenarum embryos as well as the stage-dependent susceptibility to this heavy metal, including oxygen consumption, teratogenesis, and adverse effects on cell differentiation processes. The stages evaluated were blastula (S.7), gastrula (S.11), tail bud (S.17), fin circulation (S.22), and complete operculum (S.25), in this last case by means of toxicity profile curves. Nickel increases its adverse effects gradually, with a maximum value after 96 h. The 50% lethal concentrations (LC50s) for 96, 168, and 240 h at S.25 were 1.14, 0.60, and 0.48 mg Ni²(+) /L, respectively; S.11 and S.22 were the least and most susceptible to Ni with, LC50s 96 h of 6.12 and 0.19 mg Ni²(+) /L, respectively. A reduction of approximately 25% in oxygen consumption anticipates lethal effects from S.17 onward. The main teratogenic effects were retarded growth and development, extremely severe axis incurvations, persistent yolk plug, asymmetry, microcephaly and mouth and gill agenesia, and limited neuromuscular activity. Ciliated cells were not functional. The possibility of associating the remarkable stage-dependent susceptibility to Ni with environmental changes during the evolutionary process is also considered. PMID:21312246

  6. Optimization of culturing conditions for toxicity testing with the alga Oophila sp. (Chlorophyceae), an amphibian endosymbiont.

    PubMed

    Rodríguez-Gil, José Luis; Brain, Richard; Baxter, Leilan; Ruffell, Sarah; McConkey, Brendan; Solomon, Keith; Hanson, Mark

    2014-11-01

    Eggs of the yellow-spotted salamander (Ambystoma maculatum) have a symbiotic relationship with green algae. It has been suggested that contaminants that are preferentially toxic to algae, such as herbicides, may impair the symbiont and, hence, indirectly affect the development of the salamander embryo. To enable testing under near-standard conditions for first-tier toxicity screening, the authors isolated the alga from field-collected eggs and identified conditions providing exponential growth rates in the apparent asexual phase of the alga. This approach provided a uniform, single-species culture, facilitating assessment of common toxicity end points and comparison of sensitivity relative to other species. Sequencing of the 18s ribosomal DNA indicated that the isolated alga is closely related to the recently described Oophila amblystomatis but is more similar to other known Chlamydomonas species, suggesting possible biogeographical variability in the genetic identity of the algal symbiont. After a tiered approach to culturing method refinement, a modified Bristol's media with 1 mM NH4 (+) as nitrogen source was found to provide suitable conditions for toxicity testing at 18 °C and 200 µmol m(-2) s(-1) photosynthetically active radiation (PAR) on a 24-h light cycle. The validity of the approach was demonstrated with Zn(2+) as a reference toxicant. Overall, the present study shows that screening for direct effects of contaminants on the algal symbiont without the presence of the host salamander is possible under certain laboratory conditions. PMID:25113146

  7. Toxicity testing of trinitrotoluene-contaminated soil composts

    SciTech Connect

    Honeycutt, M.E.; McFarland, V.A.; Jarvis, A.S.

    1997-10-01

    The Mutatox{trademark} assay and earthworm acute toxicity test were employed to evaluate the efficacy of composting in reducing the toxicity of TNT-contaminated soils. The Mutatox assay is a proprietary bacterial bioluminescence test that determines the mutagenic potential of sample extracts. The earthworm acute toxicity test was chosen because it exposes the organisms to the unaltered contaminant/solid matrix. Rockeye soil, a TNT-contaminated soil collected from a military installation, was composted using two methods. This yielded five samples, Rockeye, Compost A composting. Soil extracts were prepared for Mutatox using the sonification method. Ten serial dilution samples were tested soils/artificial soil were tested in the earthworm toxicity test. In the Rockeye soil samples, a toxic response was shown in both test methods. Mutatox indicated no toxicity in Composts A and B after composting but did not show a positive mutagenic response in the lower serial dilutions. The LC50s for Compost A and B after composting in the earthworm toxicity test were 35.3% and 100%, respectively. Using Mutatox and the earthworm toxicity test together provides a sensitive means of monitoring the effectiveness of various composting techniques for remediating TNT-contaminated soils.

  8. 40 CFR 797.1600 - Fish early life stage toxicity test.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... exposed during the period of their early development. (7) “Embryo cup” a small glass jar or similar container with a screened bottom in which the embryos of some species (i.e., minnow) are placed during the... of newly fertilized embryos to various concentrations of a test substance. Exposure continues for...

  9. 40 CFR 797.1600 - Fish early life stage toxicity test.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... exposed during the period of their early development. (7) “Embryo cup” a small glass jar or similar container with a screened bottom in which the embryos of some species (i.e., minnow) are placed during the... of newly fertilized embryos to various concentrations of a test substance. Exposure continues for...

  10. 40 CFR 797.1600 - Fish early life stage toxicity test.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... exposed during the period of their early development. (7) “Embryo cup” a small glass jar or similar container with a screened bottom in which the embryos of some species (i.e., minnow) are placed during the... of newly fertilized embryos to various concentrations of a test substance. Exposure continues for...

  11. 40 CFR 797.1600 - Fish early life stage toxicity test.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... exposed during the period of their early development. (7) “Embryo cup” a small glass jar or similar container with a screened bottom in which the embryos of some species (i.e., minnow) are placed during the... of newly fertilized embryos to various concentrations of a test substance. Exposure continues for...

  12. 40 CFR 797.1600 - Fish early life stage toxicity test.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... exposed during the period of their early development. (7) “Embryo cup” a small glass jar or similar container with a screened bottom in which the embryos of some species (i.e., minnow) are placed during the... of newly fertilized embryos to various concentrations of a test substance. Exposure continues for...

  13. Biologically Relevant Exposure Science for 21st Century Toxicity Testing

    EPA Science Inventory

    High visibility efforts in toxicity testing and computational toxicology including the recent NRC report, Toxicity Testing in the 21st Century: a Vision and Strategy (NRC, 2007), raise important research questions and opportunities for the field of exposure science. The authors ...

  14. Assessing contaminant sensitivity of endangered and threatened aquatic species: Part III. Effluent toxicity tests

    USGS Publications Warehouse

    Dwyer, F.J.; Hardesty, D.K.; Henke, C.E.; Ingersoll, C.G.; Whites, D.W.; Augspurger, T.; Canfield, T.J.; Mount, D.R.; Mayer, F.L.

    2005-01-01

    Toxicity tests using standard effluent test procedures described by the U.S. Environmental Protection Agency were conducted with Ceriodaphnia dubia, fathead minnows (Pimephales promelas), and seven threatened and endangered (listed) fish species from four families: (1) Acipenseridae: shortnose sturgeon (Acipenser brevirostrum); (2) Catostomidae; razorback sucker (Xyrauchen texanus); (3) Cyprinidae: bonytail chub (Gila elegans), Cape Fear shiner (Notropis mekistocholas) Colorado pikeminnow (Ptychocheilus lucius), and spotfin chub (Cyprinella monacha); and (4) Poecillidae: Gila topminnow (Poeciliopsis occidentalis). We conducted 7-day survival and growth studies with embryo-larval fathead minnows and analogous exposures using the listed species. Survival and reproduction were also determined with C. dubia. Tests were conducted with carbaryl, ammonia-or a simulated effluent complex mixture of carbaryl, copper, 4-nonylphenol, pentachlorophenol and permethrin at equitoxic proportions. In addition, Cape Fear shiners and spotfin chub were tested using diazinon, copper, and chlorine. Toxicity tests were also conducted with field-collected effluents from domestic or industrial facilities. Bonytail chub and razorback suckers were tested with effluents collected in Arizona whereas effluent samples collected from North Carolina were tested with Cape Fear shiner, spotfin chub, and shortnose sturgeon. The fathead minnow 7-day effluent test was often a reliable estimator of toxic effects to the listed fishes. However, in 21 % of the tests, a listed species was more sensitive than fathead minnows. More sensitive species results varied by test so that usually no species was always more or less sensitive than fathead minnows. Only the Gila topminnow was consistently less sensitive than the fathead minnow. Listed fish species were protected 96% of the time when results for both fathead minnows and C. dubia were considered, thus reinforcing the value of standard whole-effluent toxicity tests using those two species. If the responses of specific listed species are important for management decisions, our study supports the value in developing culture and testing procedures for those species. ?? 2005 Springer Science+Business Media, Inc.

  15. Assessing contaminant sensitivity of endangered and threatened aquatic species: part III. Effluent toxicity tests.

    PubMed

    Dwyer, F J; Hardesty, D K; Henke, C E; Ingersoll, C G; Whites, D W; Augspurger, T; Canfield, T J; Mount, D R; Mayer, F L

    2005-02-01

    Toxicity tests using standard effluent test procedures described by the U.S. Environmental Protection Agency were conducted with Ceriodaphnia dubia, fathead minnows (Pimephales promelas), and seven threatened and endangered (listed) fish species from four families: (1) Acipenseridae: shortnose sturgeon (Acipenser brevirostrum); (2) Catostomidae; razorback sucker (Xyrauchen texanus); (3) Cyprinidae: bonytail chub (Gila elegans), Cape Fear shiner (Notropis mekistocholas) Colorado pikeminnow (Ptychocheilus lucius), and spotfin chub (Cyprinella monacha); and (4) Poecillidae: Gila topminnow (Poeciliopsis occidentalis). We conducted 7-day survival and growth studies with embryo-larval fathead minnows and analogous exposures using the listed species. Survival and reproduction were also determined with C. dubia. Tests were conducted with carbaryl, ammonia--or a simulated effluent complex mixture of carbaryl, copper, 4-nonylphenol, pentachlorophenol and permethrin at equitoxic proportions. In addition, Cape Fear shiners and spotfin chub were tested using diazinon, copper, and chlorine. Toxicity tests were also conducted with field-collected effluents from domestic or industrial facilities. Bonytail chub and razorback suckers were tested with effluents collected in Arizona whereas effluent samples collected from North Carolina were tested with Cape Fear shiner, spotfin chub, and shortnose sturgeon. The fathead minnow 7-day effluent test was often a reliable estimator of toxic effects to the listed fishes. However, in 21 % of the tests, a listed species was more sensitive than fathead minnows. More sensitive species results varied by test so that usually no species was always more or less sensitive than fathead minnows. Only the Gila topminnow was consistently less sensitive than the fathead minnow. Listed fish species were protected 96% of the time when results for both fathead minnows and C. dubia were considered, thus reinforcing the value of standard whole-effluent toxicity tests using those two species. If the responses of specific listed species are important for management decisions, our study supports the value in developing culture and testing procedures for those species. PMID:15750777

  16. Arsenic (III, V), indium (III), and gallium (III) toxicity to zebrafish embryos using a high-throughput multi-endpoint in vivo developmental and behavioral assay.

    PubMed

    Olivares, Christopher I; Field, Jim A; Simonich, Michael; Tanguay, Robert L; Sierra-Alvarez, Reyes

    2016-04-01

    Gallium arsenide (GaAs), indium gallium arsenide (InGaAs) and other III/V materials are finding increasing application in microelectronic components. The rising demand for III/V-based products is leading to increasing generation of effluents containing ionic species of gallium, indium, and arsenic. The ecotoxicological hazard potential of these streams is unknown. While the toxicology of arsenic is comprehensive, much less is known about the effects of In(III) and Ga(III). The embryonic zebrafish was evaluated for mortality, developmental abnormalities, and photomotor response (PMR) behavior changes associated with exposure to As(III), As(V), Ga(III), and In(III). The As(III) lowest observable effect level (LOEL) for mortality was 500 μM at 24 and 120 h post fertilization (hpf). As(V) exposure was associated with significant mortality at 63 μM. The Ga(III)-citrate LOEL was 113 μM at 24 and 120 hpf. There was no association of significant mortality over the tested range of In(III)-citrate (56-900 μM) or sodium citrate (213-3400 μM) exposures. Only As(V) resulted in significant developmental abnormalities with LOEL of 500 μM. Removal of the chorion prior to As(III) and As(V) exposure was associated with increased incidence of mortality and developmental abnormality suggesting that the chorion may normally attenuate mass uptake of these metals by the embryo. Finally, As(III), As(V), and In(III) caused PMR hypoactivity (49-69% of control PMR) at 900-1000 μM. Overall, our results represent the first characterization of multidimensional toxicity effects of III/V ions in zebrafish embryos helping to fill a significant knowledge gap, particularly in Ga(III) and In(III) toxicology. PMID:26824274

  17. STATUS AND APPLICATIONS OF ECHINOID (PHYLUM ECHINODERMATA) TOXICITY TEST METHODS

    EPA Science Inventory

    The use of echinoderms for toxicity testing has focused primarily on sea urchins and sand dollars (Strongylocentrolus purpuratus, Arbacia punctulata, Lytechinus pictus, and Dendraster excentricus, for example). he status and relative sensitivity of various test methods are descri...

  18. Toxicity and cytochrome P450 1A mRNA induction by 6-formylindolo[3,2-b]carbazole (FICZ) in chicken and Japanese quail embryos.

    PubMed

    Jönsson, Maria E; Mattsson, Anna; Shaik, Siraz; Brunström, Björn

    2016-01-01

    The tryptophan derivative formylindolo[3,2-b]carbazole (FICZ) binds with high ligand affinity to the aryl hydrocarbon receptor (AHR) and is readily degraded by AHR-regulated cytochrome P450 family 1 (CYP1) enzymes. Whether in vivo exposure to FICZ can result in toxic effects has not been examined and the main objective of this study was to determine if FICZ is embryotoxic in birds. We examined toxicity and CYP1 mRNA induction of FICZ in embryos from chicken (Gallus domesticus) and Japanese quail (Coturnix japonica) exposed to FICZ (2-200μgkg(-1)) by yolk and air sac injections. FICZ caused liver toxicity, embryo mortality, and CYP1A4 and CYP1A5 induction in both species with similar potency. This is in stark contrast to the very large difference in sensitivity of these species to halogenated AHR agonists. We also exposed chicken embryos to a low dose of FICZ (4μgkg(-1)) in combination with a CYP inhibitor, ketoconazole (KCZ). The mixture of FICZ and KCZ was lethal while FICZ alone had no effect at 4μgkg(-1). Furthermore, mixed exposure to FICZ and KCZ caused stronger and more long-lasting hepatic CYP1A4 induction than exposure to each compound alone. These findings indicate reduced biotransformation of FICZ by co-treatment with KCZ as a cause for the enhanced effects although additive AHR activation is also possible. To conclude, FICZ is toxic to bird embryos and it seems reasonable that the toxicity by FICZ involves AHR activation. However, the molecular targets and biological events leading to hepatic damage and mortality are unknown. PMID:26456929

  19. Model validation in aquatic toxicity testing: implications for regulatory practice.

    PubMed

    McCarty, L S

    2012-08-01

    Toxicity test validity is contingent on whether models and assumptions are appropriate and sufficient. A quality control evaluation of the acute toxicity testing protocol using the US. EPA fathead minnow database focused around three key assumptions that ensure results represent valid toxicological metrics: 1, it must be possible to estimate steady-state LC50s; 2, LC50s should occur at equivalent exposure durations; 3, all substantive toxicity modifying factors should be adequately controlled. About 8% of the tests failed the first assumption and are invalid and unusable. Examination of remaining data indicated variance from unquantified effects of toxicity modifying factors remained in LC50s, thereby failing assumption three. Such flaws in toxicity data generated via recommended LC50 testing protocols means resultant data do not represent consistent, comparable measures of relative toxicity. Current regulations employing LC50 testing data are acceptable due to the use of semiquantitative, policy-driven development guidance that considers such data uncertainty. Quantitative applications such as QSARs, mixture toxicity, and regulatory chemical grouping can be compromised. These validation failures justify a formal quality control review of the LC50 toxicity testing protocol. Interim improvements in the design, execution, interpretation, and regulatory applications of LC50 and related protocols using exposure-based dose surrogates are warranted. PMID:22579501

  20. The Nicotine-Evoked Locomotor Response: A Behavioral Paradigm for Toxicity Screening in Zebrafish (Danio rerio) Embryos and Eleutheroembryos Exposed to Methylmercury

    PubMed Central

    Mora-Zamorano, Francisco X.; Svoboda, Kurt R.; Carvan, Michael J.

    2016-01-01

    This study is an adaptation of the nicotine-evoked locomotor response (NLR) assay, which was originally utilized for phenotype-based neurotoxicity screening in zebrafish embryos. Zebrafish embryos do not exhibit spontaneous swimming until roughly 4 days post-fertilization (dpf), however, a robust swimming response can be induced as early as 36 hours post-fertilization (hpf) by means of acute nicotine exposure (30–240μM). Here, the NLR was tested as a tool for early detection of locomotor phenotypes in 36, 48 and 72 hpf mutant zebrafish embryos of the non-touch-responsive maco strain; this assay successfully discriminated mutant embryos from their non-mutant siblings. Then, methylmercury (MeHg) was used as a proof-of-concept neurotoxicant to test the effectiveness of the NLR assay as a screening tool in toxicology. The locomotor effects of MeHg were evaluated in 6 dpf wild type eleutheroembryos exposed to waterborne MeHg (0, 0.01, 0.03 and 0.1μM). Afterwards, the NLR assay was tested in 48 hpf embryos subjected to the same MeHg exposure regimes. Embryos exposed to 0.01 and 0.03μM of MeHg exhibited significant increases in locomotion in both scenarios. These findings suggest that similar locomotor phenotypes observed in free swimming fish can be detected as early as 48 hpf, when locomotion is induced with nicotine. PMID:27123921

  1. Early life-stage toxicity test methods for gulf toadfish (Opsanus beta) and results using chlorpyrifos

    SciTech Connect

    Hansen, D.J.; Goodman, L.R.; Cripe, G.M.; Macauley, S.F.

    1986-02-01

    Gulf toadfish (Opsanus beta) were continuously exposed as embryos, sac fry, and juveniles to technical chlorpyrifos in two 49-day early life-stage toxicity tests. Survival was significantly (alpha = 0.05) reduced only in 150 micrograms/liter. However, toadfish exposed to chlorpyrifos concentrations from 3.7 to 150 micrograms/liter weighted significantly less than control fish: 9% lower in 3.7 micrograms/liter to 62% lower in 150 micrograms/liter. The 96-hr LC50 for juvenile fish was 520 micrograms/liter. Concentrations of chlorpyrifos in toadfish and bioconcentration factors increased with increasing exposure concentration, a condition not generally observed with other marine fishes and other test chemicals. These results demonstrated the procedures for, and the practicality of, early life-stage tests with this marine species. We recommend the use of the gulf toadfish for comparative toxicity testing and for evaluating the toxicity of substances in conjunction with ontogenetical, physiological, and histological investigations of this considerably studied genus. We do not recommend it for routine effects testing.

  2. EVALUATION OF ALTERNATIVE REFERENCE TOXICANTS FOR USE IN THE EARTHWORM TOXICITY TEST

    EPA Science Inventory

    The use of the 14-d earthworm toxicity test to aid in the evaluation of the ecological impact of contaminated soils is becoming increasingly widespread. However,the method is in need of further standardization. As part of this continuing process, the choice of reference toxicants...

  3. On the performance of acute toxicity tests using the National Reference Toxicant Database

    SciTech Connect

    Zaidhk, B.

    1995-12-31

    The US National Reference Toxicant Database was used to compile data from 158 Ceriodaphnia dubia, and 187 fathead minnow acute toxicity tests. The data are analyzed using the EPA flow-chart for acute toxicity tests to determine the distribution of test methods selected. The data are reanalyzed using maximum likelihood estimation assuming probit, logit and Gompertz tolerance distributions and non-parametrically using the Spearman-Karber method with and without trimming. The results of these analyses are compared with respect to mean square error for the parametric methods and confidence intervals for the point estimate for all analyses.

  4. The effects of 4-nonylphenol and ethanol on acute toxicity, embryo development, and reproduction in Daphnia magna.

    PubMed

    Zhang, L; Gibble, R; Baer, K N

    2003-07-01

    The mean 48-h EC(50) (n=3) of 4-nonylphenol (NP) using ethanol as the carrier solvent was 155 microg/L, compared to a mean 48-h EC(50) (n=3) of 281 microg/L without ethanol. The 96-h EC(50)'s for embryo lethality (arrested egg development) and deformities (curved or unextended shell spines and undeveloped second antennae) were 738 and 263 microg/L, respectively. Reproduction studies were conducted using conditions that stimulate male production (i.e., reduced photoperiod and food levels). An increase in neonate deformities was observed at 50 microg/L (without ethanol), but no changes were observed in fecundity or sex ratios. A decrease in sex ratios was observed at 25 and 50 microg/L (with ethanol) compared to the ethanol control. However, an increase in sex ratios was observed in the ethanol control compared to media controls. The use of ethanol as a solvent carrier confounds the effects of 4-NP on acute toxicity and male production in daphnids. PMID:12798767

  5. Developmental toxicity of a commercial herbicide mixture in mice: I. Effects on embryo implantation and litter size.

    PubMed

    Cavieres, María Fernanda; Jaeger, James; Porter, Warren

    2002-11-01

    We investigated the developmental toxicity in mice of a common commercial formulation of herbicide containing a mixture of 2,4-dichlorophenoxyacetic acid (2,4-D), mecoprop, dicamba, and inactive ingredients. Pregnant mice were exposed to one of four different doses of the herbicide mixture diluted in their drinking water, either during preimplantation and organogenesis or only during organogenesis. Litter size, birth weight, and crown-rump length were determined at birth, and pups were allowed to lactate and grow without additional herbicide exposure so that they could be subjected to additional immune, endocrine, and behavioral studies, the results of which will be reported in a separate article. At weaning, dams were sacrificed, and the number of implantation sites was determined. The data, although apparently influenced by season, showed an inverted or U-shaped dose-response pattern for reduced litter size, with the low end of the dose range producing the greatest decrease in the number of live pups born. The decrease in litter size was associated with a decrease in the number of implantation sites, but only at very low and low environmentally relevant doses. Fetotoxicity, as evidenced by a decrease in weight and crown-rump length of the newborn pups or embryo resorption, was not significantly different in the herbicide-treated litters. PMID:12417478

  6. Developmental toxicity of a commercial herbicide mixture in mice: I. Effects on embryo implantation and litter size.

    PubMed Central

    Cavieres, María Fernanda; Jaeger, James; Porter, Warren

    2002-01-01

    We investigated the developmental toxicity in mice of a common commercial formulation of herbicide containing a mixture of 2,4-dichlorophenoxyacetic acid (2,4-D), mecoprop, dicamba, and inactive ingredients. Pregnant mice were exposed to one of four different doses of the herbicide mixture diluted in their drinking water, either during preimplantation and organogenesis or only during organogenesis. Litter size, birth weight, and crown-rump length were determined at birth, and pups were allowed to lactate and grow without additional herbicide exposure so that they could be subjected to additional immune, endocrine, and behavioral studies, the results of which will be reported in a separate article. At weaning, dams were sacrificed, and the number of implantation sites was determined. The data, although apparently influenced by season, showed an inverted or U-shaped dose-response pattern for reduced litter size, with the low end of the dose range producing the greatest decrease in the number of live pups born. The decrease in litter size was associated with a decrease in the number of implantation sites, but only at very low and low environmentally relevant doses. Fetotoxicity, as evidenced by a decrease in weight and crown-rump length of the newborn pups or embryo resorption, was not significantly different in the herbicide-treated litters. PMID:12417478

  7. INTRALABORATORY PRECISION OF SALTWATER SHORT-TERM CHRONIC TOXICITY TESTS

    EPA Science Inventory

    Short-term chronic toxicity tests using five salt-water species-two fishes, a mysid, a sea urchin, and a macro-alga- were developed. opper sulfate and sodium dodecyl sulfate were used as representative toxicants. oefficients of variation for LC50 results ranged from 1.8 to 46.4%....

  8. Marine and estuarine porewater toxicity testing -- species and end point comparisons

    SciTech Connect

    Carr, R.S.; Chapman, D.C.; Biedenbach, J.M.; Robertson, L.

    1994-12-31

    As part of their continuing development and evaluation of the porewater toxicity test approach for assessing the quality of marine and estuarine sediments, a variety of studies involving species and endpoint comparisons as well as validation studies have recently been conducted. The results from numerous extensive sediment quality assessment surveys have demonstrated that porewater toxicity tests are considerably more sensitive than the standard solid-phase tests and invariably exhibit a higher degree of concordance with sediment quality assessment guidelines than the standard tests. Species that have been evaluated for use in testing marine and estuarine pore water include a life-cycle test with the polychaete Dinophilus gyrociliatus, survival and hatching success with embryo-larval stages of red drum Sciaaenops ocellatus, survival of nauplii stages of the harpacticoid copepod Longipedia sp., and three different assays (fertilization, embryological development, and cytogenetic) with the sea urchin Arbacia punctulata. The different species and end points have been compared using sediment pore water from a variety of contaminated sites. Although the results of tests with the different species and end points were often comparable, in general, the sea urchin embryological development assay appears to be the most sensitive porewater test evaluated thus far in their laboratory.

  9. Reference toxicants for toxicity testing using Caenorhabditis elegans in aquatic media

    SciTech Connect

    Cressman, C.P. III; Williams, P.L.

    1997-09-01

    Caenorhabditis elegans aquatic toxicity assays were standardized with five common reference toxicants: CdCl{sub 2}, NaCl, KCl, sodium lauryl sulfate (SLS), and sodium pentachlorophenate (PCP). Aquatic toxicity testing was conducted in 3 media: a standard C. elegans medium; EPA moderately hard reconstituted water; and EPA moderately hard mineral water. Test duration in each medium was 24h without a food source, and 24h and 48h with Escherichia coli strain OP50 as a food source. Each test was replicated three times with each replicate having 6 wells per concentration, 10 worms per well. LC{sub 50} values were calculated using probit analysis. The average LC{sub 50}s for each set of replications were compared to assess sensitivity and reproducibility of the data, identifying expected variation between replicate tests. These reference toxicants increase the database for C. elegans and provide a benchmark for further application.

  10. Sensitivity or artifact? -- IQ Toxicity Test -- effluent values

    SciTech Connect

    Hayes, K.R.; Novotny, A.N.; Batista, N.

    1995-12-31

    Several complex effluents were DAPHNIA MAGNA IQ TOXICITY TESTED -- (1.25 hours) and conventionally tested with Daphnia magna (48 hours). In many samples the IQ Technology yielded low EC50 values while the 48 hour exposures yielded no acute toxicity. Possible explanations have been suggested for this occurrence such as: genotoxicity, mutagenicity, substrate interference, and enzyme satiation. To identify the causative agent(s) of this response a Toxicity Identification Evaluation was performed on one of the samples. To define the nature of the response, THE SOS-CHROMOTEST KIT and THE MUTA-CHROMOPLATE KIT were utilized to characterize genotoxicity and mutagenicity respectively. The sample did not test positive for genotoxicity but tested positive for mutagenicity only after activation with S9 enzymes, suggesting the presence of promutagens. Additional work needs to be performed to correlate IQ TOXICITY TEST sensitivity with positive MUTA-CHROMOPLATE response.

  11. Complex mixtures: methods for in-vivo toxicity testing

    SciTech Connect

    Not Available

    1988-01-01

    The National Institute of Environmental Health Sciences asked the National Research Council (NRC) to evaluate the toxicity testing of mixtures and make recommendations for improving that testing. After reviewing both the toxicologic and epidemiologic data on the adverse health effects of mixtures, the Committee on Methods for the In vivo Toxicity Testing of Complex Mixtures of the NRC concluded that a new approach, rather than new methods, is the primary need. The report describes the rationale for this conclusion and presents a series of recommended strategies for testing mixtures. Appendices include: Origins of complex mixtures; Case studies establishing active agents and/or interactions in complex mixtures; Case studies on strategies for testing the toxicity of complex mixtures; Predicting the joint risk of a mixture in terms of the component risks; Cancer models; Developmental toxicology; and Empirical modeling of the toxicity of mixtures.

  12. The chick embryo neutralization test in the assay of meningococcal antibody*

    PubMed Central

    Ueda, K.; Diena, B. B.; Greenberg, L.

    1969-01-01

    Present methods of controlling meningococcal cerebrospinal meningitis have failed to contain the disease. This has led to the search for effective vaccines and to the development of methods for assaying the potency of these vaccines, as well as for measuring the immune response of the individual. The feasibility of using the sero-protection test in embryonated eggs for the assay of meningococcal antibody has been investigated. The paper describes detailed investigations of the infectious process established by injecting Neisseria meningitidis by various challenge routes. Embryos, even 10-12 days old, showed high susceptibility to meningococci injected via the yolksac or chorioallantoic vein, and lesser susceptibility when injected via the intra-allantoic and chorioallantoic routes. The deaths of intravenously inoculated embryos coincided with the multiplication of the organisms and ensuing septicaemia. A study of the infection in embryos which had died, or were killed, 18-24 hours after inoculation showed a specific localization of the organisms in the brain. PMID:4979390

  13. A field validation of two sediment-amphipod toxicity tests.

    PubMed

    Ferraro, Steven P; Cole, Faith A

    2002-07-01

    A field validation study of two sediment-amphipod toxicity tests was conducted using sediment samples collected subtidally in the vicinity of a polycyclic aromatic hydrocarbon (PAH)-contaminated Superfund site in Elliott Bay (WA, USA). Sediment samples were collected at 30 stations with a 0.1 m2 grab from which subsamples were taken for sediment toxicity testing and geochemical and macrofaunal analyses. Standard 10-d sediment-amphipod toxicity tests were conducted with Rhepoxynius abronius and Leptocheiros plumulosus. Sediments were analyzed for 33 PAHs, pentachlorophenol, polychlorinated biphenyls, acid-volatile sulfide, simultaneously extracted metals (Cd, Cu, Zn, Pb, Ni), total organic carbon, and grain size. Sediment temperature, oxygen-reduction potential, water depth, and interstitial water salinity were also measured. Polycyclic aromatic hydrocarbons, quantified as total PAH toxic units (TU(PAH)), were confirmed to be an important common causal agent of the changes in the two toxicity test (% survival R. abronius, % survival L. plumulosus) and five macrofaunal community (number of species, S; numerical abundance, A: total biomass, B: Swartz's dominance index, SDI; Brillouin's index, H) endpoints. Two other macrofaunal community metrics (the complement of Simpson's index, 1 - SI, and McIntosh's index, MI) were less sensitive to TU(PAH) than the two toxicity test endpoints. The sensitivities of R. abronius and L. plumulosus to TU(PAH) were statistically indistinguishable. Field validations were conducted by testing the association between or among each toxicity test endpoint, each of seven macrofaunal community metrics (S, A, B, SDI, H, 1 - SI, MI), and TU(PAH) by (1) Spearman's coefficient of rank correlation, (2) Kendall's coefficient of concordance, (3) G tests of independence, and (4) regression analysis. Some field validations based on multivariable tests of association (e.g., points 2 and 3) among toxicity test, field, and stressor endpoints produced false positive results. Both toxicity test endpoints were validated as indicators of changes in S, A, SDI, and H by all the methods tested. The resolution power of the relationships between the laboratory toxicity test and macrofaunal field endpoints was low (< or = three classes) but sufficient to discriminate ecologically important effects. We conclude that standard sediment-amphipod toxicity tests are ecologically relevant and that, under the proper conditions, their results can be used for lab-to-field extrapolation. PMID:12109743

  14. Rapid toxicity testing based on yeast respiratory activity

    SciTech Connect

    Haubenstricker, M.E. ); Meier, P.G.; Mancy, K.H. ); Brabec, M.J. )

    1990-05-01

    Rapid and economical techniques are needed to determine the effects of environmental contaminants. At present, the main methods to assess the impact of pollutants are based on chemical analysis of the samples. Invertebrate and vertebrate exposures have been used over the last two decades in assessing acute and chronic toxicities. However, these tests are labor intensive and require several days to complete. An alternative to whole organism exposure is to determine toxic effects in monocellular systems. Another approach for assessing toxicity is to monitor sensitive, nonspecific, subcellular target sites such as mitochondria. Changes in mitochondrial function which could indicate a toxic effect can be demonstrated readily after addition of a foreign substance. In initial assessments of various chemicals, rat liver mitochondria (RLM) were evaluated as a biological sensor of toxicity. False toxicity assessments will result if these ions are present even though they are generally considered nontoxic. Because of these disadvantages, an alternative mitochondrial system, such as found in bakers yeast, was evaluated.

  15. Chlorinated phenol toxicity by bacterial and biochemical tests

    SciTech Connect

    Cenci, G.; Caldini, G.; Morozzi, G.

    1987-05-01

    The aim of the present investigation, carried out using E. coli as the test organism and poly-chlorophenols as the toxic organic compounds, was to define: (i) the activity ranges of different chlorinated phenols in the series from monochlorophenol to pentachlorophenol; (ii) the effect of the above mentioned compounds on growth and viability parameters, correlating experimental results with those obtained by enzymatic activities (dehydrogenase and beta-galactosidase); (iii) the relationships between toxicity and some physico-chemical properties of the considered organic compounds. The choice of chlorophenols depends on their implication in important industrial cycles and on their high toxicity for biological systems, so that their probable presence in the final effluents can be regarded also in terms of environmental toxicity. The phenol was selected as representative organic pollutant and the toxicity of other compounds was also expressed as relative phenol toxicity.

  16. Predictive modeling of developmental toxicity using EPA’s Virtual Embryo

    EPA Science Inventory

    Standard practice in prenatal developmental toxicology involves testing chemicals in pregnant laboratory animals of two species, typically rats and rabbits, exposed during organogenesis and evaluating for fetal growth retardation, structural malformations, and prenatal death just...

  17. SHORT-TERM CHRONIC TOXICITY TEST USING 'DAPHNIA MAGNA'

    EPA Science Inventory

    A seven-day short-term chronic toxicity test using 10-day old Daphnia magna is described as a possible alternative to the Ceriodaphnia life-cycle test. Five tests using sodium pentachlorophenate, conducted during the development of the method, gave test results similar to those p...

  18. 40 CFR 797.1400 - Fish acute toxicity test.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... evaluate the toxicity of the test substance. (ii) In addition to death, any abnormal behavior such as, but..., photodegradation, chemical reaction, volatilization, or sorption. (iii) Measurement of test substance. (A) For... degradation products are not present in the test chambers during the test. (E) In addition to...

  19. 40 CFR 797.1950 - Mysid shrimp chronic toxicity test.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... toxicity. (2) “Death” means the lack of reaction of a test organism to gentle prodding. (3) “Flow-through... addition to death, any abnormal behavior or appearance shall also be reported. (vii) Test organisms shall... bias from the distributions. In addition, test chambers within the testing area shall be positioned...

  20. 40 CFR 797.1400 - Fish acute toxicity test.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... evaluate the toxicity of the test substance. (ii) In addition to death, any abnormal behavior such as, but..., photodegradation, chemical reaction, volatilization, or sorption. (iii) Measurement of test substance. (A) For... degradation products are not present in the test chambers during the test. (E) In addition to...

  1. 40 CFR 797.1950 - Mysid shrimp chronic toxicity test.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... toxicity. (2) “Death” means the lack of reaction of a test organism to gentle prodding. (3) “Flow-through... addition to death, any abnormal behavior or appearance shall also be reported. (vii) Test organisms shall... bias from the distributions. In addition, test chambers within the testing area shall be positioned...

  2. Optimization of Hyalella azteca IQ Toxicity Test{trademark} for prediction of 28-day sediment toxicity tests

    SciTech Connect

    Novotny, A.N.; Ezzard, C.L.; Douglas, W.S.; Home, M.T.

    1995-12-31

    The IQ Toxicity Test, which is a rapid screening toxicity test consisting of the observation of in-vivo inhibition of an enzymatic process using a fluorescent substrate, has proven successful for the determination of 24 and 48-hour EC50`s of D. magna, C. dubia, D. pulex and M. bahia. The application of this concept to utilize the freshwater amphipod Hyalella azteca may be an excellent way in which to reduce the standard 28-day chronic sediment toxicity test to possibly one hour`s time. This study incorporates an additive experimental design to explore the effects of and interactions between five specific variables: size of the amphipod, exposure time to the toxicant, concentration of substrate, exposure time to the substrate, and length of time starved prior to testing. The results of the IQ toxicity test were compared to those of a 28-day chronic sediment toxicity test. Preliminary data indicate that there is an optimal combination of these variables which results in a concise, reproducible toxicity test for use with Hyalella azteca, and would potentially be applicable to other freshwater amphipods in the future.

  3. Marine Complex Effluent Toxicity Program: Test sensitivity, repeatability and relevance to receiving water toxicity

    SciTech Connect

    Schimmel, S.C.; Morrison, G.E.; Heber, M.A.

    1989-01-01

    In March 1984, the U.S. Environmental Protection Agency (EPA) issued a significant change in procedures regulating toxic materials in effluents through the National Pollutant Discharge Elimination System (NPDES). Concurrent with this toxicity-based effluent control policy, the EPA established the marine/estuarine component of the Complex Effluent Toxicity Testing Program (CETTP). The CETTP was established to provide reliable, sensitive and environmentally meaningful test protocols that could be used to detect toxic industrial and municipal effluents within the NPDES. Five toxicity test methods have been developed and validated for the program since 1984 using a marine plant (Champia parvula), two invertebrate species (Arbacia punctulata and Mysidopsis bahia) and two fish species (Cyprinodon variegatus and Menidia beryllina). The laboratory precision test results for the methods were acceptable; coefficients of variation for all methods were less than 54%, averaging 34%. Numerous field tests were conducted using these methods and the results indicate that tests on receiving waters (in which effluent concentrations could be estimated through controlled dye studies) accurately reflect the toxicity of the effluents measured directly. Receiving water impacts, when observed, were generally near-field in nature.

  4. ESNATS conference - the use of human embryonic stem cells for novel toxicity testing approaches.

    PubMed

    Rovida, Costanza; Vivier, Manon; Garthoff, Bernward; Hescheler, Jürgen

    2014-05-01

    The main achievements and results of the ESNATS project (Embryonic Stem Cell-based Novel Alternative Testing Strategies) were presented at the final project conference that was held on 15 September 2013, the day before the traditional EUSAAT (European Society for Alternatives to Animal Testing) Congress in Linz, Austria. The ESNATS project was an FP7 European Integrated Project, running from 2008 to 2013, the aim of which was to develop a novel toxicity testing platform based on embryonic stem cells (ESCs), and in particular, human ESC (hESCs), to accelerate drug development, reduce related R&D costs, and propose a powerful alternative to animal tests in the spirit of the Three Rs principles. Altogether, ESNATS offered the first proof of concept that hESCs can be used to create robust, reproducible and ready-to-use test assays for predicting human toxicity. In the end, essentially five test systems were developed to an adequate level for entering possible pre-validation procedures. These methods are based on hESCs, and can be combined to study the possible effects, on the human embryo, of exposure to a chemical during the early stages of development. In addition to the presentations by the main project partners, external speakers were invited to give lectures on relevant topics, both in the field of neurotoxicity and, more generally, on the applicability of hESCs in the development of advanced in vitro tests. PMID:24901904

  5. Hypersalinity toxicity thresholds for nine California ocean plan toxicity test protocols.

    PubMed

    Voorhees, Jennifer P; Phillips, Bryn M; Anderson, Brian S; Siegler, Katie; Katz, Scott; Jennings, Lydia; Tjeerdema, Ron S; Jensen, Joanna; de la Paz Carpio-Obeso, Maria

    2013-11-01

    Currently, several desalination facilities have been proposed to operate or are actually operating in California. These facilities' use of reverse osmosis (RO) may discharge hypersaline reject brine into the marine environment. The risks, if any, this brine would pose to coastal receiving waters are unknown. To test the toxicity of hypersaline brine in the absence of any additional toxic constituents, we prepared brine and tested it with the seven toxicity test organisms listed in the 2009 California Ocean Plan. The most sensitive protocols were the marine larval development tests, whereas the most tolerant to increased salinities were the euryhaline topsmelt, mysid shrimp, and giant kelp tests. Reject brines from the Monterey Bay Aquarium's RO desalination facility were also tested with three species. The effects of the aquarium's brine effluent on topsmelt, mussels, and giant kelp were consistent with those observed in the salinity tolerance experiments. This information will be used by regulators to establish receiving water limitations for hypersaline discharges. PMID:23821235

  6. Bacterial and enzymatic bioassays for toxicity testing in the environment

    SciTech Connect

    Bitton, G.; Koopman, B. )

    1992-01-01

    Microbioassays using bacteria or enzymes are increasingly applied to measure chemical toxicity in the environment. Attractive features of these assays may include low cost, rapid response to toxicants, high sample throughput, modest laboratory equipment and space requirements, low sample volume, portability, and reproducible responses. Enzymatic tests rely on measurement of either enzyme activity or enzyme biosynthesis. Dehydrogenases are the enzymes most used in toxicity testing. Assay of dehydrogenase activity is conveniently carried out using oxidoreduction dyes such as tetrazolium salts. Other enzyme activity tests utilize ATPases, esterases, phosphatases, urease, luciferase, beta-galactosidase, protease, amylase, or beta-glucosidase. Recently, the inhibition of enzyme (beta-galactosidase, tryptophanase, alpha-glucosidase) biosynthesis has been explored as a basis for toxicity testing. Enzyme biosynthesis was found to be generally more sensitive to organic chemicals than enzyme activity.107 references.

  7. THE TOXCAST PROGRAM FOR PRIORITIZING TOXICITY TESTING OF ENVIRONMENTAL CHEMICALS

    EPA Science Inventory

    The United States Environmental Protection Agency (EPA) is developing methods for utilizing computational chemistry, high-throughput screening (HTS) and various toxicogenomic technologies to predict potential for toxicity and prioritize limited testing resources towards chemicals...

  8. UTILITY OF SHORT-TERM TESTS FOR GENETIC TOXICITY

    EPA Science Inventory

    By definition, short-term test (STTs) for genetic toxicity detect genotoxic agents, not carcinogens specifically. owever, there is ufficient evidence, based on mechanistic considerations alone, to say that genotoxic agents are potential carcinogens. TTs have high statistical powe...

  9. The Toxicity Estimation Software Tool (T.E.S.T.)

    EPA Science Inventory

    The Toxicity Estimation Software Tool (T.E.S.T.) has been developed to estimate toxicological values for aquatic and mammalian species considering acute and chronic endpoints for screening purposes within TSCA and REACH programs.

  10. Eugenic selection benefits embryos.

    PubMed

    Walker, Mark

    2014-06-01

    The primary question to be addressed here is whether pre-implantation genetic diagnosis (PGD), used for both negative and positive trait selection, benefits potential supernumerary embryos. The phrase 'potential supernumerary embryos' is used to indicate that PGD is typically performed on a set of embryos, only some of which will be implanted. Prior to any testing, each embryo in the set is potentially supernumerary in the sense that it may not be selected for implantation. Those embryos that are not selected, and hence destroyed or frozen, are 'actually supernumerary'. The argument to be advanced is hypothetical: If embryos may be said to benefit or be harmed by our actions, then PGD used to select for an embryo or embryos with the highest expected Wellbeing benefits potential supernumerary embryos. The argument shows that the 'non-identity' problem is not sufficient to show that eugenic selection does not benefit supernumerary embryos. PMID:22845885

  11. Hypoxia Suppressed Copper Toxicity during Early Development in Zebrafish Embryos in a Process Mediated by the Activation of the HIF Signaling Pathway.

    PubMed

    Fitzgerald, Jennifer A; Jameson, Hannah M; Dewar Fowler, Victoria H; Bond, Georgia L; Bickley, Lisa K; Uren Webster, Tamsyn M; Bury, Nic R; Wilson, Robert J; Santos, Eduarda M

    2016-04-19

    Hypoxia is a global and increasingly important stressor in aquatic ecosystems, with major impacts on biodiversity worldwide. Hypoxic waters are often contaminated with a wide range of chemicals but little is known about the interactions between these stressors. We investigated the effects of hypoxia on the responses of zebrafish (Danio rerio) embryos to copper, a widespread aquatic contaminant. We showed that during continuous exposures copper toxicity was reduced by over 2-fold under hypoxia compared to normoxia. When exposures were conducted during 24 h windows, hypoxia reduced copper toxicity during early development and increased its toxicity in hatched larvae. To investigate the role of the hypoxia signaling pathway on the suppression of copper toxicity during early development, we stabilized the hypoxia inducible factor (HIF) pathway under normoxia using a prolyl-4-hydroxylase inhibitor, dimethyloxalylglycine (DMOG) and demonstrated that HIF activation results in a strong reduction in copper toxicity. We also established that the reduction in copper toxicity during early development was independent of copper uptake, while after hatching, copper uptake was increased under hypoxia, corresponding to an increase in copper toxicity. These findings change our understanding of the current and future impacts of worldwide oxygen depletion on fish communities challenged by anthropogenic toxicants. PMID:27019216

  12. A COUPLED MICROSOMAL-ACTIVATING/EMBRYO CULTURE SYSTEM: TOXICITY OF REDUCED BETA-NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE (NADPH)

    EPA Science Inventory

    An NADPH-dependent microsomal-activating system has been coupled to a rat embryo culture in vitro. No embryonic morphological abnormalities or decrease in final yolk sac or embryo DNA and protein contents occurred when 0.2 mM NADPH was used in this coupled system. In contrast, 1....

  13. DEVELOPMENTAL TOXICITY OF METHANOL: PATHOGENESIS IN CD-1 AND C57BL/6J MICE EXPOSED IN WHOLE EMBRYO CULTURE

    EPA Science Inventory

    BACKGROUND: Methanol causes axial skeleton and craniofacial defects in both CD-1 and C57BL/6J mice during gastrulation, but C57BL/6J embryos are more severely affected. We evaluated methanol-induced pathogenesis in CD-1 and C57BL/6J embryos exposed during gastrulation in whole em...

  14. Rice seed toxicity tests for organic and inorganic substances

    USGS Publications Warehouse

    Wang, W.

    1994-01-01

    Plant seed toxicity tests can be used to evaluate hazardous waste sites and to assess toxicity of complex effluents and industrial chemicals. Conventional plant seed toxicity tests are performed using culture dishes containing filter paper. Some reports indicate that filter papers might interfere with the toxicity of inorganic substances. In this study, a plastic seed tray was used. Rice was used as the test species. A comparison of results in the literature and this study revealed that variation of test species, methods, exposure duration, and other factors may affect the test results. The results of this study showed that the order of decreasing toxicity of metal ions was Cu>Ag>Ni>Cd>Cr(VI)>Pb>Zn>Mn>NaF for rice. The test results were similar to those reported in the literature for lettuce Ag>Ni>Cd,Cu>Cr (VI)>Zn>Mn, millet Cu,Ni>Cd>Cr(VI)>Zn>Mn, and ryegrass Cu>Ni>Mn>>Pb>Cd>Zn> Al>Hg>Cr>Fe. The order of decreasing toxicity of organic herbicides was paraquat, 2,4-D>>glyphosate>bromacil.

  15. Gender-dependent problems in toxicity tests with Ceriodaphnia dubia

    SciTech Connect

    Haynes, G.J.; Stewart, A.J.; Harvey, B.C. )

    1989-08-01

    The 7-d, three-brood Ceriodaphnia chronic toxicity test developed by Mount and Norberg is used to assess biological quality of water and wastewaters. The test is becoming widely accepted because it involves simple procedures, uses easily measurable and biologically meaningful endpoints (survival and fecundity), and uses a microcrustacean known to be relatively sensitive to various toxicants. The test uses individual animals as true replicates, and is relatively powerful statistically. Because male and female Ceriodaphnia neonates cannot be readily distinguished from one another, a 7-d C. dubia test started with neonates produced by females from stressed cultures may include an initially indeterminant number of males. The presence of males (which can be ascertained with certainty only when the test is nearly complete) can affect interpretation of toxicity test results. In this paper the results of toxicity tests of water samples from streams near the Oak Ridge National Laboratory are used to show the extent to which the presence of male C. dubia can confound the interpretation of a 7-d Ceriodaphnia test's fecundity endpoint. The authors also show the results of short-term experiments suggesting that male C. dubia are more sensitive than females to cadmium and phenol, and discuss the statistical and practical implications of the problems caused by the presence of males in chronic toxicity tests with this species.

  16. Interpretation of coastal sediment quality based on trace metal and PAH analysis, benthic foraminifera, and toxicity tests (Sardinia, Western Mediterranean).

    PubMed

    Schintu, Marco; Buosi, Carla; Galgani, François; Marrucci, Alessandro; Marras, Barbara; Ibba, Angelo; Cherchi, Antonietta

    2015-05-15

    An integrated approach for the assessment of coastal sediment quality was utilised in three areas of Sardinia (Western Mediterranean, Italy). Sediments were analysed for trace metals and polycyclic aromatic hydrocarbons (PAHs), while benthic foraminifera were used as bioindicators. Furthermore, the embryo-toxicity test was used to provide ecologically relevant information using rapid and cost-effective screening tools. The aim was to evaluate the usefulness of coupling different analytical tools. The results revealed the presence of polluted sediments in areas exposed to petrochemical industries, smelters or military settlements. However, while foraminifera have presented similar indications for chemical analysis of contamination levels in the different areas, the toxicity test exhibited a poor relationship with the contaminants measured individually. The results raise questions concerning the bioavailability of contaminants released by sediments in the water column. Overall, the toxicity rate was significant in many samples in comparison with other sites studied in other Mediterranean regions. PMID:25795553

  17. Harmonization of standard toxicity test methods used in North America

    SciTech Connect

    Ingersoll, C.G.; Dwyer, F.J.; Ankley, G.T.

    1995-12-31

    Over the past two years, Environment Canada (EC) and the US Environmental Protection Agency (EPA) have developed standard methods for conducting toxicity and bioaccumulation tests with freshwater, estuarine, and marine sediments. Existing ASTM methods were used as a basis to harmonize these methods for conducting testing with either field-collected or laboratory-spiked sediments. For freshwater toxicity tests, methods are described by EC and EPA for the amphipod Hyalella azteca and the midges Chironomus tentans and C. riparius. Endpoints include 10- to 14-d survival of growth. Methods are also described by EPA for conducting 28-d bioaccumulation tests with the oligochaete Lumbriculus variegatus. For estuarine and marine toxicity tests, methods are described for several amphipods (i.e., Rhepoxynius abronius, Ampelisca abdita, Eohaustorius estuarius, Leptocheirus plumulosus). Endpoints include 10-d survival and reburial. EC is also developing methods for conducting toxicity tests with Atlantic, Pacific, and Arctic Canadian species of polychaetes. Methods are described by EPA for conducting 28-d bioaccumulation tests with a variety of mollusks (i.e., Macoma spp.) and polychaetes (i.e., Nereis spp.). Slight inconsistencies in methods between freshwater and estuarine/marine testing or between EC and EPA testing include: (1) static vs. flow-through conditions, (2) sieving of sediment, (3) types and quantity of food, (4) age of test organisms, or (4) duration of the test and required endpoints. Additional research is in progress to: (1) develop chronic toxicity tests with amphipods and midges measuring survival, growth, or reproduction, (2) develop whole-sediment toxicity identification evaluation (TIE) procedures, (3) refine sediment spiking procedures, and (4) field-validate laboratory tests.

  18. A New Toxicity Test Using the Freshwater Copepod Cyclops vernalis.

    PubMed

    Marus, Emma M; Elphick, James R; Bailey, Howard C

    2015-09-01

    The cladocerans Ceriodaphnia dubia and Daphnia magna are widely used in environmental toxicity testing and the test methodologies for these species are well developed. However, copepods are a much more abundant contributor to zooplankton in many lakes, but they are not routinely used in toxicity tests. Therefore, we propose toxicity test methods for the freshwater copepod, Cyclops vernalis assessing effects on its survival and growth. A case study is presented in which the proposed test was performed with a range of concentrations of total dissolved solids (TDS) and used as part of a test battery to develop a site-specific water quality objective. C. vernalis was less sensitive to TDS compared to D. magna and C. dubia, but similarly sensitive to an alga, a diatom, a rotifer, a chironomid, and two fish species. No adverse effects were observed on survival or growth of C. vernalis at TDS concentrations up to 1500 mg/L. PMID:26183385

  19. Risk of ammonia toxicity to the estuarine amphipod Leptocheirus plumulosus during sediment toxicity tests

    SciTech Connect

    Moore, D.W.; Sims, J.G.; Bridges, T.S.; Dillon, T.M.; Gray, B.R.

    1994-12-31

    High levels of ammonia in sediment toxicity tests can potentially confound test results. At issue is whether the toxicity observed is due to elevated ammonia or the presence of contaminants. To evaluate the risk of ammonia toxicity, information on (1) the probability of exposure and (2) the exposure response relationship are needed. In this study the no observable effect concentration (NOEC) for ammonia was determined in both acute and chronic exposures to ammonia spiked sediments. A clean sediment was spiked with ammonium chloride to achieve porewater ammonia concentrations encompassing natural levels and those shown to produce toxicity in the water-only exposures with L. plumulosus. Two separate tests were conducted: (1) a 10-day exposure starting with juveniles and measuring survival; (2) a 28-day exposure starting with 0-72H old neonates and measuring survival, growth, and reproduction. Risk of ammonia toxicity to L. plumulosus during sediment toxicity tests was established by comparing both acute and chronic NOECs to published porewater ammonia concentrations for field collected sediments.

  20. DEVELOPMENT OF A SPERM CELL TOXICITY TEST FOR MARINE WATERS

    EPA Science Inventory

    Preliminary methods for conducting a quick and sensitive sperm cell toxicity test for marine waters have been developed. This paper presents a simple static test in which sea urchin or sand dollar sperm cells are exposed to test or control solutions for short periods of time (typ...

  1. 40 CFR 797.1330 - Daphnid chronic toxicity test.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... for asexual (parthenogenic) reproduction. For automatic feeding devices, a suggested rate is 5 to 7 mg... cultured to produce test organisms through reproduction. (2) Chronic toxicity test means a method used to... an extended period of time. In this test guideline, mortality and reproduction (and...

  2. 40 CFR 797.1330 - Daphnid chronic toxicity test.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... for asexual (parthenogenic) reproduction. For automatic feeding devices, a suggested rate is 5 to 7 mg... cultured to produce test organisms through reproduction. (2) Chronic toxicity test means a method used to... an extended period of time. In this test guideline, mortality and reproduction (and...

  3. 40 CFR 797.1330 - Daphnid chronic toxicity test.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... for asexual (parthenogenic) reproduction. For automatic feeding devices, a suggested rate is 5 to 7 mg... cultured to produce test organisms through reproduction. (2) Chronic toxicity test means a method used to... an extended period of time. In this test guideline, mortality and reproduction (and...

  4. 40 CFR 797.1330 - Daphnid chronic toxicity test.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... for asexual (parthenogenic) reproduction. For automatic feeding devices, a suggested rate is 5 to 7 mg... cultured to produce test organisms through reproduction. (2) Chronic toxicity test means a method used to... an extended period of time. In this test guideline, mortality and reproduction (and...

  5. Test chambers and test procedures for in situ toxicity testing with zooplankton

    SciTech Connect

    Pereira, A.M.M; Da Maia Soares, A.M.V.; Goncalves, F.; Ribeiro, R.

    1999-09-01

    Ecotoxicity assessment is usually done by classical tests under controlled conditions. However, field situations may not be accurately predicted by laboratory testing. In situ testing is a pertinent way for assessing the ecological relevance of laboratory bioassays. The objectives of this study were to develop an in situ test chamber and respective test protocols suitable for ecotoxicity testing with zooplankton and to evaluate the use of laboratory protocols (water column and solid phase) by comparing results obtained under controlled conditions with in situ results at an aquatic system receiving acid mine drainage. At the most contaminated stations and at the reference stations, good agreement was found between in situ and classical setting. At intermediate stations, in situ toxicity was generally higher than that in classical tests. Sample collection and preservation probably altered actual toxicity, thus emphasizing the need for caution in estimating or extrapolating field effects from laboratory results.

  6. A new biological test of water toxicity-yeast Saccharomyces cerevisiae conductometric test.

    PubMed

    Dolezalova, Jaroslava; Rumlova, Lubomira

    2014-11-01

    This new biological test of water toxicity is based on monitoring of specific conductivity changes of yeast Saccharomyces cerevisiae suspension as a result of yeast fermentation activity inhibition in toxic conditions. The test was verified on ten substances with various mechanisms of toxic effect and the results were compared with two standard toxicity tests based on Daphnia magna mobility inhibition (EN ISO 6341) and Vibrio fischeri bioluminescence inhibition (EN ISO 11348-2) and with the results of the S. cerevisiae lethal test (Rumlova and Dolezalova, 2012). The new biological test - S. cerevisiae conductometric test - is an express method developed primarily for field conditions. It is applicable in case of need of immediate information about water toxicity. Fast completion is an advantage of this test (time necessary for test completion is about 60min), the test is simple and the test organism - dried instant yeast - belongs among its biggest advantages because of its long-term storage life and broad availability. PMID:25461558

  7. Toxicity of five antidepressant drugs on embryo-larval development and metamorphosis success in the Pacific oyster, Crassostrea gigas.

    PubMed

    Di Poi, C; Evariste, L; Serpentini, A; Halm-Lemeille, M P; Lebel, J M; Costil, K

    2014-12-01

    Unlike conventional pollutants, pharmaceutical residues are continuously discharged at low levels (low to mid ng l(-1) concentrations), which results in the chronic contamination of non-target organisms, but little is known about the effects of these residues. The purpose of this study was to provide the first assessment of the ecotoxicity of five antidepressants (selective serotonin reuptake inhibitors [SSRIs] fluoxetine and sertraline, tricyclic antidepressants [TCAs] clomipramine and amitriptyline, and serotonin norepinephrine reuptake inhibitor [SNRI] duloxetine) at a wide range of concentrations from 0.1 to 100,000 μg l(-1) on two early life stages in the Pacific oyster. The toxicity was quantified in D-shaped larvae after 36 h of exposure, and in 21-day-old pediveliger larvae after 24 h of exposure using the percentage of normal larval development and the metamorphosis rate as endpoints, respectively. The embryotoxicity assays reported that the EC50 values were within the same range of concentrations (67 to 192 μg l(-1)) for all of the tested molecules. The metamorphosis tests revealed that the antidepressants can be ranked along an increasing severity gradient: clomipramine < amitriptyline < duloxetine ~ fluoxetine. Sertraline appeared to be the less toxic molecule on this endpoint; however, a different concentration range was used. The embryotoxicity test was more sensitive than the metamorphosis bioassay for three of the five molecules tested, but the latter test showed more practical benefits. Overall, the obtained toxicity values were at least 10,000-fold higher than the reported environmental concentrations. PMID:24122267

  8. Evaluation of alternative reference toxicants for use in the earthworm toxicity test

    SciTech Connect

    Yeardley, R.B. Jr.; Lazorchak, J.M.; Pence, M.A.

    1995-07-01

    The use of the 14-d earthworm toxicity test to aid in the evaluation of the ecological impact of contaminated soils is becoming increasingly widespread. However, the method is in need of further standardization. As part of this continuing process, the choice of reference toxicants was evaluated. Reference toxicants were rated in relation to the following criteria: (a) reproducibility, (b) low human health hazard, (c) feasibility of measurement, and (d) chemical stability. Potassium chloride (KCl) and ammonium chloride (NH{sub 4}Cl) were evaluated as possible alternatives to the one currently in common use, 2-chloroacetamide. Potassium chloride rated the best for the combination of the four criteria, followed by NH{sub 4}Cl and 2-chloroacetamide. Coefficients of variation (C.V.s) from control charts of six definitive tests were use to measure reproducibility. The best reproducibility (lowest C.V.) was shown by KCl, followed by NH{sub 4}Cl and 2-chloroacetamide. Toxicants ranked KCl < NH{sub 4}Cl {much_lt} 2-chloroacetamide in terms of health hazard; and KCl = NH{sub 4}Cl > 2-chloroacetamide in terms of measurement feasibility. Both 2-chloroacetamide and NH{sub 4}Cl changed in concentration during testing. Evidence is also presented that 2-chloroacetamide degrades rapidly during testing, and that, as dead worms decay, ammonification may be adding another toxicant, ammonia, to tests.

  9. Economical test methods for developmental neurobehavioral toxicity.

    PubMed

    Bignami, G

    1996-04-01

    The assessment of behavioral changes produced by prenatal or early postnatal exposure to potentially noxious agents requires both the designing of ad hoc tests and the adaptation of tests for adult animals to the characteristics of successive developmental stages. The experience in designing tests is still more limited than in the adaptation of tests, but several tests have already proven their usefulness; some examples are the suckling test, the homing test, and evaluations of dam-pup and pup-pup interactions. Functional observational batteries can exploit the development at specified postnatal ages of several reflexes and responses that are absent at birth in altricial rodent species with a short pregnancy such as the rat and the mouse. In neonates, the assessment of early treatment effects can rely not only on deviations from normal responding but also on changes in the time of appearance of otherwise normal response patterns. The same applies to other end points such as responses to pain and various types of spontaneous motor/exploratory activities, including reactivity to a variety of drug challenges that can provide information on the regulatory systems whose development may be affected by early treatments. In particular, the analysis of ontogenetic dissociations (i.e., differential early treatment effects depending jointly on developmental stage at the time of exposure, age of testing, and response end point) can be of considerable value in the study of treatments' mechanisms of action. Overall, it appears that behavioral teratological assessments can be effectively used both proactively, i.e., in risk assessment prior to any human exposure, and reactively. In the latter case, these assessments could have special value in the face of agents suspected to produce borderline changes in developing humans, whose innocuousness or noxiousness can be difficult to establish in the absence of hard evidence of teratogenicity. PMID:9182035

  10. A COMPARISON OF BULK SEDIMENT TOXICITY TESTING METHODS AND SEDIMENT ELUTRIATE TOXICITY

    EPA Science Inventory

    Bulk sediment toxicity tests are routinely used to assess the level and extent of contamination in natural sediments. While reliable, these tests can be resource intensive, requiring significant outlays of time and materials. The purpose of this study was to compare the results ...

  11. Expression of storm water runoff toxicity in three test organisms

    SciTech Connect

    Katznelson, R.; Markel, R.P.

    1995-12-31

    Storm water toxicity was monitored at numerous locations in the San Francisco Bay Area, using the EPA protocol for short-term chronic toxicity tests with freshwater organisms in the ``screening`` mode. Toxicity to Selenastrum capricomutum and Pimephales promelas was prevalent in samples collected at industrial catchments but was rarely detected in samples collected from other land use catchments. On the other hand, Ceriodaphnia dubia responded in most samples collected from industrial, commercial, residential, and transportation corridor catchments. In further analyses of C. dubia test results, the median time to lethality (LT{sub 50}) was derived to allow comparisons of the relative intensity of toxicity, and the number of offspring per female per reproductive day (OFRD) was calculated to allow separation of mortality from reproductive effects. This approach revealed that most of the moderately toxic (LT{sub 50} of 5--7 days) storm water samples collected from catchments of mixed land use (stream stations) did not inhibit reproduction, and that most storm water samples collected from transportation corridors did inhibit reproduction. Toxicity identification evaluations (TIE) indicated that diazinon is the major cause of toxicity in many stream samples, suggesting an explanation for the differential sensitivity observed in different species.

  12. Evaluation of metals, metalloids, and ash mixture toxicity using sediment toxicity testing.

    PubMed

    Stojak, Amber; Bonnevie, Nancy L; Jones, Daniel S

    2015-01-01

    In December 2008, a release of 4.1 million m(3) of coal ash from the Tennessee Valley Authority Kingston Fossil Plant occurred. Ash washed into the Emory River and migrated downstream into the Clinch and Tennessee Rivers. A Baseline Ecological Risk Assessment evaluated risks to ecological receptors from ash in the river system post-dredging. This article describes the approach used and results from sediment toxicity tests, discussing any causal relationships between ash, metals, and toxicity. Literature is limited in the realm of aquatic coal combustion residue (CCR) exposures and the potential magnitude of effects on benthic invertebrates. Sediment samples along a spectrum of ash content were used in a tiered toxicity testing approach and included a combination of 10 day sediment toxicity acute tests and longer-term, partial life cycle "definitive" tests with 2 species (Hyalella azteca and Chironomus dilutus). Arsenic, and to a lesser extent Se, in the ash was the most likely toxicant causing observed effects in the laboratory toxicity tests. Sites in the Emory River with the greatest statistical and biologically significant effects had As concentrations in sediments twice the probable effects concentration of 33 mg/kg. These sites contained greater than 50% ash. Sites with less than approximately 50% ash in sediments exhibited fewer significant toxic responses relative to the reference sediment in the laboratory. The results discussed here present useful evidence of only limited effects occurring from a worst-case exposure pathway. These results provided a valuable line of evidence for the overall assessment of risks to benthic invertebrates and to other ecological receptors, and were crucial to risk management and development of project remediation goals. PMID:25125279

  13. Oral Toxicity Study and Skin Sensitization Test of a Cricket

    PubMed Central

    Ryu, Hyeon Yeol; Lee, Somin; Ahn, Kyu Sup; Kim, Hye Jin; Lee, Sang Sik; Ko, Hyuk Ju; Lee, Jin Kyu; Cho, Myung-Haing; Ahn, Mi Young; Kim, Eun Mi; Lim, Jeong Ho; Song, Kyung Seuk

    2016-01-01

    Crickets have been attracting considerable interest in the field of nutrition and toxicology due to the global exhaustion of food resulting from a growing population. The cricket is normally eaten in several countries after roasting, similar to the grasshopper; however, safety evaluation data on cricket powder is limited. Here, we performed general toxicity studies of cricket powder including a single, 2-week repeated dose range evaluation test, a 13-week repeated oral dose toxicity test in Sprague-Dawley rats, a single oral dose toxicity test in Beagle dogs, and a skin sensitization test in guinea pigs following the Organization for Economic Cooperation and Development test guidelines 406 and 408 in addition to Good Laboratory Practice. To investigate the NOAEL and target organs of cricket powder, Sprague-Dawley rats were allocated to 4 groups: vehicle control, 1,250 mg/kg, 2,500 mg/kg, 5,000 mg/kg dose test groups and cricket powder was administered over 13 weeks after single dose and dose range finding studies in rats based on the results of the single oral administration toxicity study in rats and Beagle dogs. The results of the study showed that the NOAEL of cricket powder was over 5,000 mg/kg for both sexes of rats without adverse effects in a 13-week repeated oral toxicity study and there was no skin hypersensitivity reaction. Therefore, our results reveal that crickets can be widely used as a new substitute food or nutrient resource. PMID:27123167

  14. Oral Toxicity Study and Skin Sensitization Test of a Cricket.

    PubMed

    Ryu, Hyeon Yeol; Lee, Somin; Ahn, Kyu Sup; Kim, Hye Jin; Lee, Sang Sik; Ko, Hyuk Ju; Lee, Jin Kyu; Cho, Myung-Haing; Ahn, Mi Young; Kim, Eun Mi; Lim, Jeong Ho; Song, Kyung Seuk

    2016-04-01

    Crickets have been attracting considerable interest in the field of nutrition and toxicology due to the global exhaustion of food resulting from a growing population. The cricket is normally eaten in several countries after roasting, similar to the grasshopper; however, safety evaluation data on cricket powder is limited. Here, we performed general toxicity studies of cricket powder including a single, 2-week repeated dose range evaluation test, a 13-week repeated oral dose toxicity test in Sprague-Dawley rats, a single oral dose toxicity test in Beagle dogs, and a skin sensitization test in guinea pigs following the Organization for Economic Cooperation and Development test guidelines 406 and 408 in addition to Good Laboratory Practice. To investigate the NOAEL and target organs of cricket powder, Sprague-Dawley rats were allocated to 4 groups: vehicle control, 1,250 mg/kg, 2,500 mg/kg, 5,000 mg/kg dose test groups and cricket powder was administered over 13 weeks after single dose and dose range finding studies in rats based on the results of the single oral administration toxicity study in rats and Beagle dogs. The results of the study showed that the NOAEL of cricket powder was over 5,000 mg/kg for both sexes of rats without adverse effects in a 13-week repeated oral toxicity study and there was no skin hypersensitivity reaction. Therefore, our results reveal that crickets can be widely used as a new substitute food or nutrient resource. PMID:27123167

  15. Interpretation of reference toxicant tests in standard test procedures for sediment: How important are they?

    SciTech Connect

    Mueller, C.A.; Thursby, G.B.; Schlekat, C.E.; Scott, K.J.

    1994-12-31

    At SAIC`s Environmental Testing Center (ETC) 10-day solid-phase sediment toxicity tests using marine and estuarine amphipods are routinely conducted with a concurrent 96-hour water-only cadmium or sodium dodecyl sulfate (SDS) reference toxicant test and with a negative control sediment collected from central Long Island Sound (LIS). Reference toxicant tests are performed to evaluate test organism sensitivity. A negative control is used to evaluate laboratory ability to obtain consistent, precise results. Analysis of data from over 100 tests revealed that four situations may arise: results obtained from both the reference toxicant test and the negative control are acceptable; results from both are unacceptable; reference toxicant tests results are acceptable but negative control survival is not; and reference toxicant test results are unacceptable but negative control survival is acceptable. The latter case is of greatest interest during data interpretation. Analyses indicate that failed reference toxicant tests rarely affect interpretation of sediment test data. Therefore, the authors recommend that the use of reference toxicant tests be restricted to documentation of ongoing laboratory performance rather than interpretation of specific test results.

  16. Environmental toxicity testing of contaminated soil based on microcalorimetry.

    PubMed

    Gruiz, K; Feigl, V; Hajdu, Cs; Tolner, M

    2010-10-01

    Contaminated site assessment and monitoring requires efficient risk-management tools including innovative environmental toxicity tests. The first application of microcalorimetry for toxicity testing draw the attention to a possible new tool to increase sensitivity, to eliminate matrix effect and to study effect-mechanism. A Thermal Activity Monitor (TAM) microcalorimeter was used for measuring the heat production of various test organisms when getting in contact with sterile toxic soils. Well known bacterial (Azomonas agilis), animal (Folsomia candida) and plant test organisms (Sinapis alba) were tested for heat production. The heat response of selected testorganisms was measured in case of metal (Cu and Zn) and organic pollutant (Diesel oil, DBNPA and PCP) contaminated soils. In addition to the quantitative determination of the heat production, the mechanism of the toxic effect can be characterized from the shape of the power-time curve (slope of the curve, height and time of the maximum). In certain concentration ranges the higher the pollutant concentration of the soil the lower the maximum of the time-heat curve. At low pollutant concentrations an increased heat production was measured in case of A. agile and 20 and 200 mg Zn kg(-1) soil. The microcalorimetric testing was more sensitive in all cases than the traditional test methods. Our results showed that the microcalorimetric test method offers a new and sensitive option in environmental toxicology, both for research and routine testing. PMID:20549622

  17. Sensitivity and accuracy of whole effluent toxicity (WET) tests

    SciTech Connect

    Chapman, G.; Lussier, S.; Norberg-King, T.; Poucher, S.; Thursby, G.

    1995-12-31

    Direct measurement of effluent toxicity is a critical tool in controlling ambient toxicity. Lack of water quality criteria for many commonly discharged chemicals and complicated toxicological interactions in complex effluents are common. Complex effluent toxicity tests should provide limits as protective as National Criteria for single chemicals. One way to evaluate WET test sensitivity and accuracy is to compare WET test results with single chemicals to the National Criteria for these chemicals. A study of eight criteria chemicals (ammonia, analine, cadmium, carbaryl, copper, lead, methyl parathion, and zinc), two freshwater WET tests (Pimephales and Ceriodaphnia), and four marine WET tests (Arbacia, Champia, Menidia, and Mysidopsis) was conducted to provide this comparison. The most sensitive of the freshwater and marine WET tests with each chemical were generally less protective than the National Final Chronic Value (FCV) concentration by factors ranging from 1.09 to 44. Less-sensitive WET tests with each chemical represented significant underestimation of chronic toxicity by factors often in the range of 100--10,000. In two instances WET test results (copper and Champia, zinc and Ceriodaphnia) were below FCV by factors of 1.85 and 3.4, respectively. It is recognized that Champia is extremely sensitive to copper and that Daphnids may not be protected by the current National zinc criterion, thus these results are not surprising. Adequate accuracy for WET tests usually requires that the most sensitive WET-test species by utilized. Although the bias of WET tests to underestimate chronic toxicity is relatively small, it should be considered in the selection of test endpoints and application of WET data.

  18. Guidance for improving comparability and relevance of oil toxicity tests.

    PubMed

    Redman, Aaron D; Parkerton, Thomas F

    2015-09-15

    The complex nature and limited aqueous solubility of petroleum substances pose challenges for consistently characterizing exposures in aquatic life hazard assessments. This paper reviews important considerations for the design, conduct and interpretation of laboratory toxicity tests with physically and chemically dispersed oils based on an understanding of the behavior and toxicity of the hydrocarbons that comprise these substances. Guiding principles are provided that emphasize the critical need to understand and, when possible, characterize dissolved hydrocarbon exposures that dictate observed toxicity in these tests. These principles provide a consistent framework for interpreting toxicity studies performed using different substances and test methods by allowing varying dissolved exposures to be expressed in terms of a common metric based on toxic units (TUs). The use of passive sampling methods is also advocated since such analyses provide an analytical surrogate for TUs. The proposed guidance is translated into a series of questions that can be used in evaluating existing data and in guiding design of future studies. Application of these questions to a number of recent publications indicates such considerations are often ignored, thus perpetuating the difficulty of interpreting and comparing results between studies and limiting data use in objective hazard assessment. Greater attention to these principles will increase the comparability and utility of oil toxicity data in decision-making. PMID:26162510

  19. In vitro developmental toxicity test detects inhibition of stem cell differentiation by silica nanoparticles

    SciTech Connect

    Park, Margriet V.D.Z. Annema, Wijtske; Salvati, Anna; Lesniak, Anna; Elsaesser, Andreas; Barnes, Clifford; McKerr, George; Howard, C. Vyvyan; Lynch, Iseult; Dawson, Kenneth A.; Piersma, Aldert H.; Jong, Wim H. de

    2009-10-01

    While research into the potential toxic properties of nanomaterials is now increasing, the area of developmental toxicity has remained relatively uninvestigated. The embryonic stem cell test is an in vitro screening assay used to investigate the embryotoxic potential of chemicals by determining their ability to inhibit differentiation of embryonic stem cells into spontaneously contracting cardiomyocytes. Four well characterized silica nanoparticles of various sizes were used to investigate whether nanomaterials are capable of inhibition of differentiation in the embryonic stem cell test. Nanoparticle size distributions and dispersion characteristics were determined before and during incubation in the stem cell culture medium by means of transmission electron microscopy (TEM) and dynamic light scattering. Mouse embryonic stem cells were exposed to silica nanoparticles at concentrations ranging from 1 to 100 {mu}g/ml. The embryonic stem cell test detected a concentration dependent inhibition of differentiation of stem cells into contracting cardiomyocytes by two silica nanoparticles of primary size 10 (TEM 11) and 30 (TEM 34) nm while two other particles of primary size 80 (TEM 34) and 400 (TEM 248) nm had no effect up to the highest concentration tested. Inhibition of differentiation of stem cells occurred below cytotoxic concentrations, indicating a specific effect of the particles on the differentiation of the embryonic stem cells. The impaired differentiation of stem cells by such widely used particles warrants further investigation into the potential of these nanoparticles to migrate into the uterus, placenta and embryo and their possible effects on embryogenesis.

  20. PREPARATION OF BENTHIC SUBSTRATES FOR SEDIMENT TOXICITY TESTING

    EPA Science Inventory

    A jar-rolling apparatus was constructed to prepare test substrates using sediments spiked with laboratory chemicals, the toxicity of which were assessed with the Rhepoxynius abronius bioassay. Test sediments were mixed by rolling them for several hours in one-gallon glass jars. M...

  1. POREWATER CHEMISTRY: EFFECTS OF SAMPLING, STORAGE, HANDLING, AND TOXICITY TESTING

    EPA Science Inventory

    As a general principle, it is nearly impossible to remove a porewater sample from sediment, use it in a toxicity testing vessel with test organisms, and prevent changes in the chemistry of the natural and anthropogenic organic and inorganic constituents. The degree of change in t...

  2. GENETIC VARIATION FOR COPPER RESISTANCE IN FATHEAD MINNOW TOXICITY TESTS

    EPA Science Inventory

    Unexplained variation in the results of aquatic organism toxicity tests is a consistently observed and troubling phenomenon. Possible sources of variation include differences in condition or nutritional status of the population prior to the test, as well as age, density and hand...

  3. PROPOSED TEST PROTOCOL TO DETERMINE TOXICANT LEACHING INTO POTABLE WATER

    EPA Science Inventory

    Research was conducted to develop a general test protocol to identify the possible deterioration of water quality as a result of leaching of toxicants from distribution/transmission pipes and tanks during distribution and storage of potable water. The developed protocol was teste...

  4. SIMPLIFIED FEEDING TECHNIQUES FOR CULTURING AND TOXICITY TESTING WITH 'CERIODAPHNIA'

    EPA Science Inventory

    Food used for Ceriodaphnia dubia in culture and toxicity testing should be uniform, adequate, and easily prepared. Using as a control the current standard food, YCTF, the authors tested a variety of foods made with algal species singly and in combination, prepared with several me...

  5. Results of chronic toxicity tests conducted on selected A-area outfalls, June-August 1996

    SciTech Connect

    Specht, W.L.

    1997-07-01

    In anticipation of possible toxicity testing requirements in the SRS`s new (1996) NPDES permit, toxicity tests were performed at selected A-Area NPDES outfalls in order to determine if the outfalls were toxic. Chronic definitive toxicity tests were conducted on Ceriodaphnia dubia using water collected from nine locations during the summer of 1996. Six of the nine locations were toxic.

  6. Differential effect of solar light in increasing the toxicity of silver and titanium dioxide nanoparticles to a fish cell line and zebrafish embryos.

    PubMed

    George, Saji; Gardner, Hannah; Seng, Eng Khuan; Chang, Hengky; Wang, Chunyan; Yu Fang, Crystal Hay; Richards, Mark; Valiyaveettil, Suresh; Chan, Woon Khiong

    2014-06-01

    The increasing use of silver (Ag) and titanium dioxide (TiO2) nanoparticles (NPs) in consumer products and their inevitable seepage into the environment prompted us to investigate their potential toxicity to a fish cell line (BF-2) and zebrafish embryos under dark and Simulated Solar Light (SSL) exposure conditions. Using high throughput screening (HTS) platforms, we showed that the oxidative stress-dependent cytotoxicity and embryonic toxicity of NPs were significantly increased upon exposure to SSL. While, the toxicity of TiO2 NPs under SSL exposure could be explained by hydroxyl radical generation, the enhanced toxicity of Ag NPs under SSL exposure was due to surface oxidation and physicochemical modification of Ag NPs and shedding of Ag+, leading to an increased bioavailability of silver. Our observations that solar light could induce physicochemical transformation of TiO2 and Ag NPs and enhance their toxic potential emphasizes the need for conducting future toxicity studies under environmentally relevant exposure conditions to guide decision making on the safe handling of NPs. PMID:24811346

  7. Comparison of ethanol toxicity to Daphnia magna and Ceriodaphnia dubia tested at two different temperatures: static acute toxicity test results

    SciTech Connect

    Takahashi, I.T.; Cowgill, U.M.; Murphy, P.G.

    1987-08-01

    Ethanol is a commonly used solvent in toxicity testing, yet there are few studies in the literature devoted to its toxicity to zooplankton. The purpose of this study was to compare the response of Daphnia magna Straus 1820 and Ceriodaphnia dubia J. Richard 1894 to ethanol. Two temperatures were selected because most toxicity data involving D. magna has been carried out at 20/sup 0/C while all discussions concerning C. dubia appear to relate to temperatures oscillating around 25/sup 0/C. Thus, the response of these two organisms to ethanol was examined at 20/sup 0/C and at 24/sup 0/C.r

  8. Sun light mediated synthesis of gold nanoparticles as carrier for 6-mercaptopurine: Preparation, characterization and toxicity studies in zebrafish embryo model

    SciTech Connect

    Ganeshkumar, Moorthy; Sastry, Thotapalli Parvathaleswara; Sathish Kumar, Muniram; Dinesh, Murugan Girija; Kannappan, Sudalyandi; Suguna, Lonchin

    2012-09-15

    Highlights: ► Gold nanoparticles prepared using eco-friendly method with good in vitro stability. ► Can be used as drug delivery system. ► Did not show any toxicity in zebrafish embryo. ► More toxic to cancer cells when compared to N-Au-Mp and Mp. -- Abstract: The objective of this study is to synthesize green chemistry based gold nanoparticles by sun light irradiation method. The prepared gold nanoparticles (AuNPs) were modified using folic acid and then coupled with 6-mercaptopurine. These modified nanoparticles were used as a tool for targeted drug delivery to treat laryngeal cancer. In the present study, novel bionanocomposites containing nutrient agar coated gold nano particles (N-AuNPs) coupled with 6-mercaptopurine (drug) (N-AuNPs-Mp), folic acid (ligand) (N-AuNPs-Mp-Fa) and rhodamine (dye) (N-AuNPs-Rd), a fluorescent agent, were prepared and characterized by IR, UV, TEM, Particle size analysis and in vitro stability. The toxicity and fluorescence of N-Au was studied using zebrafish embryo model. The in vitro cytotoxicity of free Mp, N-Au-Mp and N-Au-Mp-Fa against HEp-2 cells was compared and found that the amount of Mp required to achieve 50% of growth of inhibition (IC{sub 50}) was much lower in N-Au-Mp-Fa than in free Mp and N-Au-Mp.

  9. Using toxicity testing to evaluate electrochemical reactor operations.

    PubMed

    Saylor, Greg L; Chen, Linxi; Kupferle, Margaret J

    2012-03-01

    In the present study, the Microtox® test was used to track the toxicity of electrochemical effluents to the marine bacteria Vibrio fischeri as a function of reaction time. When electrochemistry was used to degrade aqueous phenol using different reactor configurations, two reaction pathways were identified, chlorine substitution and oxidation, depending on whether the electrolyte contained chloride. For a boron-doped diamond (BDD) anode, electrochemistry using Na₂SO₄ electrolyte produced a significantly more toxic effluent than when using NaCl electrolyte with all other conditions remaining the same. This effect is attributed to the reaction pathway, specifically the production of benzoquinone. Benzoquinone was produced only during electrochemistry using Na₂SO₄ and is the most toxic potential intermediate, having nearly 800 times more toxicity than phenol. Although the use of NaCl produced a lower toxicity effluent than Na₂SO₄, caution should be observed because of the production of chlorinated phenols, which can be of special environmental concern. When comparing graphite rod and BDD plate anodes in terms of toxicity evolution when using Na₂SO₄, BDD was found to produce a lower toxicity effluent; this is a result of the increased oxidizing power of BDD, reducing the formation of benzoquinone. In this comparison, the type of anode material/electrode configuration did not seem to affect which intermediates were detected but did affect the quantity of and rate of production of intermediates. PMID:22170029

  10. When can embryos learn? A test of the timing of learning in embryonic amphibians.

    PubMed

    Sehr, Evie K; Beasley, Lindsay N; Wilson, Kurtis W; Gall, Brian G

    2016-04-01

    Learning is crucial to the survival of organisms across their life span, including during embryonic development. We set out to determine when learning becomes possible in amphibian development by exposing spotted salamander (Ambystoma maculatum) embryos to chemical stimuli from a predator (Ambystoma opacum), nonpredator (Lithobates clamitans), or control at developmental stages 16-21 or 36-38 (Harrison 1969). Once exposures were completed and embryos hatched, we recorded the number of movements and time spent moving of individuals in both groups and all treatments. There was no significant difference in number of movements or time spent moving among any of the treatments. The groups that were exposed to predator stimuli and a blank control at stages 36-38 were also tested to determine whether there was a difference in refuge preference or difference in survivorship when exposed to a predator (marbled salamander). There was no difference in survival or refuge preference between individuals; however, all individuals preferred vegetated over open areas regardless of treatment type. We discuss hypotheses for the absence of embryonic learning in this species and suggest it may be the result of the intensity of the predator-prey interaction between the predator, large marbled salamander larvae, and the prey, spotted salamander larvae. PMID:27110353

  11. Developmental toxicity testing of biopharmaceuticals in nonhuman primates: previous experience and future directions.

    PubMed

    Martin, Pauline L; Weinbauer, Gerhard F

    2010-12-01

    Developmental toxicity studies for pharmaceutical safety testing are designed to evaluate potential adverse effects of drug treatment on pregnancy and on the developing embryo/fetus. Biopharmaceuticals present specific challenges for developmental toxicity testing because the pharmacology of these molecules, which are frequently human-specific proteins, is often restricted to humans and nonhuman primates (NHPs). For those species-restricted molecules, the only option for the evaluation of potential effects on development of the human biopharmaceutical is to use NHPs. This article reviews each of the stages of development in cynomolgus macaques (the most frequently used NHP) and the potential exposure of the embryo, fetus, and infant following administration of a biopharmaceutical during pregnancy and lactation. Because the purpose of the NHP developmental studies is to identify potential human risks, a comparison between macaque and human development and potential exposure has been made when possible. Understanding the potential exposure of the conceptus relative to critical periods in development is essential to designing a scientifically based study that adequately addresses human risks. Some options for NHP study designs, including the option of combining end points into a single study, and the pros and cons of each of the study options have been reviewed. Developmental studies for biopharmaceuticals in NHPs need to be optimally designed on a case-by-case basis taking into consideration the pharmacology of the molecule, the type of molecule (antibody or non-antibody), the potential exposure relative to the development of potential target organs, the clinical use, and the ethical considerations associated with the use of NHPs. PMID:20926830

  12. Identification of PIWIL1 Isoforms and Their Expression in Bovine Testes, Oocytes, and Early Embryos.

    PubMed

    Russell, Stewart J; Stalker, Leanne; Gilchrist, Graham; Backx, Alanna; Molledo, Gonzalo; Foster, Robert A; LaMarre, Jonathan

    2016-04-01

    PIWI proteins are members of the larger Argonaute family and bind to specific 24-32 nucleotide RNAs called PIWI-interacting RNAs (piRNAs). PIWI-interacting RNAs direct PIWI-mediated suppression of retrotransposon expression in the male germline in humans and mice, but their roles in bovine reproduction and embryogenesis are unknown. Although the majority of research in mammals has focused on the functions of PIWI proteins during spermatogenesis, this family of proteins and their associated piRNAs have recently been identified in early embryos. The goals of this study were to characterize the expression of PIWIL1 in bovine testis, oocytes, and early embryos. A full-lengthPIWIL1transcript and protein was found in the testis, specifically in the germs cells of mature seminiferous tubules. RNA-immunoprecipitation demonstrated the presence of putative piRNAs with a mean length of 30 nucleotides bound to PIWIL1 in testes. 3'-Rapid amplification of cDNA ends analysis ofPIWIL1transcripts in testes and oocytes revealed two shorter isoforms in addition to the full-length transcript that was only present in testes. TruncatedPIWIL1isoforms in oocytes and testes were confirmed through amplification of their unique intronic fragments. Expression profiling ofPIWIL1through early embryogenesis demonstrated peak mRNA expression at the 2-cell stage with decreasing levels through to the blastocyst. PIWIL1-YFP fusion plasmids were produced for each isoform and expressed in HEK 293 cells, demonstrating nuclear exclusion and size-specific banding of the different isoforms. These data represent the first comprehensive characterization of PIWIL1 in bovine, revealing functional similarities with PIWIL1 in other species and suggest tissue-specific expression of several isoforms. PMID:26911426

  13. Baker's yeast assay procedure for testing heavy metal toxicity

    SciTech Connect

    Bitton, G.; Koopman, B.; Wang, H.D.

    1984-01-01

    Baker's yeast (Saccharomyces cerevisiae) is microorganism which is commercially available and sold as packaged dry pellets in any food store at low cost. Studies have been undertaken on the effects of organic xenobiotics as well as heavy metals on yeast metabolism. This type of study has been generally useful in examining the mechanism(s) of chemical toxicity. However, a rapid and quantitative toxicity test using S. cerevisiae as the test organism has not been developed. The purpose of this study was to develop a toxicity assay for heavy metals, using commercial dry yeast as the test microorganism. This rapid and simple procedure is based on the reduction of 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT) to INT-formazan by the yeast electron transport system. The scoring of active cells following exposure to heavy metals was undertaken according to the MINT (malachite green-INT) method developed by Bitton and Koopman.

  14. Toxicity Testing in the 21st Century: Using the New Toxicity Testing Paradigm to Create a Taxonomy of Adverse Effects

    PubMed Central

    Boekelheide, Kim; Campion, Sarah N.

    2010-01-01

    Distinguishing adaptive from adverse responses is fundamental to understanding toxicity and to implementing regulatory guidelines that are protective of human health. However, what we consider to be an adverse effect may change over time as the cultural acceptance of risk alters and new knowledge and insight accumulate. The fact that the identification of an adverse effect is subject to change is obvious, necessary, and uncomfortable. In this commentary, a framework for defining adverse effects is proposed for the emerging paradigm of toxicity testing in the 21st centurya paradigm that focuses on human cells, in vitro approaches, toxicity pathways, and high-throughput techniques. The traditional meaning of an adverse effect as a change at the organismal level is not compatible with this new system of toxicity testing. Instead, based on the experience of accident investigators, we propose that a Toxicological Factors Analysis and Classification System will use the database resulting from the high-throughput toxicity testing of the future to develop a Taxonomy of Adverse Effects. Similar to an accident, predisposing latent failures identified within categories of the toxicant response database will be associated with the active failure of an adverse effect. PMID:20026472

  15. Sensitivity and precision of whole effluent toxicity (WET) tests

    SciTech Connect

    Denton, D.; Chapman, G.; Fulk, F.

    1995-12-31

    The US Environmental Protection Agency`s test method manuals recommend reference toxicant test be performed to determine test sensitivity and precision within a test and among tests over time. The levels of intra and interlaboratory precision with two reference toxicants (zinc and copper) were examined with six marine test species (Macrocystis pyrifera, Haliotis rufescens, Strongylocentrotus purpuratus, Dendraster excentricus, Mytilus spp., and Menidia beryllina) and with two freshwater test species (Ceriodaphnia dubia and Pimephales promelas). Data from the EPA`s Reference Toxicant Database were used in the analysis. Data was used if the tests met the specified test acceptability criteria. The test sensitivity was examined by calculation of the minimum significant difference (e.g. MSD) and will be discussed. Results were compared to the national final chronic value (FCV) for copper and zinc. Greater than 99% of the EC25 values were above the FCV for copper with Strongylocentrotus purpuratus, Dendraster excentricus and Macrocystis pyrifera. However, greater than 99% of the EC25 values were below the FCV for zinc with Haliotis rufescens.

  16. In vitro and in ovo effects of four brominated flame retardants on toxicity and hepatic mRNA expression in chicken embryos.

    PubMed

    Egloff, Caroline; Crump, Doug; Chiu, Suzanne; Manning, Gillian; McLaren, Kristina K; Cassone, Cristina G; Letcher, Robert J; Gauthier, Lewis T; Kennedy, Sean W

    2011-11-10

    Some currently used brominated flame retardants (BFRs), such as hexachlorocyclopentadienyl-dibromocyclooctane (HCDBCO), bis(2-ethylhexyl)tetrabromophthalate (BEHTBP), 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE) and decabromodiphenylethane (DBDPE), are persistent organic contaminants detected in various environmental matrices, including wild birds. Data on potential toxicological and molecular responses to exposure of these BFRs are lacking for avian species. A combined in vitro/in ovo approach was used to determine the concentration-dependent effects of these BFRs on overt toxicity and hepatic messenger RNA (mRNA) expression levels of 11 transcripts in (1) primary cultures of chicken embryonic hepatocytes (CEH; all four BFRs) and (2) chicken embryos (HCDBCO and BTBPE only). Neither hepatocyte viability nor embryonic pipping success were affected by the BFRs at any of the administered concentrations (CEH: 0.001-30 μM, egg injection: 0.1-10 μg/g nominal dose). In CEH, 10 μM HCDBCO induced cytochrome P450 2H1 (CYP2H1) and CYP3A37, while CYP1A4/5 were down-regulated at all tested concentrations. In contrast, only transthyretin was down-regulated by HCDBCO in embryonic liver. There was concordance between the BTBPE-induced transcriptional responses in vitro and in ovo for CYP1A4/5 (up-regulated) and type III iodothyronine 5'-deiodinase (DIO3; down-regulated). DBDPE induced CYP1A4/5 29- and 59-fold at 0.2 μM in CEH and increased DIO1. None of the gene targets were responsive to BEHTBP exposure in CEH. The multi-tiered in vitro/in ovo screening approach was effective for assessing toxicological and molecular biological effects of these BFRs in an avian species. PMID:21893176

  17. The testing and evaluation of non-toxic antifouling coatings.

    PubMed

    Swain, G W; Schultz, M P

    1996-01-01

    Field testing of non-toxic antifouling coatings has required the development of test protocols that can quantify their performance. This includes the evaluation of the biofouling communities, the measurement of biofouling adhesion using a calibrated water jet and the measurement of barnacle adhesion in shear. Data are presented for several test surfaces, and the results are discussed with respect to the coating characteristics. PMID:22115111

  18. Resolving some practical questions about Daphnia acute toxicity tests

    SciTech Connect

    Barera, Y.; Adams, W.J.

    1981-10-01

    Acute toxicity tests were performed with six age groups of Daphnia magna, ranging from less than or equal to6 h to 216 h, and with five chemicals, selected on the basis of their physical and chemical properties as well as their acute toxicity to D. magna. The age of the daphnids did not significantly alter the 48-h EC/sub 50/ values for the chemicals tested. The maximum difference observed in the 48-h EC/sub 50/ values between the 6-h and 216-h age groups was a factor of 3.9 for linear alkylbenzene sulfonate (LAS). For purposes of standardization, it appears that D. magna up to 48 h of age at the beginning of the test can be used to conduct acute toxicity tests with most chemicals. The results of static acute toxicity tests conducted with butylbenzyl phthalate (BBP) and D. magna in the presence and absence of several commonly used solvents indicate that the acute toxicity of this chemical is not altered by the use of a solvent carrier. The 48-h EC/sub 50/ value for BBP without a solvent was 1.0 mg/L, compared with a range of 1.6 to 2.2 mg/L when acetone, dimethylformamide, ethanol, or triethylene glycol were used as solvent carriers. The acute toxicities of the solvents in the absence of BBP were also determined for D. magna. The values ranged from 9.3 to 52.4 g/L. The results of static acute tests performed with D. magna and BBP in the presence of various concentrations of daphnid foods (algae or trout chow), indicate that the 48-h EC/sub 50/ values increase proportionally with an increase in food concentrations. These results suggest that acute toxicity tests with D. magna should be conducted in the presence of food with chemicals with a high Ksigma if the results are to be used to select the test concentrations for a chronic study with daphnids. The type of food and the concentration used in the acute test should be the same as those used in a chronic test.

  19. Static renewal tests using Pimephales promelas (fathead minnows). Clinch River-Environmental Restoration Program (CR-ERP) study, ambient water toxicity

    SciTech Connect

    Simbeck, D.J.

    1993-12-31

    Clinch River-Environmental Restoration Program (CR-ERP) personnel and Tennessee Valley Authority (TVA) personnel conducted a study during the week of October 21--28, 1993. The organisms specified for testing were larval fathead minnows, Pimephales promelas, and the daphnid, Ceriodaphnia dubia. Due to serious reproduction/embryo abortion problems with the TVA daphnid cultures, TVA conducted tests during this study period using only fathead minnows. Surface water samples were collected by TVA Field Engineering personnel from Poplar Creek Mile 2.9, Mile 4.3, and Mile 5.1 on October 20, 22, and 25. Samples were split and provided to the CR-ERP and TVA toxicology laboratories for testing. Exposure of test organisms to these samples resulted in no toxicity (survival or growth) in testing conducted by TVA. Attachments to this report include: Chain of custody forms -- originals; Toxicity test bench sheets and statistical analyses; and Reference toxicant test information.

  20. REPORT ON ALGAL TOXICITY TESTS ON SELECTED OFFICE OF TOXIC SUBSTANCES (OTS) CHEMICALS

    EPA Science Inventory

    Algal (Selenastrum capricornutum) assays were performed on 11 chemicals selected from a larger list of candidate chemicals provided by the EPA's Office of Toxic Substances (OTS). imited chemical analysis of those chemicals that could be tested readily at ERL-C was performed in su...

  1. Control group response variability in short-term toxicity tests

    SciTech Connect

    Gast, L.C.; Shimp, C.; Wang, Q.; Shukla, R.; Fulk, F.

    1995-12-31

    The US EPA`s National Reference Toxicant Database (NRTDB) has afforded an excellent opportunity to examine and document variability in responses within control groups (i.e. zero concentration of the toxicant.) The NRTDB has compiled acute and chronic reference toxicant test results for eight species and currently contains results for 32 laboratories and generally eight to ten tests for a species within each laboratory. The Ceriodaphnia dubia Survival and Reproduction test and the Pimephales promelas (fathead minnow) Survival and Growth test are the most frequently represented chronic tests with 331 and 144 sets of test data, respectively. For this presentation, Ceriodaphnia dubia reproduction data, expressed as total numbers of young in the test period, and fathead minnow survival and growth data were analyzed using a variance components model. The information regarding the control population is useful in examining the sources of inter and intralaboratory variability of chronic testing. In addition, this control population response variability information will be valuable for characterizing what can be termed as ``practically equivalent responses`` between a control and an effluent. The preliminary analysis indicates considerable between-test variability; however, this variability is not consistent across laboratories. Results of further exploration on this issue will be presented.

  2. Use of DNA strand damage (Comet assay) and embryo hatching effects to assess contaminant exposure in blue crab (Callinectes sapidus) embryos

    SciTech Connect

    Lee, R.F.; Steinert, S.A.; Nakayama, K.; Oshima, Y.

    1999-07-01

    After fertilization, blue crab eggs are embedded in a sponge which is attached to the female abdomen during embryo development. Embryos after 9 stages in the egg sac hatch into a swimming zoea stage (stage 10). The authors have developed a bioassay where embryo development is monitored in culture plates with and without toxicants in the water. Toxicant effects are based on determining the percentage of embryos which hatch to zoea. Hatching EC{sub 50} (toxicant concentration at which 50% of the embryos fail to hatch) for a number of pesticides, organometallics and metals were determined. The test takes from 2 to 6 days depending on the embryo stage selected for the study. In addition to embryo development effects the prevalence of DNA single-strand breaks in individual embryo cells were determined using the single cell gel electrophoresis method (Comet assay). A good correlation between DNA strand breakage and embryo defects was found after exposure to genotoxic contaminants. Thus, the bioassay linking DNA damage to embryo hatching effects is rapid, sensitive and mechanistically relevant.

  3. Guidelines for developmental toxicity testing of chemicals in Japan

    SciTech Connect

    Tanimura, T.

    1985-11-01

    With the definition of teratogenicity expanded in terms of the developmental stages when an agent acts and the types of developmental anomalies induced, the concept of developmental toxicity has been established. The examination of functional developmental disorders including behavior has become one of the most important items for the evaluation of developmental toxicity of chemicals, especially pharmaceutical drugs. The guidelines for developmental toxicity testing of drugs in Japan stress the need for examination of growth and development including behavior and fertility on the postweaning offspring. The outline of the Japanese guidelines is presented and it is emphasized that studies should be done as research and include self evaluation of the scientific truth of the experiment and extrapolation to humans. In addition, the activities of the Behavioral Teratology Meeting, a satellite meeting to the Japanese Teratology Society, are introduced and enquete surveys of the Japan Pharmaceutical Manufacturers Association and collaborative studies for the standardization of learning tests in Japan are briefly presented.

  4. Functional toxicology: tools to advance the future of toxicity testing

    PubMed Central

    Gaytán, Brandon D.; Vulpe, Chris D.

    2014-01-01

    The increased presence of chemical contaminants in the environment is an undeniable concern to human health and ecosystems. Historically, by relying heavily upon costly and laborious animal-based toxicity assays, the field of toxicology has often neglected examinations of the cellular and molecular mechanisms of toxicity for the majority of compounds—information that, if available, would strengthen risk assessment analyses. Functional toxicology, where cells or organisms with gene deletions or depleted proteins are used to assess genetic requirements for chemical tolerance, can advance the field of toxicity testing by contributing data regarding chemical mechanisms of toxicity. Functional toxicology can be accomplished using available genetic tools in yeasts, other fungi and bacteria, and eukaryotes of increased complexity, including zebrafish, fruit flies, rodents, and human cell lines. Underscored is the value of using less complex systems such as yeasts to direct further studies in more complex systems such as human cell lines. Functional techniques can yield (1) novel insights into chemical toxicity; (2) pathways and mechanisms deserving of further study; and (3) candidate human toxicant susceptibility or resistance genes. PMID:24847352

  5. Modulation of cupric ion activity by pH and fulvic acid as determinants of toxicity in Xenopus laevis embryos and larvae

    SciTech Connect

    Buchwalter, D.B. |; Linder, G.; Curtis, L.R.

    1996-04-01

    An ion-specific electrode measured cupric ion activity modulated by fulvic acid (FA) and pH in a series of modified Frog Embryo Teratogenesis Assay--Xenopus (FETAX) toxicity assays. Hydrogen ion concentration was the primary determinant of cupric ion activity, while FA played a smaller but significant role. Fulvic acid was a weak copper complexing agent at pH 5.50. At pH 5.50 there was slight reduction of ionic activity and a subsequent attenuation of copper toxicity with 5.0 mg/L FA. At pH 7.50, FA also had a mild attenuating effect on copper toxicity. At pH 6.50, copper was strongly complexed by FA at total copper (TCu) concentrations below its pH-dependent solubility limit. At TCu concentrations above the solubility limit FA enhanced toxicity. There was more cupric ion activity measured in the presence of 0.5 and 5.0 mg/L FA than without it at TCu concentrations above the solubility limit. The proposed mechanism for this behavior was FA action as a nucleation inhibitor. Under the chemical conditions of the pH 6.50 experiments, a stable supersaturation of copper was formed, resulting in a more toxic aqueous matrix.

  6. An information-rich alternative, chemicals testing strategy using a high definition toxicogenomics and zebrafish (Danio rerio) embryos.

    PubMed

    Sawle, Ashley D; Wit, Ernst; Whale, Graham; Cossins, Andrew R

    2010-11-01

    Large-scale toxicogenomic screening approaches offer great promise for generating a bias-free system-wide view of toxicological effects and modes-of-action of chemicals and ecotoxicants. However, early applications of microarray technology have identified relatively small groups of responding genes with which to define new targets for analysis by conventional means. We have trialled a more intensive approach to the design and interpretation of array experiments incorporating a balanced interwoven ANOVA design with higher levels of biological replication, a more thorough analysis of errors and false discovery rates, and an analysis of response patterns using gene network models. Zebrafish embryos were exposed from 1.5 h post-fertilization for 72 h to ecotoxicants representing different classes--2,4-dichlorophenol, 3,4-dichloroaniline, pentachlorophenol, and cadmium chloride--at low concentrations producing a developmental disturbance to 10% of embryos and half of this dose. Extracted whole embryo RNA was then analyzed on microarrays. Analysis revealed responses of 3000-5000 genes, which is 10-1000 times greater than previously reported, with significance at lower levels of fold change. Some gene responses were common to multiple toxicants, and others were restricted to just one or two toxicants. The gene expression profiles for the different toxicants were distinctive, and analysis using network-based models provided a high level of detail of affected processes, some of which were novel. This approach provides a more highly refined view of toxic effects, from which meaningful patterns of response can be discerned and related to functional deficits and from which more reliable indicators of toxicological effect can be predicted. PMID:20702589

  7. The Third FRAME Toxicity Committee: working toward greater implementation of alternatives in toxicity testing.

    PubMed

    Combes, Robert D; Balls, Michael; Bansil, Lee; Barratt, Martin; Bell, David; Botham, Philip; Broadhead, Caren; Clothier, Richard; George, Elizabeth; Fentem, Julia; Jackson, Michael; Indans, Ian; Loizou, George; Navaratnam, Vyra; Pentreath, Victor; Phillips, Barry; Stemplewski, Henry; Stewart, Jane

    2004-06-01

    FRAME (the Fund for the Replacement of Animals in Medical Experiments; http://www. frame.org.uk) is a scientific charity, which has, for over 30 years, been advocating and conducting its own research on the application of the Three Rs (reduction, refinement and replacement) to animal experimentation. FRAME develops and validates scientifically based replacement alternative methods to facilitate their acceptance by scientists and regulators. As part of these activities, FRAME established a FRAME Toxicity Committee in 1979, and a report of its work was published in 1982, and discussed in the proceedings of a subsequent meeting, published in 1983. A Second Toxicity Committee formed in 1988, reported its work in 1990, which was discussed in the proceedings of a subsequent conference, published in 1991. The work of these committees was extremely successful and influential in laying the foundation for later activities in alternatives research. A Third FRAME Toxicity Committee was formed in 1999, since much progress had been achieved in the previous decade, especially with regard to the successful validation of several non-animal replacement methods and the start of their regulatory acceptance. Moreover, some new test methods are on the point of being validated, and many new techniques and discoveries are impacting on toxicity testing. Also, interest in reduction and refinement in toxicology has increased. However, there is considerable scope and need for the further implementation of the Three Rs in toxicity testing, especially due to recent plans for the large-scale testing of high-production volume, hormonally-active and existing chemicals, and the increasing use of transgenic animal models. The new committee comprises 18 experts from industry, academia, animal welfare, legislative and regulatory bodies, with one observer from the UK Government Home Office. The main objective is to review progress made in the application of the Three Rs in the development and safety evaluation of medicines, biologicals, cosmetics, agrochemicals and other products, as well as industrial chemicals, and to make recommendations as a basis for further sensible progress according to sound scientific and ethical criteria. The main committee is to be augmented by several working parties that will focus on specific scientific issues: 1) targeted risk assessment versus hazard identification; 2) data sharing; 3) endocrine disruption; and 4) carcinogenicity testing. The Committee is also to publish a status report on the current situation regarding alternatives in toxicity testing, based on the recommendations of the Second Toxicity Committee, and will organise a conference to discuss its overall conclusions and recommendations. PMID:23581152

  8. Comparisons of Sediment Test Volumes for Freshwater Solid Phase Sediment Toxicity Tests

    EPA Science Inventory

    Laboratory tests with benthic macroinvertebrates are commonly used to assess the potential toxicity of contaminated sediments, and detailed standard test procedures have been developed for various species. For freshwater, two benthic organisms, Hyalella azteca and Chironomus dil...

  9. Developmental toxicity, EROD, and CYP1A mRNA expression in zebrafish embryos exposed to dioxin-like PCB126.

    PubMed

    Liu, Han; Nie, Fang-Hong; Lin, Hong-Ying; Ma, Yi; Ju, Xiang-Hong; Chen, Jin-Jun; Gooneratne, Ravi

    2016-02-01

    Dioxin-like PCB126 is a persistent organic pollutant that causes a range of syndromes including developmental toxicity. Dioxins have a high affinity for aryl hydrocarbon receptor (AhR) and induce cytochrome P4501A (CYP1A). However, the role of CYP1A activity in developmental toxicity is less clear. To better understand dioxin induced developmental toxicity, we exposed zebrafish (Danio rerio) embryos to PCB126 at concentrations of 0, 16, 32, 64, and 128 μg L(-1) from 3-h post-fertilization (hpf) to 168 hpf. The embryonic survival rate decreased at 144 and 168 hpf. The fry at 96 hpf displayed gross developmental malformations, including pericardial and yolk sac edema, spinal curvature, abnormal lower jaw growth, and non-inflated swim bladder. The pericardial and yolk sac edema rate significantly increased and the heart rate declined from 96 hpf compared with the controls. PCB126 did not alter the hatching rate. To elucidate the mechanism of PCB126-induced developmental toxicity, we conducted ethoxyresorufin-O-deethylase (EROD) in vivo assay to determine CYP1A enzyme activity, and real-time PCR to study the induction of CYP1A mRNA gene expression in embryo/larval zebrafish at 24, 72, 96, and 132 hpf. In vivo EROD activity was induced by PCB126 at 16 μg L(-1) concentration as early as 72 hpf but significant increases were observed only in zebrafish exposed to 64 and 128 μg L(-1) doses (p < 0.005) at 72, 96, and 132 hpf. Induction of CYP1A mRNA expression was significantly upregulated in zebrafish exposed to 32 and 64 μg L(-1) at 24, 72, 96, and 132 hpf. Overall, the severe pericardial and yolk sac edema and reduced heart rate suggest that heart defects are a sensitive endpoint, and the general trend of dose-dependent increase in EROD activity and induction of CYP1A mRNA gene expression provide evidence that the developmental toxicity of PCB126 to zebrafish embryos is mediated by activation of AhR. © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 201-210, 2016. PMID:25099626

  10. Miniaturized Embryo Array for Automated Trapping, Immobilization and Microperfusion of Zebrafish Embryos

    PubMed Central

    Akagi, Jin; Khoshmanesh, Khashayar; Evans, Barbara; Hall, Chris J.; Crosier, Kathryn E.; Cooper, Jonathan M.; Crosier, Philip S.; Wlodkowic, Donald

    2012-01-01

    Zebrafish (Danio rerio) has recently emerged as a powerful experimental model in drug discovery and environmental toxicology. Drug discovery screens performed on zebrafish embryos mirror with a high level of accuracy the tests usually performed on mammalian animal models, and fish embryo toxicity assay (FET) is one of the most promising alternative approaches to acute ecotoxicity testing with adult fish. Notwithstanding this, automated in-situ analysis of zebrafish embryos is still deeply in its infancy. This is mostly due to the inherent limitations of conventional techniques and the fact that metazoan organisms are not easily susceptible to laboratory automation. In this work, we describe the development of an innovative miniaturized chip-based device for the in-situ analysis of zebrafish embryos. We present evidence that automatic, hydrodynamic positioning, trapping and long-term immobilization of single embryos inside the microfluidic chips can be combined with time-lapse imaging to provide real-time developmental analysis. Our platform, fabricated using biocompatible polymer molding technology, enables rapid trapping of embryos in low shear stress zones, uniform drug microperfusion and high-resolution imaging without the need of manual embryo handling at various developmental stages. The device provides a highly controllable fluidic microenvironment and post-analysis eleuthero-embryo stage recovery. Throughout the incubation, the position of individual embryos is registered. Importantly, we also for first time show that microfluidic embryo array technology can be effectively used for the analysis of anti-angiogenic compounds using transgenic zebrafish line (fli1a:EGFP). The work provides a new rationale for rapid and automated manipulation and analysis of developing zebrafish embryos at a large scale. PMID:22606275

  11. A FIELD VALIDATION OF TWO SEDIMENT-AMPHIPOD TOXICITY TESTS

    EPA Science Inventory

    A field validation study of two sediment-amphipod toxicity tests was conducted using sediment samples collected subtidally in the vicinity of a polycyclic aromatic hydrocarbon (PAH)-contaminated Superfund site in Elliott Bay, WA, USA. Sediment samples were collected at 30 stati...

  12. THE FUTURE OF TOXICITY TESTING AND ASSESSMENT OF ENVIRONMENTAL AGENTS

    EPA Science Inventory

    The National Research Council's Board on Environmental Studies and Toxicology is conducting a two-part study to assess and advance current approaches to toxicity testing and assessment to meet regulatory data needs. The first part of the study was completed January 2006, in which...

  13. TOXICITY TEST USING LIFE STAGES OF 'CHAMPIA PARVULA' (RHODOPHYTA)

    EPA Science Inventory

    A static-renewal, 11- to 14-day toxicity test has been developed using the life cycle of the marine red alga Champia parvula (C. Ag.) Harv. It measures the vegetative growth, formation of tetrasporangia (meiosis), and formation of cystocarps (sexual fusion). The procedure has bee...

  14. Assessment of toxicity test endpoints for freshwater mussel larvae (glochidia).

    PubMed

    Fritts, Andrea K; Barnhart, M Christopher; Bradley, Megan; Liu, Na; Cope, W Gregory; Hammer, Edward; Bringolf, Robert B

    2014-01-01

    The objectives of the present study were to determine if the viability of freshwater mussel larvae (glochidia) is an ecologically relevant endpoint for toxicity tests and to define the appropriate duration of those tests. The authors assessed 1) how viability (the shell closure response to sodium chloride) compares with infectivity (ability to attach to a host fish and successfully metamorphose to the juvenile stage), and 2) the decline of viability and infectivity over time after glochidia were released from female mussels. Glochidia of 7 mussel species were isolated from females, placed in water, and subsampled daily for 2 d to 5 d. Viability, when ≥90%, was generally a good predictor of infectivity; however, when viability was <90%, infectivity was often disproportionately low, especially for glochidia collected near the end of the brooding period. Viability and infectivity declined more rapidly in natural water and sediment compared to reconstituted water. Following 24-h exposure to a toxicant (sodium chloride or copper), infectivity of the viable glochidia did not differ among concentrations of toxicants. The results indicate that viability is a valid proxy for infectivity and an ecologically relevant endpoint for standard toxicity tests with freshwater mussels for any test duration with control viability >90%. PMID:24122868

  15. Controlling type-1 error rates in whole effluent toxicity testing

    SciTech Connect

    Smith, R.; Johnson, S.C.

    1995-12-31

    A form of variability, called the dose x test interaction, has been found to affect the variability of the mean differences from control in the statistical tests used to evaluate Whole Effluent Toxicity Tests for compliance purposes. Since the dose x test interaction is not included in these statistical tests, the assumed type-1 and type-2 error rates can be incorrect. The accepted type-1 error rate for these tests is 5%. Analysis of over 100 Ceriodaphnia, fathead minnow and sea urchin fertilization tests showed that when the test x dose interaction term was not included in the calculations the type-1 error rate was inflated to as high as 20%. In a compliance setting, this problem may lead to incorrect regulatory decisions. Statistical tests are proposed that properly incorporate the dose x test interaction variance.

  16. Toxicogenomic responses of zebrafish embryos/larvae to tris(1,3-dichloro-2-propyl) phosphate (TDCPP) reveal possible molecular mechanisms of developmental toxicity.

    PubMed

    Fu, Jie; Han, Jian; Zhou, Bingsheng; Gong, Zhiyuan; Santos, Eduarda M; Huo, Xiaojing; Zheng, Weiling; Liu, Hongling; Yu, Hongxia; Liu, Chunsheng

    2013-09-17

    Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) is frequently present in indoor dust and can be detected in human milk. In order to evaluate the effects of TDCPP on vertebrate development, zebrafish embryos/larvae were used as an animal model to examine developmental phenotypes and explore possible mechanisms of toxicity by employing microarrays and iTRAQ labeling quantitative proteomics. The results demonstrated that treatment with TDCPP (3 μM) from 0.75 h postfertilization (hpf) inhibited cell rearrangement at 4 hpf, caused delay in epiboly at 5.7 and 8.5 hpf, and led to abnormal development (e.g., short tail, reduced body size) and lethality between 14 and 45 hpf, which might be related with altered expression of genes regulating embryogenesis. Furthermore, trunk curvature was observed as the main phenotype in 96 hpf zebrafish larvae exposed to 1 or 3 μM TDCPP, possibly by changing somite formation and expression of proteins related to fast muscle and cartilage development. Collectively, our results suggest that exposure to TDCPP causes developmental toxicity in vertebrates and warrant the need for studies to evaluate the potential health risks of TDCPP to developing human embryos/infants/children, due to its frequent presence in indoor dust and potential for human exposure. PMID:23919627

  17. Genotoxic and teratogenic effect of freshwater sediment samples from the Rhine and Elbe River (Germany) in zebrafish embryo using a multi-endpoint testing strategy.

    PubMed

    Garcia-Käufer, M; Gartiser, S; Hafner, C; Schiwy, S; Keiter, S; Gründemann, C; Hollert, H

    2015-11-01

    The embryotoxic potential of three model sediment samples with a distinct and well-characterized pollutant burden from the main German river basins Rhine and Elbe was investigated. The Fish Embryo Contact Test (FECT) in zebrafish (Danio rerio) was applied and submitted to further development to allow for a comprehensive risk assessment of such complex environmental samples. As particulate pollutants are constructive constituents of sediments, they underlay episodic source-sink dynamics, becoming available to benthic organisms. As bioavailability of xenobiotics is a crucial factor for ecotoxicological hazard, we focused on the direct particle-exposure pathway, evaluating throughput-capable endpoints and considering toxicokinetics. Fish embryo and larvae were exposed toward reconstituted (freeze-dried) sediment samples on a microcosm-scale experimental approach. A range of different developmental embryonic stages were considered to gain knowledge of potential correlations with metabolic competence during the early embryogenesis. Morphological, physiological, and molecular endpoints were investigated to elucidate induced adverse effects, placing particular emphasis on genomic instability, assessed by the in vivo comet assay. Flow cytometry was used to investigate the extent of induced cell death, since cytotoxicity can lead to confounding effects. The implementation of relative toxicity indices further provides inter-comparability between samples and related studies. All of the investigated sediments represent a significant ecotoxicological hazard by disrupting embryogenesis in zebrafish. Beside the induction of acute toxicity, morphological and physiological embryotoxic effects could be identified in a concentration-response manner. Increased DNA strand break frequency was detected after sediment contact in characteristic non-monotonic dose-response behavior due to overlapping cytotoxic effects. The embryonic zebrafish toxicity model along with the in vivo comet assay and molecular biomarker analysis should prospectively be considered to assess the ecotoxicological potential of sediments allowing for a comprehensive hazard ranking. In order to elucidate mode of action, novel techniques such as flow cytometry have been adopted and proved to be valuable tools for advanced risk assessment and management. PMID:25471716

  18. Indicators of Ceriodaphnia dubia chronic toxicity test performance and sensitivity

    SciTech Connect

    Rosebrock, M.M.; Bedwell, N.J.; Ausley, L.W.

    1994-12-31

    The North Carolina Division of Environmental Management has begun evaluation of the sensitivity of test results used for measuring chronic whole effluent toxicity in North Carolina wastewater discharges. Approximately 67% of 565 facilities required to monitor toxicity by an NPDES permit perform a Ceriodaphnia dubia chronic, single effluent concentration (pass/fail) analysis. Data from valid Ceriodaphnia dubia chronic pass/fail tests performed by approximately 20 certified biological laboratories and submitted by North Carolina NPDES permittees were recorded beginning January 1992. Control and treatment reproduction data from over 2,500 tests submitted since 1992 were analyzed to determine the minimum significant difference (MSD) at a 99% confidence level for each test and the percent reduction from the control mean that the MSD represents (%MSD) for each certified laboratory. Initial results for the 20 laboratories indicate that the average intralaboratory percent MSD ranges 12.72% (n = 367) to 34.91% (n = 7) with an average of 23.08%. Additionally, over 3,800 tests were analyzed to determine the coefficient of variation (CV) for control reproduction for each test and the average for each certified biological laboratory. Preliminary review indicates that average interlaboratory control reproduction CV values range from 10.59% (n = 367) to 31.08% (n = 572) with a mean of 20.35%. The statistics investigated are indicators of intra/interlaboratory performance and sensitivity of Ceriodaphnia chronic toxicity analyses.

  19. EP-toxicity testing of mercury removal resin grout

    SciTech Connect

    Mersman, K.E.

    1984-07-18

    To determine which category a waste will fit into, the EPA requires a classification test. The test, EP-toxicity, consists of a physical integrity test followed by an extraction. For the case of the mercury removal resin grout, the mercury concentration in the extract cannot exceed 0.2 mg/L if the waste is to be classified as ``solid waste.`` Otherwise, the waste is classified as ``hazardous.`` Simulated process solutions were used to load the mercury removal resin. The resin was solidified with the addition of cement and water using a formulation based on grout formulations typically used to solidify power reactor ion exchange resins. Envirodyne Engineers of St. Louis, Missouri, an EPA sanctioned laboratory, performed the EP-toxicity test for the two samples. One sample was a blank which was made with unloaded resin. For the formulation tested, the EP-toxicity test results showed that the mercury removal resin grout does not fit into the ``hazardous waste`` category.

  20. A Simple "in Vitro" Culture of Freshwater Prawn Embryos for Laboratory Investigations

    ERIC Educational Resources Information Center

    Porntrai, Supaporn; Damrongphol, Praneet

    2008-01-01

    Giant freshwater prawn ("Macrobrachium rosenbergii" De Man) embryos can be cultured "in vitro" to hatching in 15% (v/v) artificial seawater (ASW). This technique can be applied as a bioassay for testing toxicity or for the effects of various substances on embryo development and can be used as a simple and low-cost model for studying embryo…

  1. Sperm extracted and cryopreserved from testes several hours after death results in pregnancy following frozen embryo transfer: case report.

    PubMed

    Check, M; Summers-Chase, D; Check, J H; Choe, J; Nazari, A

    1999-01-01

    A 38-year-old male died suddenly on his honeymoon. Sperm was extracted from his testes 3 h following his death and cryopreserved. His wife had in vitro fertilization (IVF) and the eggs were inseminated by intracytoplasmic sperm injection (ICSI). None of the sperm were motile. Selection was based on softness and pliability. There were 4 embryos formed that cleaved, but only 2 were transferred on the retrieval cycle. The wife failed to conceive, but then had a second transfer of the 2 cryopreserved embryos. She achieved a chemical pregnancy with the beta-human chorionic gonadotropin level attaining a maximum level of 107 mIU/mL (rising from 19 mIU/mL). Though this retrieval cycle did not result in a successful pregnancy the achievement of a clinical pregnancy following frozen embryo transfer at least provides cautious optimism for other cases with similar conditions. PMID:10624508

  2. An evaluation of the whole effluent toxicity test method

    SciTech Connect

    Osteen, D.V.

    1999-12-17

    Whole effluent toxicity (WET) testing has become increasingly more important to the Environmental Protection Agency (EPA) and the States in the permitting of wastewater discharges from industry and municipalities. The primary purpose of the WET test is to protect aquatic life by predicting the effect of an effluent on the receiving stream. However, there are both scientific and regulatory concerns that using WET tests to regulate industrial effluents may result in either false positives and/or false negatives. In order to realistically predict the effect of an effluent on the receiving stream, the test should be as representative as possible of the conditions in the receiving stream. Studies (Rand and Petrocelli 1985) suggested several criteria for an ideal aquatic toxicity test organism, one of which is that the organism be indigenous to, or representative of, the ecosystem receiving the effluent. The other component needed in the development of a predictive test is the use of the receiving stream water or similar synthetic water as the control and dilution water in the test method. Use of an indigenous species and receiving water in the test should help reduce the variability in the method and allow the test to predict the effect of the effluent on the receiving stream. The experience with toxicity testing at the Savannah River Site (SRS) has yielded inconclusive data because of the inconsistency and unreliability of the results. The SRS contention is that the WET method in its present form does not adequately mimic actual biological/chemical conditions of the receiving streams and is neither reasonable nor accurate. This paper discusses the rationale for such a position by SRS on toxicity testing in terms of historical permitting requirements, outfall effluent test results, standard test method evaluation, scientific review of alternate test species, and concerns over the test method expressed by other organizations. This paper presents the Savannah River Site position that the EPA test is neither reasonable nor accurate and thus cannot adequately establish the impact of NPDES outfall discharges on receiving streams.

  3. Use of neomysis mercedis (crustacea: mysidacea) for estuarine toxicity tests

    SciTech Connect

    Brandt, O.M.; Fujimura, R.W.; Finlayson, B.J. )

    1993-03-01

    The mysid Neomysis mercedis was examined as a test organism for use in acute toxicity tests at intermediate salinities characteristic of estuarine waters. Several sensitive invertebrate species are available for marine assessments (mysids) and freshwater tests (cladocerans), but few are available for estuarine toxicity tests. Observations in the laboratory indicate that Neomysis mercedis can be reared successfully at a temperature of 17[degrees]C, a salinity of 2%, and a population density less than 5/L. Brine shrimp nauplii Artemia salina, algae, and commercial foods were used to sustain mysid cultures. Neomysis mercedis is vivaparous and can complete its life cycle in 3-4 months. Neomysis mercedis is as sensitive as or more sensitive to toxicants than the marine mysid Mysidopsis bahia and the freshwater cladocerans Daphnia magna, Ceriodaphnia dubia, and Simocephalus serrulatus. The mean 96-h LC50 values (concentrations lethal to half the test animals) for N. mercedis, in increasing order, were 0.20 [mu]g/L for thiobencarb, and for malathion, 14 [mu]g/L for carbofuran, 150 [mu]g/L for copper sulfate, 280 [mu]g/L for thiobencarb, and 1,600 [mu]g/L for molinate. Neonates (5 d postrelease) were generally more sensitive than older juveniles. Coefficients of variation (100[center dot]SD/mean) of LC50 values varied from 21 to 35%. 37 refs., 2 figs., 7 tabs.

  4. Effect of test conditions on relative toxicity rankings of fifteen materials

    NASA Technical Reports Server (NTRS)

    Hilado, C. J.; Cumming, H. J.

    1977-01-01

    Fifteen materials were evaluated for relative toxicity of pyrolysis effluents, using different test conditions in the USF methodology. Wool fabrics were consistently among the most toxic materials, and polystyrene and polychloroprene flexible foam were consistently among the least toxic materials.

  5. The Role of Toxicity Testing in NASA's Future Missions

    NASA Technical Reports Server (NTRS)

    Whitfield, Steve; Davis, Samuel; Wise, Harry; Moore, Robin

    2005-01-01

    NASA has deemed it necessary to perform the Toxicity Test (offgassing of toxic products) on all non-metallic materials proposed for use in habitable environments onboard the Shuttle and International Space Station flS,!J. This requirement stems from the desire to maintain a healthy, breathable atmosphere for the astronauts. As Shuttle missions have lengthened and with the habitation of the International Space Station, the need for understanding and controlling the contaminants in breathable atmospheres has increased. The increased duration of humans in space present two concerns to the astronauts with regard to their breathing air: 1. Breathing the on-board air. 2. Improved cleaning/filtering of existing air. Trends using existing toxicity data for materials and an understanding of the air cleaning/filtering capabilities in relation to Spacecraft Maximum Allowable Concentrations (SMAC) of offgassed components are explored. Recommendations are made for materials selection practices that should be followed to ensure a safe and healthy breathing environment for astronauts aboard these long term projects. The importance and relevance of Toxicity testing and materials selection in conjunction with the new NASA missions of creating a human presence on the Moon and traveling to Mars are described.

  6. Toxicity testing and instream biological monitoring in evaluating municipal effluents

    SciTech Connect

    Krier, K.; Pontasch, K.

    1995-12-31

    Twelve streams receiving municipal wastewater treatment plant effluents were evaluated in riffle areas above and below the outfall using the Environmental Protection Agency`s Rapid Bioassessment Protocols (RBPs) for benthic macroinvertebrates. Eight of the sites evaluated using RBP 1 exhibited stream health in the downstream riffles equaling or exceeding the upstream riffles. RBP 1 results suggested possible impacts at the remaining four sites, and these sites were more intensely evaluated using RBPs 2 and 3, acute effluent toxicity tests with Daphnia magna, and quantification of periphytic chlorophyll a and ash free dry weight (AFDW). Results from RBP 2 indicated three of the four sites evaluated have similar taxonomic richness above and below the outfall, while one site is heavily impacted by organic pollutants. Toxicity tests with 100% effluent resulted in no mortality with any of the four effluents tested. Relative to the respective upstream sites, chlorophyll a was significantly increased at one downstream site and significantly reduced at another. AFDW was similar above and below the outfalls in all streams. These results suggest that laboratory toxicity tests may not always be adequate predictors of instream biological effects.

  7. Hyalella IQ Toxicity Test{trademark} as a predictor of whole sediment toxicity with diversely contaminated sediments

    SciTech Connect

    Douglas, W.S.; Hayes, K.R.

    1994-12-31

    The IQ TOXICITY TEST{trademark} is a toxicity screening test that evaluates the organism`s galactosidase enzyme system functionality as a predictor of acute toxicity. Organisms are exposed to a potentially toxic solution for approximately one hour. Following the exposure, the organisms are exposed to a slurry of a galactoside sugar tagged with a fluorescent marker (methylumbelliferyl galactoside) for 15--20 minutes. A black light can then be used to examine whether the hemolymph of the organism contains free umbelliferone, which brightly fluoresces. The organisms are then scored as ``on`` or ``off`` with respect to free umbelliferone. This endpoint can then be used to calculate an EC50, which is comparable to a whole effluent, pure compound, or sediment toxicity test. Slightly different methodologies are used for different toxicity test organisms. The objective of this presentation is to discuss the use of the IQ{trademark} methodology with porewater extract exposures of the amphipod Hyalella azteca as a predictor of results of whole sediment toxicity tests. The results of over thirty 10 and 28-day whole sediment toxicity tests and the concurrent Hyalella azteca 10 TOXICITY TESTS{trademark} are compared and discussed. The use of screening tests as a reduced cost method for initial site assessment will be discussed.

  8. Non-animal Replacements for Acute Toxicity Testing.

    PubMed

    Barker-Treasure, Carol; Coll, Kevin; Belot, Nathalie; Longmore, Chris; Bygrave, Karl; Avey, Suzanne; Clothier, Richard

    2015-07-01

    Current approaches to predicting adverse effects in humans from acute toxic exposure to cosmetic ingredients still heavily necessitate the use of animals under EU legislation, particularly in the context of the REACH system, when cosmetic ingredients are also destined for use in other industries. These include the LD50 test, the Up-and-Down Procedure and the Fixed Dose Procedure, which are regarded as having notable scientific deficiencies and low transferability to humans. By expanding on previous in vitro tests, such as the animal cell-based 3T3 Neutral Red Uptake (NRU) assay, this project aims to develop a truly animal-free predictive test for the acute toxicity of cosmetic ingredients in humans, by using human-derived cells and a prediction model that does not rely on animal data. The project, funded by Innovate UK, will incorporate the NRU assay with human dermal fibroblasts in animal product-free culture, to generate an in vitro protocol that can be validated as an accepted replacement for the currently available in vivo tests. To date, the project has successfully completed an assessment of the robustness and reproducibility of the method, by using sodium lauryl sulphate (SLS) as a positive control, and displaying analogous results to those of the original studies with mouse 3T3 cells. Currently, the testing of five known ingredients from key groups (a surfactant, a preservative, a fragrance, a colour and an emulsifier) is under way. The testing consists of initial range-finding runs followed by three valid runs of a main experiment with the appropriate concentration ranges, to generate IC50 values. Expanded blind trials of 20 ingredients will follow. Early results indicate that this human cell-based test holds the potential to replace aspects of in vivo animal acute toxicity testing, particularly with reference to cosmetic ingredients. PMID:26256397

  9. Fish embryo tests with Danio rerio as a tool to evaluate surface water and sediment quality in rivers influenced by wastewater treatment plants using different treatment technologies.

    PubMed

    Thellmann, Paul; Köhler, Heinz-R; Rößler, Annette; Scheurer, Marco; Schwarz, Simon; Vogel, Hans-Joachim; Triebskorn, Rita

    2015-11-01

    In order to evaluate surface water and the sediment quality of rivers connected to wastewater treatment plants (WWTPs) with different treatment technologies, fish embryo tests (FET) with Danio rerio were conducted using native water and sediment samples collected upstream and downstream of four WWTPs in Southern Germany. Two of these WWTPs are connected to the Schussen River, a tributary of Lake Constance, and use a sand filter with final water purification by flocculation. The two others are located on the rivers Schmiecha and Eyach in the area of the Swabian Alb and were equipped with a powdered activated carbon stage 20 years ago, which was originally aimed at reducing the release of stains from the textile industry. Several endpoints of embryo toxicity including mortality, malformations, reduced hatching rate, and heart rate were investigated at defined time points of embryonic development. Higher embryotoxic potentials were found in water and sediments collected downstream of the WWTPs equipped with sand filtration than in the sample obtained downstream of both WWTPs upgraded with a powdered activated carbon stage. PMID:25391229

  10. A category approach to predicting the developmental (neuro) toxicity of organotin compounds: the value of the zebrafish (Danio rerio) embryotoxicity test (ZET).

    PubMed

    Beker van Woudenberg, Anna; Wolterbeek, André; Te Brake, Lindsey; Snel, Cor; Menke, Aswin; Rubingh, Carina; de Groot, Didima; Kroese, Dinant

    2013-11-01

    Zebrafish embryos were exposed to different organotin compounds during very early development (<100h post fertilization). Morphology, histopathology and swimming activity (in a motor activity test) were the endpoints analyzed. DBTC was, by far, the most embryotoxic compound at all time points and endpoints studied. In fact, we observed a clear concordance between the effects observed in our zebrafish embryo model, and those observed with these compounds in full rodent in vivo studies. All organotin compounds classified as developmental (neuro) toxicants in vivo, were correctly classified in the present assay. Together, our results support the ZET model as a valuable tool for providing biological verification for a grouping and a read-across approach to developmental (neuro) toxicity. PMID:23796951

  11. Using single-species toxicity tests, community-level responses, and toxicity identification evaluations to investigate effluent impacts

    SciTech Connect

    Maltby, L.; Clayton, S.A.; Yu, H.; McLoughlin, N.; Wood, R.M.; Yin, D.

    2000-01-01

    Whole effluent toxicity (WET) tests are increasingly used to monitor compliance of consented discharges, but few studies have related toxicity measured using WET tests to receiving water impacts. Here the authors adopt a four-stage procedure to investigate the toxicity and biological impact of a point source discharge and to identify the major toxicants. In stage 1, standard WET tests were employed to determine the toxicity of the effluent. This was then followed by an assessment of receiving water toxicity using in situ deployment of indigenous (Gammarus pulex) and standard (Daphnia magna) test species. The third stage involved the use of biological survey techniques to assess the impact of the discharge on the structure and functioning of the benthic macroinvertebrate community. In stage 4, toxicity identification evaluations (TIE) were used to identify toxic components in the effluent. Receiving-water toxicity and ecological impact detected downstream of the discharge were consistent with the results of WET tests performed on the effluent. Downstream of the discharge, there was a reduction in D. magna survival, in G. pulex survival and feeding rate, in detritus processing, and in biotic indices based on macroinvertebrate community structure. The TIE studies suggested that chlorine was the principal toxicant in the effluent.

  12. Applicability of subchronic toxicity test with Hyalella azteca for toxicity identification evaluation programs

    SciTech Connect

    Putt, A.E.; Jop, K.M.

    1995-12-31

    A series of screening tests including the short-term chronic exposure of Ceriodaphnia dubia to sediment pore waters, 10-day exposures of Chironomus tentans and Hyalella azteca to bulk sediments and a bioaccumulation study with Lumbriculus variegatus were performed as part of an ecological risk assessment of Plow Shop Pond, Fort Devens, Massachusetts. Chronic endpoints such as reproduction and growth indicated sediment toxicity, however, a toxicity identification evaluation program was initiated to further define the source and extent of the toxicity. A short-term chronic exposure with C. dubia was a logical choice for the TIE, however, since amphipods are epibenthic organisms, they are a better surrogate of sediment dwelling organisms than a water column species such as C. dubia. Observations performed during H. azteca culture suggested that this species of amphipod could thrive in the water column for up to three weeks. Therefore, 7-day old H. azteca were exposed to pore water samples under static-renewal conditions for 10 days. Survival and growth (i.e., dry weight) were determined at the termination of each exposure. Laboratory control group performance consistently averaged a {>=}90% survival and {>=}43 {micro}g of dry weight per amphipod. Growth of amphipods used in each exposure generally exceeded two times the initial body weight after 10 days of exposure. Previous studies have indicated that the growth and reproductive response of H. azteca are positively correlated for a given set of exposure conditions. The results of these 10-day subchronic exposures with H. azteca provide a consistent and reliable measure of the chronic sediment toxicity with a benthic invertebrate for toxicity identification evaluation programs.

  13. Study of Charge-Dependent Transport and Toxicity of Peptide-Functionalized Silver Nanoparticles Using Zebrafish Embryos and Single Nanoparticle Plasmonic Spectroscopy

    PubMed Central

    Lee, Kerry J.; Browning, Lauren M.; Nallathamby, Prakash D.; Xu, Xiao-Hong Nancy

    2013-01-01

    Nanomaterials possess unusually high surface area-to-volume ratios, and surface-determined physicochemical properties. It is essential to understand their surface-dependent toxicity in order to rationally design biocompatible nanomaterials for a wide variety of applications. In this study, we have functionalized the surfaces of silver nanoparticles (Ag NPs, 11.7 ± 2.7 nm in diameters) with three biocompatible peptides (CALNNK, CALNNS, CALNNE) to prepare positively (Ag-CALNNK NPs+ζ), negatively (Ag-CALNNS NPs−2ζ), and more negatively charged NPs (Ag-CALNNE NPs−4ζ), respectively. Each peptide differs in a single amino acid at its C-terminus, which minimizes the effects of peptide sequences and serves as a model molecule to create positive, neutral and negative charges on the surface of the NPs at pH 4–10. We have studied their charge-dependent transport into early-developing (cleavage-stage) zebrafish embryos and their effects on embryonic development using dark-field optical microscopy and spectroscopy (DFOMS). We found that all three Ag-peptide NPs passively diffused into the embryos via their chorionic pore canals, and stayed inside the embryos throughout their entire development (120 h), showing charge-independent diffusion modes and charge-dependent diffusion coefficients. Notably, the NPs create charge-dependent toxic effects on embryonic development, showing that the Ag-CALNNK NPs+ζ (positively charged) are the most biocompatible while the Ag-CALNNE NPs–4ζ (more negatively charged) are the most toxic. By comparing with our previous studies of the same sized citrated Ag and Au NPs, the Ag-peptide NPs are much more biocompatible than the citrated Ag NPs, and nearly as biocompatible as the Au NPs, showing the dependence of nanotoxicity upon the surface charges, surface functional groups and chemical compositions of the NPs. This study also demonstrates powerful applications of single NP plasmonic spectroscopy for quantitative analysis of single NPs in vivo and in tissues, and reveals the possibility of rational design of biocompatible NPs. PMID:23621491

  14. The REACH perspective: toward a new concept of toxicity testing.

    PubMed

    Schoeters, Greet

    2010-02-01

    A sustainable society and a healthy society are major goals for European policymakers. Although most Europeans live a longer healthy life than ever, there is growing concern and anxiety about unknown health risks and threats of chemicals and a strong demand for more knowledge and more control. European legislation is responding to these demands. An example is the program on Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), which came into force in 2007. It is a gigantic task for industry and for administrators to evaluate safety files of thousands of chemicals in a period of 12 years and to collect new data for chemicals not yet evaluated. Costs, number of toxicity tests, and number of animals that are needed are already well documented. REACH uses strict guidelines and focuses on apical endpoints that have been covered in the past by animal tests. Animal tests are slow, use unrealistic high doses, and have been shown to not always predict human toxicity correctly. The REACH program has made a clear opening for reduction of in vivo animal tests. Sharing toxicity data is a major improvement. For low tonnage levels, no further in vivo testing is allowed. The combination of scientifically valid information from alternative tests with available animal and human data into a weight-of-evidence approach is part of the integrated test strategy under REACH. Interpretation of this integrated information requires a high degree of expertise, flexibility, and openness toward scientific advances. This will be crucial for the success of the REACH program. It means a shift of attitude and will put a heavy responsibility on scientific experts and regulators, but it is also an opportunity for meeting the safety expectations of our modern society. PMID:20574899

  15. Produced water toxicity tests accurately measure the produced water toxicity in marine environments?

    SciTech Connect

    Douglas, W.S.; Veil, J.A.

    1996-10-01

    U.S. Environmental Protection Agency (EPA) Region VI has issued a general permit for offshore oil and gas discharges to the Gulf of Mexico that places numerical limits on whole effluent toxicity (WEI) for produced water. Recently proposed EPA general permits for other produced water discharges in Regions VI and X also include enforceable numerical limits on WET. Clearly, the industry will be conducting extensive produced water WET testing. Unfortunately, the WET test may not accurately measure the toxicity of the chemical constituents of produced water. Rather the mortality of test organisms may be attributable to (1) the high salinity of produced water, which causes salinity shock to the organisms, or (2) an ionic imbalance caused by excesses or deficiencies of one or more of seawater`s essential ions in the test chambers. Both of these effects are likely to be mitigated in actual offshore discharge settings, where the receiving water will be seawater and substantial dilution will be probable. Thus, the additional salinity of produced water will be rapidly assimilated, and the proper marine ionic balance will be quickly restored. Regulatory authorities should be aware of these factors when interpreting WET test results.

  16. Undetected Toxicity Risk in Pharmacogenetic Testing for Dihydropyrimidine Dehydrogenase

    PubMed Central

    Falvella, Felicia Stefania; Caporale, Marta; Cheli, Stefania; Martinetti, Antonia; Berenato, Rosa; Maggi, Claudia; Niger, Monica; Ricchini, Francesca; Bossi, Ilaria; Di Bartolomeo, Maria; Sottotetti, Elisa; Bernardi, Francesca Futura; de Braud, Filippo; Clementi, Emilio; Pietrantonio, Filippo

    2015-01-01

    Fluoropyrimidines, the mainstay agents for the treatment of colorectal cancer, alone or as a part of combination therapies, cause severe adverse reactions in about 10%–30% of patients. Dihydropyrimidine dehydrogenase (DPD), a key enzyme in the catabolism of 5-fluorouracil, has been intensively investigated in relation to fluoropyrimidine toxicity, and several DPD gene (DPYD) polymorphisms are associated with decreased enzyme activity and increased risk of fluoropyrimidine-related toxicity. In patients carrying non-functional DPYD variants (c.1905+1G>A, c.1679T>G, c.2846A>T), fluoropyrimidines should be avoided or reduced according to the patients’ homozygous or heterozygous status, respectively. For other common DPYD variants (c.496A>G, c.1129-5923C>G, c.1896T>C), conflicting data are reported and their use in clinical practice still needs to be validated. The high frequency of DPYD polymorphism and the lack of large prospective trials may explain differences in studies’ results. The epigenetic regulation of DPD expression has been recently investigated to explain the variable activity of the enzyme. DPYD promoter methylation and its regulation by microRNAs may affect the toxicity risk of fluoropyrimidines. The studies we reviewed indicate that pharmacogenetic testing is promising to direct personalised dosing of fluoropyrimidines, although further investigations are needed to establish the role of DPD in severe toxicity in patients treated for colorectal cancer. PMID:25906475

  17. Undetected toxicity risk in pharmacogenetic testing for dihydropyrimidine dehydrogenase.

    PubMed

    Falvella, Felicia Stefania; Caporale, Marta; Cheli, Stefania; Martinetti, Antonia; Berenato, Rosa; Maggi, Claudia; Niger, Monica; Ricchini, Francesca; Bossi, Ilaria; Di Bartolomeo, Maria; Sottotetti, Elisa; Bernardi, Francesca Futura; de Braud, Filippo; Clementi, Emilio; Pietrantonio, Filippo

    2015-01-01

    Fluoropyrimidines, the mainstay agents for the treatment of colorectal cancer, alone or as a part of combination therapies, cause severe adverse reactions in about 10%-30% of patients. Dihydropyrimidine dehydrogenase (DPD), a key enzyme in the catabolism of 5-fluorouracil, has been intensively investigated in relation to fluoropyrimidine toxicity, and several DPD gene (DPYD) polymorphisms are associated with decreased enzyme activity and increased risk of fluoropyrimidine-related toxicity. In patients carrying non-functional DPYD variants (c.1905+1G>A, c.1679T>G, c.2846A>T), fluoropyrimidines should be avoided or reduced according to the patients' homozygous or heterozygous status, respectively. For other common DPYD variants (c.496A>G, c.1129-5923C>G, c.1896T>C), conflicting data are reported and their use in clinical practice still needs to be validated. The high frequency of DPYD polymorphism and the lack of large prospective trials may explain differences in studies' results. The epigenetic regulation of DPD expression has been recently investigated to explain the variable activity of the enzyme. DPYD promoter methylation and its regulation by microRNAs may affect the toxicity risk of fluoropyrimidines. The studies we reviewed indicate that pharmacogenetic testing is promising to direct personalised dosing of fluoropyrimidines, although further investigations are needed to establish the role of DPD in severe toxicity in patients treated for colorectal cancer. PMID:25906475

  18. Studying the effect of CO2-induced acidification on sediment toxicity using acute amphipod toxicity test.

    PubMed

    Basallote, M Dolores; De Orte, Manoela R; DelValls, T ngel; Riba, Inmaculada

    2014-01-01

    Carbon capture and storage is increasingly being considered one of the most efficient approaches to mitigate the increase of CO2 in the atmosphere associated with anthropogenic emissions. However, the environmental effects of potential CO2 leaks remain largely unknown. The amphipod Ampelisca brevicornis was exposed to environmental sediments collected in different areas of the Gulf of Cdiz and subjected to several pH treatments to study the effects of CO2-induced acidification on sediment toxicity. After 10 days of exposure, the results obtained indicated that high lethal effects were associated with the lowest pH treatments, except for the Ra of Huelva sediment test. The mobility of metals from sediment to the overlying seawater was correlated to a pH decrease. The data obtained revealed that CO2-related acidification would lead to lethal effects on amphipods as well as the mobility of metals, which could increase sediment toxicity. PMID:24988484

  19. Sugars exert a major influence on the vitrification properties of ethylene glycol-based solutions and have low toxicity to embryos and oocytes.

    PubMed

    Kuleshova, L L; MacFarlane, D R; Trounson, A O; Shaw, J M

    1999-03-01

    A systematic approach was taken to assess the vitrification properties of ethylene glycol-based solutions supplemented with carbohydrates. Solutions were prepared by weight (gravimetrically) using ethylene glycol as the cryoprotectant, 0.9% NaCl in water, and six different sugars: d-glucose, d(-)-fructose, d-sorbitol, sucrose, d(+)-trehalose, and raffinose. Sugars were added on a molal basis (0. 1, 0.5, and 1 m). Characteristics of the solutions were measured during warming by differential scanning calorimetry using a cooling rate of 100 degrees C/min and a warming rate of 10 degrees C/min. In the absence of carbohydrates a 59 wt% EG-saline solution formed a stable glass. When EG was replaced by an equimolal concentration of glucose, fructose, or sorbitol (monosaccharides) at 0.1, 0.5, or 1.0 m there was no change in the total solute concentration at which vitrification occurred, but the glass transition (Tg) occurred at a higher temperature than in EG-saline alone. When EG was replaced by an equimolal concentration of sucrose or trehalose (disaccharides) both the Tg and the lowest total solute concentration required for vitrification became progressively higher as the molecular weight, or the ratio of sugar to EG in the solutions, increased. At the highest tested disaccharide concentration (1 m) vitrification was achieved at a total solute concentration of 65 wt% (sucrose) and 67 wt% (trehalose). The polysaccharide raffinose significantly modified the vitrification properties of ethylene glycol solutions. When 0.5 or 0.1 m raffinose replaced EG on an equimolal basis the glass transition point was raised more than with either the monosaccharides or the disaccharides. Raffinose allowed vitrification at a total solute concentration of 67 wt% (0.5 m) and 63 wt% (0.1 m). The maturation of immature mouse oocytes, and the development of embryos in media containing 5-7 mM of any sugar was comparable to controls, indicating that they are not toxic. Exposure of freshly collected GV or MII oocytes to sugar concentrations between 0.5 and 1.0 M, for up to 10 min had no significant effect on the proportion which subsequently formed two cells. We conclude that added sugars do contribute to a solutions overall vitrification properties, and their properties should be taken into consideration when vitrification solutions are being designed or modified. PMID:10191035

  20. VALIDATION OF EMBRYO TESTS FOR DETERMINING EFFECTS OF FUNGAL PEST CONTROL AGENTS ON NONTARGET AQUATIC ANIMALS

    EPA Science Inventory

    Developing embryos of the inland silverside fish Menidia beryllina and grass shrimp Palaemonetes pugio were exposed to conidiospores of the fungal weed control agent, Colletotrichum gloeosporioides, f. sp. aeschynomene, and the entomopathogen, Metarhizium anisopliae. nly Metarhiz...

  1. Field assessments in conjunction with whole effluent toxicity testing

    SciTech Connect

    La Point, T.W.; Waller, W.T.

    2000-01-01

    Whole effluent toxicity (WET) tests are widely used to assess potential effects of wastewater discharges on aquatic life. This paper represents a summary of chapters in a 1996 Society of Environmental Toxicology and Chemistry-sponsored workshop and a literature review concerning linkages between WET testing and associated field biomonitoring. Most published studies thus far focus primarily on benthic macroinvertebrates and on effluent-dominated stream systems in which effluents demonstrate little or no significant acute toxicity. Fewer studies examine WET test predictability in other aquatic ecosystems (e.g., wetlands, estuaries, large rivers) or deal with instream biota such as fish and primary producers. Published results indicate that standards for the usual WET freshwater test species, Ceriodaphnia dubia and Pimephales promelas, may not always protect most of the species inhabiting a receiving stream. Although WET tests are useful in predicting aquatic individual responses, they are not meant to directly measure natural population or community responses. Further, they do not address bioconcentration or bioaccumulation of hydrophobic compounds; do not assess eutrophication effects in receiving systems; and lastly, do not reflect genotoxic effects or function to test for endocrine-disrupting chemicals. Consequently, a more direct evaluation of ecosystem health, using bioassessment techniques, may be needed to properly evaluate aquatic systems affected by wastewater discharges.

  2. Evaluation of the test of significant toxicity for determining the toxicity of effluents and ambient water samples.

    PubMed

    Diamond, Jerry M; Denton, Debra L; Roberts, John W; Zheng, Lei

    2013-04-01

    The test of significant toxicity (TST) is a hypothesis-testing approach based on bioequivalence developed by the U.S. Environmental Protection Agency (U.S. EPA) for analyzing whole-effluent toxicity (WET) and ambient toxicity data. The present study compares results of acute and chronic toxicity tests of effluent, storm-water, and ambient (i.e., receiving-water) samples using both the TST and the standard no-observed-effect concentration (NOEC) approach. Valid WET data were analyzed from 890 tests provided by more than 25 dischargers in California and Washington, USA, representing the majority of test methods used in the U.S. WET program. An additional 3,201 freshwater chronic toxicity tests, obtained from ambient monitoring programs in California, were also analyzed. The TST and NOEC approaches both declared a low number (<6.5%) of tests toxic if effects were below the unacceptable toxicity regulatory management decision (RMD) of 25% effect in chronic tests or 20% effect in acute tests. However, those test methods having generally lower within-test variability and greater test power (e.g., urchin fertilization test) had a much lower percentage of tests declared toxic than the NOEC approach when effects were below the unacceptable toxicity RMD. In addition, the TST showed fewer tests to be nontoxic than NOEC if the test exhibited effects greater than the toxicity RMD (0.1 and 9.6% for TST and NOEC, respectively, for effluents and 0 and 9.5%, respectively, for ambient samples). Our results demonstrate that the TST is more likely to identify a toxic sample when effects are fairly substantial (≥ 25% effect in chronic testing and ≥ 20% effect in acute tests) and less likely to identify a sample as toxic when effects are negligible (≤ 10% effect). Furthermore, these results demonstrate that appropriate WET data interpretation benefits from having well-designed test methods with sufficient power to identify significant toxicity or biologically insignificant effects when they occur. PMID:23400869

  3. Protein expression analysis of rat testes induced testicular toxicity with several reproductive toxicants.

    PubMed

    Yamamoto, Toshinori; Fukushima, Tamio; Kikkawa, Rie; Yamada, Hiroshi; Horii, Ikuo

    2005-05-01

    The utilization of safety biomarkers to predict the possibility of compound-related toxicity provides several advantages for drug discovery and development, especially at an early stage. The objectives of this study were to investigate the effects of male reproductive toxicants on protein expression profiles in the rat testes and to identify potential biomarker candidates. Four well-known reproductive toxicants, ethylene glycol monomethyl ether (EGME), cyclophosphamide (CP), sulfasalazine (SASP) and 2,5-hexanedione (2,5-HD), were administered to male rats in a single dose, and protein expression profiles were investigated after 24 hr by two-dimensional gel electrophoresis (2DE). Histopathological examination of the testes and serum concentration analysis were also performed. From the results of the comparison of 2D-gels among different doses of a compound and among compounds, 52, 20, 24 and 111 spots were nominated as differentially expressed spots with EGME, CP, SASP and 2,5-HD treatments, respectively. Several spermatogenesis-involved proteins were identified, including glutathione S-transferase (GST), testis-specific heat shock protein 70-2 (HSP70-2), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and phosphatidylethanolamine-binding protein (PEBP). Some of them were altered by more than one compound. In summary, remarkable histopathological findings were observed only in the EGME high-dose group, and most of the protein changes were detected before histopathological changes occurred. Therefore, the proteins identified in this study could potentially serve as biomarkers to evaluate male reproductive toxicity at an early stage of drug discovery and development. PMID:15928459

  4. Toxicity of naphthenic acid fraction components extracted from fresh and aged oil sands process-affected waters, and commercial naphthenic acid mixtures, to fathead minnow (Pimephales promelas) embryos.

    PubMed

    Marentette, Julie R; Frank, Richard A; Bartlett, Adrienne J; Gillis, Patricia L; Hewitt, L Mark; Peru, Kerry M; Headley, John V; Brunswick, Pamela; Shang, Dayue; Parrott, Joanne L

    2015-07-01

    Naphthenic acids (NAs) are constituents of oil sands process-affected water (OSPW). These compounds can be both toxic and persistent and thus are a primary concern for the ultimate remediation of tailings ponds in northern Alberta's oil sands regions. Recent research has focused on the toxicity of NAs to the highly vulnerable early life-stages of fish. Here we examined fathead minnow embryonic survival, growth and deformities after exposure to extracted NA fraction components (NAFCs), from fresh and aged oil sands process-affected water (OSPW), as well as commercially available NA mixtures. Commercial NA mixtures were dominated by acyclic O2 species, while NAFCs from OSPW were dominated by bi- and tricyclic O2 species. Fathead minnow embryos less than 24h old were reared in tissue culture plates terminating at hatch. Both NAFC and commercial NA mixtures reduced hatch success, although NAFCs from OSPW were less toxic (EC50=5-12mg/L, nominal concentrations) than commercial NAs (2mg/L, nominal concentrations). The toxicities of NAFCs from aged and fresh OSPW were similar. Embryonic heart rates at 2 days post-fertilization (dpf) declined with increasing NAFC exposure, paralleling patterns of hatch success and rates of cardiovascular abnormalities (e.g., pericardial edemas) at hatch. Finfold deformities increased in exposures to commercial NA mixtures, not NAFCs. Thus, commercial NA mixtures are not appropriate surrogates for NAFC toxicity. Further work clarifying the mechanisms of action of NAFCs in OSPW, as well as comparisons with additional aged sources of OSPW, is merited. PMID:25957715

  5. Species-specific considerations in using the fish embryo test as an alternative to identify endocrine disruption.

    PubMed

    Schiller, Viktoria; Zhang, Xiaowei; Hecker, Markus; Schäfers, Christoph; Fischer, Rainer; Fenske, Martina

    2014-10-01

    A number of regulations have been implemented that aim to control the release of potentially adverse endocrine disrupters into the aquatic environment based on evidence from laboratory studies. Currently, such studies rely on testing approaches with adult fish because reliable alternatives have not been validated so far. Fish embryo tests have been proposed as such an alternative, and here we compared two species (medaka and zebrafish) to determine their suitability for the assessment of substances with estrogenic and anti-androgenic activity. Changes in gene expression (in here the phrase gene expression is used synonymously to gene transcription, although it is acknowledged that gene expression is additionally regulated, e.g., by translation and protein stability) patterns between the two species were compared in short term embryo exposure tests (medaka: 7-day post fertilization [dpf]; zebrafish: 48 and 96h post fertilization [hpf]) by using relative quantitative real-time RT-PCR. The tested genes were related to the hypothalamic-gonadal-axis and early steroidogenesis. Test chemicals included 17α-ethinylestradiol and flutamide as estrogenic and anti-androgenic reference compounds, respectively, as well as five additional substances with endocrine activities, namely bisphenol A, genistein, prochloraz, linuron and propanil. Estrogenic responses were comparable in 7-dpf medaka and 48/96-hpf zebrafish embryos and included transcriptional upregulation of aromatase b, vitellogenin 1 as well as steroidogenic genes, suggesting that both species reliably detected exposure to estrogenic compounds. However, anti-androgenic responses differed between the two species, with each species providing specific information concerning the mechanism of anti-androgenic disruption in fish embryos. Although small but significant changes in the expression of selected genes was observed in 48-hpf zebrafish embryos, exposure prolonged to 96hpf was necessary to obtain a response indicative of anti-androgenic activity. In contrast, for medaka clear anti-androgenic response, e.g. transcriptional downregulation of 11β-hydroxylase, 3β-hydroxysteroid-dehydrogenase, gonadotropin-releasing hormone receptor 2, was already observed at the pre-hatch stage. Together, this data suggests that medaka and zebrafish embryos would provide a beneficial alternative testing platform for endocrine disruption that involves additive information on interspecies and exposure time variability when using both species. PMID:24992288

  6. Bioavailability of fluoranthene in freshwater sediment toxicity tests

    SciTech Connect

    Suedel, B.C.; Rodgers, J.H. Jr. ); Clifford, P.A. )

    1993-01-01

    To examine equilibrium-partitioning model predictions of interstitial water concentrations of fluoranthene as part of the equilibrium-partitioning (EqP) approach to sediment quality criteria development, the bioavailability (toxicity) of fluoranthene-amended sediment to Hyalella azteca, Daphnia magna, and Chironomus tentans was determined. Fluoranthene was added to three freshwater sediments with similar organic carbon content. Predicted interstitial water concentrations from the equilibrium-partitioning model were similar to measured interstitial water concentrations for WRFS and TR sediment, but the model underpredicted measured values for LF sediment by a factor of two. EC50s for Daphnia magna, Hyalella azteca, and Chironomus tentans in interstitial water were a factor of two to five greater for LF than for WRFS and TR sediments. Factors other than organic carbon content of sediments probably contributed to the variability in bioavailability of fluoranthene. Based on 10-d sediment toxicity tests with Hylella azteca, Daphnia magna, and Chironomus tentans, organic carbon-normalized sediment concentrations were better predictors of toxicity than interstitial water and bulk sediment fluoranthene concentrations. In 10-d aqueous-phase tests with fluoranthene, Chironomus tentans and Hyalella azteca were twice as sensitive as Daphnia magna.

  7. 78 FR 69414 - Toxic Substances Control Act Chemical Testing; Receipt of Test Data

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-19

    ... October 21, 2011 (76 FR 65385) (FRL-8885-5) (docket ID number EPA-HQ-OPPT-2009-0112). The table in this... AGENCY Toxic Substances Control Act Chemical Testing; Receipt of Test Data AGENCY: Environmental Protection Agency (EPA). ACTION: Notice. SUMMARY: This notice announces EPA's receipt of test data on...

  8. Collection and cultivation methods of Acartia tonsa for toxicity testing

    SciTech Connect

    Hood, C.A.; Mayo, R.R.

    1995-12-31

    Acartia tonsa were located and collected from Galveston Bay, Texas in June 1995, using plankton nets and transported to the laboratory for culture. After literature searching and laboratory experimentation. A simple reliable method was designed to culture A. tonsa. This method requires a minimum of glassware and supplies. Adult A. tonsa are placed in one gallon bell jars filled with natural seawater. The jars are then maintained in a water bath at a constant temperature. Water changes are conducted twice weekly and organisms are fed daily with a mixture of algae, Skeletonema costatum, isocrysis galbana, and Thalassiosira sp. Gravid females are then isolated in generators for 24 hours to obtain known age neonates. The neonates are maintained up to a specific age and then are used in toxicity tests such as the ``Determination of the Acute Lethal Toxicity to Marine Copepods,`` required in the United Kingdom for all chemicals used for offshore drilling fluid applications.

  9. PHOTOACTIVATED POLYCYCLIC AROMATIC HYDROCARBON TOXICITY IN MEDAKA (ORYZIAS LATIPES) EMBRYOS: RELEVANCE TO ENVIRONMENTAL RISK IN CONTAMINATED SITES

    EPA Science Inventory

    The hazard for photoactivated toxicity of polycyclic aromatic hydrocarbons (PAHs) has been clearly demonstrated; however, to our knowledge, the risk in contaminated systems has not been characterized. To address this question, a median lethal dose (LD50) for fluoranthene photoa...

  10. CELLULAR TOXICITY IN CHINESE HAMSTER OVARY CELL CULTURES. 2. A STATISTICAL APPRAISAL OF SENSITIVITY WITH THE RABBIT ALVEOLAR MACROPHAGE, SYRIAN HAMSTER EMBRYO, BALB 3T3 MOUSE, AND HUMAN NEONATAL FIBROBLAST CELL SYSTEMS

    EPA Science Inventory

    Chinese hamster ovary, rabbit alveolar macrophage, Syrian Hamster embryo, mouse, and human neonatal fibroblast cells were employed in a statistical evaluation of the relative sensitivity of the cells to toxic substances. The cells were exposed to 1,2,4-trichlorobenzene, 2,4-dimet...

  11. Establishing relative sensitivities of various toxicity testing organisms to ammonia

    SciTech Connect

    Karle, L.M.; Mayhew, H.L.; Barrows, M.E.; Karls, R.K.

    1994-12-31

    The toxicity of ammonia to various organisms was examined to develop a baseline for mortality in several commonly used testing species. This baseline data will assist in choosing the proper test species and in interpreting results as they pertain to ammonia. Responses for two juvenile fish species, three marine amphipods, and two species of mysid shrimp were compared for their sensitivity to levels of ammonia. All mortality caused by ammonia in the bottom-dwelling Citharichthys stigmaeus occurred within 24 h of exposure, whereas mortality in the silverside, Menidia beryllina, occurred over the entire 96-h test duration. Responses to ammonia varied among the amphipods Rhepoxynius abronius, Ampelisca abdita, and Eohaustorius estuarius. R. abronius and A. abdita showed similar sensitivity to ammonia at lower concentrations; A. abdita appeared more sensitive than R. abronius at levels above 40 mg/L. Concentrations of ammonia required to produce significant mortality in the amphipod E. estuarius were far higher than the other species examined (> 100 mg/L NH{sub 3}). A comparison of ammonia toxicity with two commonly used invertebrates, Holmesimysis sculpts and Mysidopsis bahia, suggest that these two species of mysid have similar sensitivities to ammonia. Further studies with ammonia that examine sensitivity of different organisms should be conducted to assist regulatory and environmental agencies in determining appropriate test species and in interpreting toxicological results as they may be affected by levels of ammonia.

  12. What food and feeding rates are optimum for the Chironomus dilutus sediment toxicity test method?

    EPA Science Inventory

    Laboratory tests with benthic macroinvertebrates conducted using standard toxicity test procedures are used to assess the potential toxicity of contaminated sediments. Results are compared across sites or for batches of samples, and the performance of organisms in control treatme...

  13. An in vivo evaluation of acute toxicity of cobalt ferrite (CoFe2O4) nanoparticles in larval-embryo Zebrafish (Danio rerio).

    PubMed

    Ahmad, Farooq; Liu, Xiaoyi; Zhou, Ying; Yao, Hongzhou

    2015-09-01

    The broad spectrum applications of CoFe2O4 NPs have attracted much interest in medicine, environment and industry, resulting in exceedingly higher exposures to humans and environmental systems in succeeding days. Their health effects and potential biological impacts need to be determined for risk assessment. Zebrafish (Danio rerio) embryos were exposed to environmentally relevant doses of nano-CoFe2O4 (mean diameter of 40nm) with a concentration range of 10-500μM for 96h. Acute toxic end points were evaluated by survival rate, malformation, hatching delay, heart dysfunction and tail flexure of larvae. Dose and time dependent developmental toxicity with severe cardiac edema, down regulation of metabolism, hatching delay and tail/spinal cord flexure and apoptosis was observed. The biochemical changes were evaluated by ROS, Catalase (CAT), Lipid peroxidation (LPO), Acid phophatase (AP) and Glutatione s- transferase (GST). An Agglomeration of NPs and dissolution of ions induces severe mechanical damage to membranes and oxidative stress. Severe apoptosis of cells in the head, heart and tail region with inhibition of catalase confirms ROS induced acute toxicity with increasing concentration. Increased activity of GST and AP at lower concentrations of CoFe2O4 NPs demonstrates the severe oxidative stress. Circular dichroism (CD) spectra indicated the weak interactions of NPs with BSA and slight changes in α-helix structure. In addition, CoFe2O4 NPs at lower concentrations do not show any considerable interference with assay components and analytical instruments. The results are possible elucidation of pathways of toxicity induced by these particles, as well as contributing in defining the protocols for risk assessment of these nanoparticles. PMID:26197244

  14. Sediment toxicity testing of Lake Orta after liming

    SciTech Connect

    Baudo, R.; Beltrami, M.; Rossi, D.; Gronda, A.; Abdel-Monem, A.M.; Burton, G.A.

    1995-12-31

    Lake Orta has been severely polluted by industrial effluents containing Cu, NH{sub 3}, Cr., Ni, and Zn. Ammonia oxidation caused a pH drop below 4. More than 60 papers have described the adverse chemical and biological changes which occurred in Lake Orta in the past several decades. In 1989, CaCO{sub 3} (10,900 tons) was added to the lake as a remedial action. The treatment was effective, improving water quality and allowing some original plankton, benthic, and fish populations to return. However, Cu and Cr are still present in the top 10 cm of sediment. The toxicity of the sediment was evaluated using multiple assays: Microtox (solid phase and pore water), Brachionus calyciflorus and Thamnocephalus platyurus Toxkits (pore water), seed germination and root elongation (solid phase and pore water). In addition, in situ and laboratory tests were conducted with Hyalella azteca and an indigenous species of Daphnia. Assays responses varied yet showed surficial sediments to be toxic. Toxicity increased with sediment in pre-liming deposition zones. Recovery of the ecosystem is evident, however the contaminated sediments may impede further improvement.

  15. Discovering less toxic ionic liquids by using the Microtox® toxicity test.

    PubMed

    Hernández-Fernández, F J; Bayo, J; Pérez de los Ríos, A; Vicente, M A; Bernal, F J; Quesada-Medina, J

    2015-06-01

    New Microtox® toxicity data of 16 ionic liquids of different cationic and anionic composition were determined. The ionic liquids 1-butyl-1-methylpyrrolidinium trifluoromethanesulfonate, [BMPyr(+)][TFO(-)], 1-butyl-1-methylpyrrolidinium chloride, [BMPyr(+)][Cl(-)], hydroxypropylmethylimidazolium fluoroacetate, [HOPMIM(+)][FCH2COO(-)], and hydroxypropylmethylimidazolium glycolate [HOPMIM(+)][glycolate(-)] were found to be less toxic than conventional organic solvent such as chloroform or toluene, accoding the Microtox® toxicity assays. The toxicity of pyrrolidinium cation was lower than the imidazolium and pyridinium ones. It was found that the inclusion of an hydroxyl group in the alkyl chain length of the cation also reduce the toxicity of the ionic liquid. To sum up, the Microtox® toxicity assays can be used as screening tool to easily determined the toxicity of a wide range of ionic liquids and the toxicity data obtained could allow the obtention of structure-toxicity relationships to design less toxic ionic liquids. PMID:25748519

  16. Trend tests for proportional responses in developmental toxicity experiments

    SciTech Connect

    Chinchilli, V.M.; Clark, B.C. )

    1989-02-01

    The data from developmental toxicity experiments usually are very difficult to analyze statistically because of the lack of independence among littermates and the random nature of the litter size. Only a few of the models that have been proposed in the literature have accounted for both of these features. One of the models proposed by Van Ryzin is invoked to construct a test of trend (dose response). The construction is achieved via a statistical technique called isotonic regression, which is applied to the moment estimators derived by Van Ryzin. The trend test based on isotonic regression is relatively straightforward to calculate, and when the number of dose groups (including control) is four or less, the significance of the observed result is easily determined. An example, in which fetolethality is the end point of interest, demonstrates the test.

  17. Tests for oil/dispersant toxicity: In situ laboratory assays

    SciTech Connect

    Wright, D.A.; Coelho, G.M.; Aurand, D.V.

    1995-12-31

    As part of its readiness program in oil spill response, the Marine Pollution Control Unit (MPCU), Department of Transport, U.K. conducts annual field trials in the North Sea, approximately 30 nautical miles from the southeast coast of England. The trials take the form of controlled releases of crude oil or Medium Fuel/Gas Oil mix (MFO), with and without the application of Corexit 9527 dispersant. In 1994 and 1995 the authors conducted a series of in situ toxicity bioassays in association with these spills with included 48h LC50 tests for turbot (Scophthalmus maximus) and oyster (Crassostrea gigas) larvae, a 48 h oyster (C. gigas) embryonic development test and two full life-cycle assays using the copepods Acartia tonsa and Tisbe battagliai. Tests were also conducted in the Chesapeake Bay laboratory using estuarine species including the copepod Eurytemora affinis and the inland silverside Menidia beryllina. Here, the authors report on the results of these assays, together with 1996 in situ toxicity data resulting from Norwegian field trials in the northern North Sea.

  18. Organotypic liver culture models: Meeting current challenges in toxicity testing

    PubMed Central

    LeCluyse, Edward L.; Witek, Rafal P.; Andersen, Melvin E.; Powers, Mark J.

    2012-01-01

    Prediction of chemical-induced hepatotoxicity in humans from in vitro data continues to be a significant challenge for the pharmaceutical and chemical industries. Generally, conventional in vitro hepatic model systems (i.e. 2-D static monocultures of primary or immortalized hepatocytes) are limited by their inability to maintain histotypic and phenotypic characteristics over time in culture, including stable expression of clearance and bioactivation pathways, as well as complex adaptive responses to chemical exposure. These systems are less than ideal for longer-term toxicity evaluations and elucidation of key cellular and molecular events involved in primary and secondary adaptation to chemical exposure, or for identification of important mediators of inflammation, proliferation and apoptosis. Progress in implementing a more effective strategy for in vitro-in vivo extrapolation and human risk assessment depends on significant advances in tissue culture technology and increasing their level of biological complexity. This article describes the current and ongoing need for more relevant, organotypic in vitro surrogate systems of human liver and recent efforts to recreate the multicellular architecture and hemodynamic properties of the liver using novel culture platforms. As these systems become more widely used for chemical and drug toxicity testing, there will be a corresponding need to establish standardized testing conditions, endpoint analyses and acceptance criteria. In the future, a balanced approach between sample throughput and biological relevance should provide better in vitro tools that are complementary with animal testing and assist in conducting more predictive human risk assessment. PMID:22582993

  19. A SURROGATE SUBCHRONIC TOXICITY TEST METHOD FOR WATERS WITH HIGH TOTAL DISSOLVED SOLIDS

    EPA Science Inventory

    Total dissolved solids (TDS) are often identified as a toxicant in whole-effluent toxicity (WET) testing. The primary test organism used in WET testing, Ceriodaphnia dubia, is very sensitive to TDS ions, which can be problematic when differentiating the toxicity of TDS from those...

  20. Quantification and in situ localisation of abcb1 and abcc9genes in toxicant-exposed sea urchin embryos.

    PubMed

    Bošnjak, Ivana; Pleić, Ivana Lepen; Borra, Marco; Mladineo, Ivona

    2013-12-01

    A multixenobiotic resistance (MXR) mechanism mediated by ABC binding cassette (ABC) transport proteins is an efficient chemical defence mechanism in sea urchin embryos. The aim of our work was to evidence whether exposure to sub-lethal doses of specific contaminants (oxybenzone (OXI), mercuric chloride (HgCl2) and trybutiltin (TBT)) would induce MXR transporter activity during embryonic development (from zygote to blastula stage) in purple sea urchin (Paracentrotus lividus) embryos. Further, we present data on molecular identification, transport function, expression levels and gene localisation of two ABC efflux transporters-P-glycoprotein (ABCB1/P-gp) and sulfonylurea-receptor-like protein (ABCC9/SUR-like). Partial cDNA sequences of abcb1 and abcc9 were identified and quantitative PCR (qPCR) evidenced an increase in mRNA transcript levels of both ABC transporters during the two-cell, as well as an overall decrease during the blastulae stage. Calcein-AM efflux activity assay indicated the activation of multidrug resistance-associated protein/ABCC-like transport in the presence of HgCl2 and TBT in exposed blastulae. The in situ hybridisation of the two-cell and blastula stages showed ubiquitous localisation of both transcripts within cells, supporting qPCR data. In conclusion, ABCB1 and ABCC9 are constitutive, as are HgCl2, TBT and OXI-inducible ABC membrane transporters, coexpressed in the zygote, two-cell and blastula stages of the P. lividus. Their ubiquitous cell localisation further fortifies their protective role in early embryonic development. PMID:23690080

  1. Multiple bio-analytical methods to reveal possible molecular mechanisms of developmental toxicity in zebrafish embryos/larvae exposed to tris(2-butoxyethyl) phosphate.

    PubMed

    Han, Zhihua; Wang, Qiangwei; Fu, Jie; Chen, Hongshan; Zhao, Ye; Zhou, Bingsheng; Gong, Zhiyuan; Wei, Si; Li, Jun; Liu, Hongling; Zhang, Xiaowei; Liu, Chunsheng; Yu, Hongxia

    2014-05-01

    The flame retardant tris(2-butoxyethyl) phosphate (TBEP) is a frequently detected contaminant in the environment, wildlife and human milk. The potentially toxic effects of TBEP and their underlying molecular mechanisms have not been elucidated. Here, zebrafish embryos were exposed to different concentrations of TBEP from 4 hours of post-fertilization (hpf) to 120 hpf, and effects on embryonic development and global protein expression patterns examined. Our results demonstrate that treatment with TBEP (0.8-100mg/L) causes a concentration- and time-dependent decrease in embryonic survival and the hatching percentage. The median lethal concentration was 10.7 mg/L at 120 hpf. Furthermore, exposure to 150 or 800 μg/L TBEP inhibited the degradation and utilization of vitellogenins and down-regulated the expression of proteins related to cation binding, and lipid transport, uptake and metabolism, accompanied by a decrease in heart rate and body length. Exposure to TBEP (150 or 800 μg/L) also decreased the expression of proteins involved in cell proliferation and DNA repair, and led to an increased number of apoptotic cells in the tail region. Collectively, our results suggest that exposure to TBEP causes toxicity in the developing zebrafish by inhibiting the degradation and utilization of nutrients from the mother and inducing apoptosis. PMID:24685621

  2. Toxicity of lead and zinc to developing mussel and sea urchin embryos: critical tissue residues and effects of dissolved organic matter and salinity.

    PubMed

    Nadella, Sunita R; Tellis, Margaret; Diamond, Rachael; Smith, Scott; Bianchini, Adalto; Wood, Chris M

    2013-08-01

    Lead (Pb) EC50 values in the very sensitive early development phases (48-72h post-fertilization) of the mussels Mytilus galloprovincialis and Mytilus trossolus and sea urchin Strongylocentrotus purpuratus in 100% sea water were: M. trossolus - 45 (95% C.I.=22-72) μgL(-1); M. galloprovincialis - 63 (36-94) μgL(-1); S. purpuratus - 74 (50-101) μgL(-1). Salinity thresholds for normal development varied: M. trossolus>21ppt; M. galloprovincialis>28ppt; S. purpuratus≥30ppt. Addition of two spectroscopically distinct dissolved organic matters (DOM) from fresh water (Nordic Reservoir) and sea water (Inshore) moderately decreased the toxicity of Pb to both mussels, but not in a concentration-dependent fashion, with only an approximate doubling of EC50 over the range of 1.4-11.2mgCL(-1). Independent Pb binding capacity determinations for DOC explained the lack of a relationship between DOM concentration and toxicity. Salinity had no effect on Pb toxicity down to 21ppt in M. trossolus, and low salinity (21ppt) did not enhance the protective effect of DOC. Both DOMs increased the toxicity of Pb in developing sea urchin embryos, in contrast to mussels. Relative to Pb, the organisms were 6-9 fold less sensitive to Zn on a molar basis in 100% seawater with the following Zn EC50s: M. trossolus - 135 (103-170) μgL(-1); M. galloprovincialis - 172 (126-227) μgL(-1), S. purpuratus - 151 (129-177) μgL(-1). Nordic Reservoir and Inshore DOM (2-12mgCL(-1)) had no significant effect on Zn toxicity to mussels, in accord with voltammetry data showing an absence of any strong ligand binding for Zn by DOMs. As with Pb, DOMs increased Zn toxicity to urchin larvae. Critical Tissue Residues (CTR) based on whole body concentrations of Pb and Zn were determined for M. galloprovincialis at 48h and S. purpuratus at 72h. The median lethal CTR values (LA50s), useful parameters for development of saltwater Biotic Ligand Models (BLMs), were approximately 4-fold higher on a molar basis for Zn than for Pb. The latter were not altered by DOM exposure, despite increased EC50 values, in accord with the tenets of the BLM. PMID:23603691

  3. Non-animal methodologies within biomedical research and toxicity testing.

    TOXLINE Toxicology Bibliographic Information

    Knight A

    2008-01-01

    Laboratory animal models are limited by scientific constraints on human applicability, and increasing regulatory restrictions, driven by social concerns. Reliance on laboratory animals also incurs marked - and in some cases, prohibitive - logistical challenges, within high-throughput chemical testing programmes, such as those currently underway within Europe and the US. However, a range of non-animal methodologies is available within biomedical research and toxicity testing. These include: mechanisms to enhance the sharing and assessment of existing data prior to conducting further studies, and physicochemical evaluation and computerised modelling, including the use of structure-activity relationships and expert systems. Minimally-sentient animals from lower phylogenetic orders or early developmental vertebral stages may be used, as well as microorganisms and higher plants. A variety of tissue cultures, including immortalised cell lines, embryonic and adult stem cells, and organotypic cultures, are also available. In vitro assays utilising bacterial, yeast, protozoal, mammalian or human cell cultures exist for a wide range of toxic and other endpoints. These may be static or perfused, and may be used individually, or combined within test batteries. Human hepatocyte cultures and metabolic activation systems offer potential assessment of metabolite activity and organ-organ interaction. Microarray technology may allow genetic expression profiling, increasing the speed of toxin detection, well prior to more invasive endpoints. Enhanced human clinical trials utilising micro- dosing, staggered dosing, and more representative study populations and durations, as well as surrogate human tissues, advanced imaging modalities and human epidemiological, sociological and psycho- logical studies, may increase our understanding of illness aetiology and pathogenesis, and facilitate the development of safe and effective pharmacologic interventions. Particularly when human tissues are used, non-animal models may generate faster, cheaper results, more reliably predictive for humans, whilst yielding greater insights into human biochemical processes. Greater commitment to their development and implementation is necessary, however, to efficiently meet the needs of high-throughput chemical testing programmes, important emerging testing needs, and the ongoing development of human clinical interventions.

  4. Non-animal methodologies within biomedical research and toxicity testing.

    PubMed

    Knight, Andrew

    2008-01-01

    Laboratory animal models are limited by scientific constraints on human applicability, and increasing regulatory restrictions, driven by social concerns. Reliance on laboratory animals also incurs marked - and in some cases, prohibitive - logistical challenges, within high-throughput chemical testing programmes, such as those currently underway within Europe and the US. However, a range of non-animal methodologies is available within biomedical research and toxicity testing. These include: mechanisms to enhance the sharing and assessment of existing data prior to conducting further studies, and physicochemical evaluation and computerised modelling, including the use of structure-activity relationships and expert systems. Minimally-sentient animals from lower phylogenetic orders or early developmental vertebral stages may be used, as well as microorganisms and higher plants. A variety of tissue cultures, including immortalised cell lines, embryonic and adult stem cells, and organotypic cultures, are also available. In vitro assays utilising bacterial, yeast, protozoal, mammalian or human cell cultures exist for a wide range of toxic and other endpoints. These may be static or perfused, and may be used individually, or combined within test batteries. Human hepatocyte cultures and metabolic activation systems offer potential assessment of metabolite activity and organ-organ interaction. Microarray technology may allow genetic expression profiling, increasing the speed of toxin detection, well prior to more invasive endpoints. Enhanced human clinical trials utilising micro- dosing, staggered dosing, and more representative study populations and durations, as well as surrogate human tissues, advanced imaging modalities and human epidemiological, sociological and psycho- logical studies, may increase our understanding of illness aetiology and pathogenesis, and facilitate the development of safe and effective pharmacologic interventions. Particularly when human tissues are used, non-animal models may generate faster, cheaper results, more reliably predictive for humans, whilst yielding greater insights into human biochemical processes. Greater commitment to their development and implementation is necessary, however, to efficiently meet the needs of high-throughput chemical testing programmes, important emerging testing needs, and the ongoing development of human clinical interventions. PMID:18841317

  5. Comparison of prehatch C-start responses in rainbow trout and lake trout embryos by means of a tactile stimulus test

    USGS Publications Warehouse

    Wright, P.J.; Noltie, D.B.; Tillitt, D.E.

    2003-01-01

    The C-start in teleost fishes, a type of startle response, mediates the ability to respond to abrupt, unexpected stimuli and is characterized by a short-latency, C-type fast start acceleration. In prehatch fish embryos, the C-start appears necessary for mechanical breakdown of the egg chorion and successful hatching by way of increased embryo movement and distribution of the hatching enzymes. In later stages, the C-start plays an important role in predator avoidance. Using tactile stimulation, we evaluated the C-start response in rainbow trout Oncorhynchus mykiss at 170 degree-days, when 6.6% of embryos exhibited C-starts, and lake trout Salvelinus namaycush embryos at 320 degree-days, when 23% of embryos exhibited C-starts. Triplicate groups of embryos were later tested at three developmental stages: early (220 and 360 degree-days for rainbow trout and lake trout, respectively), middle (260 and 480 degree-days, respectively), and late (320 and 560 degree-days, respectively). The proportion of trout embryos exhibiting C-start increased through time, such that 100% had responded by the late stage, just prior to hatching. C-starts could be obtained by repeated stimulation, and the relative activity of the embryos (based on the number of flexures per stimulus) also increased over time. Rainbow trout and lake trout showed very similar C-start responses at parallel developmental stages, and these patterns of response were similar to those reported in other fish species.

  6. [Comprehensive Toxicity Evaluation and Toxicity Identification Used in Tannery and Textile Wastewaters].

    PubMed

    Huang, Li; Chen, Wen-yan; Wan, Yu-shan; Zheng, Guo-juan; Zhao, Yuan; Cai, Qiang

    2015-07-01

    To better evaluate the toxicity of tannery and textile effluents from various emission stages, the research attempted battery of toxicological bioassays and toxicological indices. The bioassays employed Microtox test, zebra fish embryo-larval test and algae (Chlorella vulgaris) test. Meanwhile, toxicological indices including Toxicity Unit (TU), Average Toxicity (AvTx), Toxic Print (TxPr), Most Sensitive Test (MST) and Potential Ecotoxic Effects Probe (PEEP) were applied. The results illustrated that PEEP was the most comprehensive index to take account of the emissions and toxic potential of effluents. PEEP values showed that the reduction rates of toxicity in tannery and textile effluents were 36. 8% and 23. 2%, respectively. Finally, based on the Microtox toxicity test, toxicants in textile effluent were identified through the toxicity identification evaluation (TIE) studies. The results indicated that the main toxicant of textile effluent was non-polar organic pollutants, followed by filterable compounds, heavy metals, oxidizing substances and volatile components. PMID:26489331

  7. Quality Testing of Three Species of Tephritid Fruit Flies After Embryo Cryopreservation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study evaluates characteristics commonly used to define insect quality or fitness by using a complement of three species of tephritid fruit flies obtained from cryopreserved embryos. The Mexican, Anastrepah ludens, Caribbean, A. suspense, and Mediterranean, Certatitis capitata, fruit flies were...

  8. Validation and sensitivity comparisons of micro-scale toxicity tests for the evaluation of freshwater sediment toxicity

    SciTech Connect

    Riebel, P.; Bureau, J.; Blaise, C.; Michaud, J.R.

    1995-12-31

    A three-year study is currently underway to develop a representative and cost-effective battery of toxicity tests for evaluating freshwater sediment and porewater toxicity. Among the tests currently being evaluated are the following: Microtox{trademark} chronic test, Microtox{trademark} solid-phase test, Microtox{trademark} liquid phase test, Thamnotoxkit F{trademark}, Rotoxkit F{trademark}, Daphnia magna IQ test{trademark}, Sediment Toxkit, SOS Chromotest, a Selenastrum capricornutum short exposure assay, and trout hepatocyte assays. Conventional sediment tests with Chironomus tentans, Hyalella azteca and Lumbriculus variegatus, as well as benthic macroinvertebrate community assessments and sediment chemical characterizations are being conducted at two contaminated sites. Toxicity test reproducibility, sensitivity, practicality, cost and ecological relevance are discussed.

  9. Effects of the Particulate Matter2.5 (PM2.5) on Lipoprotein Metabolism, Uptake and Degradation, and Embryo Toxicity

    PubMed Central

    Kim, Jae-Yong; Lee, Eun-Young; Choi, Inho; Kim, Jihoe; Cho, Kyung-Hyun

    2015-01-01

    Particulate matter2.5 (PM2.5) is notorious for its strong toxic effects on the cardiovascular, skin, nervous, and reproduction systems. However, the molecular mechanism by which PM2.5 aggravates disease progression is poorly understood, especially in a water-soluble state. In the current study, we investigated the putative physiological effects of aqueous PM2.5 solution on lipoprotein metabolism. Collected PM2.5 from Seoul, Korea was dissolved in water, and the water extract (final 3 and 30 ppm) was treated to human serum lipoproteins, macrophages, and dermal cells. PM2.5 extract resulted in degradation and aggregation of high-density lipoprotein (HDL) as well as low-density lipoprotein (LDL); apoA-I in HDL aggregated and apo-B in LDL disappeared. PM2.5 treatment (final 30 ppm) also induced cellular uptake of oxidized LDL (oxLDL) into macrophages, especially in the presence of fructose (final 50 mM). Uptake of oxLDL along with production of reactive oxygen species was accelerated by PM2.5 solution in a dose-dependent manner. Further, PM2.5 solution caused cellular senescence in human dermal fibroblast cells. Microinjection of PM2.5 solution into zebrafish embryos induced severe mortality accompanied by impairment of skeletal development. In conclusion, water extract of PM2.5 induced oxidative stress as a precursor to cardiovascular toxicity, skin cell senescence, and embryonic toxicity via aggregation and proteolytic degradation of serum lipoproteins. PMID:26615830

  10. In vitro pyrogen test for toxic or immunomodulatory drugs.

    PubMed

    Daneshian, Mardas; Guenther, Armin; Wendel, Albrecht; Hartung, Thomas; von Aulock, Sonja

    2006-06-30

    Pyrogenic contaminations of some classes of injectable drugs, e.g. toxic or immunomodulatory as well as false-positive drugs, represent a major risk which cannot yet be excluded due to the limitations of current tests. Here we describe a modification of the In vitro Pyrogen Test termed AWIPT (Adsorb, Wash, In vitro Pyrogen Test), which addresses this problem by introducing a pre-incubation step in which pyrogenic contaminations in the test sample are adsorbed to albumin-coated beads. After rinsing, the beads are incubated with human whole blood and the release of the endogenous pyrogen interleukin-1beta is measured as a marker of pyrogenic activity. Intentional contaminations with lipopolysaccharide were retrieved from the chemotherapeutic agents paclitaxel, cisplatin and liposomal daunorubicin, the antibiotic gentamicin, the antifungal agent liposomal amphotericin B, and the corticosteroid prednisolone at lower dilutions than in the standard in vitro pyrogen test. This represents a promising new approach for the detection of pyrogenic contamination in drugs or in drugs containing interfering additives and should lead to improved safety levels. PMID:16730742

  11. Thymoquinone therapy abrogates toxic effect of cadmium on rat testes.

    PubMed

    Fouad, A A; Jresat, I

    2015-05-01

    The protective effect of thymoquinone was investigated against cadmium-induced testicular toxicity in rats. Testicular toxicity was induced by a single intraperitoneal (i.p.) injection of cadmium chloride (2mgkg(-1) ). Thymoquinone treatment (10mgkg(-1) day(-1) , i.p.) was applied for five consecutive days, starting 3days before cadmium administration. Thymoquinone significantly attenuated the cadmium-induced decreases in serum testosterone, and testicular reduced glutathione and superoxide dismutase activity and significantly decreased the elevations of testicular malondialdehyde, nitric oxide and cadmium ion levels resulted from cadmium chloride administration. Also, thymoquinone ameliorated the cadmium-induced testicular tissue injury observed by histopathological examination. In addition, thymoquinone significantly decreased the cadmium-induced expression of inducible nitric oxide synthase, tumour necrosis factor-?, cyclooxygenase-2, nuclear factor-?B and caspase-3 in testicular tissue. It was concluded that thymoquinone, through its antioxidant and anti-inflammatory activities, may represent a potential candidate to protect the testes against the detrimental effect of cadmium exposure. PMID:24735446

  12. Transcriptional profiling reveals barcode-like toxicogenomic responses in the zebrafish embryo

    PubMed Central

    Yang, Lixin; Kemadjou, Jules R; Zinsmeister, Christian; Bauer, Matthias; Legradi, Jessica; Müller, Ferenc; Pankratz, Michael; Jäkel, Jens; Strähle, Uwe

    2007-01-01

    Background Early life stages are generally most sensitive to toxic effects. Our knowledge on the action of manmade chemicals on the developing vertebrate embryo is, however, rather limited. We addressed the toxicogenomic response of the zebrafish embryo in a systematic manner by asking whether distinct chemicals would induce specific transcriptional profiles. Results We exposed zebrafish embryos to a range of environmental toxicants and measured the changes in gene-expression profiles by hybridizing cDNA to an oligonucleotide microarray. Several hundred genes responded significantly to at least one of the 11 toxicants tested. We obtained specific expression profiles for each of the chemicals and could predict the identity of the toxicant from the expression profiles with high probability. Changes in gene expression were observed at toxicant concentrations that did not cause morphological effects. The toxicogenomic profiles were highly stage specific and we detected tissue-specific gene responses, underscoring the sensitivity of the assay system. Conclusion Our results show that the genome of the zebrafish embryo responds to toxicant exposure in a highly sensitive and specific manner. Our work provides proof-of-principle for the use of the zebrafish embryo as a toxicogenomic model and highlights its potential for systematic, large-scale analysis of the effects of chemicals on the developing vertebrate embryo. PMID:17961207

  13. KEPONE (TRADEMARK): CHRONIC EFFECTS ON EMBRYO, FRY, JUVENILE, AND ADULT SHEEPSHEAD MINNOWS ('CYPRINODON VARIEGATUS')

    EPA Science Inventory

    The report discusses the toxicity of Kepone to, and uptake by embryo, fry, juvenile, and adult sheepshead minnows (Cyprinodon variegatus) using intermittent-flow toxicity tests. Concentrations of Kepone and percentage of adult fish surviving in a 28-day exposure were--Control, 95...

  14. Evaluation of municipal waste incinerator fly ash toxicity and the role of cadmium by two aquatic toxicity tests

    SciTech Connect

    Kaneko, Hidehiro

    1996-12-31

    Fly ash from a municipal solid waste incinerator in Japan is regulated under the hazardous waste regulation Waste under Special Control, according to the Amendment of the Waste Disposal and Public Cleansing Law, because it contains high concentrations of heavy metals which are available for leaching. To evaluate the toxicity of fly ash, a fly ash leachate was prepared according to the Japanese standard leaching procedure. The chemical analysis of the leachate showed that possibly one of the most toxic substances was cadmium. The toxicity of the leachate and the cadmium was determined by algal assay and a Daphnia acute toxicity test. The results showed that the leachate was about seven times more toxic to the growth of algae and 20 to 30 times more toxic to the survival of Daphnia than expected from its cadmium concentration. The toxicity interaction between cadmium and the other constituents in the leachate was also examined. The toxicity of cadmium showed an additive effect with the other constituents in the leachate in algal assay. In the Daphnia test, however, cadmium showed an antagonistic effect.

  15. Formulated sediment for use in whole-sediment toxicity testing

    SciTech Connect

    Kemble, N.E.; Dwyer, F.J.; Hardesty, D.K.; Ingersoll, C.G.

    1995-12-31

    A formulated control sediment was developed to provide consistent and acceptable biological endpoints for a variety of species used in whole sediment toxicity testing. In an attempt to develop such a sediment the authors conducted multiple tests to evaluate: (1) {alpha}-cellulose as an organic carbon source, (2) various TOC concentrations, (3) various grain sizes, (4) different food types, and (5) overlying waters. Studies were conducted with the amphipod Hyalella azteca the midges Chironomus riparius, Chironomus tentans and the oligochaete Lumbriculus variegatus in 10 d exposures and H. azteca in 28 d exposures. Sediment from West Bearskin Lake Minnesota was used as a control sediment with each species in each test. Survival of test organisms in all of the 10-d experiments, with the exception of C. riparius, was above the acceptable levels for a control sediment. Survival in the formulated sediments also was not significantly different when compared to the control sediment. Amphipod survival in the 28-d exposures was low; however, the use of reconstituted water in combination with the formulated sediment may have been a problem. The authors are currently evaluating various types of overlying water with formulated sediments and sublethal endpoints in each of the exposures (i.e., growth, sexual maturation or head capsule width).

  16. AN ENVIRONMENTAL TECHNOLOGY VERIFICATION (ETV) TESTING OF SEVEN TECHNOLOGIES DETECTING TOXICITY IN DRINKING WATER (R2)

    EPA Science Inventory

    Rapid toxicity technologies can detect certain toxins and with testing it can be determined their susceptibility to interfering chemical in controlled experimental matrix. Rapid toxicity technologies do not identify or determine the concentrations of specific contaminants, but s...

  17. An interlaboratory comparison of sediment elutriate preparation and toxicity test methods

    EPA Science Inventory

    Elutriate bioassays are among numerous methods that exist for assessing the potential toxicity of sediments in aquatic systems. In this study, interlaboratory results were compared from 96-hour Ceriodaphnia dubia and Pimephales promelas static-renewal acute toxicity tests conduct...

  18. Toxicity testing of organic chemicals in groundwater polluted with landfill leachate

    SciTech Connect

    Baun, A.; Kloeft, L.; Bjerg, P.L.; Nyholm, N.

    1999-09-01

    A method for assessment of toxicity of nonvolatile organic chemicals contaminants in groundwater polluted with landfill leachate has been evaluated. The biotests utilized were composed of an algal growth inhibition test (Selenastrum capricornutum), a daphnia immobilization test (Daphnia magna), and a bacterial genotoxicity test (umuC, Salmonella typhimurium). The feasibility of the selected biotests was investigated for a series of groundwater samples collected along pollution gradients downstreams of two landfills in Jutland, Denmark. Two different approaches were used, direct toxicity testing of whole groundwater samples, and toxicity testing of concentrates obtained by solid-phase extraction. Direct testing of whole groundwater samples produced toxic responses, but the complex sample matrix masked the toxicity of the organic chemical contaminants of interest. Solid-phase extraction was used successfully as an on-site method that eliminated ion toxicity and produced biotest responses that reflected the toxicity of the nonvolatile organic chemical contaminants in the groundwater.

  19. Characterization of toxicants in urban road dust by Toxicity Identification Evaluation using ostracod Heterocypris incongruens direct contact test.

    PubMed

    Khanal, Rajendra; Furumai, Hiroaki; Nakajima, Fumiyuki

    2015-10-15

    The current study involves characterization of organic compounds, heavy metals, and ammonia as potential toxicants in one arterial road (St. 3) and two highway (St. 7 and 8) urban road dust (URD) collected in Tokyo, Japan. URD toxicity was evaluated by Toxicity Identification Evaluation using the ostracod Heterocypris incongruens direct contact test. URDs were treated with resins (10% XAD-4, 20% SIR-300 and 20% SIR-600) to determine the reduction in mortality after treatment. The mortality of ostracods exposed to St. 3 URD (baseline 80%) was significantly (p<0.05) reduced to 0% and 47% after XAD-4 and SIR-300 treatment respectively. This reduction led us to suspect hydrophobic organic compounds and heavy metals as potential toxicants. Subsequent elution of the recovered XAD-4 with polar (methanol, acetone) and non-polar (dichloromethane) solvents confirmed the dominance of relatively polar hydrophobic organic toxicants. The dissolved concentration of Cu and Zn after SIR-300 treatment exceeded the 50% lethal concentration (LC50) for ostracods, which led us to suspect Cu and Zn as other probable toxicants. The concentration of Zn in a SIR-300 acidic elutriate, recovered after the termination of toxicity test, confirmed Zn as one of the toxicants in St. 3. The baseline mortality (100%) of St. 7 was significantly reduced (23%) by SIR-300 treatment. This indicated the presence of heavy metals as the probable toxicant. However, the concentration of dissolved heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) in the overlying water was below previously reported LC50 values for each metal in St. 7. Recovery of Zn concentrations exceeding the LC50 in the St. 7 SIR-300 elutriate led us to suspect a dietary exposure route of Zn to the ostracod during the direct contact test. The overall results indicate that the toxicant types can vary widely depending on the road sampled. PMID:26026413

  20. EFFECT OF CHEMICAL CARRIERS ON AVIAN LC(50) TOXICITY TESTS

    EPA Science Inventory

    The subacute dietary (LC50) toxicity of a pesticide as prescribed by the Federal Insecticide Fungicide and Rodenticide Act and of toxic substances as defined by the Toxic Substances Control Act is a routine data point for many chemicals. The methods under which the LC50 data are ...

  1. Tigriopus fulvus: The interlaboratory comparison of the acute toxicity test.

    PubMed

    Faraponova, Olga; Giacco, Elisabetta; Biandolino, Francesca; Prato, Ermelinda; Del Prete, Francesco; Valenti, Alessandra; Sarcina, Stefania; Pasteris, Andrea; Montecavalli, Adele; Comin, Stefano; Cesca, Claudia; Francese, Marco; Cigar, Monica; Piazza, Veronica; Falleni, Fabrizio; Lacchetti, Ines

    2016-02-01

    The paper reports the results of an interlaboratory comparison involving 11 laboratories, with the objectives of apply and validate a new standardized ecotoxicological method on marine crustacean Tigriopus fulvus. Copper was chosen as reference toxicant as indicated in the official method. The results of two independent tests performed by all the participants, demonstrated that the new method is simple, fast and easy to learn. This is confirmed even by the values of z-score index calculated for each laboratory and the relative coefficient of variation (CV) which are 6.32% after 24h, 6.56 after 48h and 35.3% after 96h, mentioned in the ISO standards for the precision of interlaboratory assays. Therefore its use could be recommended in environmental studies and monitoring. PMID:26584461

  2. Toxicity testing with the marine algae, Symbiodinium kawagutii (Dinophyceae)

    SciTech Connect

    Gorrie, J.R.; Bidwell, J.R.; Rippingale, R.J.

    1994-12-31

    The dinoflagellate, Symbiodinium kawagutii, is among the algal taxa which exist in symbiosis with a range of marine invertebrates. S. kawagutii is commonly found in association with the Hawaiian stony coral, Montipora verrucosa. The algae has been successfully cultured in the laboratory using a common marine algal growth media (Guillard f/2), and sufficient cell densities were achieved in a 96-hr bioassay to allow statistical evaluation of toxicity data. A 96-hr EC{sub 50} of 6.47 mg/L (95% C.I.: 3.54--9.88 mg/L) was calculated after exposure to potassium dichromate. Wide distribution of the coral host and ecological importance of the symbiosis make S. kawagutii an excellent candidate species for hazard evaluation in tropical marine ecosystems. Continuing research will seek to further refine the bioassay, including the use of a microplate technique for more rapid testing.

  3. Lethal and sublethal toxicity of the industrial chemical epichlorohydrin on Rhinella arenarum (Anura, Bufonidae) embryos and larvae.

    PubMed

    Hutler Wolkowicz, Ianina R; Aronzon, Carolina M; Pérez Coll, Cristina S

    2013-12-15

    Lethal and sublethal toxicity of the major chemical used in epoxide compounds, epichlorohydrin (ECH) was evaluated on the early life cycle of the common South American toad, Rhinella arenarum (Anura, Bufonidae). The stages evaluated were (according to Del Conte and Sirlin): early blastula (S.3-S.4), gastrula (S.10-S.12), rotation (S.15), tail bud (S.17), muscular response (S.18), gill circulation (S.20), open mouth (S.21), opercular folds (S.23) and complete operculum (S.25). The LC50 and EC50 values for lethal and sublethal effects were calculated. The early blastula was the most sensitive stage to ECH both for continuously and pulse-exposures (LC50-24h=50.9 mg L(-1)), while S.20 was the most resistant (LC50-24h=104.9 mg L(-1)). Among sublethal effects, early blastula was also the most sensitive stage (LOEC-48 h=20 mg L(-1)) and it has a Teratogenic Index of 2.5, which indicates the teratogenic potential of the substance. The main abnormalities were persistent yolk plugs, cell dissociation, tumors, hydropsy, oral malformations, axial/tail flexures, delayed development and reduced body size. ECH also caused neurotoxicity including scarce response to stimuli, reduction in the food intake, general weakness, spasms and shortening, erratic or circular swimming. Industrial contamination is considered an important factor on the decline of amphibian populations. Considering the available information about ECH's toxicity and its potential hazard to the environment, this work shows the first results of its developmental toxicity on a native amphibian species, Rhinella arenarum. PMID:24231313

  4. Aquatic toxicity testing for multicomponent compounds with special reference to preparation of test solution

    SciTech Connect

    Tadokoro, H.; Maeda, M.; Kawashima, Y.; Kitano, M.; Hwang, D.F.; Yoshida, T. )

    1991-02-01

    An adequate method of determining the toxicity of a compound consisting of multiple components, such as creosote, coal tar, and coal tar pitch, was studied for different test solution preparation methods, i.e., direct dosing without filtration, diluting the stock solution of saturated concentration, and dispersing with acetone. Killifish, Oryzias latipes, as a freshwater fish; red sea bream, Pagrus major, as a saltwater fish; and daphnia, Daphnia magna, as a representative crustacean, were used for testing. The chemical analysis of each preparation of test solution with gas chromatography revealed an entirely different profile of the components. The highest toxicity was obtained with preparation by acetone dispersion. That was followed by the preparations with direct dosing method and with the method of dilution of saturated concentration stock solution. Considering the results obtained, the direct dosing method with a suitable settling time may provide useful information enabling extrapolation of the test results to the natural environment for complex multicomponent compounds.

  5. Towards sensible toxicity testing for nanomaterials: proposal for the specification of test design

    NASA Astrophysics Data System (ADS)

    Potthoff, Annegret; Weil, Mirco; Meißner, Tobias; Kühnel, Dana

    2015-12-01

    During the last decade, nanomaterials (NM) were extensively tested for potential harmful effects towards humans and environmental organisms. However, a sound hazard assessment was so far hampered by uncertainties and a low comparability of test results. The reason for the low comparability is a high variation in the (1) type of NM tested with regard to raw material, size and shape and (2) procedures before and during the toxicity testing. This calls for tailored, nanomaterial-specific protocols. Here, a structured approach is proposed, intended to lead to test protocols not only tailored to specific types of nanomaterials, but also to respective test system for toxicity testing. There are existing standards on single procedures involving nanomaterials, however, not all relevant procedures are covered by standards. Hence, our approach offers a detailed way of weighting several plausible alternatives for e.g. sample preparation, in order to decide on the procedure most meaningful for a specific nanomaterial and toxicity test. A framework of several decision trees (DT) and flow charts to support testing of NM is proposed as a basis for further refinement and in-depth elaboration. DT and flow charts were drafted for (1) general procedure—physicochemical characterisation, (2) choice of test media, (3) decision on test scenario and application of NM to liquid media, (4) application of NM to the gas phase, (5) application of NM to soil and sediments, (6) dose metrics, (S1) definition of a nanomaterial, and (S2) dissolution. The applicability of the proposed approach was surveyed by using experimental data retrieved from studies on nanoscale CuO. This survey demonstrated the DT and flow charts to be a convenient tool to systematically decide upon test procedures and processes, and hence pose an important step towards harmonisation of NM testing.

  6. Simple Diagnostic Tests to Detect Toxic Alcohol Intoxications

    PubMed Central

    Shin, Jai Moo; Sachs, George; Kraut, Jeffrey A.

    2008-01-01

    Methanol, ethylene glycol, and diethylene glycol intoxications can produce visual disturbances, neurological disturbances, acute renal failure, pulmonary dysfunction, cardiac dysfunction, metabolic acidosis, and death. Metabolic acidosis and an increased serum osmolality are important clues to their diagnosis. The former reflects the organic acids produced by metabolism of the parent alcohol, while the latter is due to accumulation of the offending alcohol. However, neither the clinical nor the laboratory findings are specific for toxic alcohol ingestions. The definitive diagnosis of the alcohol intoxications is commonly based on detection of the alcohol or its metabolites in blood. Early diagnosis is important, because initiation of appropriate treatment can markedly lessen their morbidity and mortality. At present detection of the parent alcohol in body fluids is inferred from its measurement in blood. This measurement is often performed by specialty laboratories using expensive equipment, and a long delay between obtaining the specimen and getting the results is not unusual. In this report, we describe liquid- based tests that detect methanol, ethylene glycol, diethylene glycol, and ethanol in saliva. The tests are sensitive and they have different specificity for each of the alcohols facilitating distinction among them. The relatively high sensitivity and specificity of the tests as a whole will facilitate the rapid diagnosis of each of these alcohol intoxications. PMID:18940722

  7. Genetically Defined Strains in Drug Development and Toxicity Testing.

    PubMed

    Festing, Michael F W

    2016-01-01

    There is growing concern about the poor quality and lack of repeatability of many pre-clinical experiments involving laboratory animals. According to one estimate as much as $28 billion is wasted annually in the USA alone in such studies. A decade ago the FDA's "Critical path" white paper noted that "The traditional tools used to assess product safety-animal toxicology and outcomes from human studies-have changed little over many decades and have largely not benefited from recent gains in scientific knowledge. The inability to better assess and predict product safety leads to failures during clinical development and, occasionally, after marketing." Repeat-dose 28-days and 90-days toxicity tests in rodents have been widely used as part of a strategy to assess the safety of drugs and chemicals but their repeatability and power to detect adverse effects have not been formally evaluated.The guidelines (OECD TG 407 and 408) for these tests specify the dose levels and number of animals per dose but do not specify the strain of animals which should be used. In practice, almost all the tests are done using genetically undefined "albino" rats or mice in which the genetic variation, a major cause of inter-individual and strain variability, is unknown and uncontrolled. This chapter suggests that a better strategy would be to use small numbers of animals of several genetically defined strains of mice or rats instead of the undefined animals used at present. Inbred strains are more stable providing more repeatable data than outbred stocks. Importantly their greater phenotypic uniformity should lead to more powerful and repeatable tests. Any observed strain differences would indicate genetic variation in response to the test substance, providing key data. We suggest that the FDA and other regulators and funding organizations should support research to evaluate this alternative. PMID:27150081

  8. The effects of Dechlorane Plus on toxicity and mRNA expression in chicken embryos: a comparison of in vitro and in ovo approaches.

    PubMed

    Crump, Doug; Chiu, Suzanne; Gauthier, Lewis T; Hickey, Nathan J; Letcher, Robert J; Kennedy, Sean W

    2011-08-01

    Dechlorane Plus (DP) is an additive chlorinated flame retardant comprising two major isomers, syn- and anti-DP, that is used in a variety of commercial/industrial products. It has been detected in biotic and abiotic matrices including the eggs of herring gulls collected from the Laurentian Great Lakes. However, data on potential toxicological and molecular responses to exposure are lacking, especially for avian species. A combined in vitro/in ovo approach was used to determine concentration-dependent effects of DP in chicken embryonic hepatocytes (CEH) and chicken embryos following injection of DP into the air cell of eggs prior to incubation. Overt toxicity (i.e. cytotoxicity and pipping success) and mRNA expression levels of transcripts previously determined to be responsive to a brominated flame retardant were assessed in CEH and hepatic tissue. DP was not cytotoxic up to a maximum concentration of 3 μM in CEH, and no effects on pipping success were observed up to the highest nominal dose group of 500 ng/g egg. A significant shift in isomeric content of syn- and anti-DP was detected between stock solutions of the commercial mixture and hepatic tissue; the proportion of the syn-DP isomer increased from 0.34 to 0.65 with a concomitant decrease of anti-DP from 0.66 to 0.35. None of the mRNA transcripts changed as a result of in vitro or in ovo exposure to DP indicating that, although there was concordance between the two approaches, DP may evoke its toxicity through other modes of action. At current environmental exposure levels, no adverse effects of DP on embryonic viability or pathways associated with the genes assessed are predicted. PMID:21539933

  9. Comparison of mathematically-predicted toxic equivalents (TEQs) and bioassay-derived dioxin-equivalents (TCDD-EQs) in heron embryos

    USGS Publications Warehouse

    Rattner, B.; Hatfield, J.; Melancon, M.; Custer, T.; Tillitt, D.

    1995-01-01

    Pipping black-crowned night-heron (Nycticorax nycticorax) embryos were collected from an uncontaminated site (Chincoteague National Wildlife Refuge,VA) and three polluted sites (Cat Island, Green Bay, WI; Bair and West Marin Islands, San Francisco Bay, CA). Hepatic microsomal monooxygenases were induced up to 85-fold relative to the reference site, and was associated with concentrations of total PCBs and 11 PCB congeners that are presumed to concern.to express toxicity through the Ah receptor. TEQs [mathematically predicted; summed product of PCB congener concentrations using 5 different sets of toxic equivalency factors (TEFs)] were compared to TCDD-EQs [derived by bioassay; ethoxyresorufin-O-dealkylase activity of treated H411E rat hepatoma cells]. Although TEQs were up to 15-fold greater than TCDD-EQs, the pattern among sites was consistent and TEQs were highly correlated with TCDD-EQs. TEFs based on single congener mammalian studies yielded TEQs that greatly exceeded values from the H411E bioassay of field sample. TEFs generated from avian egg injection studies yielded TEQs that most closely approximated bioassay-derived TCDD-EQs. Cytochrome P450 parameters were related to TEQs and TCDD-EQs; adjusted r2 often exceeded 0.5 for the relation among mathematically predicted TEQs and cytochrome P450 measurements. These data document the general predictive value of TEQs and TCDD-EQs for P450 induction in field collected samples, but also indicate the need for development of TEFs for the species and biological end point of concern.

  10. Reproducibility of a life-cycle toxicity test with Daphnia magna

    SciTech Connect

    Parkhurst, B.R.; Forte, J.L.; Wright, G.P.

    1981-01-01

    Standardized chronic life-cycle toxicity testing procedures for aquatic species are described. The reproducibility of chronic toxicity and points using the static-renewal method with Daphnia magna are investigated. The objectives were to determine if the lowest rejected concentrations tested (LRCTs) obtained for six different toxicity criteria in static-renewal tests with acridine were reproducible over time and to determine the relative sensitivity and variability of the toxicity criteria. Two of the six toxicity criteria, numbers of young per brood and the young produced per female, were found to be reliable and sensitive for estimating the LRCT for acridine to D. magna. (RJC)

  11. Test of significant toxicity: a statistical application for assessing whether an effluent or site water is truly toxic.

    PubMed

    Denton, Debra L; Diamond, Jerry; Zheng, Lei

    2011-05-01

    The U.S. Environmental Protection Agency (U.S. EPA) and state agencies implement the Clean Water Act, in part, by evaluating the toxicity of effluent and surface water samples. A common goal for both regulatory authorities and permittees is confidence in an individual test result (e.g., no-observed-effect concentration [NOEC], pass/fail, 25% effective concentration [EC25]), which is used to make regulatory decisions, such as reasonable potential determinations, permit compliance, and watershed assessments. This paper discusses an additional statistical approach (test of significant toxicity [TST]), based on bioequivalence hypothesis testing, or, more appropriately, test of noninferiority, which examines whether there is a nontoxic effect at a single concentration of concern compared with a control. Unlike the traditional hypothesis testing approach in whole effluent toxicity (WET) testing, TST is designed to incorporate explicitly both α and β error rates at levels of toxicity that are unacceptable and acceptable, given routine laboratory test performance for a given test method. Regulatory management decisions are used to identify unacceptable toxicity levels for acute and chronic tests, and the null hypothesis is constructed such that test power is associated with the ability to declare correctly a truly nontoxic sample as acceptable. This approach provides a positive incentive to generate high-quality WET data to make informed decisions regarding regulatory decisions. This paper illustrates how α and β error rates were established for specific test method designs and tests the TST approach using both simulation analyses and actual WET data. In general, those WET test endpoints having higher routine (e.g., 50th percentile) within-test control variation, on average, have higher method-specific α values (type I error rate), to maintain a desired type II error rate. This paper delineates the technical underpinnings of this approach and demonstrates the benefits to both regulatory authorities and permitted entities. PMID:21305584

  12. Environmental concentrations of the cocaine metabolite benzoylecgonine induced sublethal toxicity in the development of plants but not in a zebrafish embryo-larval model.

    PubMed

    García-Cambero, J P; García-Cortés, H; Valcárcel, Y; Catalá, M

    2015-12-30

    Several studies have found cocaine and its main active metabolite benzoylecgonine (BE) in the aquatic environment and drinking water, derived from its consumption by humans as well as the inability of water treatment processes to eliminate it. A few studies have already investigated the ecotoxicology of BE to aquatic invertebrates, but none has still addressed the effects of BE on aquatic vertebrates or vascular plants. The goal of this publication is to provide information on the toxicity of environmental concentrations of BE during animal and vascular plant development, in order to contribute to a better understanding of the potential risk of this substance for the environment. BE induced alterations in mitochondrial activity and DNA levels of fern spores at environmental concentrations (1 ng L(-1)), which could disrupt gametophyte germination. However, BE at concentrations ranging from 1 ng L(-1) to 1 mg L(-1) did not disturb morphogenesis, hatching, heartbeat rate or larval motility in a zebrafish embryo-larval model. Adverse effects on ferns agree with the allelophathic role described for alkaloids and their unspecific interference with plant germination. Therefore, the anthropogenic dispersion of alkaloid allelochemicals may pose a risk for biodiversity and irrigated food production that should be further investigated. PMID:26340554

  13. The Syrian hamster embryo cells transformation assay identifies efficiently nongenotoxic carcinogens, and can contribute to alternative, integrated testing strategies.

    PubMed

    Benigni, Romualdo; Bossa, Cecilia; Tcheremenskaia, Olga; Battistelli, Chiara Laura; Giuliani, Alessandro

    2015-02-01

    The long-term carcinogenesis bioassays have played a central role in protecting human health, but for ethical and practical reasons their use is dramatically diminishing and the genotoxicity short-term tests have taken the pivotal role in the pre-screening of chemical carcinogenicity. However, this strategy cannot detect nongenotoxic carcinogens. Since up to 25% of IARC human carcinogens are recognized to have nongenotoxic mechanisms of action, the risk they pose to human health cannot be disregarded, and it is urgent to fill the gap in the tools for alternative testing. In this paper, we analyze from different perspectives the ability of Cell Transformation Assays to identify nongenotoxic carcinogens, and we conclude that the Syrian hamster embryo cells test is able to identify nongenotoxic carcinogens with 80-90% efficiency, and thus, can play an important role in integrated, alternative testing strategies. PMID:25813724

  14. STATISTICAL APPROACH TO PREDICTING CHRONIC TOXICITY OF CHEMICALS TO FISHES FROM ACUTE TOXICITY TEST DATA

    EPA Science Inventory

    A comprehensive approach to predicting chronic toxicity from cute toxicity data was developed in which simultaneous onsideration is given to concentration, degree of response, and ime course of effect. onsistent endpoint (lethality) and degree of response (0 percent) were used to...

  15. MARINE COMPLEX EFFLUENT TOXICITY PROGRAM: TEST SENSITIVITY, REPEATAELITY AND RELEVANCE TO RECEIVING WATER TOXICITY

    EPA Science Inventory

    In March 1984, the U.S. Environmental Protection Agency (EPA) issued a significant change in procedures regulating toxic materials in effluents through the National Pollutant Discharge Elimination System (NPDES). Concurrent with this toxicity-based effluent control policy, the EP...

  16. Interlaboratory variability of amphipod sediment toxicity tests in a cooperative regional monitoring program.

    PubMed

    Bay, Steven M; Jirik, Andrew; Asato, Stanford

    2003-01-01

    Marine sediment toxicity tests are widely applied in monitoring programs, yet relatively little is known about the comparability of data from different laboratories. The need for comparability information is increased in cooperative monitoring programs, where multiple laboratories (often with variable skill levels) perform toxicity tests. An interlaboratory comparison exercise was conducted among seven laboratories in order to document the comparability of sediment toxicity measurements during the Bight '98 regional sediment survey in southern California. Sediments from four stations in Los Angeles and Long Beach Harbors were tested using a 10-day survival test of the amphipod Eohaustorius estuarius. All laboratories successfully performed the sediment test and associated reference toxicant test. Statistically significant differences were found in mean amphipod survival rates among some laboratories for the field-collected sediments, but there was little evidence of a consistent bias among laboratories. Although the reference toxicant test indicated a five-fold variation in test sensitivity among laboratories, these results were not accurate predictors of interlaboratory performance for the sediment tests. The laboratories demonstrated excellent concordance (Kendall's W = 0.91) in ranking the field-collected sediments by toxicity. Agreement on classifying the sediments into categories (nontoxic, moderately toxic, and highly toxic) based upon the percent of survival was best for highly toxic sediments. An analysis of test precision based upon the variance among replicates within a test indicated that the measured survival rate for a sample may vary by up to 12 percentage points from the actual response. PMID:12620020

  17. Results of acute and chronic toxicity tests conducted at SRS NPDES outfalls, July--October 1991

    SciTech Connect

    Specht, W.L.

    1992-01-01

    Acute (48 hour LC50) and chronic (7-day reproductive impairment) toxicity tests were conducted on Ceriodaphnia dubia in water collected from 53 NPDES outfalls. All tests were conducted at the in-stream waste concentration. only 12 of the 53 outfalls showed no evidence of toxicity. Twenty-eight of the outfalls were acutely toxic, often producing 100% mortality during the first day of exposure. Fourteen outfalls had no discharge at the time of sampling and could not be tested. Three outfalls were not tested because their toxicity has been adequately characterized in other investigations. Elevated concentrations of total residual chlorine are suspected to be responsible for the observed toxicity of many NPDES outfalls, particularly the sanitary wastewater treatment plants. Chemical data from previous studies indicate that metals may also be present in toxic concentrations at many outfalls. Toxicity identification and reduction options are discussed.

  18. Early Evolution of the Toxicity Identification Evaluation Process: Contributions from the USEPA Effluent Testing Program

    EPA Science Inventory

    As part of its whole effluent testing program, the USEPA developed an effects-directed analysis (EDA) approach to identifying the cause of toxicity in toxic effluents or ambient waters, an EDA process termed a “Toxicity Identification Evaluation” (TIE), which is the focus of this...

  19. Early Evolution of the Toxicity Identification Evaluation Process: Contributions from the USEPA Effluent Testing Program

    EPA Science Inventory

    As part of its whole effluent testing program, the USEPA developed an effects-directed analysis (EDA) approach to identifying the cause of toxicity in toxic effluents or ambient waters, an EDA process termed a Toxicity Identification Evaluation (TIE), which is the focus of this...

  20. Methods For Collecting , Culturing And Performing Toxicity Tests With Daphnia ambigua

    SciTech Connect

    Specht, Winona L.

    2005-07-01

    Toxicity tests conducted on water collected from impacted locations in SRS streams often failed chronic toxicity tests and sometimes failed acute toxicity tests (Specht 1995). These findings prompted SRS to determine the cause of the failures. Some SRS NPDES outfalls were also failing chronic toxicity tests, even though no toxicant could be identified and when TIEs were performed, none of the TIE treatments removed the toxicity. Ultimately, it was determined that the failures were due to the low hardness of SRS surface waters, rather than to the presence of a toxicant. The species of cladoceran that the EPA recommends for toxicity testing, Ceriodaphnia dubia, is stressed by the very low hardness of SRS waters. SRS developed an alternate species toxicity test that is similar to the EPA test, but uses an indigenous cladoceran, Daphnia ambigua (Specht and Harmon, 1997; Harmon et al., 2003). In 2001, SCDHEC approved the use of D. ambigua for toxicity testing at SRS, contingent upon approval by EPA Region 4. In 2002, EPA Region 4 approved the use of this species for compliance toxicity testing at SRS. Ultimately, the use of this species demonstrated that SRS effluents were not toxic, and most toxicity testing requirements were removed from the NPDES permit that was issued in December 2003, with the exception of one round of chronic definitive testing on outfalls A-01, A-11, and G-10 just before the next NPDES permit application is submitted to SCDHEC. Although the alternate species test was developed at SRS (1996-1998), the culture was transferred to a contract toxicity testing lab (ETT Environmental) located in Greer, SC in 1998. ETT Environmental became certified by SCDHEC to perform toxicity tests using D. ambigua in 2002, and at this time is the only laboratory certified by SCDHEC to perform tests with this species. Because of the expense associated with maintaining the D. ambigua culture for several years when no toxicity testing is required, SRS decided to suspend financial support associated with maintaining the cultures until testing is needed. The purpose of this document is to provide guidance on how to establish a laboratory culture of D. ambigua so that a culture can be restarted when needed.

  1. Preimplantation Genetic Diagnosis: Prenatal Testing for Embryos Finally Achieving Its Potential.

    PubMed

    Stern, Harvey J

    2014-01-01

    Preimplantation genetic diagnosis was developed nearly a quarter-century ago as an alternative form of prenatal diagnosis that is carried out on embryos. Initially offered for diagnosis in couples at-risk for single gene genetic disorders, such as cystic fibrosis, spinal muscular atrophy and Huntington disease, preimplantation genetic diagnosis (PGD) has most frequently been employed in assisted reproduction for detection of chromosome aneuploidy from advancing maternal age or structural chromosome rearrangements. Major improvements have been seen in PGD analysis with movement away from older, less effective technologies, such as fluorescence in situ hybridization (FISH), to newer molecular tools, such as DNA microarrays and next generation sequencing. Improved results have also started to be seen with decreasing use of Day 3 blastomere biopsy in favor of polar body or Day 5 trophectoderm biopsy. Discussions regarding the scientific, ethical, legal and social issues surrounding the use of sequence data from embryo biopsy have begun and must continue to avoid concern regarding eugenic or inappropriate use of this technology. PMID:26237262

  2. Preimplantation Genetic Diagnosis: Prenatal Testing for Embryos Finally Achieving Its Potential

    PubMed Central

    Stern, Harvey J.

    2014-01-01

    Preimplantation genetic diagnosis was developed nearly a quarter-century ago as an alternative form of prenatal diagnosis that is carried out on embryos. Initially offered for diagnosis in couples at-risk for single gene genetic disorders, such as cystic fibrosis, spinal muscular atrophy and Huntington disease, preimplantation genetic diagnosis (PGD) has most frequently been employed in assisted reproduction for detection of chromosome aneuploidy from advancing maternal age or structural chromosome rearrangements. Major improvements have been seen in PGD analysis with movement away from older, less effective technologies, such as fluorescence in situ hybridization (FISH), to newer molecular tools, such as DNA microarrays and next generation sequencing. Improved results have also started to be seen with decreasing use of Day 3 blastomere biopsy in favor of polar body or Day 5 trophectoderm biopsy. Discussions regarding the scientific, ethical, legal and social issues surrounding the use of sequence data from embryo biopsy have begun and must continue to avoid concern regarding eugenic or inappropriate use of this technology. PMID:26237262

  3. Effects of food and water quality on culturing and toxicity testing of Ceriodaphnia dubia: Final report

    SciTech Connect

    Cooney, J.D.; DeGraeve, G.M.; Moore, E.L.; Palmer, W.D.; Pollock, T.L.

    1988-05-01

    Effects of Food and Water Quality on Culturing and Toxicity Testing of Ceriodaphnia dubia. Tests of eight diets for culturing Ceriodaphnia have identified two foods that support high and consistent survival and reproduction. Use of these foods can contribute to the precision of EPA recommended effluent toxicity tests.

  4. SEED GERMINATION AND ROOT ELONGATION TOXICITY TESTS IN HAZARDOUS WASTE SITE EVALUATION: METHODS DEVELOPMENT AND APPLICATIONS

    EPA Science Inventory

    Seed germination tests measure soil toxicity directly, while root elongation tests consider the indirect effects of water-soluble constituents which may be present in site-samples. n the seed germination toxicity test, site-soil is mixed with a reference soil to yield exposure co...

  5. REVIEW OF THE CURRENT STATUS OF MARINE ALGAL TOXICITY TESTING IN THE UNITED STATES

    EPA Science Inventory

    Algal toxicity testing is not new, but only within the past few years have data from such testing been used to help set standards for allowable contamination. arly toxicity testing with marine algae used a few planktonic species with inhibition of growth as the primary endpoint. ...

  6. Handling nonnormality and variance heterogeneity for quantitative sublethal toxicity tests.

    PubMed

    Ritz, Christian; Van der Vliet, Leana

    2009-09-01

    The advantages of using regression-based techniques to derive endpoints from environmental toxicity data are clear, and slowly, this superior analytical technique is gaining acceptance. As use of regression-based analysis becomes more widespread, some of the associated nuances and potential problems come into sharper focus. Looking at data sets that cover a broad spectrum of standard test species, we noticed that some model fits to data failed to meet two key assumptions-variance homogeneity and normality-that are necessary for correct statistical analysis via regression-based techniques. Failure to meet these assumptions often is caused by reduced variance at the concentrations showing severe adverse effects. Although commonly used with linear regression analysis, transformation of the response variable only is not appropriate when fitting data using nonlinear regression techniques. Through analysis of sample data sets, including Lemna minor, Eisenia andrei (terrestrial earthworm), and algae, we show that both the so-called Box-Cox transformation and use of the Poisson distribution can help to correct variance heterogeneity and nonnormality and so allow nonlinear regression analysis to be implemented. Both the Box-Cox transformation and the Poisson distribution can be readily implemented into existing protocols for statistical analysis. By correcting for nonnormality and variance heterogeneity, these two statistical tools can be used to encourage the transition to regression-based analysis and the depreciation of less-desirable and less-flexible analytical techniques, such as linear interpolation. PMID:19364187

  7. Rapid toxicity testing based on mitochondrial respiratory activity

    SciTech Connect

    Haubenstricker, M.E. ); Holodnick, S.E.; Mancy, K.H. ); Brabec, M.J. )

    1990-05-01

    The need exists for rapid and inexpensive methods to determine the health effects of environmental contaminants on biological systems. One of the current research approaches for assessing cytotoxicity is to monitor the respiratory activity of the mitochondrion, a sensitive, nonspecific subcellular target site. Detected changes in mitochondrial function after the addition of a test chemical could be correlated to toxic effects. Mitochondrial respiration can be characterized by three indices: state 3 and state 4 respiratory rates, and the respiratory control ratio (RCR). State 4, the idle or resting state, results when coupled mitochondrial respire in a medium containing inorganic phosphate and a Kreb's cycle substrate in the absence of a phosphate acceptor such as adenosine diphosphate (ADP). In the presence of ADP the respiration rate increases to a maximum (state 3), accompanied by phosphorylation of ADP to adenosine triphosphate (ATP). The ratio of state 3 to state 4, or RCR, indicates how tightly the oxidative phosphorylation process is coupled. The synthesis of ATP by mitochondria is influenced by a number of compounds, most of which are either uncouplers or inhibitors.

  8. Comparison of an Ampelisca abdita growth rate test with other standard amphipod sediment toxicity tests

    SciTech Connect

    Schafer, K.; Weston, D.P.

    1995-12-31

    Amphipod crustaceans are often used to measure the toxicity of bulk sediments. Acute lethal bioassays are commonly employed, but this study investigated the potential for using a chronic growth bioassay with Ampelisca abdita. A potential advantage of this method is that the growth rate could be a more sensitive measure of contamination than mortality. Growth rates for A. abdita in sediments spiked with cadmium and crude oil were compared to mortality rates in A. abdita, Eohaustorius estuaries, and Rhepoxynius abronius in sediments with the same concentrations of contaminants. A. abdita was more sensitive to cadmium than the other two species. For crude oil, there was a significant shift in size distribution from the control even at concentrations as low as 150 mg/kg of oil. The standard acute lethal tests for all species, on the other hand, did not show significant mortality until at least 1,600 mg/kg. The results confirm that growth rates are a more sensitive indicator of toxicity, and to at least the three contaminants tested, A. abdita is as sensitive as E. estuarius and R. abronius. This study also confirmed the reported high mortality rates of E. estuaries in San Francisco Bay sediments. The causes of this high mortality are unknown but give further reason for using A. abdita for toxicity tests in this region.

  9. Field validation of 10-day freshwater sediment toxicity tests using Hyalella azteca and Chironomus tentans

    SciTech Connect

    Becker, D.S.; Bigham, G.N.

    1995-12-31

    Two of the toxicity tests commonly used to evaluate freshwater sediments are the 10-day amphipod (Hyalella azteca) and chironomid (Chironomus tentans) tests. EPA and ASTM have recently developed standardized protocols for these tests. Although both tests are considered sensitive indicators of sediment toxicity, little information exists on how well test results correspond to adverse biological effects in the field. In this study, the lethal and sublethal (i.e., biomass) responses of the two toxicity tests were compared with alterations of benthic macroinvertebrate assemblages (i.e., benthic effects) at 56 stations in Onondaga Lake, New York. The lake has received municipal and industrial discharges for more than 100 years, and sediment chemical concentrations range widely throughout the lake. Toxicity results for Onondaga Lake were compared with reference conditions using the t-test, and benthic effects were determined using classification analysis of log-transformed taxa abundances. In general, a relatively high level of agreement was found between results of the toxicity tests and alterations of benthic assemblages. Significant (P < 0.05) correlations were found between all toxicity test endpoints and taxa richness of benthic assemblages. In addition, significant concordance (P {le} 0.01, binomial test) was found between toxicity designations for the 56 stations based on toxicity tests and toxicity designations based on benthic effects. Despite the general level of agreement among the various biological indicators, chironomid biomass and benthic effects were found to be the most sensitive indicators of toxicity, whereas amphipod survival and biomass were the least sensitive indicators. This study suggests that results of the 10-day amphipod and chironomid toxicity tests are highly predictive of adverse biological effects in the field.

  10. Applicability of ambient toxicity testing to national or regional water-quality assessment

    USGS Publications Warehouse

    Elder, John F.

    1990-01-01

    Comprehensive assessment of the quality of natural waters requires a multifaceted approach. Descriptions of existing conditions may be achieved by various kinds of chemical and hydrologic analyses, whereas information about the effects of such conditions on living organisms depends on biological monitoring. Toxicity testing is one type of biological monitoring that can be used to identify possible effects of toxic contaminants. Based on experimentation designed to monitor responses of organisms to environmental stresses, toxicity testing may have diverse purposes in water-quality assessments. These purposes may include identification of areas that warrant further study because of poor water quality or unusual ecological features, verification of other types of monitoring, or assessment of contaminant effects on aquatic communities. Toxicity-test results are most effective when used as a complement to chemical analyses, hydrologic measurements, and other biological monitoring. However, all toxicity-testing procedures have certain limitations that must be considered in developing the methodology and applications of toxicity testing in any large-scale water-quality-assessment program. A wide variety of toxicity-test methods have been developed to fulfill the needs of diverse applications. The methods differ primarily in the selections made relative to four characteristics: (1) test species, (2) endpoint (acute or chronic), (3) test-enclosure type, and (4) test substance (toxicant) that functions as the environmental stress. Toxicity-test approaches vary in their capacity to meet the needs of large-scale assessments of existing water quality. Ambient testing, whereby the test organism is exposed to naturally occurring substances that contain toxicant mixtures in an organic or inorganic matrix, is more likely to meet these needs than are procedures that call for exposure of the test organisms to known concentrations of a single toxicant. However, meaningful interpretation of ambient test results depends on the existence of accompanying chemical analysis of the ambient media. The ambient test substance may be water or sediments. Sediment tests have had limited application, but they are useful because most toxicants tend to accumulate in sediments and many test species either inhabit the sediments or are in frequent contact with them. Biochemical testing methods, which have been developing rapidly in recent years, are likely to be among the most useful procedures for large-scale water-quality assessments. They are relatively rapid and simple, and more. importantly, they focus on biochemical changes that are the initial responses of virtually all organisms to environmental stimuli. Most species are sensitive to relatively few toxicants, and their sensitivities vary as conditions change. Therefore, each test method has particular uses and limitations, and no single test has universal applicability. One of the most informative approaches to toxicity testing is to combine biochemical tests with other test methods in a 'battery of tests' that is diversified enough to characterize different types of toxicants and different trophic levels. However, such an approach can be costly, and if not carefully designed, it may not yield enough additional information to warrant the additional cost. The application of toxicity tests to large-scale water-quality assessments is hampered by a number of difficulties. Toxicity tests often are not sensitive enough to enable detection of most contaminant problems in the natural environment. Furthermore, because sensitivities among different species and test conditions can be highly variable, conclusions about the toxicant problems of an ecosystem are strongly dependent on the test procedure used. In addition, the experimental systems used in toxicity tests cannot replicate the complexity or variability of natural conditions, and positive test results cannot identify the source or nature of

  11. Wheat seed embryo excision enables the creation of axenic seedlings and Koch's postulates testing of putative bacterial endophytes.

    PubMed

    Robinson, Rebekah J; Fraaije, Bart A; Clark, Ian M; Jackson, Robert W; Hirsch, Penny R; Mauchline, Tim H

    2016-01-01

    Early establishment of endophytes can play a role in pathogen suppression and improve seedling development. One route for establishment of endophytes in seedlings is transmission of bacteria from the parent plant to the seedling via the seed. In wheat seeds, it is not clear whether this transmission route exists, and the identities and location of bacteria within wheat seeds are unknown. We identified bacteria in the wheat (Triticum aestivum) cv. Hereward seed environment using embryo excision to determine the location of the bacterial load. Axenic wheat seedlings obtained with this method were subsequently used to screen a putative endophyte bacterial isolate library for endophytic competency. This absence of bacteria recovered from seeds indicated low bacterial abundance and/or the presence of inhibitors. Diversity of readily culturable bacteria in seeds was low with 8 genera identified, dominated by Erwinia and Paenibacillus. We propose that anatomical restrictions in wheat limit embryo associated vertical transmission, and that bacterial load is carried in the seed coat, crease tissue and endosperm. This finding facilitates the creation of axenic wheat plants to test competency of putative endophytes and also provides a platform for endophyte competition, plant growth, and gene expression studies without an indigenous bacterial background. PMID:27151146

  12. Wheat seed embryo excision enables the creation of axenic seedlings and Koch’s postulates testing of putative bacterial endophytes

    PubMed Central

    Robinson, Rebekah J.; Fraaije, Bart A.; Clark, Ian M.; Jackson, Robert W.; Hirsch, Penny R.; Mauchline, Tim H.

    2016-01-01

    Early establishment of endophytes can play a role in pathogen suppression and improve seedling development. One route for establishment of endophytes in seedlings is transmission of bacteria from the parent plant to the seedling via the seed. In wheat seeds, it is not clear whether this transmission route exists, and the identities and location of bacteria within wheat seeds are unknown. We identified bacteria in the wheat (Triticum aestivum) cv. Hereward seed environment using embryo excision to determine the location of the bacterial load. Axenic wheat seedlings obtained with this method were subsequently used to screen a putative endophyte bacterial isolate library for endophytic competency. This absence of bacteria recovered from seeds indicated low bacterial abundance and/or the presence of inhibitors. Diversity of readily culturable bacteria in seeds was low with 8 genera identified, dominated by Erwinia and Paenibacillus. We propose that anatomical restrictions in wheat limit embryo associated vertical transmission, and that bacterial load is carried in the seed coat, crease tissue and endosperm. This finding facilitates the creation of axenic wheat plants to test competency of putative endophytes and also provides a platform for endophyte competition, plant growth, and gene expression studies without an indigenous bacterial background. PMID:27151146

  13. Microfluidic EmbryoSort technology: towards in flow analysis, sorting and dispensing of individual vertebrate embryos

    NASA Astrophysics Data System (ADS)

    Fuad, Nurul M.; Wlodkowic, Donald

    2013-12-01

    The demand to reduce the numbers of laboratory animals has facilitated the emergence of surrogate models such as tests performed on zebrafish (Danio rerio) or African clawed frog's (Xenopus levis) eggs, embryos and larvae. Those two model organisms are becoming increasingly popular replacements to current adult animal testing in toxicology, ecotoxicology and also in drug discovery. Zebrafish eggs and embryos are particularly attractive for toxicological analysis due their size (diameter 1.6 mm), optical transparency, large numbers generated per fish and very straightforward husbandry. The current bottleneck in using zebrafish embryos for screening purposes is, however, a tedious manual evaluation to confirm the fertilization status and subsequent dispensing of single developing embryos to multitier plates to perform toxicity analysis. Manual procedures associated with sorting hundreds of embryos are very monotonous and as such prone to significant analytical errors due to operator's fatigue. In this work, we present a proofof- concept design of a continuous flow embryo sorter capable of analyzing, sorting and dispensing objects ranging in size from 1.5 - 2.5 mm. The prototypes were fabricated in polymethyl methacrylate (PMMA) transparent thermoplastic using infrared laser micromachining. The application of additive manufacturing processes to prototype Lab-on-a-Chip sorters using both fused deposition manufacturing (FDM) and stereolithography (SLA) were also explored. The operation of the device was based on a revolving receptacle capable of receiving, holding and positioning single fish embryos for both interrogation and subsequent sorting. The actuation of the revolving receptacle was performed using a DC motor and/or microservo motor. The system was designed to separate between fertilized (LIVE) and non-fertilized (DEAD) eggs, based on optical transparency using infrared (IR) emitters and receivers.

  14. Toxicity of polybrominated diphenyl ethers (de-71) in chicken (Gallus gallus), mallard (Anas platyrhynchos), and American kestrel (Falco sparverius) embryos and hatchlings

    USGS Publications Warehouse

    McKernan, M.A.; Rattner, B.A.; Hale, R.C.; Ottinger, M.A.

    2009-01-01

    Embryonic survival, pipping and hatching success, and sublethal biochemical, endocrine, and histological endpoints were examined in hatchling chickens (Gallus gallus), mallards (Anas platyrhynchos), and American kestrels (Falco sparverius) following air cell administration of a pentabrominated diphenyl ether (penta-BDE; DE-71) mixture (0.01-20 mu g/g egg) or polychlorinated biphenyl (PCB) congener 126 (3,3', 4,4', 5-pentachlorobiphenyl; 0.002 mu g/g egg). The penta-BDE decreased pipping and hatching success at concentrations of 10 and 20 mu g/g egg in kestrels but had no effect on survival endpoints in chickens or mallards. Sublethal effects in hatchling chickens included ethoxyresorufin-O-dealkylase (EROD) induction and histological changes in the bursa, but these responses were not observed in other species. Polychlorinated biphenyl congener 126 (positive control) reduced survival endpoints in chicken and kestrel embryos and caused sublethal effects (EROD induction, reduced bursal mass and follicle size) in chickens. Mallards were clearly less sensitive than the other species to administered penta-BDE and PCB 126. In a second experiment, the absorption of penta-BDE (11.1 mu g/g egg, air cell administered during early development) into the contents of chicken and kestrel eggs was determined at various intervals (24 h postinjection, midincubation, and pipping). By pipping, 29% of the penta-BDE administered dose was present in the egg contents in chickens, and 18% of the administered dose was present in kestrel egg contents. Based on uptake in kestrels, the lowest-observed-effect level on pipping and hatching success may be as low as 1.8 mu g total penta-BDE/g egg, which approaches concentrations detected in eggs of free-ranging birds. Because some penta-BDE congeners are still increasing in the environment, the toxic effects observed in the present study are cause for concern in wildlife.

  15. Pyrrolizidine Alkaloids: Testing for Toxic Constituents of Comfrey.

    ERIC Educational Resources Information Center

    Vollmer, John J.; And Others

    1987-01-01

    Discusses the possibilities of toxins present in medicinal herbs. Describes an experiment in which toxic constituents can be selectively detected by thin-layer chromatography and NMR spectroscopy. (TW)

  16. Toxicity of 8-Hydroxyquinoline in Cryprinus carpio Using the Acute Toxicity Test, Hepatase Activity Analysis and the Comet Assay.

    PubMed

    Yan, Shuaiguo; Chen, Lili; Dou, Xiaofei; Qi, Meng; Du, Qiyan; He, Qiaoqiao; Nan, Mingge; Chang, Zhongjie; Nan, Ping

    2015-08-01

    To evaluate the environmental toxicity of 8-hydroxyquinoline (8-HOQ), an important industrial raw material found in China's major ornamental fish, Cryprinus carpio, using the acute toxicity test, hepatase activity analysis and the comet assay. The results indicated that 8-HOQ had significant acute toxicity in adult C. carpio with a 96 h-LC50 of 1.15 and 0.22 mg L(-1) hepatic quinoline residues as assessed by HPLC. 8-HOQ also induced genotoxicity in the form of strand breaks in the DNA of hepatic cells as shown by the comet assay. With regard to physiological toxicity, 8-HOQ induced a decrease in the activities of hepatic GOT and GPT with increased exposure concentration and time. These data suggest that 8-HOQ may be toxic to the health of aquatic organisms when accidentally released into aquatic ecosystems. The data also suggest that the comet assay may be used in biomonitoring to determine 8-HOQ genotoxicity and hepatic GPT and GOT activities may be potential biomarkers of physiological toxicity. PMID:26067700

  17. COMPARATIVE TOXICITY TESTING OF SELECTED BENTHIC AND EPIBENTHIC ORGANISMS FOR THE DEVELOPMENT OF SEDIMENT QUALITY TEST PROTOCOLS

    EPA Science Inventory

    Sediment contamination has resulted in the need to develop an appropriate suite of toxicity tests to assess ecotoxicological impacts on estuarine ecosystems. Existing Environmental Protection Agency (EPA) protocols recommend a number of test organisms, including amphipods, polych...

  18. Triazole-induced gene expression changes in the zebrafish embryo.

    PubMed

    Hermsen, Sanne A B; Pronk, Tessa E; van den Brandhof, Evert-Jan; van der Ven, Leo T M; Piersma, Aldert H

    2012-09-01

    The zebrafish embryo is considered to provide a promising alternative test model for developmental toxicity testing. Most systems use morphological assessment of the embryos, however, microarray analyses may increase sensitivity and predictability of the test by detecting more subtle and detailed responses. In this study, we investigated the possibility of relating gene expression profiles of structurally similar chemicals tested in a single concentration, to a complete transcriptomic concentration-response of flusilazole (FLU). We tested five other triazoles, hexaconazole (HEX), cyproconazole (CYP), triadimefon (TDF), myclobutanil (MYC), and triticonazole (TTC) at equipotent concentrations based on morphological evaluation. Results showed that every compound had a different degree of regulation within their anti-fungal and developmental toxicity pathways, steroid biosynthesis and retinol metabolism, respectively. Assuming that the ratio between these pathways is relevant for efficacy compared to developmental toxicity, we found TTC was more efficient and CYP was more toxic compared to the other triazoles. With the approach used in this study we demonstrated that gene expression data allow more comprehensive assessment of compound effects by discriminating relative potencies using these specific gene sets. The zebrafish embryo model can therefore be considered a useful vertebrate model providing information of relevant pathways related to anti-fungal mechanism of action and toxicological activity. PMID:22664267

  19. Chronic toxicity bioassay with ceriodaphnia dubia': (1) An evaluation of a toxicity test-based approach for determining the sources of chronic toxicity; and (2) an evaluation of culture/dilution waters and diet as determinants of test outcomes

    SciTech Connect

    Francisco, D.E.; Elias, M.C.; LaRocca, C.A.; DiGiano, F.A.; Maerker, M.J.

    1993-08-01

    In order to limit the discharge of toxic materials in toxic amounts to the waterways of the United States, the U. S. Environmental Protection Agency (EPA) has developed a 7-day survival and reproduction toxicity bioassay test using the freshwater cladoceran, Ceriodaphnia dubia. The North Carolina Division of Environmental Management has developed a less expensive modification of this test called the NC Ceriodaphnia Mini-Chronic Pass/Fail Toxicity Test. These tests are used to monitor discharges from wastewater treatment plants. Failure to satisfy the requirements (evidence of either acute or chronic toxicity) of the discharge permit constitutes an effluent violation. Effluent violations require that studies be conducted to determine the means to eliminate the violation. The present investigation had three basic objectives: (1) to develop and test a toxicity test-based method for locating the sources of toxic materials discharged into a collection system; (2) to investigate the effect of culture/dilution water on the health of C. dubia and reproducibility of the NC Ceriodaphnia Mini-Chronic Pass/Fail Toxicity Test; and (3) to investigate the effect of 3 diets on the health and robustness of C. dubia.

  20. Evaluation of three soil toxicity tests used to monitor acceptable endpoints

    SciTech Connect

    Brinkmann, M.; Stroo, H.; Leuschner, A.; Leuteritz, D.; Stromberg, M.; Brourman, M.

    1995-12-31

    Three terrestrial toxicity tests were used to evaluate the efficacy of biological treatment of creosote and pentachlorophenol impacted soils at a Superfund site. Microtox, 5-day lettuce seed, and 14-day earthworm toxicity tests were performed on 10 soil samples at the beginning and end of 3 months of land treatment. Secondary endpoints of root length and earthworm weight loss were also evaluated. EC50 and LC50 values were calculated using a Trimmed Logit Statistical Program and compared to toxicity of 10 background samples collected from the site. Results for initial soils demonstrated toxicity with three of the five endpoints. End treatment results showed no measurable toxicity using all endpoints. Toxicity testing results are critical for obtaining regulatory approval for the full-scale treatment system. Post treatment closure requirements for the site will be based on bioassay results. Evaluation of the three tests used showed the Microtox test to be the most sensitive to this type of toxicity. Lettuce seed germination results were the least sensitive of the three primary endpoints chosen. Of the secondary endpoint criteria, root length demonstrated reliable EC50 values and showed toxicity trends similar to Microtox and earthworm tests. The earthworm weight loss endpoint was not a useful toxicity measurement at 14 days.

  1. Anodonta imbecillis copper sulfate reference toxicant test, Clinch River - Environmental Restoration Program (CR-ERP)

    SciTech Connect

    Simbeck, D.J.

    1997-06-01

    Reference toxicant testing using juvenile freshwater mussels was conducted as part of the CR-ERP biomonitoring study of Clinch River sediments to assess the sensitivity of test organisms and the overall performance of the test. Tests were conducted using moderately hard synthetic water spiked with known concentrations of copper as copper sulfate. Toxicity testing of copper sulfate reference toxicant was conducted from May 12-21, 1993. The organisms used for testing were juvenile fresh-water mussels (Anodonta imbecillis). Results from this test showed an LC{sub 50} value of 1.12 mg Cu/L which is lower than the value of 2.02 mg Cu/L obtained in a previous test. Too few tests have been conducted with copper as the toxicant to determine a normal range of values.

  2. Factors affecting toxicity test endpoints in sensitive life stages of native Gulf of Mexico species.

    PubMed

    Echols, B S; Smith, A J; Rand, G M; Seda, B C

    2015-05-01

    Indigenous species are less commonly used in laboratory aquatic toxicity tests compared with standard test species due to (1) limited availability lack of requisite information necessary for their acclimation and maintenance under laboratory conditions and (2) lack of information on their sensitivity and the reproducibility of toxicity test results. As part of the Natural Resource Damage Assessment aquatic toxicity program in response to the Deepwater Horizon Oil incident (2010), sensitive life stages of native Gulf of Mexico species were evaluated in laboratory toxicity tests to determine the potential effects of the spill. Fish (n = 5) and invertebrates (n = 2) selected for this program include the following: the Florida pompano (Trachinotus carolinus), red drum (Sciaenops ocellatus), spotted sea trout (Cynoscion nebulosus), cobia (Rachycentron canadum), red porgy (Pagrus pagrus), blue crab (Callinectes sapidus), and the common moon jellyfish (Aurelia aurita). Initially in the program, to establish part of the background information, acute tests with reference toxicants (CdCl2, KCl, CuSO4) were performed with each species to establish data on intraspecies variability and test precision as well as identify other factors that may affect toxicity results. Median lethal concentration (LC50) values were calculated for each acute toxicity test with average LC50 values ranging from 248 to 862 mg/L for fish exposures to potassium chloride. Variability between test results was determined for each species by calculating the coefficient of variation (%CV) based on LC50 values. CVs ranged from 11.2 % for pompano (96-h LC50 value) to 74.8 % for red porgy 24-h tests. Cadmium chloride acute toxicity tests with the jellyfish A. aurita had the lowest overall CV of 3.6 %. By understanding acute toxicity to these native organisms from a compound with known toxicity ranges and the variability in test results, acute tests with nonstandard species can be better interpreted and used appropriately when determining risk. PMID:25563746

  3. Strategies for integrating transcriptional profiling into high throughput toxicity testing (SOT Symposium Workshop presentation)

    EPA Science Inventory

    Presentation Description: The release of the National Research Council’s Report “Toxicity Testing in the 21st Century: A Vision and a Strategy” in 2007 initiated a broad-based movement in the toxicology community to re-think how toxicity testing and risk assessment are performed....

  4. Strategies for integrating transcriptional profiling into high throughput toxicity testing (SOT Symposium Workshop presentation)

    EPA Science Inventory

    Presentation Description: The release of the National Research Councils Report Toxicity Testing in the 21st Century: A Vision and a Strategy in 2007 initiated a broad-based movement in the toxicology community to re-think how toxicity testing and risk assessment are performed....

  5. 40 CFR 799.9355 - TSCA reproduction/developmental toxicity screening test.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 33 2013-07-01 2013-07-01 false TSCA reproduction/developmental toxicity screening test. 799.9355 Section 799.9355 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) IDENTIFICATION OF SPECIFIC CHEMICAL SUBSTANCE AND MIXTURE TESTING REQUIREMENTS Health...

  6. The Adverse Outcome Pathway: A conceptual framework to support toxicity testing in the 21st century

    EPA Science Inventory

    The field of regulatory toxicity testing is at a turning point. The U.S. National Research Council (NRC) envisioned a shift away from traditional toxicity testing and towards a focused effort to explore and understand pathways perturbed by biologically active substances or their ...

  7. PERIPHYTON PHOTOSYNTHESIS AS AN INDICATOR OF EFFLUENT TOXICITY: RELATIONSHIP TO EFFECTS ON ANIMAL TEST SPECIES

    EPA Science Inventory

    The use of aquatic plants in effluent toxicity evaluations is uncommon despite the availability of test methods and numerous recommendations for their use. It has been assumed that aquatic plants are less sensitive than animal test species and consequently, results from toxicity ...

  8. New technologies and approaches in toxicity testing and risk assessment (ESOT)

    EPA Science Inventory

    The release of the National Research Council’s Report “Toxicity Testing in the 21st Century: A Vision and a Strategy” in 2007 initiated a broad-based movement in the toxicology community to re-think how toxicity testing and risk assessment are performed. Multiple efforts in the ...

  9. USER'S GUIDE FOR CONDUCTING LIFE-CYCLE CHRONIC TOXICITY TESTS WITH FATHEAD MINNOWS (PIMEPHALES PROMELAS)

    EPA Science Inventory

    This paper represents the latest 'state-of-the-art' procedural guide for conducting life-cycle chronic toxicity tests with fathead minnow (Pimephales promelas). These new procedures are based on recent evaluations of published toxicity tests and methods used by aquatic toxicologi...

  10. A comparison of relative toxicity rankings by some small-scale laboratory tests

    NASA Technical Reports Server (NTRS)

    Hilado, C. J.; Cumming, H. J.

    1977-01-01

    Small-scale laboratory tests for fire toxicity, suitable for use in the average laboratory hood, are needed for screening and ranking materials on the basis of relative toxicity. The performance of wool, cotton, and aromatic polyamide under several test procedures is presented.

  11. TERATOGENICITY OF CYCLOPHOSPHAMIDE IN A COUPLED MICROSOMAL ACTIVATING/EMBRYO CULTURE SYSTEM

    EPA Science Inventory

    Using the coupled microsomal activating/embryo culture system, in vitro experiments were performed to establish the role of metabolism in the embryo toxicity and teratogenicity of cyclophosphamide. Cyclophosphamide in the coupled microsomal activating/embryo culture system produc...

  12. Population growth rate determinants for Arbacia: Evaluating ecological relevance of toxicity test endpoints

    SciTech Connect

    Nacci, D.; Gleason, T.; Munns, W.R. Jr.

    1995-12-31

    A population dynamics model for the sea urchin, Arbacia punctulata, was recently developed incorporating life stage endpoints frequently measured in acute and chronic toxicity studies. Model elasticity analysis was used to demonstrate that population growth rate was influenced most by adult survival and least by early life stage success, calling into question the ecological relevance of results from standardized Arbacia fertilization and larval development toxicity tests. Two approaches were used to continue this evaluation. Actual and hypothetical dose-response curves for toxicant exposures over multiple life stages were used to evaluate contributions to population growth rate of stage-specific toxicant effects. Additionally, relationships between critical life stages were developed from laboratory data for Arbacia. The results of this analysis underscore the importance of understanding both endpoint sensitivity to toxicants and sensitivity of population growth rate to test endpoints in determining the ecological relevance of toxicity tests results.

  13. Terrestrial isopods -- a good choice for toxicity testing of pollutants in the terrestrial environment

    SciTech Connect

    Drobne, D.

    1997-06-01

    Terrestrial isopods are suitable invertebrates for testing the relative toxicities of chemicals present in the terrestrial environment. Terrestrial isopods respond in numerous ways to elevated concentrations of chemicals in their food, but only a few of these responses can be used as toxicological endpoints. The most suitable are changes in reproduction, food consumption, moult cycle duration, and structure of the digestive glands. These responses are able to provide accurate indications of sublethal toxicity. Toxicity tests with terrestrial isopods could be much more reliable through the use of positive controls. A positive control with a reference toxicant could also be supplemented by a reference endpoint. The most suitable reference endpoint is change of food consumption rate. Toxicity testing with terrestrial isopods is a very promising method for fast, routine, and inexpensive laboratory determination of the relative toxicities of chemicals in the terrestrial environment.

  14. Evaluation of a chronic toxicity test based upon the use of luminescent bacteria

    SciTech Connect

    Bulich, A.; Huynh, H.; Ulitzur, S.

    1995-12-31

    The authors have previously reported on the initial development of a new chronic toxicity test which utilizes the Microtox strain of luminescent bacteria as the test organism. These freeze-dried luminescent bacteria, following inoculation into a special growth medium, initiate a series of reproductive cycles and numerous metabolic pathways resulting in the production of light. Toxic materials which inhibit any aspect of the organisms` reproductive cycle or light production are detected in low levels. Critical test parameters associated with this test were evaluated and the resulting information summarized herein. These parameters include the following: (1) Data calculation methods and software used to capture, analyze, and present test data. (2) Evaluation of test data from 70 copper sulfate experiments with resulting information about test precision, sensitivity, and variability of light production from control cuvettes. 3. An analysis and summary of comparative test data from over 50 effluent samples which were co-tested with the Ceriodaphnia dubia 7 day chronic toxicity test and the Microtox 22 hour chronic toxicity test. (4) Luminescent bacteria chronic toxicity test data from whole effluent samples compared with corresponding test data from whole sediment toxicity test methods.

  15. Development and application of a marine sediment porewater toxicity test using algal spores

    SciTech Connect

    Hooten, R.; Carr, R.S.

    1995-12-31

    An acute pore water toxicity test protocol using germination and growth of marine macroalgae as endpoints was developed to indicate the presence of toxic compounds in marine/estuarine and sediment porewater samples. Zoospores collected from Ulva fasciata and U. lactuca were used as test organisms. Preliminary results with sodium dodecyl sulfate (SDS, a reference toxicant) indicate that zoospores germination and growth of embryonic gametophytes are as sensitive as the sea urchin fertilization and embryological development toxicity tests. Algal germination and growth data for copper, mercury and other metals will be presented. The results of tests utilizing this algal assay with sediment pore water from contaminated sediments will be compared with more traditional sediment toxicity test methods.

  16. ASSESSING CONTAMINANT SENSITIVITY OF ENDANGERED AND THREATENED SPECIES: EFFLUENT TOXICITY TESTS

    EPA Science Inventory

    Toxicity tests using standard effluent test procedures were conducted (EPA 1994) with Ceriodaphnia dubia and fathead minnows and four endangered fish species: bonytail chub (Gila elegans), Colorado squawfish (Ptychocheilus lucias ), razorback sucker (Xyrauchen texanus) and Gila t...

  17. Assessing Contaminant Sensitivity of Endangered and Threatened Aquatic Species: Part III. Effluent Toxicity Tests

    EPA Science Inventory

    This paper reports on the results of chronic toxicity tests conducted with common surrogate species, and several threatened and endangered species for which there were excess artificially propagated stock to allow direct testing.

  18. ADVANTAGES OF USING REGRESSION ANALYSIS TO CALCULATE RESULTS OF CHRONIC TOXICITY TESTS

    EPA Science Inventory

    Although it is traditional to calculate results of chronic toxicity tests using hypothesis testing to detect statistically significant differences from the control, calculation of results using regression analysis offers several major advantages. Regression analysis can directly ...

  19. AN EVALUATION OF THE 7-DAY TOXICITY TEST WITH AMERICAMYSIS BAHIA (FORMERLY MYSIDOPSIS BAHIA)

    EPA Science Inventory

    The 7-d test measuring survival, growth, and fecundity of Americamysis bahia formerly Mysidopsis bahia) was developed for estimating the chronic toxicity of effluents and associated receiving waters for National Pollutant Discharge Elimination System permits. Currently, this test...

  20. Canine pituitary-testicular function in relation to toxicity testing.

    PubMed

    James, R W; Crook, D; Heywood, R

    1979-08-01

    The physiological onset and establishment of adult pituitary-testicular function has been characterised for Beagle dogs. Testicular measurements, circulating prolactin, luteinising hormone and testosterone concentrations, semen analyses and the histological appearance of testicular biopsies, suggest that male Beagles attain sexual maturity between 35 and 41 weeks of age. The assessment of testicular toxicity is discussed and attention drawn to possible approaches for investigating primary mechanisms of toxic action on the canine testis. Expected values for testicular size, hormone levels and semen characteristics are reported. PMID:524379

  1. Abamectin induces rapid and reversible hypoactivity within early zebrafish embryos.

    PubMed

    Raftery, Tara D; Volz, David C

    2015-01-01

    During early zebrafish embryogenesis, spontaneous tail contractions represent the first sign of locomotion and result from innervation of primary motoneuron axons to target axial muscles. Based on a high-content screen, we previously demonstrated that exposure of zebrafish embryos to abamectin--an avermectin insecticide--from 5-25 hours post-fertilization (hpf) abolished spontaneous activity in the absence of effects on survival and gross morphology. Therefore, the objective of this study was to begin investigating the mechanism of abamectin-induced hypoactivity in zebrafish. Similar to 384-well plates, static exposure of embryos to abamectin from 5-25 hpf in glass beakers resulted in elimination of activity at low micromolar concentrations. However, abamectin did not affect neurite outgrowth from spinal motoneurons and, compared with exposure from 5-25 hpf, embryos were equally susceptible to abamectin-induced hypoactivity when exposures were initiated at 10 and 23 hpf. Moreover, immersion of abamectin-exposed embryos in clean water resulted in complete recovery of spontaneous activity relative to vehicle controls, suggesting that abamectin reversibly activated ligand-gated chloride channels and inhibited neurotransmission. To test this hypothesis, we pretreated embryos to vehicle or non-toxic concentrations of fipronil or endosulfan--two insecticides that antagonize the γ-aminobutyric acid (GABA) receptor--from 5-23 hpf, and then exposed embryos to vehicle or abamectin from 23-25 hpf. Interestingly, activity levels within abamectin-exposed embryos pretreated with either antagonist were similar to embryos exposed to vehicle alone. Using quantitative PCR and phylogenetic analyses, we then confirmed the presence of GABA receptor α1 and β2 subunits at 5, 10, and 23 hpf, and demonstrated that zebrafish GABA receptor subunits are homologous to mammalian GABA receptor subunits. Overall, our data collectively suggest that abamectin induces rapid and reversible hypoactivity within early zebrafish embryos, an effect that may be mediated through the GABA receptor. PMID:25733401

  2. Effect of Calcium Chloride on the Permeation of the Cryoprotectant Dimethyl Sulfoxide to Japanese Whiting Sillago japonica Embryos

    NASA Astrophysics Data System (ADS)

    Rahman, Sk. Mustafizur; Majhi, Sullip Kumar; Suzuki, Toru; Strussmann, Carlos Augusto; Watanabe, Manabu

    Cryopreservation of fish eggs and embryos is a highly desired tool to promote aquaculture production and fisheries resource management, but it is still not technically feasible. The failure to develop successful cryopreservation protocols for fish embryos is largely attributed to poor cryoprotectant permeability. The purpose of this study was to test the effectiveness of CaCl2 to enhance cryoprotectant uptake by fish embryos. In this study, embryos (somites and tail elongation stages) of Japanese whiting Sillago japonica were exposed to 10 and 15% dimethyl sulfoxide (DMSO) in artificial sea water (ASW) or a solution of 0.125M CaCl2 in distilled water for 20 min at 24°C. The toxicity of all solutions was estimated from the hatching rates of the embryos and High Performance Liquid Chromatography was used to determine the amount of DMSO taken up during impregnation. The results showed that DMSO incorporation into the embryos was greatly (›50%) enhanced in the presence of CaCl2 compared to ASW. CaCl2 itself was not toxic to the embryos but, probably as a result of the enhanced DMSO uptake, caused decreases in survival of about 14-44% relative to ASW. Somites stage embryos were more tolerant than tail elongation ones to DMSO both as ASW and CaCl2 solutions. The use of CaCl2 as a vehicle for DMSO impregnation could be a promising aid for the successful cryopreservation of fish embryos.

  3. Documentation of toxicity testing results on increased supernate treatment rate of 2700 gallons/batch

    SciTech Connect

    Pickett, J.B.

    1992-07-06

    In February 1991, Reactor Materials increased the rate of supernate treatment in the M-Area Dilute Effluent Treatment Facility (DETF) from 1800 gallons to [approximately]2700 gallons of supernate per 36,000 gallon dilute wastewater batch. The first release of the treated effluent began on March 3, 1991. A series of whole effluent toxicity tests was conducted on the DETF effluent to determine if the increased supernate concentration would result in any chronic toxicity affects in the receiving stream (Tims Branch). The toxicity tests were conducted at instream concentrations equivalent to DETF release rates of 5, 10, 15, 20, and 25 gallons/min. The test results, based on 7-day Ceriodaphnia dubia chronic toxicity, indicated no toxicity effects at any concentration tested. Supernate treatment in DETF continued at the higher concentration.

  4. Pilot study for ambient toxicity testing in Chesapeake bay. Year two report

    SciTech Connect

    Hall, L.W.; Ziegenfuss, M.C.; Fischer, S.A.; Anderson, R.D.; Killen, W.D.

    1992-11-01

    The primary goal of the ambient toxicity testing pilot study was to identify toxic areas in living resource habitats of the Chesapeake Bay watershed by using a battery of standardized, directly modified or recently developed water column, sediment and suborganismal toxicity tests. Tests were conducted twice at the following stations: Potomac River-Morgantown, Potomac River-Dahlgren, Patapsco River and Wye River. A suite of inorganic and organic contaminants was evaluated in the water column and sediment during these tests. Standard water quality conditions were also evaluated in water and sediment from all stations.

  5. ExpoCast: Exposure Science for Prioritization and Toxicity Testing

    EPA Science Inventory

    The US EPA is completing the Phase I pilot for a chemical prioritization research program, called ToxCastTM. Here EPA is developing methods for using computational chemistry, high-throughput screening, and toxicogenomic technologies to predict potential toxicity and prioritize l...

  6. Comparison of mouse and chick embryo liver and hepatoma cell lines as model systems used for enzymological estimation of toxicity potentials of organic pollutants.

    PubMed

    Machala, M; Mátlová, L; Nezveda, K; Turánek, J; Horavová, P; Granátová, M; Nevoránková, Z

    1995-10-01

    Cytochromes P450-dependent monooxygenase activities were determined and compared in mouse liver microsomes and in hepatoma cell homogenates after exposure to prototype inducers of individual P450 enzymes. In vivo inductions of levels of mouse hepatic monooxygenase activities have been found as effective biochemical markers of toxicity potentials of a series of classes of xenobiotics (CYP1A induction for toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin, coplanar polychlorinated biphenyls, polycyclic aromatic hydrocarbons and related pollutants; CYP2E induction for dialkylnitrosamines and organic solvents, e.g. acetone and ethanol; CYP2B and CYP3A induction for phenobarbital- and dexamethasone-type of xenobiotics). A specific induction of CYP1A-dependent O-dealkylase activities by TCDD was found in Hepa-1 and Hep G2 cell cultures, but no in vitro induction of other P450 enzymes was found after the treatment with phenobarbital, acetone or dexamethasone. Therefore, mouse liver is a suitable in vivo system for the testing of inducing effects of xenobiotics on all relevant P450 forms, while hepatoma cell cultures are usable only for the bioassay of TCDD-like toxicity. PMID:8659083

  7. The sensitivity of relative toxicity rankings by the USF/NASA test method to some test variables

    NASA Technical Reports Server (NTRS)

    Hilado, C. J.; Labossiere, L. A.; Leon, H. A.; Kourtides, D. A.; Parker, J. A.; Hsu, M.-T. S.

    1976-01-01

    Pyrolysis temperature and the distance between the source and sensor of effluents are two important variables in tests for relative toxicity. Modifications of the USF/NASA toxicity screening test method to increase the upper temperature limit of pyrolysis, reduce the distance between the sample and the test animals, and increase the chamber volume available for animal occupancy, did not significantly alter rankings of relative toxicity of four representative materials. The changes rendered some differences no longer significant, but did not reverse any rankings. The materials studied were cotton, wool, aromatic polyamide, and polybenzimidazole.

  8. Sediment toxicity testing with the amphipod Ampelisca abdita in Calcasieu Estuary, Louisiana

    USGS Publications Warehouse

    Redmond, M.S.; Crocker, P.A.; McKenna, K.M.; Petrocelli, E.A.; Scott, K.J.; Demas, C.R.

    1996-01-01

    Discharges from chemical and petrochemical manufacturing facilities have contaminated portions of Louisiana's Calcasieu River estuary with a variety of organic and inorganic contaminants. As part of a special study, sediment toxicity testing was conducted to assess potential impact to the benthic community. Ten-day flow-through sediment toxicity tests with the amphipod Ampelisca abdita revealed significant toxicity at 68% (26 of 38) of the stations tested. A. abdita mortality was highest in the effluent-dominated bayous, which are tributaries to the Calcasieu River. Mortality was correlated with total heavy metal and total organic compound concentrations in the sediments. Ancillary experiments showed that sediment interstitial water salinity as low as 2.5 o/o-o did not significantly affect A. abdita's, response in the flow-through system; sediment storage for 7 weeks at 4??C did not significantly affect toxicity. Sediment toxicity to A. abdita was more prevalent than receiving water toxicity using three short-term chronic bioassays. Results suggest that toxicity testing using this amphipod is a valuable tool when assessing sediments containing complex contaminant mixtures and for assessing effects of pollutant loading over time. In conjunction with chemical analyses, the testing indicated that the effluent-dominated, brackish bayous (Bayou d'Inde and Bayou Verdine) were the portions of the estuary most impacted by toxicity.

  9. 40 CFR 797.1330 - Daphnid chronic toxicity test.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... laboratory procedures. Cleaning of test chambers may be necessary during the testing period. (3) Test... SUBSTANCES CONTROL ACT (CONTINUED) ENVIRONMENTAL EFFECTS TESTING GUIDELINES Aquatic Guidelines § 797.1330... distributions. In addition, test chambers within the testing area shall be positioned in a random manner as in...

  10. 40 CFR 797.1930 - Mysid shrimp acute toxicity test.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...) Summary of the test. In preparation for the test, test chambers are filled with appropriate volumes of... test substance in a sample shall be validated before beginning the test by appropriate laboratory... periods. (iv) Feeding. Mysids should be fed during testing. Any food utilized should support...

  11. 40 CFR 797.1950 - Mysid shrimp chronic toxicity test.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... the test. (i) In preparation for the test, the flow of test solution through each chamber is adjusted... the amount of test substance in a sample shall be validated before beginning the test by appropriate...) Feeding. Mysids should be fed during testing. Any food utilized should support survival, growth...

  12. Non-animal alternative toxicity tests for detergents: genuine replacements or mere prescreens?

    PubMed

    Balls, M; Reader, S; Atkinson, K; Tarrant, J; Clothier, R

    1991-01-01

    Non-animal toxicity tests and testing strategies for use in identifying the potential toxic hazard of chemicals and products, and in providing information for use in risk and safety assessment, are in the course of development, validation and evaluation. Possible replacement alternatives to the rabbit Draize eye irritancy test are discussed, and results obtained for two in-vitro tests developed by FRAME (the kenacid blue test and the neutral red release test), applied to 19 surfactants and 32 formulations, are discussed. It is concluded that there are now legal and moral requirements that relevant and reliable non-animal tests should be developed and accepted for use whenever possible. PMID:1367188

  13. Evaluation of Daphnia ambigua for Routine Aquatic Toxicity Testing at the Savannah River Site

    SciTech Connect

    Specht, W.L.; Harmon, S.M.

    1997-09-01

    Short-term whole effluent toxicity testing, which is currently a requirement of the U.S. EPA`s National Pollution Discharge Elimination System (NPDES), commonly uses the cladoceran species Ceriodaphnia dubia. Despite the advantages to using a common test species to model the toxic effects of effluents, it could be argued that toxicity test results would be more meaningful if a wider variety of test organisms were commonly used. One particular argument against C. dubia is that tests conducted with this species do not always reflect local, site-specific conditions. The careful selection and use of an indigenous test species would produce a more realistic model of local instream effects and would account for regional differences in water quality. Permitted effluent discharges from Savannah River Site (SRS), a government weapons facility operated by the U.S. Department of Energy, require toxicity testing with C. dubia. However, water quality in these receiving streams is markedly different (lower pH and hardness) from standard laboratory water used for the culturing and testing of C. dubia, and it has been shown that this receiving water presents varying degrees of toxicity to C. dubia. Based on these results, it is possible that toxic effects observed during an effluent study could be the result of test organism stress from the dilution water and not the effects of SRS effluents. Therefore, this study addressed the substitution of C. dubia with an indigenous cladoceran species, Daphnia ambigua for routine regulatory testing at SRS. Given the indigenous nature of this species, combined with the fact that it has been successfully cultured by other investigators, D. ambigua was ideal for consideration as a replacement for C. dubia, but further study of the overall success and sensitivity of laboratory-reared D. ambigua was required. This investigation determined that D. ambigua could be laboratory cultured with only minimal changes to established regulatory protocol and that the life-cycle characteristics of this species were conducive to traditional acute and chronic aquatic toxicity test methods. Acute toxicity tests showed that when comparing LC50 values for C. dubia and D. ambigua, D. ambigua was less sensitive to some toxicants (sodium chloride, copper sulfate, and sodium lauryl sulfate) while more sensitive to others (chlorpyrifos). Results of chronic tests with copper sulfate and sodium chloride resulted in the same NOEC/LOEC values for both species. When exposed to unaltered SRS stream water, C. dubia demonstrated a `toxic` response for two of the three streams tested, while reproduction for D. ambigua was higher in all stream samples. Acute toxicity tests with sodium chloride in stream water, generally followed the sensitivity trend noted in tests conducted with regular laboratory water.

  14. Polychloroprene flexible foam as a reference material. [for fire toxicity tests

    NASA Technical Reports Server (NTRS)

    Hilado, C. J.; Cumming, H. J.; Morford, R. H.

    1977-01-01

    Polychloroprene flexible foam was evaluated as a reference material for fire toxicity tests. A commercial sample was evaluated using nine different test conditions of the USF methodology. The material exhibited a wide range of relative toxicity depending on the test conditions. Times to incapacitation and times to death were shortest at 16 ml/sec (1 l/min) air flow, at any fixed heating rate and upper limit temperature.

  15. An interlaboratory comparison of sediment elutriate preparation and toxicity test methods.

    PubMed

    Haring, Herman J; Smith, Mark E; Lazorchak, James M; Crocker, Philip A; Euresti, Abel; Blocksom, Karen; Wratschko, Melissa C; Schaub, Michael C

    2012-12-01

    Elutriate bioassays are among numerous methods that exist for assessing the potential toxicity of sediments in aquatic systems. In this study, interlaboratory results were compared from 96-h Ceriodaphnia dubia and Pimephales promelas static-renewal acute toxicity tests conducted independently by two laboratories using elutriate samples prepared from the same sediment. The goal of the study was to determine if the results from the elutriate tests were comparable between two U.S. Environmental Protection Agency (USEPA) laboratories when different elutriate preparation procedures were employed by each lab. Complete agreement in site characterization was attained in 22 of the 25 samples for both bioassays amongst each lab. Of the 25 samples analyzed, 10 were found to be toxic to at least one of the species tested by either laboratory. The C. dubia elutriate tests conducted by the National Exposure Research Laboratory (NERL) indicated that 7 of the 25 sediment samples were toxic, while 8 sediment samples were characterized as such in testing conducted by USEPA Region 6 (Region 6). The P. promelas elutriate tests conducted by NERL determined 8 samples as toxic, while Region 6 tests displayed toxicity in 5 of the samples. McNemar's test of symmetry for C. dubia (S = 0.33, p = 0.5637) and P. promelas (S = 3.0, p = 0.0833) tests indicated no significant differences in designating a site toxic between NERL and Region 6 laboratories. Likewise, Cohen's kappa test revealed significant agreement between NERL and Region 6 C. dubia (K = 0.7148, p < 0.01) and P. promelas (K = 0.6939, p < 0.01) elutriate tests. The authors conclude that differences in interlaboratory elutriate preparation procedures have no bearing on the ability of either the C. dubia or P. promelas bioassay testing methods to detect toxicity while yielding similar results. PMID:22278676

  16. Assessment of four different test designs for Hyalella azteca 10 days sediment toxicity test

    SciTech Connect

    Ramirez-Romero, P.; Oris, J.T.; Bailer, J.; DePoy, M.

    1995-12-31

    The purpose of this study was to assess the adequacy of four experimental designs of the Hyalella azteca 10 days sediment toxicity test. The authors conducted a series of sediment toxicity tests using an EPA recommended experimental design (8 chambers with 10 organisms per treatment) and three other designs. These had the same total number of organisms (80) per treatment and the same sediment:water ratio (1:1.5) but different number of chambers (4,2,1). The number of organisms recovered, the time to sort and count the animals, as well as the time to make a water change were compared for these four designs. Logistic regression was used to analyze the recovery data while one-way analysis of variance methods were used to analyze the time responses. The results showed that the four treatments were comparable in terms of proportion of organisms recovered. However, the sorting time and the water change time decreased as the number of chambers decreased, making those designs with less chambers more desirable.

  17. [Subchronic toxicity testing of mold-ripened cheese].

    PubMed

    Schoch, U; Lüthy, J; Schlatter, C

    1984-08-01

    The biological effects of known mycotoxins of Penicillium roqueforti or P. camemberti and other still unknown, but potentially toxic metabolites in mould ripened cheese (commercial samples of Blue- and Camembert cheese) were investigated. High amounts of mycelium (equivalents of 100 kg cheese/man and day) were fed to mice in a subchronic feeding trial. The following parameters were determined: development of body weight, organ weights, hematology, blood plasma enzymes. No signs of adverse effects produced by cheese mycotoxins could be detected after 28 days. No still unknown toxic metabolites could be demonstrated. From these results no health hazard from the consumption of mould ripened cheese, even in high amounts, appears to exist. PMID:6485557

  18. Concentration-time data in toxicity tests and resulting relationships

    NASA Technical Reports Server (NTRS)

    Hilado, C. J.; Brauer, D. P.

    1979-01-01

    Periodic analyses for carbon monoxide and methane in the animal exposure chamber during pyrolysis of polyethylene at 800 C without forced air flow showed that the concentrations of these compounds increased with exposure time. These observations, and similar observations for polyurethane flexible foam, permitted the calculation of carbon monoxide toxicity in terms of a DP (Death Product Concentration) value, in addition to flammability in terms of HC (Hydro Carbon) value. Observed DP values exceeding the critical DP(CO) value of 47,200 ppm-min for carbon monoxide may indicate that lethal exposures were reached earlier but not immediately manifested because of the time delay involved in physiological processes. On the basis of this DP(CO) value, carbon monoxide could have been the sole toxicant in the case of polyethylene, polypropylene, polyoxymethylene, polystyrene, polycarbonate, ethylene propylene diene rubber, and wood.

  19. Alternative testing systems for evaluating noncarcinogenic, hematologic toxicity.

    PubMed Central

    Parchment, R E

    1998-01-01

    Hematopoietic tissues are the targets of numerous xenobiotics. Clinical hematotoxicity is either a decrease or an increase in peripheral blood cell counts in one or more cell lineages--a cytopenia or a cytosis, respectively--that carries a risk of an adverse clinical event. The purpose of in vitro hematotoxicology is the prediction of these adverse hematologic effects from the effect