DEVELOPMENT OF MONOCLONAL ANTIBODIES AGAINST FATHEAD MINNOW (PIMEPHALES PROMELAS) VITELLOGENIN

Science.gov (United States)

We have obtained a panel of monoclonal antibodies directed against fathead minnow vitellogenin (Vtg) for use in sensitive ELISAs to quantify the response of exposure in vivo to estrogen or estrogen mimics.

Adaptive Response in Female Fathead Minnows Exposed to an Aromatase Inhibitor: Computational Modeling of the Hypothalamic-Pituitary-Gonadal Axis

Science.gov (United States)

Exposure to endocrine disrupting chemicals can affect reproduction and development in both humans and wildlife. We are developing a mechanistic computational model of the hypothalamic-pituitary-gonadal (HPG) axis in female fathead minnows to predict dose-response and time-course ...

The effect of chronic exposure to 238Pu(IV) citrate on the embryonic development of carp and fathead minnow eggs

International Nuclear Information System (INIS)

Till, J.E.

1978-01-01

This study focused on the effects of plutonium to the developing fish egg. A quantitative analysis of the uptake of 238 Pu(IV) citrate by carp eggs indicated that plutonium is accumulated in the egg and reaches a concentration factor of approx 4 at hatching (72hr after exposure begins). Autoradiographs made from carp eggs that had been exposed to 238 Pu revealed that, although some plutonium was concentrated on the egg chorion, plutonium that penetrated the chorion was uniformly distri-buted throughout the perivitelline fluid, embryo and yolk sac. Using the uptake data for carp eggs, dose conversion factors were calculated for both carp eggs and fathead minnow eggs exposed to 238 Pu during embryological development. These values were 2100 rad/μCi/ml and 7500 rad/μCi/ml for carp and fathead minnows, respectively. Experiments were conducted in which both carp and fathead minnow eggs were exposed to plutonium in solution during embryogenesis; the percentage of eggs hatching, the number of abnormal larvae produced, and the survival of larvae were used as indicators of radiation toxicity. Concentrations in excess of 1 μCi/ml were required to prevent both species of eggs from hatching. It is concluded that fish eggs developing in natural aquatic ecosystems contaminated with plutonium probably do not receive a significant dose from the plutonium alpha radioactivity. Future analysis of fecundity and the production of abnormalities in the offspring of larvae exposed during embryological development would provide an additional degree of sensitivity to the toxicity test system used in this study. (author)

Computational Modeling of Hypothalamic-Pituitary-Gonadal Axis to Predict Adaptive Responses in Female Fathead Minnows Exposed to an Aromatase Inhibitor

Science.gov (United States)

Exposure to endocrine disrupting chemicals can affect reproduction and development in both humans and wildlife. We are developing a mechanistic computational model of the hypothalamic-pituitary-gonadal (HPG) axis in female fathead minnows to predict dose response and time-course...

Effects of spray-dried Pseudomonas fluorescens, strain CL145A (Zequanox®) on reproduction and early development of the fathead minnow (Pimephales promelas).

Science.gov (United States)

Waller, Diane L.; Luoma, James A.

2016-01-01

The biopesticide, Zequanox®, is registered for dreissenid mussel control in open water systems. Previous toxicity trials with nontarget organisms, including young-of-the year of several fish species and invertebrates, demonstrated selectivity of Zequanox for dreissenids. However, data are lacking on its safety to reproductive and early life stages of fish. The present study evaluated the effects of Zequanox on spawning and early life stages of the fathead minnow, Pimephales promelas, at the maximum approved concentration (100 mg Zequanox active ingredient /L) and exposure duration (8 h) for open water application. The results showed no significant effect of Zequanox on survival, condition, or cumulative egg deposition (21 d) in adult fathead minnow. Eggs (<24-h old) exposed to Zequanox developed to the eyed-stage at a similar rate to that of unexposed eggs. Additionally, Zequanox did not have a significant effect on survival and growth (90 d) of newly hatched fry (<24-h old). The results indicate that Zequanox treatment will not affect survival, spawning, and early life development of fathead minnows when applied at the recommended treatment regime.

Taxonomy Icon Data: fathead minnow [Taxonomy Icon

Lifescience Database Archive (English)

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A study of temporal effects of the model anti-androgen flutamide on components of the hypothalamic-pituitary-gonadal axis in adult fathead minnows

Science.gov (United States)

The aim of this study was to investigate temporal changes in the hypothalamic-pituitary-gonadal (HPG) axis of fathead minnow (Pimephales promelas) treated with the model androgen receptor (AR) antagonist, flutamide. Reproductively-mature fish were exposed in a flow-through, meas...

Sex-specific gonadal and gene expression changes throughout development in fathead minnow

Science.gov (United States)

Although fathead minnows (Pimephales promelas) are commonly used as a model fish in endocrine disruption studies, none have characterized sex-specific baseline expression of genes involved in sex differentiation during development in this species. Using a sex-linked DNA marker t...

Transcriptional and physiological response of fathead minnows (Pimephales promelas) exposed to urban waters entering into wildlife protected areas

International Nuclear Information System (INIS)

Rodriguez-Jorquera, Ignacio A.; Kroll, Kevin J.; Toor, Gurpal S.; Denslow, Nancy D.

2015-01-01

The mission of protected areas is to conserve biodiversity and improve human welfare. To assess the effect of urban waters entering into protected areas, we performed 48-h whole-effluent exposures with fathead minnows, analyzing changes in steady state levels of mRNAs in the livers of exposed fish. Raw wastewater, treated city wastewater, and treated wastewater from a university were collected for exposures. All exposed fish showed altered mRNA levels of DNA damage-repair genes. Fish exposed to raw and treated wastewaters showed down-regulation of transcripts for key intermediates of cholesterol biosynthesis and elevated plasma cholesterol. The type of wastewater treatment influenced the response of gene transcription. Because of the relevance of some of the altered cellular pathways, we suggest that these effluents may cause deleterious effects on fish inside protected areas that receive these waters. Inclusion of research and mitigation efforts for this type of threat in protected areas management is advised. - Highlights: • Wastewater entering wildlife preserves alters gene expression in exposed fish. • DNA repair mechanisms and cholesterol metabolism were altered in fish. • Effects on cholesterol genes were in agreement with fish hypercholesterolemia. - Urban wastewaters released into protected areas altered gene transcription of key genes such as DNA repair and cholesterol biosynthesis and produced hypercholesterolemia in fish

A new bacilliform fathead minnow rhabdovirus that produces syncytia in tissue culture.

Science.gov (United States)

Iwanowicz, L R; Goodwin, A E

2002-05-01

A pathogenic bacilliform virus 130-180 nm in length and 31-47 nm in diameter was isolated from moribund fathead minnows (Pimephales promelas) exhibiting hemorrhages in their eyes and skin. A cytopathic effect of multifocal syncytia was observed in the epithelioma papulosum cyprini cell line after a 48 h incubation at 20 degrees C. A similar cytopathic effect was also observed in other cell lines tested, but not in bluegill fry, koi fin, or Chinook salmon embryo cells. The filterable agent was inactivated by exposure to 50 degrees C for 10 min, 20% ether, 2 and 50% chloroform, pH 3, and pH 10, was unaffected by 5'-iodo-2 deoxyuridine, and appeared bacilliform and occasionally bullet-shaped by electron microscopy. These results are consistent with those of rhabdoviruses. Immunodot blots performed with antisera against selected fish rhabdoviruses, an aquareovirus, and a birnavirus were all negative. River's postulates were fulfilled in fathead minnows, but the agent did not replicate or cause disease in other cyprinids or salmonids during challenge experiments. Hepatic, splenic, and renal lesions were observed during histological analysis of diseased fish from viral challenges and from the original case. Structural proteins resolved via SDS-PAGE had molecular weights similar to those reported in lyssaviruses of the family Rhabdoviridae; however, syncytia formation is not a typical cytopathic effect of rhabdoviruses. This virus, has tentatively been named the fathead minnow rhabdovirus (FHMRV) and is most similar to the members of the family Rhabdoviridae, but atypical properties like syncytia formation may justify the assignment to a novel taxon.

SEQUENCING AND DE NOVO DRAFT ASSEMBLIES OF A FATHEAD MINNOW (Pimpehales promelas) reference genome

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U.S. Environmental Protection Agency — The dataset provides the URLs for accessing the genome sequence data and two draft assemblies as well as fathead minnow genotyping data associated with estimating...

Toward an adverse outcome pathway for impaired growth: Mitochondrial dysfunction impairs growth in early life stages of the fathead minnow (Pimephales promelas).

Science.gov (United States)

Bolser, Derek G; Dreier, David A; Li, Erchao; Kroll, Kevin J; Martyniuk, Christopher J; Denslow, Nancy D

2018-07-01

Chemical contaminants present in the environment can affect mitochondrial bioenergetics in aquatic organisms and can have substantial effects on individual fitness. As early life stages of fish are particularly vulnerable to environmental contaminants, they are ideal models for examining the relationship between impaired mitochondrial bioenergetics (ATP-dependent respiration, basal oxidative respiration) and apical endpoints such as growth. Here, early life stages of the fathead minnow (Pimephales promelas), an ecologically relevant North American species, were used to investigate the relationship between mitochondrial bioenergetics and growth following perturbation with model mitochondrial toxicants 2,4-dinitrophenol and octylamine. Fathead minnows were exposed to 2,4-dinitrophenol and octylamine at 3 concentrations for 24 h and endpoints related to mitochondrial bioenergetics were measured with the Agilent Seahorse XFe24 Bioanalyzer. In order to link changes in mitochondrial bioenergetics to growth, fathead minnows were exposed to the same chemical contaminants for 7-14 days and growth was measured by measuring total length on a weekly basis. There was a significant correlation between decrease in average length at 14 days and basal respiration (r = 0.997, p = 0.050, n = 3), as well as maximal respiration (r = 0.998, p-value = 0.043, n = 3) for embryos exposed to 2,4 dinitrophenol. For octylamine, ATP production was highly correlated with average length at 7 days (p-value = 0.1) and spare respiratory capacity and average length at 14 days were highly correlated (p-value = 0.1). These data improve understanding of how mitochondrial toxicants impair growth in fish larvae and may be useful for developing an adverse outcome pathway for growth. Copyright © 2018 Elsevier Inc. All rights reserved.

Extended fish short term reproduction assays with the fathead minnow and Japanese medaka: No evidence of impaired fecundity from exposure to atrazine.

Science.gov (United States)

Brain, Richard A; Schneider, Suzanne Z; Anderson, Julie C; Knopper, Loren D; Wolf, Jeffrey C; Hanson, Mark L

2018-08-01

Short-term reproduction assays were conducted with fathead minnow (Pimephales promelas) and Japanese medaka (Oryzias latipes) to evaluate responses from atrazine exposure at environmentally relevant concentrations and above. Breeding groups of fish with multiple males and females were exposed to atrazine under flow-through conditions. Fathead minnows were exposed to mean measured concentrations of 1.0, 10, 26, 52, and 105 μg atrazine/L for 28 days. Medaka were exposed to mean measured concentrations of 9.4, 48, 74, 97, and 244 μg atrazine/L for 28 or 29 days. Fish were evaluated for survival, fecundity, fertility, total length, wet weight, secondary sex characteristics, gonadosomatic index (GSI) (P. promelas only), plasma or hepatic vitellogenin (VTG), and histopathology of gonads. General observations of health and behaviour were also conducted. There were no statistically significant effects (i.e., p < 0.05) of atrazine on survival, size, reproduction, behaviour, GSI, VTG, or secondary sex characteristics in either species at any exposure level. In fathead minnows, there were no histopathological findings associated with atrazine exposure in male fish, but there was an increased proportion of Stage 4.0 ovaries accompanied by an increase in proportion of Grade 3 post-ovulatory follicles in females of the 105 μg/L treatment group. Without a concomitant increase in oocyte atresia, neither of these findings are considered adverse for the health of the fish. In medaka, there were no significant effects of atrazine exposure on histopathology in either sex. These data support current weight-of-evidence assessments that atrazine does not cause direct adverse effects on fish reproduction at environmentally realistic concentrations. Copyright © 2018 Elsevier Ltd. All rights reserved.

Pathway-based approaches for assessment of real-time exposure to an estrogenic wastewater treatment plant effluent on fathead minnow reproduction

Science.gov (United States)

Cavallin, Jenna E.; Jensen, Kathleen M.; Kahl, Michael D.; Villeneuve, Daniel L.; Lee, Kathy E.; Schroeder, Anthony L.; Mayasich, Joe; Eid, Evan P.; Nelson, Krysta R.; Milsk, Rebecca Y.; Blackwell, Brett R.; Berninger, Jason P.; LaLone, Carlie A.; Blanskma, Chad; Jicha, Terri M.; Elonen, Colleen M.; Johnson, Rodney C.; Ankley, Gerald T.

2016-01-01

Wastewater treatment plant (WWTP) effluents are known contributors of chemical mixtures into the environment. Of particular concern are endocrine-disrupting compounds, such as estrogens, which can affect the hypothalamic-pituitary-gonadal axis function in exposed organisms. The present study examined reproductive effects in fathead minnows exposed for 21 d to a historically estrogenic WWTP effluent. Fathead minnow breeding pairs were held in control water or 1 of 3 effluent concentrations (5%, 20%, and 100%) in a novel onsite, flow-through system providing real-time exposure. The authors examined molecular and biochemical endpoints representing key events along adverse outcome pathways linking estrogen receptor activation and other molecular initiating events to reproductive impairment. In addition, the authors used chemical analysis of the effluent to construct a chemical-gene interaction network to aid in targeted gene expression analyses and identifying potentially impacted biological pathways. Cumulative fecundity was significantly reduced in fish exposed to 100% effluent but increased in those exposed to 20% effluent, the approximate dilution factor in the receiving waters. Plasma vitellogenin concentrations in males increased in a dose-dependent manner with effluent concentration; however, male fertility was not impacted. Although in vitro analyses, analytical chemistry, and biomarker responses confirmed the effluent was estrogenic, estrogen receptor agonists were unlikely the primary driver of impaired reproduction. The results provide insights into the significance of pathway-based effects with regard to predicting adverse reproductive outcomes.

Chronic nitrate exposure alters reproductive physiology in fathead minnows.

Science.gov (United States)

Kellock, Kristen A; Moore, Adrian P; Bringolf, Robert B

2018-01-01

Nitrate is a ubiquitous aquatic pollutant that is commonly associated with eutrophication and dead zones in estuaries around the world. At high concentrations nitrate is toxic to aquatic life but at environmental concentrations it has also been purported as an endocrine disruptor in fish. To investigate the potential for nitrate to cause endocrine disruption in fish, we conducted a lifecycle study with fathead minnows (Pimephales promelas) exposed to nitrate (0, 11.3, and 56.5 mg/L (total nitrate-nitrogen (NO 3 -N)) from nitrate-exposed males both 11-KT and vitellogenin were significantly induced when compared with controls. No significant differences occurred for body mass, condition factor, or GSI among males and intersex was not observed in any of the nitrate treatments. Nitrate-exposed females also had significant increases in vitellogenin compared to controls but no significant differences for mass, condition factor, or GSI were observed in nitrate exposed groups. Estradiol was used as a positive control for vitellogenin induction. Our findings suggest that environmentally relevant nitrate levels may disrupt steroid hormone synthesis and/or metabolism in male and female fish and may have implications for fish reproduction, watershed management, and regulation of nutrient pollution. Copyright © 2017 Elsevier Ltd. All rights reserved.

FATHEAD MINNOW VITELLOGENIN: CDNA SEQUENCE AND MRNA AND PROTEIN EXPRESSION AFTER 17 BETA-ESTRADIOL TREATMENT

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In the present study, a sensitive ribonuclease protection assay (RPA) for VTG mRNA was developed for the fathead minnow (Pimephales promelas), a species proposed for routine endocrine-disrupting chemical (EDC) screening.

In Silico analysis of perturbed steroidogenesis and gonad growth in fathead minnows (P. promelas) exposed to 17α-ethynylestradiol.

Science.gov (United States)

Hala, David; Petersen, Lene H; Martinović, Dalma; Huggett, Duane B

2015-06-01

The multi-factorial nature of adverse reproductive effects mediated by endocrine disrupting compounds (or EDCs) makes understanding the mechanistic basis of reproductive dysfunction a highly pertinent area of research. As a consequence, a main motivator for continued research is to integrate 'multi-leveled' complexity (i.e., from genes to phenotype) using mathematical methods capable of encapsulating properties of physiological relevance. In this study, an in silico stoichiometric model of piscine steroidogenesis was augmented with a 'biomass' reaction associating the underlying stoichiometry of steroidogenesis with a reaction representative of gonad growth. The ability of the in silico model to predict perturbed steroidogenesis and subsequent effects on gonad growth was tested by exposing reproductively active male and female fathead minnows (Pimephales promelas) to 88 ng/L of the synthetic estrogen, 17α-ethynylestradiol (EE2). The in silico model was parameterized (or constrained) with experimentally quantified concentrations of selected steroid hormones (using mass spectrometry) and fold changes in gene expression (using RT-qPCR) for selected steroidogenic enzyme genes, in gonads of male and female fish. Once constrained, the optimization framework of flux balance analysis (FBA) was used to calculate an optimal flux through the biomass reaction (analogous to gonad growth) and associated steroidogenic flux distributions required to generate biomass. FBA successfully predicted effects of EE2 exposure on fathead minnow gonad growth (%gonadosomatic index or %GSI) and perturbed production of steroid hormones. Specifically, FBA accurately predicted no effects of exposure on male %GSI and a significant reduction for female %GSI. Furthermore, in silico simulations accurately identified disrupted reaction fluxes catalyzing productions of androgens (in male fish) and progestogens (in female fish), an observation which agreed with in vivo experimentation. The analyses

Alternative methods for toxicity assessments in fish: comparison of the fish embryo toxicity and the larval growth and survival tests in zebrafish and fathead minnows.

Science.gov (United States)

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

2014-11-01

An increased demand for chemical toxicity evaluations has resulted in the need for alternative testing strategies that address animal welfare concerns. The fish embryo toxicity (FET) test developed for zebrafish (Danio rerio) is one such alternative, and the application of the FET test to other species such as the fathead minnow (Pimephales promelas) has been proposed. In the present study, the performances of the FET test and the larval growth and survival (LGS; a standard toxicity testing method) test in zebrafish and fathead minnows were evaluated. This required that testing methods for the fathead minnow FET and zebrafish LGS tests be harmonized with existing test methods and that the performance of these testing strategies be evaluated by comparing the median lethal concentrations of 2 reference toxicants, 3,4-dicholoraniline and ammonia, obtained via each of the test types. The results showed that procedures for the zebrafish FET test can be adapted and applied to the fathead minnow. Differences in test sensitivity were observed for 3,4-dicholoraniline but not ammonia; therefore, conclusions regarding which test types offer the least or most sensitivity could not be made. Overall, these results show that the fathead minnow FET test has potential as an alternative toxicity testing strategy and that further analysis with other toxicants is warranted in an effort to better characterize the sensitivity and feasibility of this testing strategy. © 2014 SETAC.

Examining waterborne and dietborne routes of exposure and their contribution to biological response patterns in fathead minnow (Pimephales promelas)

International Nuclear Information System (INIS)

Rozon-Ramilo, Lisa D.; Dubé, Monique G.; Squires, Allison J.; Niyogi, Som

2011-01-01

Highlights: Objective (i) was achieved by isolating each route of exposure (diet, water and the combination of the two) and comparing it to a control. The metal mining effluent studied encompassed several different effluent types including: mine water, mill facility, tailing impoundment, treatment pond, treatment facility, seepage and surface drainage effluent. Experiment 1 examined exposure pathways by isolating effluent exposure through the diet, water or the combination of the two. Experiment 2 examined whether differences in Fathead minnow responses occurred when fish were fed a live or frozen (dead) diet of chironomids. Multiple trophic levels (consumers and carnivores) were placed in each artificial stream simultaneously. Primary route of exposure to Fathead minnow when held in control water and fed effluent-exposed chironomid larvae. - Abstract: The objectives of the current study were: (i) to gain a better understanding of the relative importance of water and diet as routes of exposure causing toxicity in fathead minnow (FHM) exposed to metal mining effluents (MME) using a full factorial water/food experimental design (Experiment 1), and (ii) to assess differences in the effects of food quality on toxicity by comparing FHM fed both a live and frozen diet of Chironomus dilutus (Experiment 2). The results showed significant increases in general water quality parameters (e.g., hardness, conductivity) and various metals in the effluent treatment waters compared to control waters, with maximum increase seen in the multi-trophic streams. Metals accumulation (Rb, Al, Se, Sr, Tl, Ce, Co, Cu, Pb) effects of both waterborne and multi-trophic exposures were significant in one or more fathead minnow tissue type (muscle, gonads, liver, larvae) relative to those in the control systems. Condition factor and liver somatic index (LSI) of FHM were also significantly affected in both exposures by one or both routes of exposure (water and/or diet). In addition, cumulative

Predator avoidance performance of larval fathead minnows (Pimephales promelas) following short-term exposure to estrogen mixtures

Science.gov (United States)

McGee, M.R.; Julius, M.L.; Vajda, A.M.; Norris, D.O.; Barber, L.B.; Schoenfuss, H.L.

2009-01-01

Aquatic organisms exposed to endocrine disrupting compounds (EDCs) at early life-stages may have reduced reproductive fitness via disruption of reproductive and non-reproductive behavioral and physiological pathways. Survival to reproductive age relies upon optimal non-reproductive trait expression, such as adequate predator avoidance responses, which may be impacted through EDC exposure. During a predator–prey confrontation, larval fish use an innate C-start escape behavior to rapidly move away from an approaching threat. We tested the hypotheses that (1) larval fathead minnows exposed to estrogens, a primary class of EDCs, singularly or in mixture, suffer a reduced ability to perform an innate C-start behavior when faced with a threat stimulus; (2) additive effects will cause greater reductions in C-start behavior; and (3) effects will differ among developmental stages. In this study, embryos (post-fertilization until hatching) were exposed for 5 days to environmentally relevant concentrations of estrone (E1), 17β-estradiol (E2), and 17α-ethinylestradiol (EE2) singularly and in mixture. Exposed embryos were allowed to hatch and grow in control well water until 12 days old. Similarly, post-hatch fathead minnows were exposed for 12 days to these compounds. High-speed (1000 frames/s) video recordings of escape behavior were collected and transferred to National Institutes of Health Image for frame-by-frame analysis of latency period, escape velocity, and total escape response (combination of latency period and escape velocity). When tested 12 days post-hatch, only E1 adversely affected C-start performance of larvae exposed as embryos. Conversely, larvae exposed for 12 days post-hatch did not exhibit altered escape responses when exposed to E1, while adverse responses were seen in E2 and the estrogen mixture. Ethinylestradiol exposure did not elicit changes in escape behaviors at either developmental stage. The direct impact of reduced C-start performance on

Sequencing and De novo Draft Assemblies of the Fathead Minnow (Pimphales promelas)Reference Genome

Science.gov (United States)

This study was undertaken to develop genome-scale resources for the fathead minnow (Pimphales promelas) an important model organism widely used in both aquatic ecotoxicology research and in regulatory toxicity testing. We report on the first sequencing and two draft assemblies fo...

Tissue lead concentration during chronic exposure of Pimephales promelas (fathead minnow) to lead nitrate in aquarium water.

Science.gov (United States)

Spokas, Eric G; Spur, Bernd W; Smith, Holly; Kemp, Francis W; Bogden, John D

2006-11-01

The fathead minnow is a useful species for evaluating the toxicity of wastewater effluents. While this fish is widely used for "survival" studies of metal toxicity, little or no work has been done on the tissue distribution of metals in fathead minnows. To determine the distribution of tissue lead, aquarium studies were conducted for several weeks with fish maintained in soft synthetic freshwater. Lead- (II) nitrate was added to three aquaria attaining concentrations of 20-30 ppb (aquarium B), 100-140 ppb (aquarium C), and roughly 200 ppb (aquarium D). Results were compared to controls (aquarium A). During the initial week, the majority of aquarium D fish died, whereas few deaths occurred in the other groups. Lead accumulation was dose- and tissue-dependent, with highest uptake by the gills. Gill concentrations of aquarium D fish averaged about 4-fold higherthan in skeleton or skin and muscle. In vitro, lead (2.5-25 ppm) caused dose-dependent reductions in the ratio of reduced glutathione/oxidized glutathione (GSH/GSSG) in gills incubated in physiological buffer. These findings demonstrate that fathead minnow gills bind and accumulate waterborne lead rapidly and preferentially and raise the possibility that gill lipid peroxidation contributes to lead toxicity at low water hardness.

Laboratory technique for obtaining fathead minnow eggs for use in toxicity experiments

International Nuclear Information System (INIS)

Till, J.E.

1977-01-01

A laboratory technique for collecting freshly fertilized eggs of the fathead minnow (Pimephales promelas) has been developed for use in toxicity testing of radioactive materials or other environmental pollutants. The fathead minnow egg was selected as a test organism because the period of embryogenesis is relatively short, the number of eggs per spawning is sufficiently large to provide a test group and a control group, and the size of the parent fish is compatible with laboratory facilities. Under a controlled laboratory environment, eggs are available all year. Nesting sites were constructed by cutting plastic pipe in half and covering the inside of the pipe with 1-mm-thick translucent polyethylene. The polyethylene with the eggs attached was removed from the shelter and cut into two pieces, each piece containing approximately the same number of eggs. One piece of polyethylene was floated in the test solution, and the other in spring water as a control. Hatching occurs in 168 h at 24 0 C. Toxicity of the test solution may be scored on the percentage of eggs hatching, the number of abnormal larvae produced, and larval survival

Fathead minnows (Pimephales promelas) and goldfish (Carassius auratus) as standard fish in bioassays and their reaction to potential reference toxicants

Energy Technology Data Exchange (ETDEWEB)

Adelman, I.R.; Smith, L.L. Jr.

1976-02-01

Fathead minnows (Pimephales promelas) and goldfish (Carassius auratus) were compared for their suitability as standard bioassay fish. Both species showed the same variability of bioassay results when tested with four toxicants. Fathead minnows are recommended on the basis of their small size and on their capability for use in complete life cycle tests. On the basis of minimum variability of bioassay results, sodium chloride was superior for use as a reference toxicant. Both sodium chloride and pentachlorophenol seemed capable of detecting abnormal fish. On the basis of seven listed criteria either sodium chloride or pentachlorophenol would be acceptable as a reference toxicant.

Predicting Adaptive Response to Fadrozole Exposure:Computational Model of the Fathead MinnowsHypothalamic-Pituitary-Gonadal Axis

Science.gov (United States)

Exposure to endocrine disrupting chemicals can affect reproduction and development in both humans and wildlife. We are developing a mechanistic mathematical model of the hypothalamic-pituitary-gonadal (HPG) axis in female fathead minnows to predict doseresponse and time-course (...

Predicting Adaptive Response to Fadrozole Exposure: Computational Model of the Fathead Minnow Hypothalamic-Pituitary-Gonadal Axis

Science.gov (United States)

Exposure to endocrine disrupting chemicals can affect reproduction and development in both humans and wildlife. We are developing a mechanistic mathematical model of the hypothalamic-pituitary-gonadal (HPG) axis in female fathead minnows to predict dose-response and time-course (...

Chronic effect of low pH on fathead minnow survival, growth, and reproduction

Energy Technology Data Exchange (ETDEWEB)

Mount, D I

1973-01-01

Fathead minnows (Pimephales promelas rafinesque) were continuously exposed to reduced pH levels of 4.5, 5.2, 5.9, 6.6, and 7.5 (control) during a 13-month, one-generation test. Survival was not affected, even at the lowest pH tested. Fish behavior was abnormal, and fish were deformed at pH 4.5 and 5.2. Egg production and egg hatchability were reduced at pH 5.9 and lower, and all eggs were abnormal. A pH of 6.6 was marginal for vital life functions, but safe for continuous exposure. Free carbon dioxide, liberated by the addition of sulfuric acid to reduce the pH, may have had an unknown effect. The fish did not become acclimiated to low pH levels.

Effects of lipid-lowering pharmaceuticals bezafibrate and clofibric acid on lipid metabolism in fathead minnow (Pimephales promelas).

Science.gov (United States)

Weston, Anna; Caminada, Daniel; Galicia, Hector; Fent, Karl

2009-12-01

The lipid-lowering agents bezafibrate and clofibric acid, which occur at concentrations up to 3.1 and 1.6 microg/L, respectively, are among the most frequently found human pharmaceuticals in the aquatic environment. In contrast to knowledge about their environmental occurrence, little is known about their effects in the environment. The aim of the present study was to analyze effects of these lipid-lowering agents in fish by focusing on their modes of action, lipid metabolism. Fathead minnows were exposed in aquaria to measured concentrations of 0.1, 1.27, 10.18, 101.56, and 106.7 mg/L bezafibrate and to 1.07, 10.75, and 108.91 mg/L clofibric acid for 14 and 21 d, respectively. After exposure, fish liver was analyzed for expression of peroxisome proliferator-activated receptor alpha (PPARalpha) by quantitative polymerase chain reaction (PCR), and the PPAR-regulated enzyme fatty acyl-coenzyme-A oxidase (FAO) involved in fatty acid oxidation. Bezafibrate had no effect, either on PPARalpha expression or on FAO activity, at all concentrations. In contrast, clofibric acid induced FAO activity in male fathead minnows at 108.91 mg/L. No increase in expression of PPARalpha messenger ribonucleic acid was observed. Egg production was apparently decreased after 21 d of exposure to 108.91 mg/L clofibric acid. The present study demonstrates that bezafibrate has very little or no effect on PPARalpha expression and FAO activity, but clofibric acid affects FAO activity.

The influence of food quantity on metal bioaccumulation and reproduction in fathead minnows (Pimephales promelas) during chronic exposures to a metal mine effluent.

Science.gov (United States)

Ouellet, Jacob D; Dubé, Monique G; Niyogi, Som

2013-05-01

Metal mine effluents can impact fish in the receiving environment via both direct effects from exposure as well as indirect effects via food web. The main objective of the present study was to assess whether an indirect effect such as reduced food (prey) availability could influence metal accumulation and reproductive capacity in fish during chronic exposure to a metal mine effluent. Breeding pairs of fathead minnows (Pimephales promelas) were exposed to either reference water (RW) or an environmentally relevant metal mine effluent [45 percent process water effluent (PWE)] for 21 days and fed either low food quantities [LF (a daily ration of 6-10 percent body weight)] or normal food quantities [NF (a daily ration of 20-30 percent body weight)] in artificial stream systems. Fish in RW treatments were fed Chironomus dilutus larvae cultured in RW (Treatments: RW-NF or RW-LF), while fish in PWE treatments were fed C. dilutus larvae cultured in PWE (Treatments: PWE-NF or PWE-LF). Tissue-specific (gill, liver, gonad and carcass) metal accumulation, egg production, and morphometric parameters in fish were analyzed. Fathead minnows that were exposed to LF rations had significantly smaller body, gonad and liver sizes, and were in a relatively poor condition compared to fathead minnows exposed to NF rations, regardless of the treatment water type (RW or PWE) (two-way ANOVA; pmetal-contaminated food in the PWE-NF treatment, tissue metal accumulation pattern were almost similar between the PWE-NF and PWE-LF treatments, except for higher liver barium, cobalt and manganese concentrations in the latter treatment. This indicated that a higher food ration could help reduce the tissue burden of at least some metals and thereby ameliorate the toxicity of metal-mine effluents in fish. More importantly, cumulative egg production in fish was found to be lowest in the PWE-LF treatment, whereas fish egg production in the PWE-NF treatment was not impacted. Overall, these findings suggest

EFFECTS OF THE MAMMALIAN ANTIANDROGEN VINCLOZOLIN ON DEVELOPMENT AND REPRODUCTION OF THE FATHEAD MINNOW

Science.gov (United States)

Previous work with the chlorinated fungicide vinclozolin and its metabolites, 2-{[(3,5-dichlorophenyl)-carbamoyl]oxy}-2-methyl-3-butenoic acid (M1) and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2), indicated antiandrogenic properties expressed in vivo as abnormalities in...

A computational model linking oocyte growth and spawning to the hypothalamic-pituitary-gonadal axis in fathead minnow (Pimephales promelas)

Science.gov (United States)

Reproduction is vital to the survival of all living organisms, and reproductive toxicity is an important outcome in determining the ecological risks of chemicals in the environment. To evaluate reproductive toxicity, fathead minnow fecundity, as measured by the average number of...

Environmentally relevant exposure to 17{alpha}-ethinylestradiol affects the telencephalic proteome of male fathead minnows

Energy Technology Data Exchange (ETDEWEB)

Martyniuk, Christopher J., E-mail: cmartyn@unb.ca [Department of Physiological Sciences and Center for Environmental and Human Toxicology, University of Florida, Gainesville, FL, 32611 (United States); Kroll, Kevin J.; Doperalski, Nicholas J.; Barber, David S.; Denslow, Nancy D. [Department of Physiological Sciences and Center for Environmental and Human Toxicology, University of Florida, Gainesville, FL, 32611 (United States)

2010-07-15

Estrogens are key mediators of neuronal processes in vertebrates. As such, xenoestrogens present in the environment have the potential to alter normal central nervous system (CNS) function. The objectives of the present study were (1) to identify proteins with altered abundance in the male fathead minnow telencephalon as a result of low-level exposure to17{alpha}-ethinylestradiol (EE{sub 2}), and (2) to better understand the underlying mechanisms of 17{beta}-estradiol (E{sub 2}) feedback in this important neuroendocrine tissue. Male fathead minnows exposed to a measured concentration of 5.4 ng EE{sub 2}/L for 48 h showed decreased plasma E{sub 2} levels of approximately 2-fold. Of 77 proteins that were quantified statistically, 14 proteins were down-regulated after EE{sub 2} exposure, including four histone proteins, ATP synthase, H+ transporting subunits, and metabolic proteins (lactate dehydrogenase B4, malate dehydrogenase 1b). Twelve proteins were significantly induced by EE{sub 2} including microtubule-associated protein tau (Mapt), astrocytic phosphoprotein, ependymin precursor, and calmodulin. Mapt showed an increase in protein abundance but a decrease in mRNA expression after EE{sub 2} exposure{sub ,} suggesting there may be a negative feedback response in the telencephalon to decreased mRNA transcription with increasing Mapt protein abundance. These results demonstrate that a low, environmentally relevant exposure to EE{sub 2} can rapidly alter the abundance of proteins involved in cell differentiation and proliferation, neuron network morphology, and long-term synaptic potentiation. Together, these findings provide a better understanding of the molecular responses underlying E{sub 2} feedback in the brain and demonstrate that quantitative proteomics can be successfully used in ecotoxicology to characterize affected cellular pathways and endocrine physiology.

Modulation of estrogenic exposure effects via alterations in salinity and dissolved oxygen in male fathead minnows, Pimephales promelas

Science.gov (United States)

Laboratory exposure data indicate that estrogens and estrogen mimics can cause endocrine disruption in male fathead minnows (Pimephales promelas). In the wild, conditions are not static as is often the case in the laboratory. Changes in water quality parameters, such as salinity influx due to road s...

Larval exposure to environmentally relevant mixtures of alkylphenolethoxylates reduces reproductive competence in male fathead minnows

Science.gov (United States)

Bistodeau, T.J.; Barber, L.B.; Bartell, S.E.; Cediel, R.A.; Grove, K.J.; Klaustermeier, J.; Woodard, J.C.; Lee, K.E.; Schoenfuss, H.L.

2006-01-01

The ubiquitous presence of nonylphenolethoxylate/octylphenolethoxylate (NPE/OPE) compounds in aquatic environments adjacent to wastewater treatment plants (WWTP) warrants an assessment of the endocrine disrupting potential of these complex mixtures on aquatic vertebrates. In this study, fathead minnow larvae were exposed for 64 days to a mixture of NPE/OPE, which closely models the NPE/OPE composition of a major metropolitan WWTP effluent. Target exposure concentrations included a total NPE/OPE mixture load of 200% of the WWTP effluent concentration (148 ??g/L), 100% of the WWTP effluent concentration (74 ??g/L) and 50% of the WWTP effluent concentration (38 ??g/L). The NPE/OPE mixture contained 0.2% 4-t-octylphenol, 2.8% 4-nonylphenol, 5.1% 4-nonylphenolmonoethoxylate, 9.3% 4-nonylphenoldiethoxylate, 0.9% 4-t-octylphenolmonoethoxylate, 3.1% 4-t-octylphenoldiethoxylate, 33.8% 4-nonylphenolmonoethoxycarboxylate, and 44.8% 4-nonylphenoldiethoxycarboxylate. An additional exposure of 5 ??g/L 4-nonylphenol (nominal) was conducted. The exposure utilized a flow-through system supplied by ground water and designed to deliver consistent concentrations of applied chemicals. Following exposure, larvae were raised to maturity. Upon sexual maturation, exposed male fish were allowed to compete with control males in a competitive spawning assay. Nest holding ability of control and exposed fish was carefully monitored for 7 days. All male fish were then sacrificed and analyzed for plasma vitellogenin, developmental changes in gonadal tissues, alterations in the development of secondary sexual characters, morphometric changes, and changes to reproductive behavior. When exposed to the 200% NPE/OPE treatment most larvae died within the first 4 weeks of exposure. Both the 100% and 50% NPE/OPE exposures caused a significant decrease in reproductive behavior, as indicated by an inability of many of the previously exposed males to acquire and hold a nest site required for reproduction

A study of temporal effects of the model anti-androgen flutamide on components of the hypothalamic-pituitary-gonadal axis in adult fathead minnows

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U.S. Environmental Protection Agency — The aim of this study was to investigate temporal changes in the hypothalamic-pituitary-gonadal axis of fathead minnow treated with the model androgen receptor (AR)...

A Time-course Analysis of Effects of the Steroidogenesis Inhibitor Ketoconazole on Components of the Hypothalamic-pituitary-gonadal Axis of Fathead Minnows

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The objective of this study was to evaluate temporal effects of the model steroidogenesis inhibitor ketoconazole (KTC) on aspects of reproductive endocrine function controlled by the hypothalamic-pituitary-gonadal (HPG) axis in the fathead minnow (Pimephales promelas). Ketoconazo...

Effects-based monitoring in the lower Green Bay/Fox River and Milwaukee Estuary Areas of Concern using caged Fathead Minnows (Pimephales promelas)

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Within the Great Lakes there is an increased focus on contaminants of emerging concern (CECs) and their potential effects on aquatic organisms, including adverse reproductive effects. To further characterize the utility of caged fathead minnows (Pimephales promelas) for effects-b...

Effects of a Short-term Exposure to the Fungicide Prochloraz on Endocrine Function and Gene Expression in Female Fathead Minnows (Pimephales promelas)

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Prochloraz is a fungicide known to cause endocrine disruption through effects on the hypothalamic-pituitary-gonadal (HPG) axis. To determine the short-term impacts of prochloraz on gene expression and steroid production, adult female fathead minnows (Pimephales promelas) were exp...

A time-course analysis of effects of the steroidogenesis inhibitor ketoconazole on components of the hypothalamic-pituitary-gonadal axis of fathead minnows (Presentation)

Science.gov (United States)

The objective of this study was to evaluate temporal effects of the model steroidogenesis inhibitor ketoconazole (KTC) on aspects of reproductive endocrine function controlled by the hypothalamic-pituitary-gonadal (HPG) axis in the fathead minnow (Pimephales promelas). Ketoconazo...

EFFECTS OF THE MAMMALIAN ANTIANDROGEN VINCLOZOLIN ON DEVELOPMENT AND REPRODUCTION OF THE FATHEAD MINNOW (PIMEPHALES PROMELAS)

Science.gov (United States)

Previous work with the chlorinated fungicide vinclozolin and its metabolites, 2-{[(3,5-dichloropheny1]-carbamoyl]oxy}-2-methyl-3-butenoic acid (M1) and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2), indicated antiandrogenic properties expressed in vivo as abnormalities in...

Preliminary studies into the effects of the human pharmaceutical Clofibric acid on sperm parameters in adult Fathead minnow.

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Runnalls, Tamsin J; Hala, David N; Sumpter, John P

2007-08-15

The effects of Clofibric acid (a persistent environmental metabolite of Clofibrate, a human pharmaceutical), on Fathead minnows were studied. Fibrates are used to prevent cardiovascular disease through their antilipidemic activity. In a series of experiments, in which fish were exposed to waterborne Clofibric acid, no convincing, reproducible antilipidemic effects were observed. In contrast, in three separate experiments, Clofibric acid affected the reproductive axis of fish. Spermatogenesis was apparently impaired, leading to a marked reduction in sperm count in two of the three experiments. Various measures of sperm motility were also reduced, although only significantly so at the highest concentration of Clofibric acid tested (1mg/L). There were also indications that plasma androgen concentrations were reduced. These effects of Clofibric acid on the reproductive axis of fish are similar to those that occur in some mammals as a side-effect of the drug. Taken together, a weight-of-evidence argument would suggest that the main discernable effect of Clofibric acid on fish is likely to be a reproductive, not an antilipidemic, one. Although some of these reproductive effects of Clofibric acid occurred only at a high concentration (1mg/L), others occurred at lower concentrations (microg/litre), near or similar to those reported in the aquatic environment (ng to low microg/litre range). Although we recognise that this is not a definitive study of the effects of Clofibric acid on fish reproduction, the results strongly suggest that Clofibric acid could adversely affect sperm parameters and androgen concentrations in adult Fathead minnows. Further studies are warranted. This may be an example of a drug in which an accidentally discovered side-effect found in mammals turns out to be the most important effect in a different vertebrate group, namely fish.

CDNA CLONING OF FATHEAD MINNOW (PIMEPHALES PROMELAS) ESTROGEN AND ANDROGEN RECEPTORS FOR USE IN STEROID RECEPTOR EXTRAPOLATION STUDIES FOR ENDOCRINE DISRUPTING CHEMICALS

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cDNA Cloning of Fathead minnow (Pimephales promelas) Estrogen and Androgen Receptors for Use in Steroid Receptor Extrapolation Studies for Endocrine Disrupting Chemicals. Wilson, V.S.1,, Korte, J.2, Hartig P. 1, Ankley, G.T.2, Gray, L.E., Jr 1, , and Welch, J.E.1. 1U.S...

Ammonia causes decreased brain monoamines in fathead minnows (Pimephales promelas)

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Ronan, Patrick J.; Gaikowski, Mark P.; Hamilton, Steven J.; Buhl, Kevin J.; Summers, Cliff H.

2007-01-01

Hyperammonemia, arising from variety of disorders, leads to severe neurological dysfunction. The mechanisms of ammonia toxicity in brain are not completely understood. This study investigated the effects of ammonia on monoaminergic systems in brains of fathead minnows (Pimephales promelas). Fish serve as a good model system to investigate hyperammonemic effects on brain function since no liver manipulations are necessary to increase endogenous ammonia concentrations. Using high performance liquid chromatography with electrochemical detection, monoamines and some associated metabolites were measured from whole brain homogenate. Adult males were exposed for 48 h to six different concentrations of ammonia (0.01–2.36 mg/l unionized) which bracketed the 96-h LC50 for this species. Ammonia concentration-dependent decreases were found for the catecholamines (norepinephrine and dopamine) and the indoleamine serotonin (5-HT). After an initial increase in the 5-HT precursor 5-hydroxytryptophan it too decreased with increasing ammonia concentrations. There were also significant increases in the 5-HIAA/5-HT and DOPAC/DA ratios, often used as measures of turnover. There were no changes in epinephrine (Epi) or monoamine catabolites (DOPAC, 5-HIAA) at any ammonia concentrations tested. Results suggest that ammonia causes decreased synthesis while also causing increased release and degradation. Increased release may underlie behavioral reactions to ammonia exposure in fish. This study adds weight to a growing body of evidence demonstrating that ammonia leads to dysfunctional monoaminergic systems in brain which may underlie neurological symptoms associated with human disorders such as hepatic encephalopathy.

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

Energy Technology Data Exchange (ETDEWEB)

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

2012-05-01

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.

Acute toxicity of sodium chloride, pentachlorophenol, Guthion, and hexavalent chromium to fathead minnows (Pimephales promelas) and goldfish (Carassius auratus)

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Adelman, I.R.; Smith, L.L. Jr.; Siesennop, G.D.

1976-02-01

The 96-h LC50's for sodium chloride were 7650 and 7341 mg/liter, for pentachlorophenol 0.21 and 0.22 mg/liter, for Guthion 1.9 and 2.4 mg/liter, and for hexavalent chromium 48 and 120 mg/liter, for fathead minnows (Pimephales promelas) and goldfish (Carassius auratus), respectively. Threshold LC50's were reached in 6 days for sodium chloride (7650 and 7322 mg/liter for fathead minnows and goldfish, respectively), and pentachlorophenol (0.21 and 0.21 mg/liter), but were not attained in 11 days (termination of testing) with Guthion (0.76 and 0.80 mg/liter) and hexavalent chromium (18 and 33 mg/liter). With pentachlorophenol and Guthion goldfish were initially more resistant, but by termination there was no significant difference in LC50's between the two species. With hexavalent chromium the goldfish were more resistant throughout the 11-day test, and with sodium chloride goldfish were initially more resistant but at attainment of a threshold LC50 were less resistant. Use of toxicity curves for assessment of acute mortality permits interpretation not possible in 96-h tests where LC50's are computed at 24-h intervals.

High-resolution mass spectrometry of skin mucus for monitoring physiological impacts and contaminant biotransformation products in fathead minnows exposed to wastewater effluent.

Science.gov (United States)

Mosley, Jonathan D; Ekman, Drew R; Cavallin, Jenna E; Villeneuve, Daniel L; Ankley, Gerald T; Collette, Timothy W

2018-03-01

High-resolution mass spectrometry is advantageous for monitoring physiological impacts and contaminant biotransformation products in fish exposed to complex wastewater effluent. We evaluated this technique using skin mucus from male and female fathead minnows (Pimephales promelas) exposed to control water or treated wastewater effluent at 5, 20, and 100% levels for 21 d, using an on-site, flow-through system providing real-time exposure. Both sex-specific and non-sex-specific responses were observed in the mucus metabolome, the latter suggesting the induction of general compensatory pathways for xenobiotic exposures. Altogether, 85 statistically significant treatment-dependent metabolite changes were observed out of the 310 total endogenous metabolites that were detected (156 of the 310 were annotated). Partial least squares-regression models revealed strong covariances between the mucus metabolomes and up-regulated hepatic messenger ribonucleic acid (mRNA) transcripts reported previously for these same fish. These regression models suggest that mucus metabolomic changes reflected, in part, processes by which the fish biotransformed xenobiotics in the effluent. In keeping with this observation, we detected a phase II transformation product of bisphenol A in the skin mucus of male fish. Collectively, these findings demonstrate the utility of mucus as a minimally invasive matrix for simultaneously assessing exposures and effects of environmentally relevant mixtures of contaminants. Environ Toxicol Chem 2018;37:788-796. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Selective uptake and biological consequences of environmentally relevant antidepressant pharmaceutical exposures on male fathead minnows

Science.gov (United States)

Schultz, Melissa M.; Painter, Meghan M.; Bartell, Stephen E.; Logue, Amanda; Furlong, Edward T.; Werner, Stephen L.; Schoenfuss, Heiko L.

2011-01-01

Antidepressant pharmaceuticals have been reported in wastewater effluent at the nanogram to low microgram-per-liter range, and include bupropion (BUP), fluoxetine (FLX), sertraline (SER), and venlafaxine (VEN). To assess the effects of antidepressants on reproductive anatomy, physiology, and behavior, adult male fathead minnows (Pimeplwles promelas) were exposed for 21 days either to a single concentration of the antidepressants FLX, SER, VEN, or BUP, or to an antidepressant mixture. The data demonstrated that exposure to VEN (305 ng/L and 1104 ng/L) and SER (5.2 ng/L) resulted in mortality. Anatomical alterations were noted within the testes of fish exposed to SER and FLX, both modulators of the neurotransmitter serotonin. Additionally, FLX at 28 ng/L induced vitellogenin in male fish—a common endpoint for estrogenic endocrine disruption. Significant alterations in male secondary sex characteristics were noted with single exposures. Effects of single compound exposures neither carried over, nor became additive in the antidepressant mixtures, and reproductive behavior was not affected. Analysis of brain tissues from the exposed fish suggested increased uptake of FLX, SER and BUP and minimal uptake of VEN when compared to exposure water concentrations. Furthermore, the only metabolite detected consistently in the brain tissues was norfluoxetine. Similar trends of uptake by brain tissue were observed when fish were exposed to antidepressant mixtures. The present study demonstrates that anatomy and physiology, but not reproductive behavior, can be disrupted by exposure to environmental concentrations of some antidepressants. The observation that antidepressant uptake into fish tissues is selective may have consequences on assessing the mode-of-action and effects of these compounds in future studies.

The interactive effects of multiple stressors on physiological stress responses and club cell investment in fathead minnows

International Nuclear Information System (INIS)

Manek, Aditya K.; Ferrari, Maud C.O.; Niyogi, Som; Chivers, Douglas P.

2014-01-01

Anthropogenic activities have dramatically increased over the past decades, with the consequence that many organisms are simultaneously exposed to multiple stressors. Understanding how organisms respond to these stressors is a key focus for scientists from many disciplines. Here we investigated the interactive effects of two stressors, UV radiation (UVR) and cadmium (Cd) exposure on a common freshwater fish, fathead minnow (Pimephales promelas). UVR is known to influence the density of epidermal club cells (ECCs), which are not only a key component of the innate immune system of fishes, but are also the source of chemical alarm cues that serve to warn other fishes of nearby predators. In contrast, Cd impairs the physiological stress response and ability of fish to respond to alarm cues. We used an integrative approach to examine physiological stress response as well as investment in ECCs. Fish exposed to UVR had higher levels of cortisol than non-exposed controls, but Cd reduced cortisol levels substantially for fish exposed to UVR. Fish exposed to UVR, either in the presence or absence of Cd, showed consistent decreases in ECC investment compared to non-exposed controls. Despite differences in ECC number, there was no difference in the potency of alarm cues prepared from the skin of UVR and Cd exposed or non-exposed fish indicating that UVR and Cd exposure combined may have little influence on chemically-mediated predator–prey interactions. - Highlights: • UV radiation caused a physiological stress response (cortisol release) in fish. • Cd reduced cortisol levels substantially for fish exposed to UV. • Fish exposed to UV, with or without Cd, showed decreases in club cell investment. • There was no difference in alarm cues potency from UV and Cd exposed fish. • Our work highlights the difficulty of untangling effects of multiple stressors

The interactive effects of multiple stressors on physiological stress responses and club cell investment in fathead minnows

Energy Technology Data Exchange (ETDEWEB)

Manek, Aditya K., E-mail: aditya.manek@usask.ca [Department of Biology, University of Saskatchewan, Saskatoon, S7N 5E2 SK (Canada); Ferrari, Maud C.O. [Department of Biomedical Sciences, WCVM, University of Saskatchewan, Saskatoon, SK S7N 5B4 (Canada); Niyogi, Som; Chivers, Douglas P. [Department of Biology, University of Saskatchewan, Saskatoon, S7N 5E2 SK (Canada)

2014-04-01

Anthropogenic activities have dramatically increased over the past decades, with the consequence that many organisms are simultaneously exposed to multiple stressors. Understanding how organisms respond to these stressors is a key focus for scientists from many disciplines. Here we investigated the interactive effects of two stressors, UV radiation (UVR) and cadmium (Cd) exposure on a common freshwater fish, fathead minnow (Pimephales promelas). UVR is known to influence the density of epidermal club cells (ECCs), which are not only a key component of the innate immune system of fishes, but are also the source of chemical alarm cues that serve to warn other fishes of nearby predators. In contrast, Cd impairs the physiological stress response and ability of fish to respond to alarm cues. We used an integrative approach to examine physiological stress response as well as investment in ECCs. Fish exposed to UVR had higher levels of cortisol than non-exposed controls, but Cd reduced cortisol levels substantially for fish exposed to UVR. Fish exposed to UVR, either in the presence or absence of Cd, showed consistent decreases in ECC investment compared to non-exposed controls. Despite differences in ECC number, there was no difference in the potency of alarm cues prepared from the skin of UVR and Cd exposed or non-exposed fish indicating that UVR and Cd exposure combined may have little influence on chemically-mediated predator–prey interactions. - Highlights: • UV radiation caused a physiological stress response (cortisol release) in fish. • Cd reduced cortisol levels substantially for fish exposed to UV. • Fish exposed to UV, with or without Cd, showed decreases in club cell investment. • There was no difference in alarm cues potency from UV and Cd exposed fish. • Our work highlights the difficulty of untangling effects of multiple stressors.

A computational model of the hypothalamic - pituitary - gonadal axis in female fathead minnows (Pimephales promelas exposed to 17α-ethynylestradiol and 17β-trenbolone

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Lazorchak James M

2011-05-01

Full Text Available Abstract Background Endocrine disrupting chemicals (e.g., estrogens, androgens and their mimics are known to affect reproduction in fish. 17α-ethynylestradiol is a synthetic estrogen used in birth control pills. 17β-trenbolone is a relatively stable metabolite of trenbolone acetate, a synthetic androgen used as a growth promoter in livestock. Both 17α-ethynylestradiol and 17β-trenbolone have been found in the aquatic environment and affect fish reproduction. In this study, we developed a physiologically-based computational model for female fathead minnows (FHM, Pimephales promelas, a small fish species used in ecotoxicology, to simulate how estrogens (i.e., 17α-ethynylestradiol or androgens (i.e., 17β-trenbolone affect reproductive endpoints such as plasma concentrations of steroid hormones (e.g., 17β-estradiol and testosterone and vitellogenin (a precursor to egg yolk proteins. Results Using Markov Chain Monte Carlo simulations, the model was calibrated with data from unexposed, 17α-ethynylestradiol-exposed, and 17β-trenbolone-exposed FHMs. Four Markov chains were simulated, and the chains for each calibrated model parameter (26 in total converged within 20,000 iterations. With the converged parameter values, we evaluated the model's predictive ability by simulating a variety of independent experimental data. The model predictions agreed with the experimental data well. Conclusions The physiologically-based computational model represents the hypothalamic-pituitary-gonadal axis in adult female FHM robustly. The model is useful to estimate how estrogens (e.g., 17α-ethynylestradiol or androgens (e.g., 17β-trenbolone affect plasma concentrations of 17β-estradiol, testosterone and vitellogenin, which are important determinants of fecundity in fish.

Effects of a homologous series of linear alcohol ethoxylate surfactants on fathead minnow early life stages.

Science.gov (United States)

Lizotte, R E; Wong, D C; Dorn, P B; Rodgers, J H

1999-11-01

Effects of a homologous series of three primarily linear alcohol ethoxylate surfactants were studied in laboratory flow-through 28-day early-life-stage tests with fathead minnow (Pimephales promelas Rafinesque). Surfactants were a C(9-11), C(12-13), and C(14-15) with an average of 6, 6.5, and 7 ethylene oxide units per mole of alcohol, respectively. Average measured surfactant recoveries were 103%, 81%, and 79% of nominal concentrations for the C(9-11) EO 6, C(12-13) EO 6.5, and C(14-15) EO 7 studies, respectively. Embryo survival at 48 h was not adversely affected at any of the concentrations tested. Impaired hatching and deformed fry were observed only in the C(12-13) EO 6.5 study. The 28-day LC50 values were 4.87, 2.39, and 1.02 mg/L for the C(9-11) EO 6, C(12-13) EO 6.5, and C(14-15) EO 7 surfactants, respectively. The corresponding NOECs for survival were 1.01, 1.76, and 0.74 mg/L. Posthatch fry growth was more sensitive than survival for the C(12-13) EO 6.5 and C(14-15) EO 7 surfactants. Survival of posthatch fry decreased with increasing surfactant alkyl chain length. Twenty-eight-day laboratory data were compared to 96-h laboratory, 10-day laboratory and 30-day stream mesocosm data for fathead minnow previously determined for these surfactants. Survival endpoints from the different exposures were comparable and only varied within a factor of two. Similarity of results suggests that it is possible to effectively use 96-h, 10-day, or 28-day laboratory data to predict environmental effects concentrations of these surfactants for fish. http://link.springer-ny. com/link/service/journals/00244/bibs/37n4p536.html

Influence of elevated alkalinity and natural organic matter (NOM) on tissue-specific metal accumulation and reproductive performance in fathead minnows during chronic, multi-trophic exposures to a metal mine effluent.

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Ouellet, Jacob D; Dubé, Monique G; Niyogi, Som

2013-09-01

Metal bioavailability in aquatic organisms is known to be influenced by various water chemistry parameters. The present study examined the influence of alkalinity and natural organic matter (NOM) on tissue-specific metal accumulation and reproductive performance of fathead minnows (Pimephales promelas) during environmentally relevant chronic exposures to a metal mine effluent (MME). Sodium bicarbonate (NaHCO3) or NOM (as commercial humic acid) were added to a Canadian MME [45 percent process water effluent (PWE)] in order to evaluate whether increases in alkalinity (3-4 fold) or NOM (~1.5-3mg/L dissolved organic carbon) would reduce metal accumulation and mitigate reproductive toxicity in fathead minnows during a 21-day multi-trophic exposure. Eleven metals (barium, boron, cobalt, copper, lithium, manganese, molybdenum, nickel, rubidium, selenium, and strontium) were elevated in the 45 percent PWE relative to the reference water. Exposure to the unmodified 45 percent PWE resulted in a decrease of fathead minnow egg production (~300 fewer eggs/pair) relative to the unmodified reference water, over the 21-day exposure period. Water chemistry modifications produced a modest decrease in free ion activity of some metals (as shown by MINTEQ, Version 3) in the 45 percent PWE exposure water, but did not alter the metal burden in the treatment-matched larval Chironomus dilutus (the food source of fish during exposure). The tissue-specific metal accumulation increased in fish exposed to the 45 percent PWE relative to the reference water, irrespective of water chemistry modifications, and the tissue metal concentrations were found to be similar between fish in the unmodified and modified 45 percent PWE (higher alkalinity or NOM) treatments. Interestingly however, increased alkalinity and NOM markedly improved fish egg production both in the reference water (~500 and ~590 additional eggs/pair, respectively) and 45 percent PWE treatments (~570 and ~260 additional eggs

GENE ARRAY ANALYSIS OF THE VENTRAL PROSTATE IN RATS EXPOSED TO EITHER VINCLOZOLIN OR PROCYMIDONE

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GENE ARRAY ANALYSIS OF THE VENTRAL PROSTATE IN RATS EXPOSED TO EITHER VINCLOZOLIN OR PROCYMIDONE. MB Rosen, VS Wilson, JE Schmid, and LE Gray Jr. US EPA, ORD, NHEERL, RTP, NC.Vinclozolin (Vi) and procymidone (Pr) are antiandrogenic fungicides. While changes in gene expr...

Assessing the sublethal effects of in-river concentrations of parameters contributing to cumulative effects in the Athabasca river basin using a fathead minnow bioassay.

Science.gov (United States)

Squires, Allison J; Dubé, Monique G; Rozon-Ramilo, Lisa D

2013-03-01

The Athabasca River basin, located in Alberta, Canada, covers 157, 000 km(2) and holds significant cultural and economic importance. Recent research assessed changes in several water quality and quantity parameters that have changed both spatially (along the river continuum) and temporally (pre-development and present day) in the Athabasca River Basin. In particular, parameters such as salinity and dissolved sulphate have changed significantly across the Athabasca River mainstem over the past five decades. Further laboratory testing has linked concentrations of these parameters to changes in fathead minnow reproduction. Research is required to determine whether these changes observed in the laboratory can be applied to actual in-river conditions. The objectives of the present study were to twofold: assess changes in fathead minnow response metrics (i.e., condition, liver and gonad size, egg production, and gill histology) associated with increasing concentrations of salinity and dissolved sulphate and determine whether sublethal effect thresholds established in laboratory experiments correspond to actual in-river concentrations using water from the mouth and headwaters of the Athabasca River. Three dose-response experiments (NaCl, SO4, and water sampled from the mouth of the Athabasca River) were conducted at Jasper National Park, Alberta, Canada. Significant increases in mean eggs per female per day occurred at the 50% treatment for the mouth experiment and thresholds previously developed in the laboratory were verified. Copyright © 2012 SETAC.

Molecular initiating events of the intersex phenotype: Low-dose exposure to 17α-ethinylestradiol rapidly regulates molecular networks associated with gonad differentiation in the adult fathead minnow testis

International Nuclear Information System (INIS)

Feswick, April; Loughery, Jennifer R.; Isaacs, Meghan A.; Munkittrick, Kelly R.; Martyniuk, Christopher J.

2016-01-01

Highlights: • Male fathead minnow were exposed to 17alpha ethinylestradiol (EE2). • Both 11-ketotestosterone and testosterone production was decreased relative to controls. • A gene network associated with doublesex and mab-3 related transcription factor 1 were suppressed. • Genes involved in granulosa cell development were increased and sensitive to EE2 exposure. • Molecular initiating events that may be related to the intersex condition were identified. - Abstract: Intersex, or the presence of oocytes in the testes, has been documented in fish following exposure to wastewater effluent and estrogenic compounds. However, the molecular networks underlying the intersex condition are not completely known. To address this, we exposed male fathead minnows to a low, environmentally-relevant concentration of 17alpha-ethinylestradiol (EE2) (15 ng/L) and measured the transcriptome response in the testis after 96 h to identify early molecular initiating events that may proceed the intersex condition. The short-term exposure to EE2 did not affect gonadosomatic index and proportion of gametes within the testes. However, the production of 11-ketotestosterone and testosterone from the testis in vitro was decreased relative to controls. Expression profiling using a 8 × 60 K fathead minnow microarray identified 10 transcripts that were differentially expressed in the testes, the most dramatic change being that of coagulation factor XIII A chain (20-fold increase). Transcripts that included guanine nucleotide binding protein (Beta Polypeptide 2), peroxisome proliferator-activated receptor delta, and WNK lysine deficient protein kinase 1a, were down-regulated by EE2. Subnetwork enrichment analysis revealed that EE2 suppressed transcriptional networks associated with steroid metabolism, hormone biosynthesis, and sperm mobility. Most interesting was that gene networks associated with doublesex and mab-3 related transcription factor 1 (dmrt1) were suppressed in the adult

Molecular initiating events of the intersex phenotype: Low-dose exposure to 17α-ethinylestradiol rapidly regulates molecular networks associated with gonad differentiation in the adult fathead minnow testis

Energy Technology Data Exchange (ETDEWEB)

Feswick, April; Loughery, Jennifer R.; Isaacs, Meghan A.; Munkittrick, Kelly R.; Martyniuk, Christopher J., E-mail: cmartyni@yahoo.ca

2016-12-15

Highlights: • Male fathead minnow were exposed to 17alpha ethinylestradiol (EE2). • Both 11-ketotestosterone and testosterone production was decreased relative to controls. • A gene network associated with doublesex and mab-3 related transcription factor 1 were suppressed. • Genes involved in granulosa cell development were increased and sensitive to EE2 exposure. • Molecular initiating events that may be related to the intersex condition were identified. - Abstract: Intersex, or the presence of oocytes in the testes, has been documented in fish following exposure to wastewater effluent and estrogenic compounds. However, the molecular networks underlying the intersex condition are not completely known. To address this, we exposed male fathead minnows to a low, environmentally-relevant concentration of 17alpha-ethinylestradiol (EE2) (15 ng/L) and measured the transcriptome response in the testis after 96 h to identify early molecular initiating events that may proceed the intersex condition. The short-term exposure to EE2 did not affect gonadosomatic index and proportion of gametes within the testes. However, the production of 11-ketotestosterone and testosterone from the testis in vitro was decreased relative to controls. Expression profiling using a 8 × 60 K fathead minnow microarray identified 10 transcripts that were differentially expressed in the testes, the most dramatic change being that of coagulation factor XIII A chain (20-fold increase). Transcripts that included guanine nucleotide binding protein (Beta Polypeptide 2), peroxisome proliferator-activated receptor delta, and WNK lysine deficient protein kinase 1a, were down-regulated by EE2. Subnetwork enrichment analysis revealed that EE2 suppressed transcriptional networks associated with steroid metabolism, hormone biosynthesis, and sperm mobility. Most interesting was that gene networks associated with doublesex and mab-3 related transcription factor 1 (dmrt1) were suppressed in the adult

High-resolution Mass Spectrometry of Skin Mucus for Monitoring Physiological Impacts in Fish Exposed to Wastewater Effluent at a Great Lakes AOC

Science.gov (United States)

High-resolution mass spectrometry is advantageous for monitoring physiological impacts and contaminant biotransformation products in fish exposed to complex wastewater effluent. We evaluated this technique using skin mucus from male and female fathead minnows (Pimephales promela...

Screening differentially expressed genes in an amphipod (Hyalella azteca) exposed to fungicide vinclozolin by suppression subtractive hybridization.

Science.gov (United States)

Wu, Yun H; Wu, Tsung M; Hong, Chwan Y; Wang, Yei S; Yen, Jui H

2014-01-01

Vinclozolin, a dicarboximide fungicide, is an endocrine disrupting chemical that competes with an androgenic endocrine disruptor compound. Most research has focused on the epigenetic effect of vinclozolin in humans. In terms of ecotoxicology, understanding the effect of vinclozolin on non-target organisms is important. The expression profile of a comprehensive set of genes in the amphipod Hyalella azteca exposed to vinclozolin was examined. The expressed sequence tags in low-dose vinclozolin-treated and -untreated amphipods were isolated and identified by suppression subtractive hybridization. DNA dot blotting was used to confirm the results and establish a subtracted cDNA library for comparing all differentially expressed sequences with and without vinclozolin treatment. In total, 494 differentially expressed genes, including hemocyanin, heatshock protein, cytochrome, cytochrome oxidase and NADH dehydrogenase were detected. Hemocyanin was the most abundant gene. DNA dot blotting revealed 55 genes with significant differential expression. These genes included larval serum protein 1 alpha, E3 ubiquitin-protein ligase, mitochondrial cytochrome c oxidase, mitochondrial protein, proteasome inhibitor, hemocyanin, zinc-finger-containing protein, mitochondrial NADH-ubiquinone oxidoreductase and epididymal sperm-binding protein. Vinclozolin appears to upregulate stress-related genes and hemocyanin, related to immunity. Moreover, vinclozolin downregulated NADH dehydrogenase, related to respiration. Thus, even a non-lethal concentration of vinclozolin still has an effect at the genetic level in H. azteca and presents a potential risk, especially as it would affect non-target organism hormone metabolism.

Warmed Winter Water Temperatures Alter Reproduction in Two Fish Species

Science.gov (United States)

Firkus, Tyler; Rahel, Frank J.; Bergman, Harold L.; Cherrington, Brian D.

2018-02-01

We examined the spawning success of Fathead Minnows ( Pimephales promelas) and Johnny Darters ( Etheostoma nigrum) exposed to elevated winter water temperatures typical of streams characterized by anthropogenic thermal inputs. When Fathead Minnows were exposed to temperature treatments of 12, 16, or 20 °C during the winter, spawning occurred at 16 and 20 °C but not 12 °C. Eggs were deposited over 9 weeks before winter spawning ceased. Fathead Minnows from the three winter temperature treatments were then exposed to a simulated spring transition. Spawning occurred at all three temperature treatments during the spring, but fish from the 16° and 20 °C treatment had delayed egg production indicating a latent effect of warm winter temperatures on spring spawning. mRNA analysis of the egg yolk protein vitellogenin showed elevated expression in female Fathead Minnows at 16 and 20 °C during winter spawning that decreased after winter spawning ceased, whereas Fathead Minnows at 12 °C maintained comparatively low expression during winter. Johnny Darters were exposed to 4 °C to represent winter temperatures in the absence of thermal inputs, and 12, 16, and 20 °C to represent varying degrees of winter thermal pollution. Johnny Darters spawned during winter at 12, 16, and 20 °C but not at 4 °C. Johnny Darters at 4 °C subsequently spawned following a simulated spring period while those at 12, 16, and 20 °C did not. Our results indicate elevated winter water temperatures common in effluent-dominated streams can promote out-of-season spawning and that vitellogenin expression is a useful indicator of spawning readiness for fish exposed to elevated winter temperatures.

Evaluation of potential mechanisms of atrazine-induced reproductive impairment in fathead minnow (Pimephales promelas) and Japanese medaka (Oryzias latipes)

Science.gov (United States)

Richter, Cathy; Papoulias, Diana M.; Whyte, Jeffrey J.; Tillitt, Donald E.

2016-01-01

Atrazine has been implicated in reproductive dysfunction of exposed organisms, and previous studies documented decreased egg production in Japanese medaka (Oryzias latipes) and fathead minnows (Pimephales promelas) during 30-d to 38-d exposures to 0.5 µg/L, 5 µg/L, and 50 µg/L atrazine. The authors evaluated possible mechanisms underlying the reduction in egg production. Gene expression in steroidogenesis pathways and the hypothalamus–pituitary–gonad axis of male and female fish was measured. Atrazine did not significantly induce gonad aromatase (cyp19a1a) expression. An atrazine-induced shift in the number of females in an active reproductive state was observed. Expression of the egg maturation genes vitellogenin 1 (vtg1) and zona pellucida glycoprotein 3.1 (zp3.1) in medaka females was correlated and had a bimodal distribution. In both species, females with low vtg1 or zp3.1 expression also had low expression of steroidogenesis genes in the gonad, estrogen receptor in the liver, and gonadotropins in the brain. In the medaka, the number of females per tank that had high expression of zp3.1 was significantly correlated with egg production per tank. The number of medaka females with low expression of zp3.1 increased significantly with atrazine exposure. Thus, the decline in egg production observed in response to atrazine exposure may be the result of a coordinated downregulation of genes required for reproduction in a subset of females.

Warmed Winter Water Temperatures Alter Reproduction in Two Fish Species.

Science.gov (United States)

Firkus, Tyler; Rahel, Frank J; Bergman, Harold L; Cherrington, Brian D

2018-02-01

We examined the spawning success of Fathead Minnows (Pimephales promelas) and Johnny Darters (Etheostoma nigrum) exposed to elevated winter water temperatures typical of streams characterized by anthropogenic thermal inputs. When Fathead Minnows were exposed to temperature treatments of 12, 16, or 20 °C during the winter, spawning occurred at 16 and 20 °C but not 12 °C. Eggs were deposited over 9 weeks before winter spawning ceased. Fathead Minnows from the three winter temperature treatments were then exposed to a simulated spring transition. Spawning occurred at all three temperature treatments during the spring, but fish from the 16° and 20 °C treatment had delayed egg production indicating a latent effect of warm winter temperatures on spring spawning. mRNA analysis of the egg yolk protein vitellogenin showed elevated expression in female Fathead Minnows at 16 and 20 °C during winter spawning that decreased after winter spawning ceased, whereas Fathead Minnows at 12 °C maintained comparatively low expression during winter. Johnny Darters were exposed to 4 °C to represent winter temperatures in the absence of thermal inputs, and 12, 16, and 20 °C to represent varying degrees of winter thermal pollution. Johnny Darters spawned during winter at 12, 16, and 20 °C but not at 4 °C. Johnny Darters at 4 °C subsequently spawned following a simulated spring period while those at 12, 16, and 20 °C did not. Our results indicate elevated winter water temperatures common in effluent-dominated streams can promote out-of-season spawning and that vitellogenin expression is a useful indicator of spawning readiness for fish exposed to elevated winter temperatures.

CHRONIC EFFECTS OF THE HERBICIDE DIURON ON FRESHWATER CLADOCERANS,AMPHIPODS,MIDGES,MINNOWS,WORMS, AND SNAILS

Science.gov (United States)

The chronic effects of the herbicide diuron on survival and reproduction of Daphnia pulex, and survival and growth of the amphipod Hyalella azteca, the midge Chironomus tentans, juvenile and embro/larval fathead minnows, Pimephales promelas, annelid worms, Lumbriculus variegatus,...

Expression profiling and gene ontology analysis in fathead minnow (Pimephales promelas) liver following exposure to pulp and paper mill effluents

Energy Technology Data Exchange (ETDEWEB)

Costigan, Shannon L.; Werner, Julieta; Ouellet, Jacob D.; Hill, Lauren G. [Department of Biology, Lakehead University, 955 Oliver Road, Ontario P7B 5E1, (Canada); Law, R. David, E-mail: dlaw@lakeheadu.ca [Department of Biology, Lakehead University, 955 Oliver Road, Ontario P7B 5E1, (Canada)

2012-10-15

Many studies link pulp and paper mill effluent (PPME) exposure to adverse effects in fish populations present in the mill receiving environments. These impacts are often characteristic of endocrine disruption and may include impaired reproduction, development and survival. While these physiological endpoints are well-characterized, the molecular mechanisms causing them are not yet understood. To investigate changes in gene transcription induced by exposure to a PPME at several stages of treatment, male and female fathead minnows (FHMs) were exposed for 6 days to 25% (v/v) secondary (biologically) treated kraft effluent (TK) or 100% (v/v) combined mill outfall (CMO) from a mill producing both kraft pulp and newsprint. The gene expression changes in the livers of these fish were analyzed using a 22 K oligonucleotide microarray. Exposure to TK or CMO resulted in significant changes in the expression levels of 105 and 238 targets in male FHMs and 296 and 133 targets in females, respectively. Targets were then functionally analyzed using gene ontology tools to identify the biological processes in fish hepatocytes that were affected by exposure to PPME after its secondary treatment. Proteolysis was affected in female FHMs exposed to both TK and CMO. In male FHMs, no processes were affected by TK exposure, while sterol, isoprenoid, steroid and cholesterol biosynthesis and electron transport were up-regulated by CMO exposure. The results presented in this study indicate that short-term exposure to PPMEs affects the expression of reproduction-related genes in the livers of both male and female FHMs, and that secondary treatment of PPMEs may not neutralize all of their metabolic effects in fish. Gene ontology analysis of microarray data may enable identification of biological processes altered by toxicant exposure and thus provide an additional tool for monitoring the impact of PPMEs on fish populations.

Expression profiling and gene ontology analysis in fathead minnow (Pimephales promelas) liver following exposure to pulp and paper mill effluents

International Nuclear Information System (INIS)

Costigan, Shannon L.; Werner, Julieta; Ouellet, Jacob D.; Hill, Lauren G.; Law, R. David

2012-01-01

Many studies link pulp and paper mill effluent (PPME) exposure to adverse effects in fish populations present in the mill receiving environments. These impacts are often characteristic of endocrine disruption and may include impaired reproduction, development and survival. While these physiological endpoints are well-characterized, the molecular mechanisms causing them are not yet understood. To investigate changes in gene transcription induced by exposure to a PPME at several stages of treatment, male and female fathead minnows (FHMs) were exposed for 6 days to 25% (v/v) secondary (biologically) treated kraft effluent (TK) or 100% (v/v) combined mill outfall (CMO) from a mill producing both kraft pulp and newsprint. The gene expression changes in the livers of these fish were analyzed using a 22 K oligonucleotide microarray. Exposure to TK or CMO resulted in significant changes in the expression levels of 105 and 238 targets in male FHMs and 296 and 133 targets in females, respectively. Targets were then functionally analyzed using gene ontology tools to identify the biological processes in fish hepatocytes that were affected by exposure to PPME after its secondary treatment. Proteolysis was affected in female FHMs exposed to both TK and CMO. In male FHMs, no processes were affected by TK exposure, while sterol, isoprenoid, steroid and cholesterol biosynthesis and electron transport were up-regulated by CMO exposure. The results presented in this study indicate that short-term exposure to PPMEs affects the expression of reproduction-related genes in the livers of both male and female FHMs, and that secondary treatment of PPMEs may not neutralize all of their metabolic effects in fish. Gene ontology analysis of microarray data may enable identification of biological processes altered by toxicant exposure and thus provide an additional tool for monitoring the impact of PPMEs on fish populations.

Chronic effects assessment and plasma concentrations of the {beta}-blocker propranolol in fathead minnows (Pimephales promelas)

Energy Technology Data Exchange (ETDEWEB)

Giltrow, Emma [Institute for the Environment, Brunel University, Uxbridge, Middlesex UB8 3PH (United Kingdom); Eccles, Paul D. [Institute for the Environment, Brunel University, Uxbridge, Middlesex UB8 3PH (United Kingdom); Biosciences, School of Health Sciences and Social Care, Brunel University, Uxbridge, Middlesex UB8 3PH (United Kingdom); Winter, Matthew J.; McCormack, Paul J. [AstraZeneca Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA (United Kingdom); Rand-Weaver, Mariann [Biosciences, School of Health Sciences and Social Care, Brunel University, Uxbridge, Middlesex UB8 3PH (United Kingdom); Hutchinson, Thomas H. [Natural Environmental Research Council, Plymouth Marine Laboratory, Prospect Place, The Hoe, Plymouth PL1 3DH (United Kingdom); Sumpter, John P., E-mail: john.sumpter@brunel.ac.uk [Institute for the Environment, Brunel University, Uxbridge, Middlesex UB8 3PH (United Kingdom)

2009-11-27

The presence of many human pharmaceuticals in the aquatic environment is now a worldwide concern, yet little is known of the chronic effects that these bioactive substances may be having on aquatic organisms. Propranolol, a non-specific beta adrenoreceptor blocker ({beta}-blocker), is used to treat high blood pressure and heart disease in humans. Propranolol has been found in surface waters worldwide at concentrations ranging from 12 to 590 ng/L. To test the potential for ecologically relevant effects in fish in receiving waters, short-term (21 days) adult reproduction studies were conducted, in which fathead minnows were exposed to nominal concentrations of propranolol hydrochloride [CAS number 318-98-9] ranging from 0.001 to 10 mg/L (measured concentrations typically from 78 to 130%). Exposure of fish to 3.4 mg/L (measured) over 3 days caused 100% mortality or severe toxicity requiring euthanasia. The most sensitive endpoints from the studies were a decrease in hatchability (with regard to the number of days to hatch) and a concentration-related increase in female gonadal somatic index (GSI), giving LOEC{sup hatchability} and LOEC{sup female} {sup GSI} values of 0.1 mg/L. Concentration-related decreases in weights of male fish were also observed, with LOEC{sup m}ale wet weight value of 1.0 mg/L, and the LOEC{sup r}eproduction value was 1.0 mg/L. Collectively, these data do not suggest that propranolol was acting as a reproductive toxin. Plasma concentrations of propranolol in male fish exposed to nominal concentrations of 0.1 and 1.0 mg/L were 0.34 and 15.00 mg/L, respectively, which constitutes 436 and 1546% of measured water concentrations. These compare with predicted concentrations of 0.07 and 0.84 mg/L, and thus to a degree support the use of partition coefficient models for predicting concentrations in plasma in fish. In addition, propranolol plasma concentrations in fish exposed to water concentrations of 0.1 and 1.0 mg/L were greater than the human

Chronic effects assessment and plasma concentrations of the β-blocker propranolol in fathead minnows (Pimephales promelas)

International Nuclear Information System (INIS)

Giltrow, Emma; Eccles, Paul D.; Winter, Matthew J.; McCormack, Paul J.; Rand-Weaver, Mariann; Hutchinson, Thomas H.; Sumpter, John P.

2009-01-01

The presence of many human pharmaceuticals in the aquatic environment is now a worldwide concern, yet little is known of the chronic effects that these bioactive substances may be having on aquatic organisms. Propranolol, a non-specific beta adrenoreceptor blocker (β-blocker), is used to treat high blood pressure and heart disease in humans. Propranolol has been found in surface waters worldwide at concentrations ranging from 12 to 590 ng/L. To test the potential for ecologically relevant effects in fish in receiving waters, short-term (21 days) adult reproduction studies were conducted, in which fathead minnows were exposed to nominal concentrations of propranolol hydrochloride [CAS number 318-98-9] ranging from 0.001 to 10 mg/L (measured concentrations typically from 78 to 130%). Exposure of fish to 3.4 mg/L (measured) over 3 days caused 100% mortality or severe toxicity requiring euthanasia. The most sensitive endpoints from the studies were a decrease in hatchability (with regard to the number of days to hatch) and a concentration-related increase in female gonadal somatic index (GSI), giving LOEC hatchability and LOEC female GSI values of 0.1 mg/L. Concentration-related decreases in weights of male fish were also observed, with LOEC m ale wet weight value of 1.0 mg/L, and the LOEC r eproduction value was 1.0 mg/L. Collectively, these data do not suggest that propranolol was acting as a reproductive toxin. Plasma concentrations of propranolol in male fish exposed to nominal concentrations of 0.1 and 1.0 mg/L were 0.34 and 15.00 mg/L, respectively, which constitutes 436 and 1546% of measured water concentrations. These compare with predicted concentrations of 0.07 and 0.84 mg/L, and thus to a degree support the use of partition coefficient models for predicting concentrations in plasma in fish. In addition, propranolol plasma concentrations in fish exposed to water concentrations of 0.1 and 1.0 mg/L were greater than the human therapeutic plasma concentration

Effects of chronic exposure to waterborne copper and nickel in binary mixture on tissue-specific metal accumulation and reproduction in fathead minnow (Pimephales promelas).

Science.gov (United States)

Driessnack, Melissa K; Jamwal, Ankur; Niyogi, Som

2017-10-01

The current study evaluated the interactive effects of chronic waterborne copper (Cu) and nickel (Ni) exposure on tissue-specific metal accumulation and reproductive performance in fathead minnow (Pimephales promelas). Fish trios (1 male: 2 female; n = 5-6) were exposed for 21 days to: (i) control (no added Cu or Ni), (ii) waterborne Cu (45 μg/L), (iii) waterborne Ni (270 μg/L), and (iv) binary mixture of waterborne Cu and Ni (45 and 270 μg/L, respectively). Fish fecundity (cumulative egg production) was found to be the most sensitive reproductive endpoint, and the interaction of Cu and Ni elicited an additive effect on egg production. Tissue-specific accumulation of both metals was not influenced by the interaction of Cu and Ni, except an increased Cu and Ni burden in the carcass and ovary, respectively, were recorded. The expressions of hepatic estrogen receptor genes (ER-α and ER-β) and the circulating estradiol level in females were also not affected by the metal-mixture treatment. However, co-exposure to waterborne Cu and Ni resulted in a significant downregulation of the hepatic vitellogenin gene in females, which was associated with the maximum upregulation of the hepatic metallothionein gene. In addition, a significant alteration of ovarian histopathology (decreased abundance of post-vitellogenic follicles, and increased follicular atresia) was also observed only in females exposed to Cu and Ni in mixture. Collectively, these observations suggest that chronic waterborne exposure to Cu and Ni in binary mixture may impair fish reproductive capacity by inducing histopathological damage in ovarian tissue, and disrupting of energy homeostasis in fish. Copyright © 2017 Elsevier Ltd. All rights reserved.

Fathead minnow steroidogenesis: in silico analyses reveals tradeoffs between nominal target efficacy and robustness to cross-talk

Directory of Open Access Journals (Sweden)

Villeneuve Daniel L

2010-06-01

Full Text Available Abstract Background Interpreting proteomic and genomic data is a major challenge in predictive ecotoxicology that can be addressed by a systems biology approach. Mathematical modeling provides an organizational platform to consolidate protein dynamics with possible genomic regulation. Here, a model of ovarian steroidogenesis in the fathead minnow, Pimephales promelas, (FHM is developed to evaluate possible transcriptional regulation of steroid production observed in microarray studies. Results The model was developed from literature sources, integrating key signaling components (G-protein and PKA activation with their ensuing effect on steroid production. The model properly predicted trajectory behavior of estradiol and testosterone when fish were exposed to fadrozole, a specific aromatase inhibitor, but failed to predict the steroid hormone behavior occurring one week post-exposure as well as the increase in steroid levels when the stressor was removed. In vivo microarray data implicated three modes of regulation which may account for over-production of steroids during a depuration phase (when the stressor is removed: P450 enzyme up-regulation, inhibin down-regulation, and luteinizing hormone receptor up-regulation. Simulation studies and sensitivity analysis were used to evaluate each case as possible source of compensation to endocrine stress. Conclusions Simulation studies of the testosterone and estradiol response to regulation observed in microarray data supported the hypothesis that the FHM steroidogenesis network compensated for endocrine stress by modulating the sensitivity of the ovarian network to global cues coming from the hypothalamus and pituitary. Model predictions of luteinizing hormone receptor regulation were consistent with depuration and in vitro data. These results challenge the traditional approach to network elucidation in systems biology. Generally, the most sensitive interactions in a network are targeted for further

Temperature and metal exposure affect membrane fatty acid composition and transcription of desaturases and elongases in fathead minnow muscle and brain.

Science.gov (United States)

Fadhlaoui, Mariem; Pierron, Fabien; Couture, Patrice

2018-02-01

In this study, we tested the hypothesis that metal exposure affected the normal thermal response of cell membrane FA composition and of elongase and desaturase gene transcription levels. To this end, muscle and brain membrane FA composition and FA desaturase (fads2, degs2 and scd2) and elongase (elovl2, elovl5 and elovl6) gene transcription levels were analyzed in fathead minnows (Pimephales promelas) acclimated for eight weeks to 15, 25 or 30°C exposed or not to cadmium (Cd, 6μg/l) or nickel (Ni, 450 6μg/l). The response of membrane FA composition to temperature variations or metal exposure differed between muscle and brain. In muscle, an increase of temperature induced a decrease of polyunsaturated FA (PUFA) and an increase of saturated FA (SFA) in agreement with the current paradigm. Although a similar response was observed in brain between 15 and 25°C, at 30°C, brain membrane unsaturation was higher than predicted. In both tissues, metal exposure affected the normal thermal response of membrane FA composition. The transcription of desaturases and elongases was higher in the brain and varied with acclimation temperature and metal exposure but these variations did not generally reflect changes in membrane FA composition. The mismatch between gene transcription and membrane composition highlights that several levels of control other than gene transcription are involved in adjusting membrane FA composition, including post-transcriptional regulation of elongases and desaturases and de novo phospholipid biosynthesis. Our study also reveals that metal exposure affects the mechanisms involved in adjusting cell membrane FA composition in ectotherms. Copyright © 2017 Elsevier Inc. All rights reserved.

Assessing effects of a mining and municipal sewage effluent mixture on fathead minnow (Pimephales promelas) reproduction using a novel, field-based trophic-transfer artificial stream.

Science.gov (United States)

Rickwood, Carrie J; Dubé, Monique G; Weber, Lynn P; Lux, Sarah; Janz, David M

2008-01-31

The Junction Creek watershed, located in Sudbury, ON, Canada receives effluent from three metal mine wastewater treatment plants, as well as a municipal wastewater (MWW) discharge. Effects on fish have been documented within the creek (decreased egg size and increased metal body burdens). It has been difficult to identify the cause of the effects observed due to the confounded nature of the creek. The objectives of this investigation were to assess the: (1) effects of a mine effluent and municipal wastewater (CCMWW) mixture on fathead minnow (FHM; Pimephales promelas) reproduction in an on-site artificial stream and (2) importance of food (Chironomus tentans) as a source of exposure using a trophic-transfer system. Exposures to CCMWW through the water significantly decreased egg production and spawning events. Exposure through food and water using the trophic-transfer system significantly increased egg production and spawning events. Embryos produced in the trophic-transfer system showed similar hatching success but increased incidence and severity of deformities after CCMWW exposure. We concluded that effects of CCMWW on FHM were more apparent when exposed through the water. Exposure through food and water may have reduced effluent toxicity, possibly due to increased nutrients and organic matter, which may have reduced metal bioavailability. More detailed examination of metal concentrations in the sediment, water column, prey (C. tentans) and FHM tissues is recommended to better understand the toxicokinetics of potential causative compounds within the different aquatic compartments when conducting exposures through different pathways.

Effect of the anti-androgenic endocrine disruptor vinclozolin on embryonic testis cord formation and postnatal testis development and function.

Science.gov (United States)

Uzumcu, Mehmet; Suzuki, Hiroetsu; Skinner, Michael K

2004-01-01

Vinclozolin is a systemic dicarboximide fungicide that is used on fruits, vegetables, ornamental plants, and turf grass. Vinclozolin and its metabolites are known to be endocrine disruptors and act as androgen receptor antagonists. The hypothesis tested in the current study is that transient embryonic exposure to an anti-androgenic endocrine disruptor at the time of testis determination alters testis development and subsequently influences adult spermatogenic capacity and male reproduction. The effects of vinclozolin on embryonic testicular cord formation in vitro were examined, as well as the effects of transient in utero vinclozolin exposure on postnatal testis development and function. Embryonic day 13 (E13, sperm-positive vaginal smear day = E0) gonads were cultured in the absence or presence of vinclozolin (50-500microM). Vinclozolin treated gonads had significantly fewer cords (P vinclozolin (100 mg/kg/day) between embryonic days 8 and 14 (E8-E14) of development. Testis morphology and function were analyzed from postnatal day (P) 0, pubertal P20, and adult P60. No significant effect of vinclozolin on testis histology or germ cell viability was observed in P0 testis. The pubertal P20 testis from vinclozolin exposed animals had significantly higher numbers of apoptotic germ cells (P vinclozolin exposed males (P vinclozolin exposed animals was higher in adult P60 animals. Observations demonstrate that vinclozolin can effect embryonic testicular cord formation in vitro and that transient in utero exposure to vinclozolin increases apoptotic germ cell numbers in the testis of pubertal and adult animals. This correlated to reduced sperm motility in the adult. In conclusion, transient exposure to vinclozolin during the time of testis differentiation (i.e. cord formation) alters testis development and function. Observations indicate that transient exposure to an anti-androgenic endocrine disruptor during embryonic development causes delayed effects later in adult life

Social hierarchy modulates responses of fish exposed to contaminants of emerging concern

Science.gov (United States)

Many organisms, including the fathead minnow (Pimephales promelas), a toxicological model organism, establish social hierarchies. The social rank of each male in a population is under the control of the hypothalamic-pituitary-gonadal (HPG) axis mainly through regulation of circul...

Short and long term bystander effect induction by fathead minnows (Pimephales promelas, Rafinesque, 1820) injected with environmentally relevant whole body doses of 226Ra

International Nuclear Information System (INIS)

Smith, Richard W.; Seymour, Colin B.; Mothersill, Carmel E.

2013-01-01

Bystander effect induction by fathead minnows injected with environmentally relevant doses of 226 Ra was investigated. Twenty four h and 6 months after injection with a single dose of 21, 210 or 2100 μBq, fin tissue samples emitted a pro-apoptotic signal, which reduced the clonogenic survival of an apoptosis sensitive reporter cell line. Twenty four h and 10 weeks after injection explants from non-injected bystander fish, swum with the injected fish, also emitted a pro-apoptotic signal. However 6 months after injection the bystander fish to 21 and 210 μBq injected fish emitted an anti-apoptotic signal. This demonstrates that extremely low dose irradiation can have effects outside of the irradiated fish. This has implications for population and ecosystem responses to contamination. -- Highlights: • Non-injected fish, swum with injected fish, gave the same response for up to 10 weeks. • After 6 months this response by non-injected fish changed to an anti-apoptotic signal. • This shows that 226 Ra contamination can have effects outside of the irradiated fish

Comparative anti-androgenic actions of vinclozolin and flutamide on transgenerational adult onset disease and spermatogenesis

OpenAIRE

Anway, Matthew D.; Rekow, Stephen S.; Skinner, Michael K.

2008-01-01

Exposure of gestating female rats to the anti-androgenic endocrine disruptor vinclozolin has been shown to induce transgenerational adult onset disease phenotypes. The current study, was designed to compare the actions of vinclozolin to the known anti-androgenic compound flutamide. The gestating female rats were exposed to intraperitoneal injections during embryonic day 8–14 (E8–E14) to 100 mg/kg/day vinclozolin or flutamide at either 5mg or 20 mg/kg/day. As previously observed, vinclozolin i...

Do laboratory species protect endangered species? Interspecies variation in responses to 17β-estradiol, a model endocrine active compound

Science.gov (United States)

Jorgenson, Zachary G.; Buhl, Kevin J.; Bartell, Stephen E.; Schoenfuss, Heiko L.

2015-01-01

Although the effects of estrogens on model laboratory species are well documented, their utility as surrogates for other species, including those listed as endangered, are less clear. Traditionally, conservation policies are evaluated based on model organism responses but are intended to protect all species in an environment. We tested the hypothesis that the endangered Rio Grande silvery minnow (Hybognathus amarus) is more vulnerable to endocrine disruption—as assessed through its larval predator-escape performance, survival, juvenile sex ratios, and whole-body vitellogenin concentration—than the commonly used toxicological model species fathead minnow (Pimephales promelas) and the bluegill sunfish (Lepomis macrochirus). Fish were exposed concurrently for 21 days to the model endocrine active compound (EAC) 17ß-estradiol (E2) at 10 ng E2/L and 30 ng E2/L in a flow-through system using reconstituted water that simulated the physicochemical conditions of the Middle Rio Grande in New Mexico, USA. No significant differences were observed between the fathead and silvery minnow in larval predator-escape response or juvenile sex ratio. Rio Grande silvery minnow survival decreased significantly at day 14 compared with the other two species; by day 21, both cyprinid species (silvery minnow and fathead minnow) exhibited a significant decrease in survival compared with bluegill sunfish, a member of the family Centrarchidae. Male Rio Grande silvery minnow showed a significant increase in whole-body vitellogenin concentration in the 10 ng/L treatment, whereas fathead minnow and bluegill sunfish showed no significant increases in vitellogenin concentrations across treatments. Our study showed response differences to estrogen exposures between the two cyprinid species and further divergence in responses between the families Cyprinidae and Centrarchidae. These results suggest that commonly used laboratory model organisms may be less sensitive to EACs than the endangered

Do laboratory species protect endangered species? Interspecies variation in responses to 17β-estradiol, a model endocrine active compound.

Science.gov (United States)

Jorgenson, Z G; Buhl, K; Bartell, S E; Schoenfuss, H L

2015-01-01

Although the effects of estrogens on model laboratory species are well documented, their utility as surrogates for other species, including those listed as endangered, are less clear. Traditionally, conservation policies are evaluated based on model organism responses but are intended to protect all species in an environment. We tested the hypothesis that the endangered Rio Grande silvery minnow (Hybognathus amarus) is more vulnerable to endocrine disruption-as assessed through its larval predator-escape performance, survival, juvenile sex ratios, and whole-body vitellogenin concentration-than the commonly used toxicological model species fathead minnow (Pimephales promelas) and the bluegill sunfish (Lepomis macrochirus). Fish were exposed concurrently for 21 days to the model endocrine active compound (EAC) 17ß-estradiol (E2) at 10 ng E2/L and 30 ng E2/L in a flow-through system using reconstituted water that simulated the physicochemical conditions of the Middle Rio Grande in New Mexico, USA. No significant differences were observed between the fathead and silvery minnow in larval predator-escape response or juvenile sex ratio. Rio Grande silvery minnow survival decreased significantly at day 14 compared with the other two species; by day 21, both cyprinid species (silvery minnow and fathead minnow) exhibited a significant decrease in survival compared with bluegill sunfish, a member of the family Centrarchidae. Male Rio Grande silvery minnow showed a significant increase in whole-body vitellogenin concentration in the 10 ng/L treatment, whereas fathead minnow and bluegill sunfish showed no significant increases in vitellogenin concentrations across treatments. Our study showed response differences to estrogen exposures between the two cyprinid species and further divergence in responses between the families Cyprinidae and Centrarchidae. These results suggest that commonly used laboratory model organisms may be less sensitive to EACs than the endangered Rio

A mixture of an environmentally realistic concentration of a phthalate and herbicide reduces testosterone in male fathead minnow (Pimephales promelas) through a novel mechanism of action

Energy Technology Data Exchange (ETDEWEB)

Crago, Jordan, E-mail: jcrago@uwm.edu [Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI 53204 (United States); Klaper, Rebecca, E-mail: rklaper@uwm.edu [School of Freshwater Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI 53204 (United States)

2012-04-15

Several chemicals that are used by humans, such as pesticides and plastics, are released into the aquatic environment through wastewater and runoff and have been shown to be potent disruptors of androgen synthesis at high concentrations. Although many of these chemicals have been studied in isolation, a large amount of uncertainty remains over how fish respond to low concentrations of anti-androgenic mixtures, which more accurately reflects how such chemicals are present in the aquatic environment. In this study male fathead minnows (FHM) (Pimephales promelas) were exposed to environmentally relevant concentrations of two anti-androgens, the herbicide linuron, and the plasticizer di(2-ethylhexyl) phthalate (DEHP) individually and as part of a mixture of the two for a 28-day period. At the end of this period there was a reduction in plasma testosterone (T) concentrations in male FHM exposed to the mixture, but not in FHM exposed individually to linuron or DEHP or the control FHM. There was also a significant reduction in 17{beta}-estradiol (E2) in the DEHP-only and mixture exposed groups as compared to the control. Contrary to what has been previously published for these two chemicals in mammals, the lower plasma T concentrations in male FHM exposed to the mixture was not a result of the inhibition of genes involved in steroidogenesis; nor due to an increase in the expression of genes associated with peroxisome proliferation. Rather, an increase in relative transcript abundance for CYP3A4 in the liver and androgen- and estrogen-specific SULT2A1 and SULT1st2 in the testes provides evidence that the decrease in plasma T and E2 may be linked to increased steroid catabolism. Feedback from the pituitary is not repressed as the relative expression of follicle stimulating hormone {beta}-subunit mRNA transcript levels in the brain was significantly higher in both DEHP and mixture exposed FHM. In addition, luteinizing hormone {beta}-subunit mRNA transcript levels increased

A mixture of an environmentally realistic concentration of a phthalate and herbicide reduces testosterone in male fathead minnow (Pimephales promelas) through a novel mechanism of action

International Nuclear Information System (INIS)

Crago, Jordan; Klaper, Rebecca

2012-01-01

Several chemicals that are used by humans, such as pesticides and plastics, are released into the aquatic environment through wastewater and runoff and have been shown to be potent disruptors of androgen synthesis at high concentrations. Although many of these chemicals have been studied in isolation, a large amount of uncertainty remains over how fish respond to low concentrations of anti-androgenic mixtures, which more accurately reflects how such chemicals are present in the aquatic environment. In this study male fathead minnows (FHM) (Pimephales promelas) were exposed to environmentally relevant concentrations of two anti-androgens, the herbicide linuron, and the plasticizer di(2-ethylhexyl) phthalate (DEHP) individually and as part of a mixture of the two for a 28-day period. At the end of this period there was a reduction in plasma testosterone (T) concentrations in male FHM exposed to the mixture, but not in FHM exposed individually to linuron or DEHP or the control FHM. There was also a significant reduction in 17β-estradiol (E2) in the DEHP-only and mixture exposed groups as compared to the control. Contrary to what has been previously published for these two chemicals in mammals, the lower plasma T concentrations in male FHM exposed to the mixture was not a result of the inhibition of genes involved in steroidogenesis; nor due to an increase in the expression of genes associated with peroxisome proliferation. Rather, an increase in relative transcript abundance for CYP3A4 in the liver and androgen- and estrogen-specific SULT2A1 and SULT1st2 in the testes provides evidence that the decrease in plasma T and E2 may be linked to increased steroid catabolism. Feedback from the pituitary is not repressed as the relative expression of follicle stimulating hormone β-subunit mRNA transcript levels in the brain was significantly higher in both DEHP and mixture exposed FHM. In addition, luteinizing hormone β-subunit mRNA transcript levels increased but were not

Effects of the antiandrogens, vinclozolin and cyproterone acetate on gonadal development in the Japanese medaka (Oryzias latipes).

Science.gov (United States)

Kiparissis, Yiannis; Metcalfe, Tracy L; Balch, Gordon C; Metcalfe, Chris D

2003-05-29

This study was focused on determining the effects of exposure to antiandrogens on the gonadal development of Japanese medaka (Oryzias latipes). Test compounds included the fungicide, vinclozolin and the clinical antiandrogen, cyproterone acetate. Newly hatched medaka were exposed to aqueous solutions of vinclozolin (2500 microg/l) and the vinclozolin fungicide formulation, Ronilan (1000 and 5000 microg/l) and cyproterone acetate (1 and 10 microg/l), for 3 months. Histological evaluation of the gonadal tissues of exposed fish indicated that the 5000 microg/l concentration of the vinclozolin formulation (Ronilan) induced a low incidence of intersex (i.e. testis-ova) and the 2500 microg/l concentration of vinclozolin-affected spermatogenesis in males. Also, the vinclozolin treatments induced moderate ovarian atresia. Cyproterone acetate also induced a low incidence of testis-ova, but in contrast to the vinclozolin treatment the amount of ovarian tissue in the testis-ova was equal to or greater than the amount of testicular tissue. In the cyproterone acetate treatments, both oogenesis and spermatogenesis were moderately inhibited at all test concentrations. The results of this study indicate that antiandrogens have the potential to alter testicular development and gametogenesis in fish. However, research is needed to determine the mechanisms by which antiandrogens affect fish.

Study of morbidity of personnel with potential exposure to vinclozolin.

Science.gov (United States)

Zober, A; Hoffmann, G; Ott, M G; Will, W; Germann, C; van Ravenzwaay, B

1995-01-01

OBJECTIVES--To examine internal exposure and targeted health outcomes of employees exposed to 3-(3,5-dichlorophenyl)-5-methyl-5-vinyl-1,3-oxazolidine-2,4-dione; chemical abstracts service (CAS) number: 50471-44-8 (vinclozolin). METHODS--A cross sectional study of 67 men exposed to vinclozolin for one to 13 years during synthesis and formulation operations and 52 controls. Biomonitoring was based on determination of urinary metabolites that contained a 3,5-dichloroaniline (3,5-DCA) moiety. Targeted health endpoints were the same as in previous subchronic and chronic animal studies--namely, reversible changes in the concentrations of hormones of the adrenocorticotrophic and gonadotrophic feedback systems, signs of liver injury, haemolytic anaemia, cataract formation (uniquely in rats), and hormonally induced hyperplasia and tumours at high doses. The clinical investigation consisted of a medical and occupational history questionnaire, physical examination, laboratory determinations (including testosterone, LH, and FSH measurements), ultrasonography of the liver and prostate, a detailed eye examination, and routine spirometry. RESULTS--The mean 3,5-DCA concentration for two thirds of the study group exceeded an equivalent of the vinclozolin acceptable daily intake (ADI) used for consumer regulatory purposes. Even the highest concentrations were, however, at least 10 times below the no observable adverse effect level (NOAEL) based on animal studies. Analysis of physical examination and laboratory data provided no evidence of hormonal responses induced by vinclozolin. Furthermore, no evidence of liver injury, prostate changes, cataract formation, or haemolytic anaemia was found. CONCLUSION--There was no evidence of any health effects induced by vinclozolin among employees with potential long term exposure. In particular, no antiandrogenic effects were found. PMID:7795738

Impaired swim bladder inflation in early-life stage fathead ...

Science.gov (United States)

The present study investigated whether inhibition of deiodinase, the enzyme which converts thyroxine (T4) to the more biologically-active form, 3,5,3'-triiodothyronine (T3), would impact inflation of the posterior and/or anterior chamber of the swim bladder, processes previously demonstrated to be thyroid-hormone regulated. Two experiments were conducted using a model deiodinase inhibitor, iopanoic acid (IOP). In the first study, fathead minnow (Pimephales promelas) embryos were exposed to 0.6, 1.9, or 6.0 mg IOP/L or control water in a flow-through system until reaching 6 days post-fertilization (dpf) at which time posterior swim bladder inflation was assessed. To examine effects on anterior swim bladder inflation, a second study was conducted with 6 dpf larvae exposed to the same IOP concentrations until reaching 21 dpf. Fish from both studies were sampled for T4/T3 measurements, gene transcription analyses, and thyroid histopathology. In the embryo study, incidence and length of inflated posterior swim bladders were significantly reduced in the 6.0 mg/L treatment at 6 dpf. Incidence of inflation and length of anterior swim bladder in larval fish were significantly reduced in all IOP treatments at 14 dpf, but inflation recovered by 18 dpf. Throughout the larval study, whole body T4 concentrations were significantly increased and T3 concentrations were significantly decreased in all IOP treatments. Consistent with hypothesized compensatory responses, sig

Endocrine disruptors and hypospadias: role of genistein and the fungicide vinclozolin.

Science.gov (United States)

Vilela, Marcelo L B; Willingham, Emily; Buckley, Jill; Liu, Ben Chun; Agras, Koray; Shiroyanagi, Yoshiyuki; Baskin, Laurence S

2007-09-01

The phytoestrogen (plant estrogen) genistein, present in soy products, is of interest because in utero exposure to genistein can cause hypospadias in our mouse model and maternal consumption of soy is prevalent in human populations. Another compound of interest is the fungicide vinclozolin, which also causes hypospadias in the mouse and rat and can occur concurrently with genistein in the diet as a residue on exposed foods. A study in the United Kingdom found no relationship between a maternal organic vegetarian diet and hypospadias frequency, but women who consumed nonorganic vegetarian diets had a greater percentage of sons with hypospadias. Because nonorganic diets can include residues of pesticides such as vinclozolin, we sought to assess the interaction of realistic daily exposures to genistein and vinclozolin and their effects on the incidence of hypospadias. Pregnant mice were fed a soy-free diet and orally gavaged from gestational days 13 to 17 with 0.17 mg/kg/day of genistein, 10 mg/kg/day of vinclozolin, or genistein and vinclozolin together at the same doses, all in 100 microL of corn oil. The controls received the corn oil vehicle. The male fetuses were examined at gestational day 19 for hypospadias, both macroscopically and histologically. We identified no hypospadias in the corn oil group. The incidence of hypospadias was 25% with genistein alone, 42% with vinclozolin alone, and 41% with genistein and vinclozolin together. These findings support the idea that exposure to these compounds during gestation could contribute to the development of hypospadias.

EVALUATION OF THE ANTIANDROGENIC PROPERTIES OF VINCLOZOLIN IN THE FATHEAD MINNOW

Science.gov (United States)

Anti-androgens are an important class of endocrine-disrupting chemicals, however, little is known concerning their effects in fish. This presentation will detail the effects of a known mammalian anti-androgen, vincolozin, on reproduction in a model small fish species, the fathea...

Prioritization of contaminants of emerging concern in wastewater treatment plant discharges using chemical: Gene interactions in caged fish

Science.gov (United States)

We examined whether contaminants present in surface waters could be prioritized for further assessment by linking the presence of specific chemicals to gene expression changes in exposed fish. Fathead minnows were deployed in cages for 2, 4, or 8 days at three locations near two ...

Vinclozolin, a widely used fungizide, enhanced BaP-induced micronucleus formation in human derived hepatoma cells by increasing CYP1A1 expression.

Science.gov (United States)

Wu, Xin-Jiang; Lu, Wen-qing; Roos, Peter H; Mersch-Sundermann, Volker

2005-10-15

Vinclozolin, a widely used fungicide, can be identified as a residue in numerous vegetable and fruit samples. To get insight in its genetic toxicity, we investigated the genotoxic effect of vinclozolin in the human derived hepatoma cell line HepG2 using the micronucleus (MN) assay. Additionally, to evaluate the co- or anti-mutagenic potency of vinclozolin, we treated HepG2 cells with different concentrations of vinclozolin for 24 h. Subsequently, the cells were exposed to benzo[a]pyrene (BaP) for 1h. Exposure of HepG2 cells to 50-400 microM vinclozolin alone did not cause any induction of micronuclei. However, a pronounced co-mutagenic effect was observed. MN frequencies caused by BaP increased by 30.6%, 52.8% and 65.3% after pretreatment of the cell cultures with 50, 100 and 200 microM vinclozolin, respectively. The highest concentration (400 microM) of vinclozolin tested caused cytotoxicity. Therefore, micronuclei were not considered for that concentration. To clarify the mechanism of cogenotoxicity, we assayed cytochrome P450 1A1 (CYP1A1), which plays a pivotal role in activation of BaP. Cells exposed to vinclozolin led to significant increase of CYP1A1 expression in Western blot. The result suggested that induction of CYP1A1 by vinclozolin account for its enhancing effect on genotoxicity caused by BaP.

Acclimation-induced changes in toxicity and induction of metallothionein-like proteins in the fathead minnow following sublethal exposure to cobalt, silver, and zinc

International Nuclear Information System (INIS)

Hobson, J.F.

1986-01-01

Increases in tolerance and resistance to metal toxicity by aquatic organisms have been linked to elevated levels of low-molecular-weight metal-binding proteins (e.g., metallothioneins). Acclimation-induced changes in toxic response and the concentration of metallothionein-like proteins (MTP) were studied in laboratory populations of the fathead minnow, Pimephales promelas, following sublethal exposure to Co, Ag, and Zn. Following 7 and 14 days of sublethal exposure, tolerance and resistance, as measured by acute toxicity values, were altered in a dose dependent fashion. Acute toxicity values returned to control levels after 21 days of continuous exposure. Tolerance and resistance of Co- and Zn-acclimated animals were depressed after a 7-day post-acclimation period in control water. Tolerance and resistance of Ag-acclimated animals were temporarily enhanced after 7 days post-acclimation and returned to control levels after 14 days. Accumulation of Co, Ag, and Zn measured as wholebody residues appeared to be regulated in 4 of 6 exposure regimes with residues reaching stable levels after 7 to 14 days of exposure. MTP was induced by exposure to 1.8 mg Zn/L and 0.01 mg Ag/L, however, no sustained (i.e., post 21 days) tolerance or resistance were observed at these dose levels indicating that these two biological responses may not be directly related

Estimating the effects of 17α-ethinylestradiol on stochastic population growth rate of fathead minnows: a population synthesis of empirically derived vital rates

Science.gov (United States)

Schwindt, Adam R.; Winkelman, Dana L.

2016-01-01

Urban freshwater streams in arid climates are wastewater effluent dominated ecosystems particularly impacted by bioactive chemicals including steroid estrogens that disrupt vertebrate reproduction. However, more understanding of the population and ecological consequences of exposure to wastewater effluent is needed. We used empirically derived vital rate estimates from a mesocosm study to develop a stochastic stage-structured population model and evaluated the effect of 17α-ethinylestradiol (EE2), the estrogen in human contraceptive pills, on fathead minnow Pimephales promelas stochastic population growth rate. Tested EE2 concentrations ranged from 3.2 to 10.9 ng L−1 and produced stochastic population growth rates (λ S ) below 1 at the lowest concentration, indicating potential for population decline. Declines in λ S compared to controls were evident in treatments that were lethal to adult males despite statistically insignificant effects on egg production and juvenile recruitment. In fact, results indicated that λ S was most sensitive to the survival of juveniles and female egg production. More broadly, our results document that population model results may differ even when empirically derived estimates of vital rates are similar among experimental treatments, and demonstrate how population models integrate and project the effects of stressors throughout the life cycle. Thus, stochastic population models can more effectively evaluate the ecological consequences of experimentally derived vital rates.

Impact of natural organic matter and increased water hardness on DGT prediction of copper bioaccumulation by yellow lampmussel (Lampsilis cariosa) and fathead minnow (Pimephales promelas).

Science.gov (United States)

Philipps, Rebecca R; Xu, Xiaoyu; Mills, Gary L; Bringolf, Robert B

2018-06-01

We conducted an exposure experiment with Diffusive Gradients in Thin- Films (DGT), fathead minnow (Pimephales promelas), and yellow lampmussel (Lampsilis cariosa) to estimate bioavailability and bioaccumulation of Cu. We hypothesized that Cu concentrations measured by DGT can be used to predict Cu accumulation in aquatic animals and alterations of water chemistry can affect DGT's predict ability. Three water chemistries (control soft water, hard water, and addition of natural organic matter (NOM)) and three Cu concentrations (0, 30, and 60 μg/L) were selected, so nine Cu-water chemistry combinations were used. NOM addition treatments resulted in decreased concentrations of DGT-measured Cu and free Cu ion predicted by Biotic Ligand Model (BLM). Both hard water and NOM addition treatments had reduced concentrations of Cu ion and Cu-dissolved organic matter complexes compared to other treatments. DGT-measured Cu concentrations were linearly correlated to fish accumulated Cu, but not to mussel accumulated Cu. Concentrations of bioavailable Cu predicted by BLM, the species complexed with biotic ligands of aquatic organisms and, was highly correlated to DGT-measured Cu. In general, DGT-measured Cu fit Cu accumulations in fish, and this passive sampling technique is acceptable at predicting Cu concentrations in fish in waters with low NOM concentrations. Copyright © 2018 Elsevier Ltd. All rights reserved.

Evaluation of fish models of soluble epoxide hydrolase inhibition.

Science.gov (United States)

Newman, J W; Denton, D L; Morisseau, C; Koger, C S; Wheelock, C E; Hinton, D E; Hammock, B D

2001-01-01

Substituted ureas and carbamates are mechanistic inhibitors of the soluble epoxide hydrolase (sEH). We screened a set of chemicals containing these functionalities in larval fathead minnow (Pimphales promelas) and embryo/larval golden medaka (Oryzias latipes) models to evaluate the utility of these systems for investigating sEH inhibition in vivo. Both fathead minnow and medaka sEHs were functionally similar to the tested mammalian orthologs (murine and human) with respect to substrate hydrolysis and inhibitor susceptibility. Low lethality was observed in either larval or embryonic fish exposed to diuron [N-(3,4-dichlorophenyl), N'-dimethyl urea], desmethyl diuron [N-(3,4-dichlorophenyl), N'-methyl urea], or siduron [N-(1-methylcyclohexyl), N'-phenyl urea]. Dose-dependent inhibition of sEH was a sublethal effect of substituted urea exposure with the potency of siduron diuron = diuron, differing from the observed in vitro sEH inhibition potency of siduron > desmethyl diuron > diuron. Further, siduron exposure synergized the toxicity of trans-stilbene oxide in fathead minnows. Medaka embryos exposed to diuron, desmethyl diuron, or siduron displayed dose-dependent delays in hatch, and elevated concentrations of diuron and desmethyl diuron produced developmental toxicity. The dose-dependent toxicity and in vivo sEH inhibition correlated, suggesting a potential, albeit undefined, relationship between these factors. Additionally, the observed inversion of in vitro to in vivo potency suggests that these fish models may provide tools for investigating the in vivo stability of in vitro inhibitors while screening for untoward effects. PMID:11171526

Comparative anti-androgenic actions of vinclozolin and flutamide on transgenerational adult onset disease and spermatogenesis.

Science.gov (United States)

Anway, Matthew D; Rekow, Stephen S; Skinner, Michael K

2008-10-01

Exposure of gestating female rats to the anti-androgenic endocrine disruptor vinclozolin has been shown to induce transgenerational adult onset disease phenotypes. The current study, was designed to compare the actions of vinclozolin to the known anti-androgenic compound flutamide. The gestating female rats were exposed to intraperitoneal injections during embryonic day 8-14 (E8-E14) to 100mg/kg/day vinclozolin or flutamide at either 5mg or 20mg/kg/day. As previously observed, vinclozolin induced a transgenerational testis phenotype of increased spermatogenic cell apoptosis and decreased epididymal sperm number. In contrast, the flutamide exposures resulted in a testis phenotype of increased spermatogenic cell apoptosis and decreased epididymal sperm numbers in the F1 generation only, and not the F2 and F3 generation adult males. Interestingly, some of the low dose (5mg/kg) flutamide F2 generation offspring developed spinal agenesis and supernummery development (polymelia) of limbs. Although the actions of vinclozolin and flutamide appear similar in the F1 generation males, the transgenerational effects of vinclozolin do not appear to be acting through the same anti-androgenic mechanism as flutamide.

Metabolite profiles of striped marsh frog (Limnodynastes peronii) larvae exposed to the anti-androgenic fungicides vinclozolin and propiconazole are consistent with altered steroidogenesis and oxidative stress.

Science.gov (United States)

Melvin, Steven D; Leusch, Frederic D L; Carroll, Anthony R

2018-06-01

Amphibians use wetlands in urban and agricultural landscapes for breeding, growth and development. Fungicides and other pesticides used in these areas have therefore been identified as potential threats that could contribute towards amphibian population declines. However, relatively little is known about how such chemicals influence sensitive early life-stages or how short episodic exposures influence sub-lethal physiological and metabolic pathways. The present study applied untargeted metabolomics to evaluate effects in early post-hatch amphibian larvae exposed to the anti-androgenic fungicides vinclozolin and propiconazole. Recently hatched (Gosner developmental stage 25) striped marsh frog (Limnodynastes peronii) larvae were exposed for 96 h to vinclozolin at 17.5, 174.8 and 1748.6 nM and propiconazole at 5.8, 58.4 and 584.4 nM. Nuclear Magnetic Resonance (NMR) spectroscopy was performed on polar metabolites obtained from whole-body extracts. Both fungicides altered metabolite profiles compared to control animals at all concentrations tested, and there were notable differences between the two chemicals. Overall responses were consistent with altered steroidogenesis and/or cholesterol metabolism, with inconsistent responses between the two fungicides likely reflecting minor differences in the mechanisms of action of these chemicals. Broad down-regulation of the tricarboxylic acid (TCA) cycle was also observed and is indicative of oxidative stress. Interestingly, formic acid was significantly increased in larvae exposed to vinclozolin but not propiconazole, suggesting this metabolite may serve as a useful biomarker of exposure to androgen-receptor binding anti-androgenic contaminants. This study demonstrates the power of untargeted metabolomics for distinguishing between similarly acting, but distinct, pollutants and for unraveling non-endocrine responses resulting from exposure to known endocrine active contaminants. Copyright © 2018 Elsevier B.V. All

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

Science.gov (United States)

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

Complex mixtures, complex responses: Assessing pharmaceutical mixtures using field and laboratory approaches

Science.gov (United States)

Schoenfuss, Heiko L.; Furlong, Edward T.; Phillips, Patrick J.; Scott, Tia-Marie; Kolpin, Dana W.; Cetkovic-Cvrlje, Marina; Lesteberg, Kelsey E.; Rearick, Daniel C.

2016-01-01

Pharmaceuticals are present in low concentrations (pharmaceutical formulation facilities. Using existing concentration data, the authors assessed pharmaceuticals in laboratory exposures of fathead minnows (Pimephales promelas) and added environmental complexity through effluent exposures. In the laboratory, larval and mature minnows were exposed to a simple opioid mixture (hydrocodone, methadone, and oxycodone), an opioid agonist (tramadol), a muscle relaxant (methocarbamol), a simple antidepressant mixture (fluoxetine, paroxetine, venlafaxine), a sleep aid (temazepam), or a complex mixture of all compounds. Larval minnow response to effluent exposure was not consistent. The 2010 exposures resulted in shorter exposed minnow larvae, whereas the larvae exposed in 2012 exhibited altered escape behavior. Mature minnows exhibited altered hepatosomatic indices, with the strongest effects in females and in mixture exposures. In addition, laboratory-exposed, mature male minnows exposed to all pharmaceuticals (except the selective serotonin reuptake inhibitor mixture) defended nest sites less rigorously than fish in the control group. Tramadol or antidepressant mixture exposure resulted in increased splenic T lymphocytes. Only male minnows exposed to whole effluent responded with increased plasma vitellogenin concentrations. Female minnows exposed to pharmaceuticals (except the opioid mixture) had larger livers, likely as a compensatory result of greater prominence of vacuoles in liver hepatocytes. The observed alteration of apical endpoints central to sustaining fish populations confirms that effluents containing waste streams from pharmaceutical formulation facilities can adversely impact fish populations but that the effects may not be temporally consistent. The present study highlights the importance of including diverse biological endpoints spanning levels of biological organization and life stages when assessing contaminant interactions.

GENE EXPRESSION ANALYSIS IN THE VENTRAL PROSTATE OF RATS EXPOSED TO VINCLOZOLIN OR PROCYMIDONE

Science.gov (United States)

Vinclozolin and procymidone are antiandrogens that are thought to share a common androgen receptor (AR) mediated mechanism of action. This assessment is based primarily on morphological, AR binding, and in vitro transcriptional activation studies. Studies designed to evaluate t...

Vinclozolin exposure in utero induces postpubertal prostatitis and reduces sperm production via a reversible hormone-regulated mechanism.

Science.gov (United States)

Cowin, Prue A; Gold, Elspeth; Aleksova, Jasna; O'Bryan, Moira K; Foster, Paul M D; Scott, Hamish S; Risbridger, Gail P

2010-02-01

Vinclozolin is an endocrine-disrupting chemical (EDC) that binds with high affinity to the androgen receptor (AR) and blocks the action of gonadal hormones on male reproductive organs. An alternative mechanism of action of Vinclozolin involves transgenerational effects on the male reproductive tract. We previously reported in utero Vinclozolin exposure-induced prostatitis (prostate inflammation) in postpubertal rats concurrent with down-regulation of AR and increased nuclear factor-kappaB activation. We postulated the male reproductive abnormalities induced by in utero Vinclozolin exposure could be reversed by testosterone supplementation, in contrast to the permanent modifications involving DNA methyltransferases (Dnmts) described by others. To test this hypothesis, we administered high-dose testosterone at puberty to Vinclozolin-treated rats and determined the effect on anogenital distance (AGD); testicular germ cell apoptosis, concentration of elongated spermatids, and the onset of prostatitis. Concurrently we examined Dnmt1, -3A, -3B, and -3L mRNA expression. Consistent with previous reports, in utero exposure to Vinclozolin significantly reduced AGD, increased testicular germ cell apoptosis 3-fold, reduced elongated spermatid number by 40%, and induced postpubertal prostatitis in 100% of exposed males. Administration of high-dose testosterone (25 mg/kg) at puberty normalized AGD, reduced germ cell apoptosis, and restored elongated spermatid number. Testosterone restored AR and nuclear factor-kappaB expression in the prostate and abolished Vinclozolin-induced prostatitis. Altered Dnmt expression was evident with in utero Vinclozolin exposure and was not normalized after testosterone treatment. These data demonstrate in utero Vinclozolin-induced male reproductive tract abnormalities are AR mediated and reversible and involve a mechanism independent of Dnmt expression.

Organizational effects of the antiandrogen, Vinclozolin, on penis development in the mouse.

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Amato, Ciro M; Boyd, Morgan; Yang, Joshua; McCoy, Krista A

2018-04-14

Endocrine disrupting chemicals (EDCs) are pollutants found throughout the environment that disrupt normal endocrine processes. In mice, penis development is thought to be most susceptible to EDCs during a critical developmental window occurring on embryonic days (E) 15.5-17.5. However, androgen signaling begins on E13.5 when Androgen Receptor (AR) protein is found in the genitalia and testosterone is circulating. We hypothesize that disrupting androgen signaling prior to the established critical window sensitizes the penis to future androgen disruption. To test this hypothesis, CD1 dams were exposed to Vinclozolin or a corn oil solvent control on E13.5 and E14.5 and AR levels were measured with immunohistochemistry on E14.5. Early antiandrogen exposure reduced AR within nuclei and decreased intensity of AR expression within E14.5 genitalia. To evaluate the influence of antiandrogen exposure before the known critical window of penis development, two groups of pregnant dams (n = 3) were exposed to Vinclozolin starting at either E13.5 or E14.5 and continued exposure through E16.5. Histology and M.O.U.S.E. scoring were used to quantify penis abnormalities. To account for differences in total doses mice experienced due to differences in length of dosing time, we compared animals that received the same total doses. Exposure to antiandrogens on E13.5 exacerbated malformations when exposure was continued through sexually dimorphic development. Both exposure time and Vinclozolin dose are important for severity of Vinclozolin-induced penis abnormalities in mice. This work shows, antiandrogen exposure prior to sensitive periods can exacerbate the effects of later antiandrogen exposure on reproductive development.

Alterations in sperm DNA methylation, non-coding RNA expression, and histone retention mediate vinclozolin-induced epigenetic transgenerational inheritance of disease.

Science.gov (United States)

Ben Maamar, Millissia; Sadler-Riggleman, Ingrid; Beck, Daniel; McBirney, Margaux; Nilsson, Eric; Klukovich, Rachel; Xie, Yeming; Tang, Chong; Yan, Wei; Skinner, Michael K

2018-04-01

Epigenetic transgenerational inheritance of disease and phenotypic variation can be induced by several toxicants, such as vinclozolin. This phenomenon can involve DNA methylation, non-coding RNA (ncRNA) and histone retention, and/or modification in the germline (e.g. sperm). These different epigenetic marks are called epimutations and can transmit in part the transgenerational phenotypes. This study was designed to investigate the vinclozolin-induced concurrent alterations of a number of different epigenetic factors, including DNA methylation, ncRNA, and histone retention in rat sperm. Gestating females (F0 generation) were exposed transiently to vinclozolin during fetal gonadal development. The directly exposed F1 generation fetus, the directly exposed germline within the fetus that will generate the F2 generation, and the transgenerational F3 generation sperm were studied. DNA methylation and ncRNA were altered in each generation rat sperm with the direct exposure F1 and F2 generations being distinct from the F3 generation epimutations. Interestingly, an increased number of differential histone retention sites were found in the F3 generation vinclozolin sperm, but not in the F1 or F2 generations. All three different epimutation types were affected in the vinclozolin lineage transgenerational sperm (F3 generation). The direct exposure generations (F1 and F2) epigenetic alterations were distinct from the transgenerational sperm epimutations. The genomic features and gene pathways associated with the epimutations were investigated to help elucidate the integration of these different epigenetic processes. Our results show that the three different types of epimutations are involved and integrated in the mediation of the epigenetic transgenerational inheritance phenomenon.

Development of a New Technique to Assess Susceptibility to Predation Resulting from Sublethal Stresses (Indirect Mortality)

Energy Technology Data Exchange (ETDEWEB)

Cada, G.F.

2003-08-25

from controls at 1-min post-exposure and for fathead minnows at 1- and 5-min post-exposure. The greatest effects occurred with exposure to the fish anesthetic; in fathead minnows all of the recorded measures were significantly different from controls at 1-min and 5-min post-exposure at the 100 mg/L dose. For striped shiners all recorded behavioral measures were significantly different from controls at 1-min at the 200 and 100 mg/L doses and for selected behavioral measures at 5-min. Turbulence also had significant effects on striped shiner startle responses following 20- and 30-min exposures for all behavioral measures at 1-min. The patterns suggest that any effects on startle response due to turbulence or low doses of anesthetic are short-lived, but can be evaluated using the escape behavior technique. The most useful indication of changes in escape behavior in these tests was the simple reaction/no reaction to the startle stimulus. The startle response occurred reliably among unstressed fish, and was frequently reduced or eliminated in fish exposed to turbulence or anesthesia. The other behavioral parameters observed were often altered by the sublethal stresses as well. A standard predator preference test was also conducted with largemouth bass as the predators and fathead minnows as prey. In this test design, groups of 10 unstressed fish (controls) and 10 stressed fish were put in a tank with a predator. The stressed fathead minnows were exposed to turbulence or fish anesthetic. The predator was allowed to eat half of the prey, and the data were evaluated to determine whether predators consumed greater proportions of stressed minnows than control minnows. The predation test indicated that exposure to MS-222 resulted in significant predation in fathead minnows, but exposure to turbulence did not. This pattern was the same as seen in fathead minnows using the startle response (escape behavior) test. For the sublethal stresses we applied, evaluation of changes in fish

Changes in gene transcription and whole organism responses in larval fathead minnow (Pimephales promelas) following short-term exposure to the synthetic pyrethroid bifenthrin.

Science.gov (United States)

Beggel, Sebastian; Connon, Richard; Werner, Inge; Geist, Juergen

2011-09-01

The combination of molecular and whole-organism endpoints in ecotoxicology provides valuable information about the ecological relevance of sublethal stressor effects in aquatic ecosystems such as those caused by the use of insecticides and translocation of their residues into surface waters. This study contributes knowledge about the sublethal effects of a common use insecticide, the synthetic pyrethroid bifenthrin, on larval fathead minnow (Pimephales promelas). Transcriptomic responses, assessed by quantitative real-time PCR, combined with individual effects on swimming performance were used to estimate the ecological relevance of insecticide impacts. Significant transcriptomic responses were observed at 0.07 μg L(-1) bifenthrin (lowest observed effect concentration, LOEC) but mostly followed a biphasic rather than a linear dose-response with increasing concentration. Transcript patterns for genes involved in detoxification, neuromuscular function and energy metabolism were linked to an impairment of swimming performance at ≥0.14 μg L(-1) bifenthrin. With increasing treatment concentration, a significant down-regulation was observed for genes coding for cyp3a, aspartoacylase, and creatine kinase, whereas metallothionein was up-regulated. Additionally, bifenthrin induced endocrine responses as evident from a significant up-regulation of vitellogenin and down-regulation of insuline-like growth factor transcripts. Recovery occurred after 6 days and was dependent on the magnitude of the initial stress. During the recovery period, down-regulation of vitellogenin was observed at lowest exposure concentrations. The data presented here emphasize that links can be made between gene transcription changes and behavioral responses which is of great value for the evaluation and interpretation of biomarker responses. Copyright © 2011 Elsevier B.V. All rights reserved.

Embryonic exposure to the fungicide vinclozolin causes virilization of females and alteration of progesterone receptor expression in vivo: an experimental study in mice.

Science.gov (United States)

Buckley, Jill; Willingham, Emily; Agras, Koray; Baskin, Laurence S

2006-02-21

Vinclozolin is a fungicide that has been reported to have anti-androgenic effects in rats. We have found that in utero exposure to natural or synthetic progesterones can induce hypospadias in mice, and that the synthetic progesterone medroxyprogesterone acetate (MPA) feminizes male and virilizes female genital tubercles. In the current work, we selected a relatively low dose of vinclozolin to examine its in utero effects on the development of the genital tubercle, both at the morphological and molecular levels. We gave pregnant dams vinclozolin by oral gavage from gestational days 13 through 17. We assessed the fetal genital tubercles from exposed fetuses at E19 to determine location of the urethral opening. After determination of gonadal sex, either genital tubercles were harvested for mRNA quantitation, or urethras were injected with a plastic resin for casting. We analyzed quantified mRNA levels between treated and untreated animals for mRNA levels of estrogen receptors alpha and beta, progesterone receptor, and androgen receptor using nonparametric tests or ANOVA. To determine effects on urethral length (males have long urethras compared to females), we measured the lengths of the casts and performed ANOVA analysis on these data. Our morphological results indicated that vinclozolin has morphological effects similar to those of MPA, feminizing males (hypospadias) and masculinizing females (longer urethras). Because these results reflected our MPA results, we investigated the effects of in utero vinclozolin exposure on the mRNA expression levels of androgen, estrogen alpha and beta, and progesterone receptors. At the molecular level, vinclozolin down-regulated estrogen receptor alpha mRNA in females and up-regulated progesterone receptor mRNA. Vinclozolin-exposed males exhibited up-regulated estrogen receptor alpha and progesterone receptor mRNA, effects we have also seen with exposure to the synthetic estrogen, ethinyl estradiol. The results suggest that

Embryonic exposure to the fungicide vinclozolin causes virilization of females and alteration of progesterone receptor expression in vivo: an experimental study in mice

Directory of Open Access Journals (Sweden)

Baskin Laurence S

2006-02-01

Full Text Available Abstract Background Vinclozolin is a fungicide that has been reported to have anti-androgenic effects in rats. We have found that in utero exposure to natural or synthetic progesterones can induce hypospadias in mice, and that the synthetic progesterone medroxyprogesterone acetate (MPA feminizes male and virilizes female genital tubercles. In the current work, we selected a relatively low dose of vinclozolin to examine its in utero effects on the development of the genital tubercle, both at the morphological and molecular levels. Methods We gave pregnant dams vinclozolin by oral gavage from gestational days 13 through 17. We assessed the fetal genital tubercles from exposed fetuses at E19 to determine location of the urethral opening. After determination of gonadal sex, either genital tubercles were harvested for mRNA quantitation, or urethras were injected with a plastic resin for casting. We analyzed quantified mRNA levels between treated and untreated animals for mRNA levels of estrogen receptors α and β, progesterone receptor, and androgen receptor using nonparametric tests or ANOVA. To determine effects on urethral length (males have long urethras compared to females, we measured the lengths of the casts and performed ANOVA analysis on these data. Results Our morphological results indicated that vinclozolin has morphological effects similar to those of MPA, feminizing males (hypospadias and masculinizing females (longer urethras. Because these results reflected our MPA results, we investigated the effects of in utero vinclozolin exposure on the mRNA expression levels of androgen, estrogen α and β, and progesterone receptors. At the molecular level, vinclozolin down-regulated estrogen receptor α mRNA in females and up-regulated progesterone receptor mRNA. Vinclozolin-exposed males exhibited up-regulated estrogen receptor α and progesterone receptor mRNA, effects we have also seen with exposure to the synthetic estrogen, ethinyl

Identifying non-point sources of endocrine active compounds and their biological impacts in freshwater lakes

Science.gov (United States)

Baker, Beth H.; Martinovic-Weigelt, Dalma; Ferrey, Mark L.; Barber, Larry B.; Writer, Jeffrey H.; Rosenberry, Donald O.; Kiesling, Richard L.; Lundy, James R.; Schoenfuss, Heiko L.

2014-01-01

Contaminants of emerging concern, particularly endocrine active compounds (EACs), have been identified as a threat to aquatic wildlife. However, little is known about the impact of EACs on lakes through groundwater from onsite wastewater treatment systems (OWTS). This study aims to identify specific contributions of OWTS to Sullivan Lake, Minnesota, USA. Lake hydrology, water chemistry, caged bluegill sunfish (Lepomis macrochirus), and larval fathead minnow (Pimephales promelas) exposures were used to assess whether EACs entered the lake through OWTS inflow and the resultant biological impact on fish. Study areas included two OWTS-influenced near-shore sites with native bluegill spawning habitats and two in-lake control sites without nearby EAC sources. Caged bluegill sunfish were analyzed for plasma vitellogenin concentrations, organosomatic indices, and histological pathologies. Surface and porewater was collected from each site and analyzed for EACs. Porewater was also collected for laboratory exposure of larval fathead minnow, before analysis of predator escape performance and gene expression profiles. Chemical analysis showed EACs present at low concentrations at each study site, whereas discrete variations were reported between sites and between summer and fall samplings. Body condition index and liver vacuolization of sunfish were found to differ among study sites as did gene expression in exposed larval fathead minnows. Interestingly, biological exposure data and water chemistry did not match. Therefore, although results highlight the potential impacts of seepage from OWTS, further investigation of mixture effects and life history factor as well as chemical fate is warranted.

Evaluation of fish models of soluble epoxide hydrolase inhibition.

OpenAIRE

Newman, J W; Denton, D L; Morisseau, C; Koger, C S; Wheelock, C E; Hinton, D E; Hammock, B D

2001-01-01

Substituted ureas and carbamates are mechanistic inhibitors of the soluble epoxide hydrolase (sEH). We screened a set of chemicals containing these functionalities in larval fathead minnow (Pimphales promelas) and embryo/larval golden medaka (Oryzias latipes) models to evaluate the utility of these systems for investigating sEH inhibition in vivo. Both fathead minnow and medaka sEHs were functionally similar to the tested mammalian orthologs (murine and human) with respect to substrate hydrol...

Zebrafish (Danio rerio) androgen receptor: sequence homology and up-regulation by the fungicide vinclozolin.

Science.gov (United States)

Smolinsky, Amanda N; Doughman, Jennifer M; Kratzke, Liên-Thành C; Lassiter, Christopher S

2010-03-01

Steroid hormones regulate gene expression in organisms by binding to receptor proteins. These hormones include the androgens, which signal through androgen receptors (ARs). Endocrine disrupters (EDCs) are chemicals in the environment that adversely affect organisms by binding to nuclear receptors, including ARs. Vinclozolin, a fungicide used on fruit and vegetable crops, is a known anti-androgen, a type of EDC that blocks signals from testosterone and its derivatives. In order to better understand the effects of EDCs, further research on androgen receptors and other hormone signaling pathways is necessary. In this study, we demonstrate the evolutionary conservation between the genomic structure of the human and zebrafish ar genes and find that ar mRNA expression increases in zebrafish embryos exposed to vinclozolin, which may be evolutionarily conserved as well. At 48 and 72 h post-fertilization, vinclozolin-treated embryos express ar mRNA 8-fold higher than the control level. These findings suggest that zebrafish embryos attempt to compensate for the presence of an anti-androgen by increasing the number of androgen receptors available.

Tissue explant coculture model of the hypothalamic-pituitary-gonadal-liver axis of the fathead minnow (Pimephales promelas) as a predictive tool for endocrine disruption.

Science.gov (United States)

Johnston, Theresa K; Perkins, Edward; Ferguson, Duncan C; Cropek, Donald M

2016-10-01

Endocrine-disrupting compounds (EDCs) can impact the reproductive system by interfering with the hypothalamic-pituitary-gonadal (HPG) axis. Although in vitro testing methods have been developed to screen chemicals for endocrine disruption, extrapolation of in vitro responses to in vivo action shows inconsistent accuracy. The authors describe a tissue coculture of the fathead minnow (Pimephales promelas) HPG axis and liver (HPG-L) as a tissue explant model that mimics in vivo results. Brain (hypothalamus), pituitary, gonad, and liver tissue explants from adult fish were examined for function both individually and in coculture to determine combinations and conditions that could replicate in vivo behavior. Only cocultures had the ability to respond to an EDC, trenbolone, similarly to in vivo studies, based on estradiol, testosterone, and vitellogenin production trends, where lower exposure doses suppressed hormone production but higher doses increased production, resulting in distinctive U-shaped curves. These data suggest that a coculture system with all components of the HPG-L axis can be used as a link between in vitro and in vivo studies to predict endocrine system disruption in whole organisms. This tissue-based HPG-L system acts as a flexible deconstructed version of the in vivo system for better control and examination of the minute changes in system operation and response on EDC exposure with options to isolate, interrogate, and recombine desired components. Environ Toxicol Chem 2016;35:2530-2541. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America.

Assessing water quality suitability for shortnose sturgeon in the Roanoke River, North Carolina, USA with an in situ bioassay approach

Science.gov (United States)

Cope, W.G.; Holliman, F.M.; Kwak, T.J.; Oakley, N.C.; Lazaro, P.R.; Shea, D.; Augspurger, T.; Law, J.M.; Henne, J.P.; Ware, K.M.

2011-01-01

The aim of this study was to determine the suitability of water quality in the Roanoke River of North Carolina for supporting shortnose sturgeon Acipenser brevirostrum, an endangered species in the United States. Fathead minnows Pimephales promelas were also evaluated alongside the sturgeon as a comparative species to measure potential differences in fish survival, growth, contaminant accumulation, and histopathology in a 28-day in situ toxicity test. Captively propagated juvenile shortnose sturgeon (total length 49??8mm, mean??SD) and fathead minnows (total length 39??3mm, mean??SD) were used in the test and their outcomes were compared to simultaneous measurements of water quality (temperature, dissolved oxygen, pH, conductivity, total ammonia nitrogen, hardness, alkalinity, turbidity) and contaminant chemistry (metals, polycyclic aromatic hydrocarbons, organochlorine pesticides, current use pesticides, polychlorinated biphenyls) in river water and sediment. In the in situ test, there were three non-riverine control sites and eight riverine test sites with three replicate cages (25??15-cm (OD) clear plexiglass with 200-??m tear-resistant Nitex?? screen over each end) of 20 shortnose sturgeon per cage at each site. There was a single cage of fathead minnows also deployed at each site alongside the sturgeon cages. Survival of caged shortnose sturgeon among the riverine sites averaged 9% (range 1.7-25%) on day 22 of the 28-day study, whereas sturgeon survival at the non-riverine control sites averaged 64% (range 33-98%). In contrast to sturgeon, only one riverine deployed fathead minnow died (average 99.4% survival) over the 28-day test period and none of the control fathead minnows died. Although chemical analyses revealed the presence of retene (7-isopropyl-1-methylphenanthrene), a pulp and paper mill derived compound with known dioxin-like toxicity to early life stages of fish, in significant quantities in the water (251-603ngL-1) and sediment (up to 5000ngg-1

Microbial biodegradation and toxicity of vinclozolin and its toxic metabolite 3,5-dichloroaniline.

Science.gov (United States)

Lee, Jung-Bok; Sohn, Ho-Yong; Shin, Kee-Sun; Kim, Jong-Sik; Jo, Min-Sub; Jeon, Chun-Pyo; Jang, Jong-Ok; Kim, Jang-Eok; Kwon, Gi-Seok

2008-02-01

Vinclozolin, an endocrine disrupting chemical, is a chlorinated fungicide widely used to control fungal diseases. However, its metabolite 3,5-dichloroaniline is more toxic and persistent than the parent vinclozolin. For the biodegradation of vinclozolin, vinclozolin- and/or 3,5-dichloroaniline-degrading bacteria were isolated from pesticide-polluted agriculture soil. Among the isolated bacteria, a Rhodococcus sp. was identified from a 16S rDNA sequence analysis and named Rhodococcus sp. T1-1. The degradation ratios for vinclozolin or 3,5- dichloroaniline in a minimal medium containing vinclozolin (200 microg/ml) or 3,5-dichloroaniline (120 microg/ml) were 90% and 84.1%, respectively. Moreover, Rhodococcus sp. T1-1 also showed an effective capability to biodegrade dichloroaniline isomers on enrichment cultures in which they were contained. Therefore, these results suggest that Rhodococcus sp. T1-1 can bioremediate vinclozolin as well as 3,5-dichloroaniline.

SURROGATE SPECIES IN ASSESSING CONTAMINANT RISK FOR ENDANGERED FISHES

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Rainbow trout, fathead minnows, and sheepshead minnows were tested as surrogate species to assess contaminant risk for 17 endangered fishes and one toad species. Acute toxicity tests were conducted with carbaryl, copper, 4-nonylphenol, pentachlorophenol, and permethrin in accord...

ALTERATIONS IN THE DEVELOPING TESTIS TRANSCRIPTOME FOLLOWING EMBRYONIC VINCLOZOLIN EXPOSURE

OpenAIRE

Clement, Tracy M.; Savenkova, Marina I.; Settles, Matthew; Anway, Matthew D.; Skinner, Michael K.

2010-01-01

The current study investigates the direct effects of in utero vinclozolin exposure on the developing F1 generation rat testis transcriptome. Previous studies have demonstrated that exposure to vinclozolin during embryonic gonadal sex determination induces epigenetic modifications of the germ line and transgenerational adult onset disease states. Microarray analyses were performed to compare control and vinclozolin treated testis transcriptomes at embryonic day 13, 14 and 16. A total of 576 di...

Modified expression of several sperm proteins after chronic exposure to the antiandrogenic compound vinclozolin.

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Auger, Jacques; Eustache, Florence; Maceiras, Paula; Broussard, Cédric; Chafey, Philippe; Lesaffre, Corinne; Vaiman, Daniel; Camoin, Luc; Auer, Jana

2010-10-01

Little is known about the molecular impact of in vivo exposure to endocrine disruptors (EDs) on sperm structures and functions. We recently reported that the lifelong exposure of rats to the antiandrogenic compound vinclozolin results in low epididymal weight, changes in sperm kinematic parameters, and immature sperm chromatin condensation, together with the impairment of several fertility end points. These results led us to focus specifically on possible molecular abnormalities in sperm. Sperm samples were recovered from the frozen epididymides of rats exposed during the previous study. The proteins present in the samples from six exposed and six control rats were analyzed in pairs, by two-dimensional fluorescence difference gel electrophoresis, to investigate possible exposure-induced changes to sperm protein profiles. Twelve proteins, from the 380 matched spots observed in at least five gels, were present in larger or smaller amounts after vinclozolin exposure. These proteins were identified by mass spectrometry, and several are known to play a crucial role in the sperm fertilizing ability, among which, two mitochondrial enzymes, malate dehydrogenase 2 and aldehyde dehydrogenase (both of which were present in smaller amounts after treatment) and A-kinase anchor protein 4 (larger amounts of precursor after treatment). Finally, Ingenuity Pathway Analysis revealed highly significant interactions between proteins over- and underexpressed after treatment. This is the first study to show an association between in vivo exposure to an ED and changes to the sperm protein profile. These modifications may be at least partly responsible for the reproductive abnormalities and impaired fertility recently reported in this rat model of vinclozolin exposure.

Environmental Sciences Division Toxicology Laboratory standard operating procedures

International Nuclear Information System (INIS)

Kszos, L.A.; Stewart, A.J.; Wicker, L.F.; Logsdon, G.M.

1989-09-01

This document was developed to provide the personnel working in the Environmental Sciences Division's Toxicology Laboratory with documented methods for conducting toxicity tests. The document consists of two parts. The first part includes the standard operating procedures (SOPs) that are used by the laboratory in conducting toxicity tests. The second part includes reference procedures from the US Environmental Protection Agency document entitled Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, upon which the Toxicology Laboratory's SOPs are based. Five of the SOPs include procedures for preparing Ceriodaphnia survival and reproduction test. These SOPs include procedures for preparing Ceriodaphnia food (SOP-3), maintaining Ceriodaphnia cultures (SOP-4), conducting the toxicity test (SOP-13), analyzing the test data (SOP-13), and conducting a Ceriodaphnia reference test (SOP-15). Five additional SOPs relate specifically to the fathead minnow (Pimephales promelas) larval survival and growth test: methods for preparing fathead minnow larvae food (SOP-5), maintaining fathead minnow cultures (SOP-6), conducting the toxicity test (SOP-9), analyzing the test data (SOP-12), and conducting a fathead minnow reference test (DOP-14). The six remaining SOPs describe methods that are used with either or both tests: preparation of control/dilution water (SOP-1), washing of glassware (SOP-2), collection and handling of samples (SOP-7), preparation of samples (SOP-8), performance of chemical analyses (SOP-11), and data logging and care of technical notebooks (SOP-16)

Transgenerational effect of the endocrine disruptor vinclozolin on male spermatogenesis.

Science.gov (United States)

Anway, Matthew D; Memon, Mushtaq A; Uzumcu, Mehmet; Skinner, Michael K

2006-01-01

The current study was designed to examine the actions of a model endocrine disruptor on embryonic testis development and male fertility. Pregnant rats (F0) that received a transient embryonic exposure to an environmental endocrine disruptor, vinclozolin, had male offspring (F1) with reduced spermatogenic capacity. The reduced spermatogenetic capacity observed in the F1 male offspring was transmitted to the subsequent generations (F2-F4). The administration of vinclozolin, an androgen receptor antagonist, at 100 mg/kg/day from embryonic day 8-14 (E8-E14) of pregnancy to only the F0 dam resulted in a transgenerational phenotype in the subsequent male offspring in the F1-F4 generations. The litter size and male/female sex ratios were similar in controls and the vinclozolin generations. The average testes/body weight index of the postnatal day 60 (P60) males was not significantly different in the vinclozolin-treated generations compared to the controls. However, the testicular spermatid number, as well as the epididymal sperm number and motility, were significantly reduced in the vinclozolin generations compared to the control animals. Postnatal day 20 (P20) testis from the vinclozolin F2 generation had no morphological abnormalities, but did have an increase in spermatogenic cell apoptosis. Although the P60 testis morphology was predominantly normal, the germ cell apoptosis was significantly increased in the testes cross sections of animals from the vinclozolin generations. The increase in apoptosis was stage-specific in the testis, with tubules at stages IX-XIV having the highest increase in apoptotic germ cells. The tubules at stages I-V also had an increase in apoptotic germ cells compared to the control samples, but tubules at stages VI-VIII had no increase in apoptotic germ cells. An outcross of a vinclozolin generation male with a wild-type female demonstrated that the reduced spermatogenic cell phenotype was transmitted through the male germ line. An outcross

Computational Model of the Fathead Minnow Hypothalamic-Pituitary-Gonadal Axis: Incorporating Protein Synthesis in Improving Predictability of Responses to Endocrine Active Chemicals

Science.gov (United States)

There is international concern about chemicals that alter endocrine system function in humans and/or wildlife and subsequently cause adverse effects. We previously developed a mechanistic computational model of the hypothalamic-pituitary-gonadal (HPG) axis in female fathead minno...

Progesterone increases ex vivo testosterone production and decreases the expression of progestin receptors and steroidogenic enzymes in the fathead minnow (Pimephales promelas) ovary.

Science.gov (United States)

Chishti, Yasmin Z; Feswick, April; Martyniuk, Christopher J

2014-04-01

Progesterone (P4) is a metabolic precursor for a number of steroids, including estrogens and androgens. P4 also has diverse roles within the vertebrate ovary that include oocyte growth and development. The objectives of this study were to measure the effects of P4 on testosterone (T) and 17β-estradiol (E2) production in the fathead minnow (FHM) ovary and on the mRNA abundance of transcripts involved in steroidogenesis and steroid receptor signaling. Ovary explants were treated with P4 (10(-6)M) for 6 and 12h. P4 administration significantly increased T production ∼3-fold at both 6 and 12h, whereas E2 production was not affected, consistent with the hypothesis that excess P4 is not converted to terminal estrogens in the mature ovary. Nuclear progesterone receptor mRNA was decreased at 6h and membrane progesterone receptor gamma-2 mRNA was significantly down-regulated at both 6 and 12h; however there was no change in membrane progesterone receptor alpha or beta mRNA levels. Androgen receptor (ar) and estrogen receptor 2a (esr2a) mRNA were significantly reduced at 6h with P4 treatment, but there was no change in esr2b mRNA at either time point. Transcripts for enzymes in the steroid pathway (star, hsd11b2) were significantly lower at 6h compared to controls, whereas cyp17a and cyp19a mRNA abundance did not change with treatments at either time point. These data suggest that P4 incubation can lead to increased T production in the FHM ovary without a concomitant change in E2, and that the membrane bound progestin receptors are differentially regulated by P4 in the teleost ovary. As environmental progestins have received increased attention due to their suspected role as endocrine disruptors, mechanistic data on the role of exogenous P4 treatments in the male and female gonad is warranted. Copyright © 2014 Elsevier Inc. All rights reserved.

InP/ZnS QDs exposure induces developmental toxicity in rare minnow (Gobiocypris rarus) embryos.

Science.gov (United States)

Chen, Yao; Yang, Yang; Ou, Fang; Liu, Li; Liu, Xiao-Hong; Wang, Zhi-Jian; Jin, Li

2018-04-05

We investigated the in vivo toxicity of InP/ZnS quantum dots (QDs) in Chinese rare minnow (Gobiocypris rarus) embryos. The 72 h post-fertilization (hpf) LC 50 (median lethal concentration) was 1678.007 nmol/L. Rare minnows exposed to InP/ZnS QDs exhibited decreased spontaneous movement, decreased survival and hatchability rates, and an increased malformation rate. Pericardial edema, spinal curvature, bent tails and vitelline cysts were observed. Embryonic Wnt8a and Mstn mRNA levels were significantly up-regulated after InP/ZnS QDs treatment at 48 hpf (200 nmol/L) (p InP/ZnS QDs treatments did not significantly change the Olive tail moments (p > 0.05). Thus, InP/ZnS QDs caused teratogenic effects and death during the development of Chinese rare minnow embryos, but InP/ZnS QDs did not cause significant genetic toxicity during Chinese rare minnow development. Copyright © 2018 Elsevier B.V. All rights reserved.

Assessment of combinations of antiandrogenic compounds vinclozolin and flutamide in a yeast based reporter assay.

Science.gov (United States)

Kolle, Susanne N; Melching-Kollmuss, Stephanie; Krennrich, Gerhard; Landsiedel, Robert; van Ravenzwaay, Bennard

2011-08-01

Humans are exposed to a combination of various substances such as cosmetic ingredients, drugs, biocides, pesticides and natural-occurring substances in food. The combined toxicological effects of two or more substances can simply be additive on the basis of response-addition, or it can be greater (synergistic) or smaller (antagonistic) than this. The need to assess combined effects of compounds with endocrine activity is currently discussed for regulatory risk assessment. We have used a well described yeast based androgen receptor transactivation assay YAS to assess the combinatorial effects of vinclozolin and flutamide; both mediating antiandrogenicity via the androgen receptor. Both vinclozolin and flutamide were antiandrogens of similar potency in the YAS assay. In the concentration range tested the two antiandrogens vinclozolin and flutamide did not act synergistically. Concentration additivity was observed in the linear, non-receptor-saturated concentration range. At high concentrations of one of the two substances tested the contribution of the second at lower concentration levels was less than additive. The combined response of both compounds at high concentration levels was also less than additive (saturation effect). At concentration levels which did not elicit a response of the individual compounds, the combination of these compounds also did not elicit a response. Copyright © 2011 Elsevier Inc. All rights reserved.

Ecotoxicological effects extrapolation models

Energy Technology Data Exchange (ETDEWEB)

Suter, G.W. II

1996-09-01

One of the central problems of ecological risk assessment is modeling the relationship between test endpoints (numerical summaries of the results of toxicity tests) and assessment endpoints (formal expressions of the properties of the environment that are to be protected). For example, one may wish to estimate the reduction in species richness of fishes in a stream reach exposed to an effluent and have only a fathead minnow 96 hr LC50 as an effects metric. The problem is to extrapolate from what is known (the fathead minnow LC50) to what matters to the decision maker, the loss of fish species. Models used for this purpose may be termed Effects Extrapolation Models (EEMs) or Activity-Activity Relationships (AARs), by analogy to Structure-Activity Relationships (SARs). These models have been previously reviewed in Ch. 7 and 9 of and by an OECD workshop. This paper updates those reviews and attempts to further clarify the issues involved in the development and use of EEMs. Although there is some overlap, this paper does not repeat those reviews and the reader is referred to the previous reviews for a more complete historical perspective, and for treatment of additional extrapolation issues.

Abnormal peripubertal development of the rat mammary gland following exposure in utero and during lactation to a mixture of genistein and the food contaminant vinclozolin.

Science.gov (United States)

El Sheikh Saad, H; Meduri, G; Phrakonkham, P; Bergès, R; Vacher, S; Djallali, M; Auger, J; Canivenc-Lavier, M C; Perrot-Applanat, M

2011-07-01

The impact of early exposure to endocrine disruptor mixtures on mammary gland development is poorly known. Here, we identify the effects of a conception to weaning exposure of rats to the phytoestrogen genistein (G) and/or the antiandrogen vinclozolin (V) at 1mg/kg-d, alone or in association. Using several approaches, we found that G- and GV-exposed rats displayed significantly greater epithelial branching and proliferation, wider terminal end buds than controls at PND35, as well as ductal hyperplasia and periductal fibrosis. Focal branching defects were present in V-exposed rats. An increased ER and AR expression was observed in G- and GV- as compared to V-exposed rats at PND35. Surprisingly, a significant number of GV- and to a lesser extent, V-exposed animals displayed abnormal hyperplasic alveolar structures at PND50. Thus, gestational and lactational exposure to low doses of genistein plus vinclozolin may seriously affect peripubertal development of the rat mammary gland. Copyright © 2011 Elsevier Inc. All rights reserved.

Alterations in the developing testis transcriptome following embryonic vinclozolin exposure.

Science.gov (United States)

Clement, Tracy M; Savenkova, Marina I; Settles, Matthew; Anway, Matthew D; Skinner, Michael K

2010-11-01

The current study investigates the direct effects of in utero vinclozolin exposure on the developing F1 generation rat testis transcriptome. Previous studies have demonstrated that exposure to vinclozolin during embryonic gonadal sex determination induces epigenetic modifications of the germ line and transgenerational adult onset disease states. Microarray analyses were performed to compare control and vinclozolin treated testis transcriptomes at embryonic days 13, 14 and 16. A total of 576 differentially expressed genes were identified and the major cellular functions and pathways associated with these altered transcripts were examined. The sets of regulated genes at the different development periods were found to be transiently altered and distinct. Categorization by major known functions of altered genes was performed. Specific cellular process and pathway analyses suggest the involvement of Wnt and calcium signaling, vascular development and epigenetic mechanisms as potential mediators of the direct F1 generation actions of vinclozolin. Copyright © 2010 Elsevier Inc. All rights reserved.

Use of the Keele injector for sample introduction for gas chromatographic analysis of vinclozolin in lettuces.

Science.gov (United States)

Shim, J H; Lee, Y S; Kim, M R; Lee, C J; Kim, I S

2003-10-10

We examined a Keele injector for sample introduction for gas chromatographic analysis of vinclozolin treated in lettuces. Samples in milligram quantity were introduced into a glass tube in a Keele injector at a gas chromatograph injection port. The glass tube was then crushed to allow the sample to carry onto a capillary column in a normal manner. The standard calibration curve for quantitative detection of vinclozolin was obtained by determining vinclozolin spiked in samples at variable concentrations. The calibration curve showed a linear response to vinclozolin ranging from 0.05 to 1.0 microg/g, giving a slope value of 174.8, the y-intercept value of -2.8146 and the mean r2-value of 0.9994. Limit of quantification for vinclozolin was 0.05 microg/g by this method, comparable to 0.01 microg/g by a normal injector. When samples treated previously with vinclozolin were determined by the Keele injector, vinclozolin was found to be about 30% lower as compared to a normal method, suggesting about 70% recovery of the spiked vinclozolin by the Keele injector. From these results, the Keele injector was suggested to be potential for sample introduction in gas chromatographic analysis of vinclozolin in lettuce samples.

Effects of anti-androgens cyproterone acetate, linuron, vinclozolin, and p,p'-DDE on the reproductive organs of the copepod Acartia tonsa

DEFF Research Database (Denmark)

Watermann, Burkard T.; Albanis, Triantafyllos A.; Galassi, Silvana

2016-01-01

The study was performed to detect the effects of anti-androgenic compounds on the reproduction. In this paper alterations observed in the marine calanoid copepod Acartia tonsa exposed to environmental concentrations of cyproterone acetate (CPA), linuron (LIN), vinclozolin (VIN), and 1,1-dichloro-2...

Products and kinetics of the heterogeneous reaction of suspended vinclozolin particles with ozone.

Science.gov (United States)

Gan, Jie; Yang, Bo; Zhang, Yang; Shu, Xi; Liu, Changgeng; Shu, Jinian

2010-11-25

Vinclozolin is a widely used fungicide that can be released into the atmosphere via application and volatilization. This paper reports an experimental investigation on the heterogeneous ozonation of vinclozolin particles. The ozonation of vinclozolin adsorbed on azelaic acid particles under pseudo-first-order conditions is investigated online with a vacuum ultraviolet photoionization aerosol time-of-flight mass spectrometer (VUV-ATOFMS). The ozonation products are analyzed with a combination of VUV-ATOFMS and GC/MS. Two main ozonation products are observed. The formation of the ozonation products results from addition of O(3) on the C-C double bond of the vinyl group. The heterogeneous reactive rate constant of vinclozolin particles under room temperature is (2.4 ± 0.4) × 10(-17) cm(3) molecules(-1) s(-1), with a corresponding lifetime at 100 ppbv O(3) of 4.3 ± 0.7 h, which is almost comparable with the estimated lifetime due to the reaction with atmospheric OH radicals (∼1.7 h). The reactive uptake coefficient for O(3) on vinclozolin particles is (6.1 ± 1.0) × 10(-4).

Metabolomics for Informing Adverse Outcome Pathways: Androgen Receptor Activation and the Pharmaceutical Spironolactone

Data.gov (United States)

U.S. Environmental Protection Agency — Metabolite Input Files for Determining Biochemical Pathways Impacted by Spironolactone Exposures of Fathead Minnows (Pimephales promelas) Using the Mummichog...

Molecular characterization of cytochrome P450 1A and 3A and the effects of perfluorooctanoic acid on their mRNA levels in rare minnow (Gobiocypris rarus) gills

Energy Technology Data Exchange (ETDEWEB)

Liu Yong; Wang Jianshe; Wei Yanhong; Zhang Hongxia; Liu Yang [Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Datun Road, Beijing 100101 (China); Dai Jiayin [Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Datun Road, Beijing 100101 (China)], E-mail: daijy@ioz.ac.cn

2008-07-07

Perfluorooctanoic acid (PFOA), a potentially toxic perfluorinated compound (PFC), has been widely disseminated in the environment. In the present study, rare minnows (Gobiocypris rarus) exposed to PFOA exhibited histopathological gill damage, including epithelial hyperplasia of the lamellae, inflammatory cell infiltration, and lamellar fusion. Cytochrome P450s (CYPs) play a central role in the metabolism and biotransformation of a wide range of endogenous substrates and foreign compounds. Thus, we studied the CYPs and the effects of waterborne PFOA on their corresponding mRNA levels in the gills of rare minnows. Two novel CYP cDNAs (CYP1A and CYP3A) were identified in rare minnow and their mRNAs were ubiquitously expressed in all tissues examined. Upregulation of CYP3A mRNA was observed in the gills of male rare minnows exposed to 30 mg/L PFOA, while no significant changes occurred in exposed females. In contrast, down regulation of CYP1A mRNA was detected in the gills of male and female minnows exposed to PFOA. However, the effect of PFOA on gill mRNA levels of their potential regulators, aryl hydrocarbon receptor (AhR) for CYP1A, and pregnane X receptor (PXR) for CYP3A, were not consistent with the observed effects of PFOA on the corresponding CYP mRNA concentrations. This suggests a different or more complex transcriptional regulation of CYP expression following PFOA exposure.

Steroid receptor profiling of vinclozolin and its primary metabolites

International Nuclear Information System (INIS)

Molina-Molina, Jose-Manuel; Hillenweck, Anne; Jouanin, Isabelle; Zalko, Daniel; Cravedi, Jean-Pierre; Fernandez, Mariana-Fatima; Pillon, Arnaud; Nicolas, Jean-Claude; Olea, Nicolas; Balaguer, Patrick

2006-01-01

Several pesticides and fungicides commonly used to control agricultural and indoor pests are highly suspected to display endocrine-disrupting effects in animals and humans. Endocrine disruption is mainly caused by the interference of chemicals at the level of steroid receptors: it is now well known that many of these chemicals can display estrogenic effects and/or anti-androgenic effects, but much less is known about the interaction of these compounds with other steroid receptors. Vinclozolin, a dicarboximide fungicide, like its primary metabolites 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3-butenoic acid (M1), and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2), is known to bind androgen receptor (AR). Although vinclozolin and its metabolites were characterized as anti-androgens, relatively little is known about their effects on the function of the progesterone (PR), glucocorticoid (GR), mineralocorticoid (MR) or estrogen receptors (ERα and ERβ). Objectives of the study were to determine the ability of vinclozolin and its two primary metabolites to activate AR, PR, GR, MR and ER. For this purpose, we used reporter cell lines bearing luciferase gene under the control of wild type or chimeric Gal4 fusion AR, PR, GR, MR or ERs. We confirmed that all three were antagonists for AR, whereas only M2 was found a partial agonist. Interestingly, M2 was also a PR, GR and MR antagonist (MR >> PR > GR) while vinclozolin was an MR and PR antagonist. Vinclozolin, M1 and M2 were agonists for both ERs with a lower affinity for ERβ. Although the potencies of the fungicide and its metabolites are low when compared to natural ligands, their ability to act via more than one mechanism and the potential for additive or synergistic effect must be taken into consideration in the risk assessment process

Steroid receptor profiling of vinclozolin and its primary metabolites.

Science.gov (United States)

Molina-Molina, José-Manuel; Hillenweck, Anne; Jouanin, Isabelle; Zalko, Daniel; Cravedi, Jean-Pierre; Fernández, Mariana-Fátima; Pillon, Arnaud; Nicolas, Jean-Claude; Olea, Nicolás; Balaguer, Patrick

2006-10-01

Several pesticides and fungicides commonly used to control agricultural and indoor pests are highly suspected to display endocrine-disrupting effects in animals and humans. Endocrine disruption is mainly caused by the interference of chemicals at the level of steroid receptors: it is now well known that many of these chemicals can display estrogenic effects and/or anti-androgenic effects, but much less is known about the interaction of these compounds with other steroid receptors. Vinclozolin, a dicarboximide fungicide, like its primary metabolites 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3-butenoic acid (M1), and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2), is known to bind androgen receptor (AR). Although vinclozolin and its metabolites were characterized as anti-androgens, relatively little is known about their effects on the function of the progesterone (PR), glucocorticoid (GR), mineralocorticoid (MR) or estrogen receptors (ERalpha and ERbeta). Objectives of the study were to determine the ability of vinclozolin and its two primary metabolites to activate AR, PR, GR, MR and ER. For this purpose, we used reporter cell lines bearing luciferase gene under the control of wild type or chimeric Gal4 fusion AR, PR, GR, MR or ERs. We confirmed that all three were antagonists for AR, whereas only M2 was found a partial agonist. Interestingly, M2 was also a PR, GR and MR antagonist (MR>PR>GR) while vinclozolin was an MR and PR antagonist. Vinclozolin, M1 and M2 were agonists for both ERs with a lower affinity for ERbeta. Although the potencies of the fungicide and its metabolites are low when compared to natural ligands, their ability to act via more than one mechanism and the potential for additive or synergistic effect must be taken into consideration in the risk assessment process.

Disruption of sexual function, FSH secretion, and spermiogenesis in rabbits following developmental exposure to vinclozolin, a fungicide.

Science.gov (United States)

Veeramachaneni, D N R; Palmer, J S; Amann, R P; Kane, C M; Higuchi, T T; Pau, K-Y F

2006-04-01

We studied sequelae of prenatal plus infantile exposure of male rabbits to vinclozolin, because it is ingested by women and children. Female Dutch-Belted rabbits (7-10/group) were treated daily per orum from gestation day 15 through post-natal week 4 to provide 0, 7.2, or 72 mg vinclozolin/kg dam's body weight/day. Vinclozolin had no effect on maintenance of pregnancy, growth of pups, age at testicular descent or weight of organs. Concentrations of serum LH or testosterone at 6, 12, or 24 weeks of age were unaffected. However, FSH was lower (P vinclozolin groups at all three ages. Following injection of GnRH at 12 or 24 weeks, the increase in FSH was less (P vinclozolin groups, as was testosterone at 12 weeks of age. After full sexual maturity, 2 of 7 low dose rabbits were uninterested in female or male teasers and never achieved erection or ejaculation. Overall, rates of ejaculation failure were: control 0% (0/48), low dose 29% (12/42), and high dose 5% (3/60). Daily sperm production per gram of testis and total number of sperm per ejaculate in both vinclozolin groups were similar (P > 0.1) to controls. However, semen from vinclozolin rabbits contained over two times more (P vinclozolin rabbits than in controls. Lesions included syncytia of spherical spermatids and desquamation of germ cells. Hence, developmental exposure to vinclozolin caused presumably permanent changes in copulatory ability, secretion of FSH, and spermiogenesis.

Biological effects of tritium on fish cells in the concentration range of international drinking water standards.

Science.gov (United States)

Stuart, Marilyne; Festarini, Amy; Schleicher, Krista; Tan, Elizabeth; Kim, Sang Bog; Wen, Kendall; Gawlik, Jilian; Ulsh, Brant

2016-10-01

To evaluate whether the current Canadian tritium drinking water limit is protective of aquatic biota, an in vitro study was designed to assess the biological effects of low concentrations of tritium, similar to what would typically be found near a Canadian nuclear power station, and higher concentrations spanning the range of international tritium drinking water standards. Channel catfish peripheral blood B-lymphoblast and fathead minnow testis cells were exposed to 10-100,000 Bq l(-1) of tritium, after which eight molecular and cellular endpoints were assessed. Increased numbers of DNA strand breaks were observed and ATP levels were increased. There were no increases in γH2AX-mediated DNA repair. No differences in cell growth were noted. Exposure to the lowest concentrations of tritium were associated with a modest increase in the viability of fathead minnow testicular cells. Using the micronucleus assay, an adaptive response was observed in catfish B-lymphoblasts. Using molecular endpoints, biological responses to tritium in the range of Canadian and international drinking water standards were observed. At the cellular level, no detrimental effects were noted on growth or cycling, and protective effects were observed as an increase in cell viability and an induced resistance to a large challenge dose.

Cytotoxicity of the dicarboximide fungicides, vinclozolin and iprodione, in rat hepatoma-derived Fa32 cells.

Science.gov (United States)

Dierickx, Paul J

2004-10-01

Dicarboximide fungicides are widely used to control various fungal species. Their primary action is not known, due to a lack of knowledge concerning the mechanism of action of the dicarboximide group. The cytotoxicities of vinclozolin and iprodione in rat hepatoma-derived Fa32 cells were investigated. Cytotoxicity was measured by neutral red uptake inhibition after treatment for 24 hours. Iprodione was more toxic than vinclozolin. Vinclozolin was less toxic in glutathione-depleted cells than in control cells. This was also true for iprodione at lower concentrations, but iprodione became more toxic at higher concentrations. Both the fungicides increased the endogenous glutathione content by 20% after 1 hour. After 24 hours, the glutathione content was doubled by vinclozolin, but was not affected by iprodione. No effect on glutathione S-transferase activity or reactive oxygen species formation could be observed. Cytochrome P450-dependent ethoxyresorufin-O-deethylase and pentoxyresorufin-O-depentylase activities were moderately activated by iprodione and strongly activated by vinclozolin. A glutathione-related cytochrome P450-dependent metabolic attack of vinclozolin and iprodione could be responsible for their cytotoxicity in Fa32 cells. Further research is needed to fully elucidate these (or other) mechanisms.

Evaluation of soil bioassays for use at Washington state hazardous waste sites: A pilot study

International Nuclear Information System (INIS)

Blakley, N.; Norton, D.; Stinson, M.; Boyer, R.

1994-01-01

The Washington State Department of Ecology (Ecology) is developing guidelines to assess soil toxicity at hazardous waste sites being investigated under the Washington Model Toxics Control Act Cleanup Regulation. To evaluate soil toxicity, Ecology selected five bioassay protocols -- Daphnia, Earthworm, Seedling, Fathead Minnow, and Frog Embryo Teratogenesis Assay Xenopus (FETAX) -- for use as screening level assessment tools at six State hazardous waste sites. Sites contained a variety of contaminants including metals, creosote, pesticides, and petroleum products (leaking underground storage tanks). Three locations, representing high, medium, and low levels of contamination, were samples at each site. In general, the high contaminant samples resulted in the highest toxic response in all bioassays. The order of site toxicity, as assessed by overall toxic response, is creosote, petroleum products, metals, and pesticides. Results indicate that human health standards, especially for metals, may not adequately protect some of the species tested. The FETAX bioassay had the greatest overall number of toxic responses and lowest variance. The seedling and Daphnia bioassays had lower and similar overall toxic response results, followed by the earthworm and fathead minnow. Variability was markedly highest for the seedling. The Daphnia and fathead minnow variability were similar to the FETAX level, while the earthworm variability was slightly higher

Sequelae in male rabbits following developmental exposure to p,p'-DDT or a mixture of p,p'-DDT and vinclozolin: cryptorchidism, germ cell atypia, and sexual dysfunction.

Science.gov (United States)

Veeramachaneni, D N R; Palmer, J S; Amann, R P; Pau, K-Y F

2007-01-01

Rabbit does (7-9 per group) were treated daily per orum from gestation day 15 through post-natal week 4 to provide per kg body wt 25 micaromol (low) or 250 micromol (high) p,p'-DDT or a mixture of DDT and vinclozolin (12.5 and 125 micromol each). Developmental as well as post-pubertal reproductive sequelae of male progeny were studied. Testicular descent in some pups was impaired by DDT. Serum LH or testosterone was not affected. FSH was lower in mixture- but not in DDT-exposed rabbits. Lack of sexual interest, penile erection and ejaculation were observed in some mixture rabbits. Sperm counts were unaffected, but morphologically normal spermatozoa were fewer; nuclear and acrosomal morphogenesis was disrupted. Atypical germ cells resembling carcinoma in situ were found. Also considering data for vinclozolin [Veeramachaneni DNR, Palmer JS, Amann RP, Kane CM, Higuchi TT, Pau K-YF. Disruption of sexual function, FSH secretion, and spermiogenesis in rabbits following developmental exposure to vinclozolin, a fungicide. Reproduction 2006;131:805-16], we concluded that DDT causes cryptorchidism and germ cell atypia, vinclozolin permanently disrupts FSH secretion and sexual function, and the mixture causes the full spectrum of dysgenesis.

Bisphenol A induces spermatocyte apoptosis in rare minnow Gobiocypris rarus

International Nuclear Information System (INIS)

Zhang, Yingying; Cheng, Mengqian; Wu, Lang; Zhang, Guo; Wang, Zaizhao

2016-01-01

Highlights: • Adult male G. rarus were exposed to 225 μg/L BPA for 7, 21 and 63 days. • BPA could induce spermatocyte apoptosis in rare minnow testis. • The mitochondrial apoptotic pathway participated in the germ cell apoptosis. • The spermatocyte apoptosis was likely initiated by BPA induced meiosis arrest. - Abstract: Bisphenol A (BPA) is an endocrine disruptor, and could induce germ cells apoptosis in the testis of mammals. But whether it could affect fish in the same mechanism has not’ been studied till now. In the present study, to investigate the influence of BPA on testis germ cells in fish, adult male rare minnow Gobiocypris rarus were exposed to 225 μg L"−"1 (0.99 μM) BPA for 1, 3 and 9 weeks. Through TdT-mediated dUTP nick end labeling (TUNEL) and transmission electron microscope (TEM) analysis, we found that the amount of apoptotic spermatocytes significantly increased in a time dependent manner following BPA exposure. Western Blot results showed that the ratio of Bcl2/Bax, the important apoptosis regulators in intrinsic mitochondrial apoptotic pathway, was significantly decreased. qPCR showed that mRNA expression of several genes in mitochondrial apoptotic pathway including bcl2, bax, casp9, cytc and mcl1b were significantly changed following BPA exposure. In addition, mRNA expression of meiosis regulation genes (kpna7 and wee2), and genes involved in both apoptosis and meiosis (birc5, ccna1, and gsa1a) were also affected by BPA. Taken together, the present study demonstrated that BPA could induce spermatocytes apoptosis in rare minnow testis, and the apoptosis was probably under regulation of intrinsic mitochondrial apoptotic pathway. Moreover, the spermatocyte apoptosis was likely initiated by BPA induced meiosis arrest.

Bisphenol A induces spermatocyte apoptosis in rare minnow Gobiocypris rarus

Energy Technology Data Exchange (ETDEWEB)

Zhang, Yingying; Cheng, Mengqian; Wu, Lang; Zhang, Guo; Wang, Zaizhao, E-mail: zzwang@nwsuaf.edu.cn

2016-10-15

Highlights: • Adult male G. rarus were exposed to 225 μg/L BPA for 7, 21 and 63 days. • BPA could induce spermatocyte apoptosis in rare minnow testis. • The mitochondrial apoptotic pathway participated in the germ cell apoptosis. • The spermatocyte apoptosis was likely initiated by BPA induced meiosis arrest. - Abstract: Bisphenol A (BPA) is an endocrine disruptor, and could induce germ cells apoptosis in the testis of mammals. But whether it could affect fish in the same mechanism has not’ been studied till now. In the present study, to investigate the influence of BPA on testis germ cells in fish, adult male rare minnow Gobiocypris rarus were exposed to 225 μg L{sup −1} (0.99 μM) BPA for 1, 3 and 9 weeks. Through TdT-mediated dUTP nick end labeling (TUNEL) and transmission electron microscope (TEM) analysis, we found that the amount of apoptotic spermatocytes significantly increased in a time dependent manner following BPA exposure. Western Blot results showed that the ratio of Bcl2/Bax, the important apoptosis regulators in intrinsic mitochondrial apoptotic pathway, was significantly decreased. qPCR showed that mRNA expression of several genes in mitochondrial apoptotic pathway including bcl2, bax, casp9, cytc and mcl1b were significantly changed following BPA exposure. In addition, mRNA expression of meiosis regulation genes (kpna7 and wee2), and genes involved in both apoptosis and meiosis (birc5, ccna1, and gsa1a) were also affected by BPA. Taken together, the present study demonstrated that BPA could induce spermatocytes apoptosis in rare minnow testis, and the apoptosis was probably under regulation of intrinsic mitochondrial apoptotic pathway. Moreover, the spermatocyte apoptosis was likely initiated by BPA induced meiosis arrest.

Ancestral vinclozolin exposure alters the epigenetic transgenerational inheritance of sperm small noncoding RNAs.

Science.gov (United States)

Schuster, Andrew; Skinner, Michael K; Yan, Wei

Exposure to the agricultural fungicide vinclozolin during gestation promotes a higher incidence of various diseases in the subsequent unexposed F3 and F4 generations. This phenomenon is termed epigenetic transgenerational inheritance and has been shown to in part involve alterations in DNA methylation, but the role of other epigenetic mechanisms remains unknown. The current study investigated the alterations in small noncoding RNA (sncRNA) in the sperm from F3 generation control and vinclozolin lineage rats. Over 200 differentially expressed sncRNAs were identified and the tRNA-derived sncRNAs, namely 5' halves of mature tRNAs (5' halves), displayed the most dramatic changes. Gene targets of the altered miRNAs and tRNA 5' halves revealed associations between the altered sncRNAs and differentially DNA methylated regions. Dysregulated sncRNAs appear to correlate with mRNA profiles associated with the previously observed vinclozolin-induced disease phenotypes. Data suggest potential connections between sperm-borne RNAs and the vinclozolin-induced epigenetic transgenerational inheritance phenomenon.

Prenatal exposure to vinclozolin disrupts selective aspects of the gonadotropin-releasing hormone neuronal system of the rabbit

Science.gov (United States)

Wadas, B.C.; Hartshorn, C.A.; Aurand, E.R.; Palmer, J.S.; Roselli, C.E.; Noel, M.L.; Gore, A.C.; Veeramachaneni, D.N.R.; Tobet, S.A.

2010-01-01

Developmental exposure to the agricultural fungicide vinclozolin can impair reproductive function in male rabbits and was previously found to decrease the number of immunoreactive-gonadotropin-releasing hormone (ir-GnRH) neurons in the region of the organum vasculosum of the lamina terminalis (OVLT) and rostral preoptic area (rPOA) by postnatal week (PNW) 6. To further examine the disruption of GnRH neurons by fetal vinclozolin exposure, in the current study, pregnant rabbits were dosed orally with vinclozolin, flutamide, or carrot paste vehicle for the last two weeks of gestation. Offspring were euthanized at birth (males and females), PNW6 (females), PNW26 (adult males), or PNW30 (adult females) of age. At birth and in adults, brains were sectioned and processed for immunoreactive GnRH. The numbers of immunoreactive GnRH neuronal perikarya were significantly decreased in vinclozolin-treated rabbits at birth and in adult littermates. By contrast, there was an increase in GnRH immunoreactivity in the terminals in the region of the median eminence. Analysis of PNW6 female brains by radioimmunoassay (RIA) revealed a two-fold increase in GnRH peptide content in the mediobasal hypothalamus in vinclozolin-treated rabbits. This finding was complemented by immunofluorescence analyses that showed a 2.8-fold increase in GnRH immunoreactivity in the median eminence of vinclozolin compared to vehicle-treated females at PNW30. However, there was no difference between treatment groups in the measures of reproduction that were evaluated: ejaculation latency, conception rates or litter size. These results indicate that subacute, prenatal vinclozolin treatment is sufficient to create perdurable alterations in the GnRH neuronal network that forms an important input into the reproductive axis. Finally, the effect of vinclozolin on the GnRH neuronal network was not comparable to that of flutamide, suggesting that vinclozolin was not acting through anti-androgenic mechanisms. PMID

Prenatal exposure to vinclozolin disrupts selective aspects of the gonadotrophin-releasing hormone neuronal system of the rabbit.

Science.gov (United States)

Wadas, B C; Hartshorn, C A; Aurand, E R; Palmer, J S; Roselli, C E; Noel, M L; Gore, A C; Veeramachaneni, D N R; Tobet, S A

2010-06-01

Developmental exposure to the agricultural fungicide vinclozolin can impair reproductive function in male rabbits and was previously found to decrease the number of immunoreactive-gonadotrophin-releasing hormone (GnRH) neurones in the region of the organum vasculosum of the lamina terminalis and rostral preoptic area by postnatal week (PNW) 6. In the present study, in an aim to further examine the disruption of GnRH neurones by foetal vinclozolin exposure, pregnant rabbits were dosed orally with vinclozolin, flutamide or carrot paste vehicle for the last 2 weeks of gestation. Offspring were euthanised at birth (males and females), PNW 6 (females), PNW 26 (adult males) or PNW 30 (adult females) of age. At birth and in adults, brains were sectioned and processed for immunoreactive GnRH. The numbers of immunoreactive GnRH neuronal perikarya were significantly decreased in vinclozolin-treated rabbits at birth and in adult littermates. By contrast, there was an increase in GnRH immunoreactivity in the terminals in the region of the median eminence. Analysis of PNW 6 female brains by radioimmunoassay revealed a two-fold increase in GnRH peptide content in the mediobasal hypothalamus in vinclozolin-treated rabbits. This finding was complemented by immunofluorescence analyses, which revealed a 2.8-fold increase in GnRH immunoreactivity in the median eminence of vinclozolin compared to vehicle-treated females at PNW 30. However, there was no difference between treatment groups in the measures of reproduction that were evaluated: ejaculation latency, conception rates or litter size. These results indicate that sub-acute, prenatal vinclozolin treatment is sufficient to create perdurable alterations in the GnRH neuronal network that forms an important input into the reproductive axis. Finally, the effect of vinclozolin on the GnRH neuronal network was not comparable to that of flutamide, suggesting that vinclozolin was not acting through anti-androgenic mechanisms.

Transgenerational epigenetic effects of the endocrine disruptor vinclozolin on pregnancies and female adult onset disease.

Science.gov (United States)

Nilsson, Eric E; Anway, Matthew D; Stanfield, Jacob; Skinner, Michael K

2008-05-01

Endocrine disruptor exposure during gonadal sex determination was previously found to induce male rat adult onset transgenerational disease (F1-F4 generation), and this was associated with an alteration in the epigenetic (i.e., DNA methylation) programming of the male germ line. The current study was designed to characterize the transgenerational disease phenotypes of the female adult offspring. Pregnant rats (F0 generation) were treated transiently with vinclozolin (i.e., fungicide with anti-androgenic activity) on embryonic (E) days E8-E14 of gestation. F1 control and vinclozolin generation offspring from different litters were mated to produce F2 offspring, and similarly F2 generation animals produced F3 generation offspring. Observations demonstrated that 9 out of 105 pregnant rats (8.6%) from the vinclozolin F1-F3 generations exhibited uterine hemorrhage and/or anemia late in pregnancy. None (0 out of 82) of the control F1-F3 generation females had similar pregnancy problems. Complete blood cell counts and serum chemistry profiles demonstrated that selected vinclozolin generation animals, but not controls, exhibited marked regenerative anemia in late pregnancy. Examination of kidney histology revealed moderate or severe glomerular abnormalities in 67% of the vinclozolin F2 and F3 generation adult females compared with 18% of the controls. Adult female vinclozolin generation animals also developed various types of tumors in 6.5% of the animals (11 out of 170), while 2% of control-line animals (3 out of 151) developed mammary tumors. Observations demonstrate that vinclozolin exposure during gonadal sex determination promotes a transgenerational increase in pregnancy abnormalities and female adult onset disease states.

Effect of sublethal levels of ionizing radiation on a predator-prey interaction

International Nuclear Information System (INIS)

Chee, P.C.

1976-01-01

The predator-prey interaction studied was that between the largemouth bass (Micropterus salmoides) and the fathead minnow (Pimephales promelas) in an artificial test environment. Experiments were first conducted to determine the 50% lethal dose at 30 days of the minnow. Three different dose rates were used to test the effect of dose rate on the 50% lethal dose value. After the 50% lethal dose was determined the predator-prey interaction experiment was conducted using 30% of the 50% lethal dose as the highest radiation dose, this dose being considered the upper limit to sublethal radiation levels. A 4 x 4 Latin square design was chosen for the experiment, with four treatment levels (control plus three radiation levels) and four replicates. In each test 10 prey minnow were offered to one predator bass and the number of prey left after 14 days was the parameter of interest. A predator-prey interaction experiment using a single high level of radiation and two types of controls as conducted to ascertain the ability of the test environment to detect changes in the predator-prey interaction. The two types of controls were irradiated prey not exposed to predation and non-irradiated prey exposed to predation. An experiment was also conducted to test the correlation between the physical activity patterns of minnow and different doses of radiation. At a dose rate of 37.8 rad/min the 50% lethal dose at 30 days for minnow was found to be 2650 rad. It was found that dose rate had a strong influence on the 50% lethal dose. In the predator-prey interaction test it was found that the 14-day survival rate of prey was unaffected by sublethal levels of ionizing radiation. No significant correlation was detected between the physical activity patterns of minnow and radiation dose

Sex differences and the development of the rabbit brain: effects of vinclozolin.

Science.gov (United States)

Bisenius, Erin S; Veeramachaneni, D N Rao; Sammonds, Ginger E; Tobet, Stuart

2006-09-01

The preoptic/anterior hypothalamic area (POA/AH) is one of the most sexually dimorphic areas of the vertebrate brain and plays a pivotal role in regulating male sexual behavior. Vinclozolin is a fungicide thought to be an environmental antiandrogen, which disrupts masculine sexual behavior when administered to rabbits during development. In this study, we examined several characteristics of the rabbit POA/AH for sexual dimorphism and endocrine disruption by vinclozolin. Pregnant rabbits were dosed orally with vinclozolin (10 mg/kg body weight) or carrot paste vehicle once daily for 6 wk beginning at midgestation and continuing through nursing until Postpartum Week 4. At 6 wk, offspring were perfused with 4% paraformaldehyde and brains processed for immunocytochemical localization of tyrosine hydroxylase, calbindin, gonadotropin-releasing hormone (GnRH), or Nissl stain. There were significant sex differences in the distribution of calbindin in the POA/AH and the size of cells in the dorsal POA/AH (values greater in females than in males), but not in the number or distribution of tyrosine hydroxylase or GnRH neurons. In both sexes, exposure to vinclozolin significantly increased calbindin expression in the ventral POA/AH and significantly decreased number of GnRH neurons selectively in the region of the organum vasculosum of the lamina terminalis (OVLT) but not more caudally in the POA/AH. This is the first documentation of a sexually dimorphic region in the rabbit brain, and further supports the use of this species as a model for studying the influence of vinclozolin on reproductive development with potential application to human systems.

The combined effects of vinclozolin and procymidone do not deviate from expected additivity in vitro and in vivo

DEFF Research Database (Denmark)

Nellemann, Christine Lydia; Dalgaard, Majken; Lam, Henrik Rye

2003-01-01

The combination effects of the well-known antiandrogenic fungicides, vinclozolin and procymidone, were tested both in vitro and in vivo. In vitro both vinclozolin and procymidone significantly inhibited the binding of agonist to the androgen receptor with the concentration that resulted in 50......% inhibition (IC50) values of 0.1 and 0.6 muM, respectively. By applying the isobole method, the effect of combining the two pesticides in vitro was found to be additive. In castrated testosterone-treated rats the administration of vinclozolin starting at 10 mg/kg led to a decrease in organ weight of all...... tested reproductive organs. The levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH) were increased significantly with doses of 100 mg/kg vinclozolin and above. Expression of the androgen-responsive gene, TRPM-2, was increased starting at 100 mg/kg vinclozolin. For procymidone...

Fate of vinclozolin, thiabendazole and dimethomorph during storage, handling and forcing of chicory.

Science.gov (United States)

Spanoghe, Pieter; Ryckaert, Bert; Van Gheluwe, Cindy; Van Labeke, Marie-Christine

2010-02-01

As part of ongoing research for a sustainable production of Belgian endives, the fate of three fungicides during storage, handling and forcing of witloof chicory roots was investigated. Storage roots are protected against Sclerotinia sp. Fuckel and Phoma exigua var. exigua Desm. by means of vinclozolin and thiabendazole respectively. During hydroponic forcing, the most imminent pathogen is Phytophthora cryptogea Pethybr. & Laff., which is controlled by the use of dimethomorph. Vinclozolin and thiabendazole concentrations on roots remained constant during storage at -1 degrees C. Dermal exposure of the workers in hydroponics was exceeded. Vinclozolin and thiabendazole residues were not detected 2 weeks after hydroponic forcing; dimethomorph was still detected at harvest. At harvest, the vinclozolin concentration in the chicory heads was below the maximum residue limit, but the chicory roots contained residues much above the thiabendazole and dimethomorph maximum residue level. Vinclozolin and thiabendazole residues applied before storage are still present on the roots at the start of the forcing cycle. During the set-up of chicory roots, preventive measures are recommended, as effects of repeated human exposure to low doses of applied fungicides cannot be excluded. Dimethomorph applied at the start of the hydroponic forcing is the only pesticide detected in the drainage water at harvest. The chicory heads were safe for human consumption. However, more attention should be paid to the residues of fungicides in the roots used for cattle feeding.

Vinclozolin Exposure in Utero Induces Postpubertal Prostatitis and Reduces Sperm Production via a Reversible Hormone-Regulated Mechanism

OpenAIRE

Cowin, Prue A.; Gold, Elspeth; Aleksova, Jasna; O'Bryan, Moira K.; Foster, Paul M. D.; Scott, Hamish S.; Risbridger, Gail P.

2010-01-01

Vinclozolin is an endocrine-disrupting chemical (EDC) that binds with high affinity to the androgen receptor (AR) and blocks the action of gonadal hormones on male reproductive organs. An alternative mechanism of action of Vinclozolin involves transgenerational effects on the male reproductive tract. We previously reported in utero Vinclozolin exposure-induced prostatitis (prostate inflammation) in postpubertal rats concurrent with down-regulation of AR and increased nuclear factor-κB activat...

Epigenetic transgenerational inheritance of vinclozolin induced mouse adult onset disease and associated sperm epigenome biomarkers.

Science.gov (United States)

Guerrero-Bosagna, Carlos; Covert, Trevor R; Haque, Md M; Settles, Matthew; Nilsson, Eric E; Anway, Matthew D; Skinner, Michael K

2012-12-01

The endocrine disruptor vinclozolin has previously been shown to promote epigenetic transgenerational inheritance of adult onset disease in the rat. The current study was designed to investigate the transgenerational actions of vinclozolin on the mouse. Transient exposure of the F0 generation gestating female during gonadal sex determination promoted transgenerational adult onset disease in F3 generation male and female mice, including spermatogenic cell defects, testicular abnormalities, prostate abnormalities, kidney abnormalities and polycystic ovarian disease. Pathology analysis demonstrated 75% of the vinclozolin lineage animals developed disease with 34% having two or more different disease states. Interestingly, the vinclozolin induced transgenerational disease was observed in the outbred CD-1 strain, but not the inbred 129 mouse strain. Analysis of the F3 generation sperm epigenome identified differential DNA methylation regions that can potentially be utilized as epigenetic biomarkers for transgenerational exposure and disease. Copyright © 2012 Elsevier Inc. All rights reserved.

Cumulative and antagonistic effects of a mixture of the antiandrogens vinclozolin and iprodione in the pubertal male rat.

Science.gov (United States)

Blystone, Chad R; Lambright, Christy S; Cardon, Mary C; Furr, Johnathan; Rider, Cynthia V; Hartig, Phillip C; Wilson, Vickie S; Gray, Leon E

2009-09-01

Vinclozolin and iprodione are dicarboximide fungicides that display antiandrogenic effects in the male rat, which suggests that a mixture would lead to cumulative effects on androgen-sensitive end points. Iprodione is a steroid synthesis inhibitor, but androgen receptor antagonist activity, which is displayed by vinclozolin, has not been fully evaluated. Here, we demonstrate that iprodione binds to the human androgen receptor (IC(50) = 86.0 microM), reduces androgen-dependent gene expression, and reduces androgen-sensitive tissue weights in castrated male rats (Hershberger assay). Since vinclozolin and iprodione affect common targets in the pubertal male rat, we tested the hypothesis that a mixture would have cumulative antiandrogenic effects. An iprodione dose, that does not significantly affect androgen-dependent morphological end points, was combined with vinclozolin doses (2 x 5 factorial design). Sprague-Dawley rats were dosed by gavage with vinclozolin at 0, 10, 30, 60, and 100 mg/kg/day with and without 50 mg iprodione/kg/day from postnatal day (PND) 23 to 55-57 (n = 8 per group). The age at puberty (preputial separation [PPS]), organ weights, serum hormones, and ex vivo testis steroid hormone production were measured. Vinclozolin delayed PPS, reduced androgen-sensitive organ weights, and increased serum testosterone. The addition of iprodione enhanced the vinclozolin inhibition of PPS (PND 47.5 vs.49.1; two-way ANOVA: iprodione main effect p = 0.0002). The dose response for several reproductive and nonreproductive organ weights was affected in a cumulative manner. In contrast, iprodione antagonized the vinclozolin-induced increase in serum testosterone. These results demonstrate that these fungicides interact on common targets in a tissue-specific manner when coadministered to the pubertal male rat.

Developing Predictive Approaches to Characterize Adaptive Responses of the Reproductive Endocrine Axis to Aromatase Inhibition II: Computational Modeling

Science.gov (United States)

ABSTRACT Exposure to endocrine disrupting chemicals can affect reproduction and development in both humans and wildlife. We developed a mechanistic mathematical model of the hypothalamic­ pituitary-gonadal (HPG) axis in female fathead minnows to predic...

Assessing the efficacy of corn-based bait containing antimycin-a to control common carp populations using laboratory and pond experiments

Science.gov (United States)

Poole, Joshua R.; Sauey, Blake W.; Amberg, Jon J.; Bajer, Przemyslaw G.

2018-01-01

Strategic use of oral toxicants could allow for practical and sustainable control schemes for the invasive common carp (Cyprinus carpio, or ‘carp’) if a toxicant selectively targeted carp and not native species. In this study, we incorporated antimycin-a (ANT-A), a known fish toxicant, into a corn-based bait and conducted a series of experiments to determine its toxicity, leaching rate, and species-specificity. Our results showed that ANT-A was lethal to carp at doses ≥ 4 mg/kg and that the amount of ANT-A that leached out of the bait in 72 h was not lethal to carp or bluegill (Lepomis macrochirus). Species-specificity trials were conducted in 227 L tanks, in which carp were stocked with three native species representing families that occur sympatrically with carp in our study region: the fathead minnow (Pimephales promelas), yellow perch (Perca flavescens) and bluegill. These trials showed high mortality of carp (46%) and fathead minnows (76%) but no significant mortality of perch or bluegill. Finally, a pond study, which used the same species composition except for fathead minnows, resulted in 37% morality among adult carp and no mortality among perch or bluegill. Our results suggest that corn-based bait that contains ANT-A could be used to selectively control carp in ecosystems dominated by percids or centrarchids, such as lakes across the Great Plains ecoregion of North America, where carp are especially problematic.

Sexually dimorphic effects of ancestral exposure to vinclozolin on stress reactivity in rats.

Science.gov (United States)

Gillette, Ross; Miller-Crews, Isaac; Nilsson, Eric E; Skinner, Michael K; Gore, Andrea C; Crews, David

2014-10-01

How an individual responds to the environment depends upon both personal life history as well as inherited genetic and epigenetic factors from ancestors. Using a 2-hit, 3 generations apart model, we tested how F3 descendants of rats given in utero exposure to the environmental endocrine-disrupting chemical (EDC) vinclozolin reacted to stress during adolescence in their own lives, focusing on sexually dimorphic phenotypic outcomes. In adulthood, male and female F3 vinclozolin- or vehicle-lineage rats, stressed or nonstressed, were behaviorally characterized on a battery of tests and then euthanized. Serum was used for hormone assays, and brains were used for quantitative PCR and transcriptome analyses. Results showed that the effects of ancestral exposure to vinclozolin converged with stress experienced during adolescence in a sexually dimorphic manner. Debilitating effects were seen at all levels of the phenotype, including physiology, behavior, brain metabolism, gene expression, and genome-wide transcriptome modifications in specific brain nuclei. Additionally, females were significantly more vulnerable than males to transgenerational effects of vinclozolin on anxiety but not sociality tests. This fundamental transformation occurs in a manner not predicted by the ancestral exposure or the proximate effects of stress during adolescence, an interaction we refer to as synchronicity.

Residuen van iprodion, procymidon en vinclozolin in knolvenkel

NARCIS (Netherlands)

Goewie; C.E.; Hogendoorn; E.A.

1986-01-01

In het kader van veldproeven werden residuen van de fungiciden iprodion, procymidon en vinclozolin bepaald in knolvenkel. Hiertoe werd een vloeistofchromatografische analysemethode ontwikkeld, welke tevens een volautomatische clean-up stap omvat. Deze methode is een verbeterde versie van de

Toxicity of contaminants in lagoons and pannes of the Indiana Dunes National Lakeshore

International Nuclear Information System (INIS)

Gillespie, R.; Speelman, J.; Stewart, P.M.

1995-01-01

Contaminants in water and sediments of lagoons and pannes were 2--90 times greater at sites adjacent to slag and coal piles than those at reference sites. One site (Lagoon-US5) had sediments with very high concentrations of toxic organics (e.g. naphthalene, phenanthrene, dibenzofuran). Although analyses indicated a gradient of contaminant concentration with distance from their sources, toxicity assays were somewhat equivocal. With the exception of less reproduction in Ceriodaphnia at one lagoon site (US3 = 0.55 of reference), survival of fathead minnows and reproduction in Ceriodaphnia in lagoon and panne waters varied independently of the contaminant concentration. In fact, there was better Ceriodaphnia reproduction in water from two contaminated sites (Lagoon-US5, Panne-WP1) than in water from reference sites. Fathead minnow survival, Ceriodaphnia survival, Ceriodaphnia reproduction, amphipod survival, and amphipod growth varied among sites in toxicity assays with sediments, 100% mortality of fatheads at Lagoon-US5, 100% mortality of Ceriodaphnia at Lagoon-US3, and less survival of fathead minnows at Lagoon-US3 indicate possible toxicity from contaminants in sediments at these sites. Of all organisms and end-points tested, Ceriodaphnia survival seemed to be most closely associated with concentrations of contaminants in lagoon water and sediments. Amphipod survival also varied with contaminants in sediments, however, survival in sediments of contaminated sites ranged only from 0.90--0.93 of reference sites. Although the results are not consistent among organisms, toxicity assays indicate that sediments from the lagoon site with the highest contaminants (Lagoon-US5) and possibly those from another contaminated lagoon site (Lagoon-US3) could be toxic to aquatic organisms. Water and sediments from contaminated panne sites do not appear to be toxic to aquatic test organisms

Effects of the UV-filter 2-ethyl-hexyl-4-trimethoxycinnamate (EHMC) on expression of genes involved in hormonal pathways in fathead minnows (Pimephales promelas) and link to vitellogenin induction and histology

International Nuclear Information System (INIS)

Christen, Verena; Zucchi, Sara; Fent, Karl

2011-01-01

UV-filters are increasingly used in cosmetics and in the protection of materials against UV-irradiation, and ultimately they reach aquatic systems. The lipophilic UV-filter 2-ethyl-hexyl-4-trimethoxycinnamate (EHMC) belongs to one of the most frequently used UV-filters and accumulates in aquatic animals. Despite its ubiquitous presence in water and biota, very little is known about its potential hormonal effects on aquatic organisms. In our study, we evaluated the effects of measured water concentration of 5.4, 37.5, 244.5 and 394 μg/L EHMC on the expression of genes involved in hormonal pathways in the liver, testis and brain of male and female fathead minnows (Pimephales promelas). We compare the transcription profile with the plasma vitellogenin (VTG) content, secondary sex characteristics, and gonad histology. Transcripts of the androgen receptor (ar) were significantly down-regulated in the liver of females at 37.5, 244.5 μg/L and 394 μg/L EHMC. Additionally, the 3β-hydroxysteroid dehydrogenase (3β-HSD) transcript was significantly decreased in the liver of males at 37.5, 244.5 and 394 μg/L EHMC, and at 244.5 and 394 μg/L EHMC in females. The expressional changes were tissue-specific in most cases, being most significant in the liver. Vitellogenin plasma concentration was significantly increased at 244.5 μg/L EHMC in males. EHMC induced significant histological changes in testes and ovaries at 394 μg/L. Testes displayed a decrease in spermatocytes, and ovaries a decrease in previtellogenic oocytes. The induction of VTG plasma concentration and the histological changes in gonads suggest an estrogenic and/or antiandrogenic activity of EHMC. On the other hand, the gene expression profile shows an antiestrogenic (e.g.: down-regulation of esr1) activity of EHMC. In conclusion, our data demonstrate that EHMC displays low but multiple hormonal activities in fish.

Effects of the UV-filter 2-ethyl-hexyl-4-trimethoxycinnamate (EHMC) on expression of genes involved in hormonal pathways in fathead minnows (Pimephales promelas) and link to vitellogenin induction and histology

Energy Technology Data Exchange (ETDEWEB)

Christen, Verena; Zucchi, Sara [University of Applied Sciences Northwestern Switzerland, School of Life Sciences, Gruendenstrasse 40, CH-4132 Muttenz (Switzerland); Fent, Karl, E-mail: karl.fent@fhnw.ch [University of Applied Sciences Northwestern Switzerland, School of Life Sciences, Gruendenstrasse 40, CH-4132 Muttenz (Switzerland); Swiss Federal Institute of Technology Zuerich (ETH Zuerich), Department of Environmental Sciences, 8092 Zuerich (Switzerland)

2011-04-15

UV-filters are increasingly used in cosmetics and in the protection of materials against UV-irradiation, and ultimately they reach aquatic systems. The lipophilic UV-filter 2-ethyl-hexyl-4-trimethoxycinnamate (EHMC) belongs to one of the most frequently used UV-filters and accumulates in aquatic animals. Despite its ubiquitous presence in water and biota, very little is known about its potential hormonal effects on aquatic organisms. In our study, we evaluated the effects of measured water concentration of 5.4, 37.5, 244.5 and 394 {mu}g/L EHMC on the expression of genes involved in hormonal pathways in the liver, testis and brain of male and female fathead minnows (Pimephales promelas). We compare the transcription profile with the plasma vitellogenin (VTG) content, secondary sex characteristics, and gonad histology. Transcripts of the androgen receptor (ar) were significantly down-regulated in the liver of females at 37.5, 244.5 {mu}g/L and 394 {mu}g/L EHMC. Additionally, the 3{beta}-hydroxysteroid dehydrogenase (3{beta}-HSD) transcript was significantly decreased in the liver of males at 37.5, 244.5 and 394 {mu}g/L EHMC, and at 244.5 and 394 {mu}g/L EHMC in females. The expressional changes were tissue-specific in most cases, being most significant in the liver. Vitellogenin plasma concentration was significantly increased at 244.5 {mu}g/L EHMC in males. EHMC induced significant histological changes in testes and ovaries at 394 {mu}g/L. Testes displayed a decrease in spermatocytes, and ovaries a decrease in previtellogenic oocytes. The induction of VTG plasma concentration and the histological changes in gonads suggest an estrogenic and/or antiandrogenic activity of EHMC. On the other hand, the gene expression profile shows an antiestrogenic (e.g.: down-regulation of esr1) activity of EHMC. In conclusion, our data demonstrate that EHMC displays low but multiple hormonal activities in fish.

Effects of the UV-filter 2-ethyl-hexyl-4-trimethoxycinnamate (EHMC) on expression of genes involved in hormonal pathways in fathead minnows (Pimephales promelas) and link to vitellogenin induction and histology.

Science.gov (United States)

Christen, Verena; Zucchi, Sara; Fent, Karl

2011-04-01

UV-filters are increasingly used in cosmetics and in the protection of materials against UV-irradiation, and ultimately they reach aquatic systems. The lipophilic UV-filter 2-ethyl-hexyl-4-trimethoxycinnamate (EHMC) belongs to one of the most frequently used UV-filters and accumulates in aquatic animals. Despite its ubiquitous presence in water and biota, very little is known about its potential hormonal effects on aquatic organisms. In our study, we evaluated the effects of measured water concentration of 5.4, 37.5, 244.5 and 394 μg/L EHMC on the expression of genes involved in hormonal pathways in the liver, testis and brain of male and female fathead minnows (Pimephales promelas). We compare the transcription profile with the plasma vitellogenin (VTG) content, secondary sex characteristics, and gonad histology. Transcripts of the androgen receptor (ar) were significantly down-regulated in the liver of females at 37.5, 244.5 μg/L and 394 μg/L EHMC. Additionally, the 3β-hydroxysteroid dehydrogenase (3β-HSD) transcript was significantly decreased in the liver of males at 37.5, 244.5 and 394 μg/L EHMC, and at 244.5 and 394 μg/L EHMC in females. The expressional changes were tissue-specific in most cases, being most significant in the liver. Vitellogenin plasma concentration was significantly increased at 244.5 μg/L EHMC in males. EHMC induced significant histological changes in testes and ovaries at 394 μg/L. Testes displayed a decrease in spermatocytes, and ovaries a decrease in previtellogenic oocytes. The induction of VTG plasma concentration and the histological changes in gonads suggest an estrogenic and/or antiandrogenic activity of EHMC. On the other hand, the gene expression profile shows an antiestrogenic (e.g.: down-regulation of esr1) activity of EHMC. In conclusion, our data demonstrate that EHMC displays low but multiple hormonal activities in fish. Copyright © 2011 Elsevier B.V. All rights reserved.

Chiral separation and enantioselective degradation of vinclozolin in soils.

Science.gov (United States)

Liu, Hui; Liu, Donghui; Shen, Zhigang; Sun, Mingjing; Zhou, Zhiqiang; Wang, Peng

2014-03-01

Vinclozolin is a chiral fungicide with potential environmental problems. The chiral separation of the enantiomers and enantioselective degradation in soil were investigated in this work. The enantiomers were separated by high-performance liquid chromatography (HPLC) on Chiralpak IA, IB, and AZ-H chiral columns under normal phase and the influence of the mobile phase composition on the separation was also studied. Complete resolutions were obtained on all three chiral columns under optimized conditions with the same elution order of (+)/(-). The residual analysis of the enantiomers in soil was conducted using accelerate solvent extraction followed by HPLC determination. The recoveries of the enantiomers ranged from 85.7-105.7% with relative standard deviation (SD) of 0.12-3.83%, and the limit of detection (LOD) of the method was 0.013 µg/g. The results showed that the degradations of vinclozolin enantiomers in the soils followed first-order kinetics. Preferential degradation of the (-)-enantiomer was observed only in one soil with the largest |ES| value of 0.047, and no obvious enantioselective degradation was observed in other soils. It was found that the persistence of vinclozolin in soil was related to pH values based on the half-lives. The two enantiomers disappeared about 8 times faster in basic soils than that in neutral or acidic soils. © 2014 Wiley Periodicals, Inc.

Vinclozolin alters the expression of hormonal and stress genes in the midge Chironomus riparius.

Science.gov (United States)

Aquilino, Mónica; Sánchez-Argüello, Paloma; Martínez-Guitarte, José-Luis

2016-05-01

Vinclozolin is a fungicide used in agriculture that can reach aquatic ecosystems and affect the organisms living there. Its effects have been intensively studied in vertebrates, where it acts as an antiandrogen, but there is a lack of information about its mechanistic effects on invertebrates. In this work, we analyzed the response of genes related to the endocrine system, the stress response, and the detoxification mechanisms of Chironomus riparius fourth instar larvae after 24h and 48h exposures to 20 (69.9nM), 200 (699nM), and 2000μg/L (6.99μM) of Vinclozolin. Survival analysis showed that this compound has low toxicity, as it was not lethal for this organism at the concentrations used. However, this fungicide was shown to modify the transcriptional activity of the ecdysone response pathway genes EcR, E74, and Kr-h1 by increasing their mRNA levels. While no changes were observed in disembodied, a gene related with the ecdysone synthesis metabolic pathway, Cyp18A1, which is involved in the inactivation of the active form of ecdysone, was upregulated. Additionally, the expression of two genes related to other hormones, FOXO and MAPR, did not show any changes when Vinclozolin was present. The analysis of stress response genes showed significant changes in the mRNA levels of Hsp70, Hsp24, and Gp93, indicating that Vinclozolin activates the cellular stress mechanisms. Finally, the expressions of the genes Cyp4G and GstD3, which encode enzymes involved in phase I and phase II detoxification, respectively, were analyzed. It was found that their mRNA levels were altered by Vinclozolin, suggesting their involvement in the degradation of this compound. For the first time, these results show evidence that Vinclozolin can modulate gene expression, leading to possible significant endocrine alterations of the insect endocrine system. These results also offer new clues about the mode of action of this compound in invertebrates. Copyright © 2016 Elsevier B.V. All rights

Chronic dietary exposure to a low-dose mixture of genistein and vinclozolin modifies the reproductive axis, testis transcriptome, and fertility.

Science.gov (United States)

Eustache, Florence; Mondon, Françoise; Canivenc-Lavier, Marie Chantal; Lesaffre, Corinne; Fulla, Yvonne; Berges, Raymond; Cravedi, Jean Pierre; Vaiman, Daniel; Auger, Jacques

2009-08-01

The reproductive consequences and mechanisms of action of chronic exposure to low-dose endocrine disruptors are poorly understood. We assessed the effects of a continuous, low-dose exposure to a phytoestrogen (genistein) and/or an antiandrogenic food contaminant (vinclozolin) on the male reproductive tract and fertility. Male rats were exposed by gavage to genistein and vinclozolin from conception to adulthood, alone or in combination, at low doses (1 mg/kg/day) or higher doses (10 and 30 mg/kg/day). We studied a number of standard reproductive toxicology end points and also assessed testicular mRNA expression profiles using long-oligonucleotide microarrays. The low-dose mixture and high-dose vinclozolin produced the most significant alterations in adults: decreased sperm counts, reduced sperm motion parameters, decreased litter sizes, and increased post implantation loss. Testicular mRNA expression profiles for these exposure conditions were strongly correlated. Functional clustering indicated that many of the genes induced belong to the "neuroactive ligand-receptor interactions" family encompassing several hormonally related actors (e.g., follicle-stimulating hormone and its receptor). All exposure conditions decreased the levels of mRNAs involved in ribosome function, indicating probable decreased protein production. Our study shows that chronic exposure to a mixture of a dose of a phytoestrogen equivalent to that in the human diet and a low dose-albeit not environmental-of a common anti-androgenic food contaminant may seriously affect the male reproductive tract and fertility.

Developing Predictive Approaches to Characterize Adaptive Responses of the Reproductive Endocrine Axis to Aromatase Inhibition: Computational Modeling

Science.gov (United States)

Exposure to endocrine disrupting chemicals can affect reproduction and development in both humans and wildlife. We developed a mechanistic mathematical model of the hypothalamic-pituitary-gonadal (HPG) axis in female fathead minnows to predict dose-response and time-course (DRTC)...

Adaptive Response in Female Modeling of the Hypothalamic-pituitary-gonadal Axis

Science.gov (United States)

Exposure to endocrine disrupting chemicals can affect reproduction and development in both humans and wildlife. We are developing a mechanistic computational model of the hypothalamic-pituitary-gonadal (HPG) axis in female fathead minnows to predict dose-response and time-course ...

Proton-induced accelerated decay of the fungicide, vinclozolin, on TiO2 surface under solar irradiation: Experimental and DFT study.

Science.gov (United States)

Osipov, Ivan; Gorbachev, Mikhail Y; Gorinchoy, Natalia N

2018-04-03

The photochemical degradation of vinclozolin by addition of titanium dioxide on silica support has been examined both experimental and quantum-chemically. Solar irradiation of vinclozolin on silica with and without addition of titanium dioxide for 6 h resulted in 21% and 97.8% vinclozolin residues, respectively. In both these cases, phototransformation leads to the formation of (3,5-dichlorophenyl isocyanate) and (3,5-dichloroaniline). The presence of the intermediary product resulted from opening of the 2,4-oxazolidine-dione ring is also confirmed by GS-MS and LC-MS chromatography. The proton-induced mechanism of vinclozolin decay at the above experimental conditions is clarified on the base of DFT calculations.

SORPTION OF VINCLOZOLIN AND ATRAZINE ON FOUR GEOSORBENTS

Science.gov (United States)

The objectives of this study were to evaluate the magnitude and kinetics of vinclozolin and atrazine sorption on one surface soil and three freshwater sediments using batch and column techniques. Data from miscible displacement column studies were analyzed using a two-domain, fir...

Sexually Dimorphic Effects of Ancestral Exposure to Vinclozolin on Stress Reactivity in Rats

OpenAIRE

Gillette, Ross; Miller-Crews, Isaac; Nilsson, Eric E.; Skinner, Michael K.; Gore, Andrea C.; Crews, David

2014-01-01

How an individual responds to the environment depends upon both personal life history as well as inherited genetic and epigenetic factors from ancestors. Using a 2-hit, 3 generations apart model, we tested how F3 descendants of rats given in utero exposure to the environmental endocrine-disrupting chemical (EDC) vinclozolin reacted to stress during adolescence in their own lives, focusing on sexually dimorphic phenotypic outcomes. In adulthood, male and female F3 vinclozolin- or vehicle-linea...

METABOLISM AND DOSIMETRY OF VINCLOZOLIN IN RAT

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Vinclozolin (V) is an agricultural fungicide. V administered to rats is hydrolyzed to 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3-butenoic acid (M1) and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2). V, M1and M2 are antiandrogenic by interacting with the androgen r...

Impact of three endocrine disruptors, Bisphenol A, Genistein and Vinclozolin on female rat enamel.

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Jedeon, K; Berdal, A; Babajko, A

2016-06-28

Concerns about the potential adverse effectsof endocrine disruptors (EDs) have been increasingover the last three decades. BisphenolA (BPA), genistein (G) and vinclozolin (V) arethree widely used EDs sharing similar effects.Since populations are exposed to many diverseEDs simultaneously, we demonstratedrecently their impact alone or combined onmale rat tooth enamel. The purpose of thisstudy was therefore to assess their effects onfemale rat tooth enamel in order to understandwhy they are differentially sensitive. Ratswere exposed daily in utero and after birth tolow doses of EDs during the critical fetal andsuckling periods when amelogenesis takesplace. Enamel of rats exposed to EDs presentedopaque areas of hypomineralization. Theproportion of affected rats was the highestin the groups of rats treated with BPA aloneand higher in males than in females (in all thegroups). Comparison of enamel key gene expressionlevels showed modulations of Klk4and Enamelin in males but no significant variationsin females. These findings show thatfemale rats are less affected than males bythe three EDs chosen in this study and suggestthat enamel hypomineralization may differbetween males and females.

DEGRADATION AND MIGRATION OF VINCLOZOLIN IN SAND AND SOIL

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The migration of the dicarboximide fungicide vinclozolin and its principal degradation products through porous media was experimentally determined by simulating pesticide applications to a 23-30 mesh Ottawa sand and a North Carolina Piedmont, aquic hapludult soil in laboratory ...

Antagonistic effects of gestational dietary exposure to low-dose vinclozolin and genistein on rat fetal germ cell development.

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Lehraiki, Abdelali; Messiaen, Sébastien; Berges, Raymond; Canivenc-Lavier, Marie-Chantal; Auger, Jacques; Habert, René; Levacher, Christine

2011-05-01

Continuous, low-dose exposure to a phytoestrogen (1 mg/kg/day genistein) and/or to an antiandrogenic food contaminant (1 mg/kg/day vinclozolin) has been recently reported to affect male reproductive tract and fertility [1] in adults. We investigated whether alterations of the testis are already present at the end of in utero exposure using the same rat model and doses following exposure from conception to delivery. After vinclozolin exposure, we observed in the neonate a slight but significant alteration of steroidogenesis and gametogenesis with a reduction of testosterone secretion and of the number of gonocytes. In contrast, genistein exposure had no effect. While the vinclozolin-genistein mixture acts in a synergistic manner to induce the most significant alterations in the adult, interestingly, genistein antagonized the deleterious effect of vinclozolin on germ cells in the neonate. This difference emphasizes the importance of studying the effects of endocrine disruptors during various developmental stages to understand their effects. Copyright © 2011 Elsevier Inc. All rights reserved.

The impact of long term exposure to phthalic acid esters on reproduction in Chinese rare minnow (Gobiocypris rarus)

International Nuclear Information System (INIS)

Guo, Yongyong; Yang, Yuanjin; Gao, Yong; Wang, Xianfeng; Zhou, Bingsheng

2015-01-01

The environmental risk of phthalic acid esters (PAEs) is of great concern. We investigated the reproductive impairment of di-(2-ethylhexyl)-phthalate (DEHP) on Chinese rare minnow, an endemic fish inhabiting the upper streams of the Yangtze River. Chinese rare minnow larvae were exposed to environmentally relevant concentrations of DEHP (0, 4.2, 13.3, and 40.8 μg/L) for 6 months. Plasma testosterone and 17β-estradiol levels decreased in females, accompanied by downregulation of cyp19a and cyp17 gene transcription in ovary. Increases in plasma testosterone concentration were observed in males, accompanied by downregulation of cyp19a gene transcription in testes. Hepatic VTG gene transcription was upregulated in males and females. Exposure to DEHP reduced egg production and inhibited oocyte maturation in females and retarded spermiation in males. Decreased egg protein content was measured in F1 embryos. These results indicate that long-term exposure to low concentrations of DEHP (13.3 μg/L) causes endocrine disruption and impairs fish reproduction. - Highlights: • PAEs in the aquatic environment may pose risk to endemic fish species. • Long-term exposure to DEHP affected sex hormone levels in rare minnow. • DEHP affected gonad development. • Long-term exposure caused reduction of fecundity, but not sex ratio. • Environmentally relevant concentrations of DEHP impair fish reproduction. - Long-term exposure to low concentrations of DEHP adversely impact Chinese rare minnow reproduction

Transcripts of genes encoding reproductive neuroendocrine hormones and androgen receptor in the brain and testis of goldfish exposed to vinclozolin, flutamide, testosterone, and their combinations.

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Golshan, Mahdi; Habibi, Hamid R; Alavi, Sayyed Mohammad Hadi

2016-08-01

Vinclozolin (VZ) is a pesticide that acts as an anti-androgen to impair reproduction in mammals. However, VZ-induced disruption of reproduction is largely unknown in fish. In the present study, we have established a combination exposure in which adult goldfish were exposed to VZ (30 and 100 μg/L), anti-androgen flutamide (Flu, 300 μg/L), and androgen testosterone (T, 1 μg/L) to better understand effects of VZ on reproductive endocrine system. mRNA levels of kisspeptin (kiss-1 and kiss-2) and its receptor (gpr54), salmon gonadotropin-releasing hormone (gnrh3) and androgen receptor (ar) in the mid-brain, and luteinizing hormone receptor (lhr) in the testis were analyzed and compared with those of control following 10 days of exposure. kiss-1 mRNA level was increased in goldfish exposed to 100 µg/L VZ and to Flu, while kiss-2 mRNA level was increased following exposure to Flu and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. gpr54 mRNA level was increased in goldfish exposed to Flu and to combination of 30 µg/L VZ with Flu and 100 µg/L VZ with T. gnrh3 mRNA level was increased in goldfish exposed to 100 µg/L VZ, to Flu, and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. The mid-brain ar mRNA level was increased in goldfish exposed to Flu and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. Testicular lhr mRNA level was increased in goldfish exposed to Flu and to combination of 30 µg/L VZ with Flu. These results suggest that VZ and Flu are capable of interfering with kisspeptin and GnRH systems to alter pituitary and testicular horonal functions in adult goldfish and the brain ar mediates VZ-induced disruption of androgen production.

Simultaneous determination of vinclozolin and detection of its degradation products in mouse plasma, serum and urine, and from rabbit bile, by high-performance liquid chromatography.

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Dhananjeyan, Mugunthu R; Erhardt, Paul W; Corbitt, Cynthia

2006-05-19

A specific high-performance liquid chromatography method has been developed for simultaneous detection of vinclozolin and its degradation products (M1, M2, and M3). The method has been validated according to ICH guidelines and can be extended to quantitation of vinclozolin. A base-line separation of vinclozolin and its degradation products was found with symmetrical peak shapes on an XTerra MS C18 column using 10 mM ammonium bicarbonate at pH 9.2 and acetonitrile as mobile phase. The retention times of vinclozolin, M1, M2, and M3 were 12.8, 8.1, 11.6, and 11.1 min, respectively. A linear calibration curve was obtained across a range from 5 to 200 microM for vinclozolin. The intra- and inter-day relative standard deviations (%RSD) were vinclozolin from bio-fluids including mouse plasma, serum and urine, and rabbit bile, were obtained in a single step with a single solvent.

Field and laboratory tests on acute toxicity of cadmium to freshwater crayfish

Energy Technology Data Exchange (ETDEWEB)

1986-09-01

Environmental regulatory standards for cadmium (EPA 1980), like those for most pollutants, are based on acute, laboratory toxicity tests of single species. Such tests can be conducted rapidly and inexpensively in comparison to acute or chronic field studies, but their validity has often been questioned. Laboratory-based criteria are subject to two criticisms: (1) chemical and physical conditions differ greatly in degree and variability from laboratory to field, and (2) species are not isolated, but live in an ecosystem of interacting taxa and biofeedback. To investigate the validity of basing field toxicity standards on laboratory data, the authors subjected the freshwater crayfish Orconectes immunis for 96 h to various levels of cadmium in laboratory aquaria and experimental ponds. The study was designed to evaluate in part the first criticism of lab-based criteria. The studies were conducted concurrently with similar short-term experiments on the fathead minnow, Pimephales promelas, and coincided with studies of chronic cadmium stress on fathead minnows in experimental ponds.

TRANSFORMATION AND TRANSPORT OF VINCLOZOLIN FROM SOIL TO AIR

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A laboratory chamber was designed and used to determine the headspace flux of the fungicide vinclozolin (3-(3,5-dichlorophenyl)-5-methyl-5-vinyl-oxzoli-dine-2.4-dione) and its three degradation products from chamber surfaces, 20-30 mesh Ottawa sand, and sterilized and nonsteril...

The combined effects of vinclozolin and procymidone do not deviate from expected additivity in vitro and in vivo.

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Nellemann, Christine; Dalgaard, Majken; Lam, Henrik Rye; Vinggaard, Anne Marie

2003-02-01

The combination effects of the well-known antiandrogenic fungicides, vinclozolin and procymidone, were tested both in vitro and in vivo. In vitro both vinclozolin and procymidone significantly inhibited the binding of agonist to the androgen receptor with the concentration that resulted in 50% inhibition (IC(50)) values of 0.1 and 0.6 micro M, respectively. By applying the isobole method, the effect of combining the two pesticides in vitro was found to be additive. In castrated testosterone-treated rats the administration of vinclozolin starting at 10 mg/kg led to a decrease in organ weight of all tested reproductive organs. The levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH) were increased significantly with doses of 100 mg/kg vinclozolin and above. Expression of the androgen-responsive gene, TRPM-2, was increased starting at 100 mg/kg vinclozolin. For procymidone, reproductive organ weights were diminished at 10 mg/kg and LH was increased at a concentration of 25 mg/kg and above, compared to the testosterone-treated controls. FSH was significantly increased only at 25 mg/kg procymidone. The studied gene expressions were changed by 100 mg/kg procymidone. Dosing the animals with a combination of a 1:1 mixture of vinclozolin and procymidone resulted in a weight reduction in the reproductive organs and an increase of serum LH and FSH as early as with 10 mg/kg combined dose. The relative expressions of TRPM-2 and PBP C3 were changed compared to controls at 100 mg/kg. The level of 5-HT in the rat brain was increased after a dose of 10 mg/kg. Using the isobole method, comparisons of the observed and predicted effects assuming additivity on reproductive organ weights, hormone levels, and gene expression showed agreement and thus the combination effects are suggested to be additive in vivo as well as in vitro.

Epigenetic transgenerational actions of vinclozolin on promoter regions of the sperm epigenome.

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Carlos Guerrero-Bosagna

2010-09-01

Full Text Available Previous observations have demonstrated that embryonic exposure to the endocrine disruptor vinclozolin during gonadal sex determination promotes transgenerational adult onset disease such as male infertility, kidney disease, prostate disease, immune abnormalities and tumor development. The current study investigates genome-wide promoter DNA methylation alterations in the sperm of F3 generation rats whose F0 generation mother was exposed to vinclozolin. A methylated DNA immunoprecipitation with methyl-cytosine antibody followed by a promoter tilling microarray (MeDIP-Chip procedure was used to identify 52 different regions with statistically significant altered methylation in the sperm promoter epigenome. Mass spectrometry bisulfite analysis was used to map the CpG DNA methylation and 16 differential DNA methylation regions were confirmed, while the remainder could not be analyzed due to bisulfite technical limitations. Analysis of these validated regions identified a consensus DNA sequence (motif that associated with 75% of the promoters. Interestingly, only 16.8% of a random set of 125 promoters contained this motif. One candidate promoter (Fam111a was found to be due to a copy number variation (CNV and not a methylation change, suggesting initial alterations in the germline epigenome may promote genetic abnormalities such as induced CNV in later generations. This study identifies differential DNA methylation sites in promoter regions three generations after the initial exposure and identifies common genome features present in these regions. In addition to primary epimutations, a potential indirect genetic abnormality was identified, and both are postulated to be involved in the epigenetic transgenerational inheritance observed. This study confirms that an environmental agent has the ability to induce epigenetic transgenerational changes in the sperm epigenome.

Epigenetic transgenerational actions of vinclozolin on promoter regions of the sperm epigenome.

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Guerrero-Bosagna, Carlos; Settles, Matthew; Lucker, Ben; Skinner, Michael K

2010-09-30

Previous observations have demonstrated that embryonic exposure to the endocrine disruptor vinclozolin during gonadal sex determination promotes transgenerational adult onset disease such as male infertility, kidney disease, prostate disease, immune abnormalities and tumor development. The current study investigates genome-wide promoter DNA methylation alterations in the sperm of F3 generation rats whose F0 generation mother was exposed to vinclozolin. A methylated DNA immunoprecipitation with methyl-cytosine antibody followed by a promoter tilling microarray (MeDIP-Chip) procedure was used to identify 52 different regions with statistically significant altered methylation in the sperm promoter epigenome. Mass spectrometry bisulfite analysis was used to map the CpG DNA methylation and 16 differential DNA methylation regions were confirmed, while the remainder could not be analyzed due to bisulfite technical limitations. Analysis of these validated regions identified a consensus DNA sequence (motif) that associated with 75% of the promoters. Interestingly, only 16.8% of a random set of 125 promoters contained this motif. One candidate promoter (Fam111a) was found to be due to a copy number variation (CNV) and not a methylation change, suggesting initial alterations in the germline epigenome may promote genetic abnormalities such as induced CNV in later generations. This study identifies differential DNA methylation sites in promoter regions three generations after the initial exposure and identifies common genome features present in these regions. In addition to primary epimutations, a potential indirect genetic abnormality was identified, and both are postulated to be involved in the epigenetic transgenerational inheritance observed. This study confirms that an environmental agent has the ability to induce epigenetic transgenerational changes in the sperm epigenome.

Biotransformation of vinclozolin in rat precision-cut liver slices: comparison with in vivo metabolic pattern.

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Bursztyka, Julian; Debrauwer, Laurent; Perdu, Elisabeth; Jouanin, Isabelle; Jaeg, Jean-Philippe; Cravedi, Jean-Pierre

2008-06-25

Vinclozolin is a dicarboxymide fungicide that presents antiandrogenic properties through its two hydrolysis products M1 and M2, which bind to the androgen receptor. Because of the lack of data on the biotransformation of vinclozolin, its metabolism was investigated in vitro in precision-cut rat liver slices and in vivo in male rat using [ (14)C]-vinclozolin. Incubations were performed using different concentrations of substrate, and the kinetics of formation of the major metabolites were studied. Three male Wistar rats were fed by gavage with [ (14)C]-VZ. Urine was collected for 24 h and analyzed by radio-HPLC for metabolic profiling. Metabolite identification was carried out on a LCQ ion trap mass spectrometer. In rat liver slices and in vivo, the major primary metabolite has been identified as 3',5'-dichloro-2,3,4-trihydroxy-2-methylbutyranilide (M5) and was mainly present as glucuronoconjugates. M5 is produced by dihydroxylation of the vinyl group of M2. Other metabolites have been identified as 3-(3,5-dichlorophenyl)-5-methyl-5-(1,2-dihydroxyethyl)-1,3-oxazolidine-2,4-dione (M4), a dihydroxylated metabolite of vinclozolin, which undergoes further conjugation to glucuronic acid, and 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3,4-dihydroxy-butanoic acid (M6), a dihydroxylated metabolite of M1.

Quantitative (q)AOP for aromatase inhibition as case study to advance qAOP development practices

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Here we describe how “read across” of a quantitative adverse outcome pathway (qAOP) developed with data for one chemical can be used to screen impacts of other chemicals. We developed a qAOP starting with inhibition of CYP19A (aromatase) in fathead minnows (FHM) as th...

Verification of responses of Japanese medaka (Oryzias latipes) to anti-androgens, vinclozolin and flutamide, in short-term assays.

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Nakamura, Ataru; Takanobu, Hitomi; Tamura, Ikumi; Yamamuro, Masumi; Iguchi, Taisen; Tatarazako, Norihisa

2014-05-01

Various testing methods for the detection of the endocrine disruptive activities of chemicals have been developed in freshwater fish species. However, a few relatively easier specific methods for detecting anti-androgenic activities are available for fish. The aim of this study was to verify the papillary process in Japanese medaka (Oryzias latipes) as an indicator of the anti-androgenic activity of chemicals. Japanese medaka were exposed to two types of anti-androgenic compounds, vinclozolin and flutamide, using two short-term assays; one was conformed to the existing short-term reproduction assay using adult fish (adult test) and the other was a test based on the same methods but using juvenile fish at the beginning of exposure (juvenile test). Significant decreases in male papillary processes were observed in the juvenile test treated with the highest concentration of both antiandrogens (640 µg l(-1) vinclozolin and 1000 µg l(-1) flutamide); however, no significant effects were observed in the adult test. Consequently, our results indicate that papillary processes in Japanese medaka can be used as the end-point for screening the anti-androgenic activity of chemicals using juvenile fish for a specific period based on the existing short-term reproduction assay. Copyright © 2013 John Wiley & Sons, Ltd.

Anti-androgen vinclozolin impairs sperm quality and steroidogenesis in goldfish.

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Hatef, Azadeh; Alavi, Sayyed Mohammad Hadi; Milla, Sylvain; Křišťan, Jiří; Golshan, Mahdi; Fontaine, Pascal; Linhart, Otomar

2012-10-15

In mammals, vinclozolin (VZ) is known as anti-androgen, which causes male infertility via androgen receptor (AR) antagonism. In aquatic animals, the VZ effects on reproductive functions are largely unknown and results are somewhat contradictory. To understand VZ adverse effects on male reproduction, mature goldfish (Carassius auratus) were exposed to three nominal VZ concentrations (100, 400, and 800 μg/L) and alternations in gonadosomatic (GSI) and hepatosomatic indices (HSI), 17β-estradiol (E(2)), 11-ketotestosterone (11-KT) and sperm quality were investigated compared to the solvent control. One group was exposed to E(2) (nominal concentration of 5 μg/L), an estrogenic compound, as a negative control. Following one month exposure, GSI and HSI were unchanged in all VZ treated groups compared to solvent control. Sperm volume, motility and velocity were reduced in fish exposed to 800 μg/L VZ. This was associated with the decrease in 11-KT level, suggesting direct VZ effects on testicular androgenesis and sperm functions. In goldfish exposed to 100 μg/L VZ, 11-KT was increased but E(2) remained unchanged. This is, probably, the main reason for unchanged sperm quality at 100 μg/L VZ. In goldfish exposed to E(2), GSI and 11-KT were decreased, E(2) was increased and no sperm was produced. The present study shows different dose-dependent VZ effects, which lead to impairment in sperm quality via disruption in steroidogenesis. In addition to VZ effects through competitive binding to AR, our data suggests potential effects of VZ by direct inhibition of 11-KT biosynthesis in fish as well as abnormalities in sperm morphology. Copyright © 2012 Elsevier B.V. All rights reserved.

Ra-226 bioaccumulation and growth indices in fish.

Science.gov (United States)

Shi, Xiaopei; Smith, Richard; Seymour, Colin; Mothersill, Carmel

2017-06-01

To determine the accumulated activity of Ra-226 in fathead minnows fed with environmentally relevant levels of Ra-226 for 5 months in water at 20 °C, and to evaluate the influence of this level of Ra-226 on the growth of fathead minnows. Fathead minnows were fed with fish food containing 10-10,000 mBq/g Ra-226 for 5 months. At the end of the experiment, the fish were sacrificed, flash frozen in liquid nitrogen and kept at -20 °C. Longitudinal sections of 40 μm thickness were cut at the middle of the fish body using a cryostat. The activity of Ra-226 in each section was determined using autoradiography with a nuclear track detector CR-39. According to the weight and the width of the fish, the activity of Ra-226 in the whole fish body could be estimated. In addition, the length and the weight of the fish were measured and the condition factor was calculated to evaluate the growth and fitness of the fish. There is a positive but non-linear relationship between the accumulated activity of Ra-226 in fish body and the concentration of Ra-226 in fish food. The highest activity of Ra-226 accumulated in fish body was found from fish fed with 10,000 mBq/g Ra-226 food. This was calculated as 256.4 ± 49.1 mBq/g, p fish fed with food containing lower concentration of Ra-226 (up to 1000 mBq/g), the bioaccumulation of Ra-226 in the body saturated. The Ra-226 concentration factor (CF) for fish was inversely proportional to the Ra-226 activity in food, and the highest CF value was 2.489, obtained from the lowest dietary Ra-226 activity (10 mBq/g). In addition, condition factors (K) of fish in all Ra-226-treated groups were significantly lower than those of the controls. The results show that the bioaccumulation of Ra-226 in fish is not simply related to the dietary Ra-226 activity, and has a saturation value when the dietary activity is low. In addition, the environmental level of Ra-226 in the fish food has a small adverse effect on the growth and fitness of fathead

EFFECT OF THE ANTI-ANDROGENIC ENDOCRINE DISRUPTOR VINCLOZOLIN ON EMBRYONIC TESTIS CORD FORMATION AND POSTNATAL TESTIS DEVELOPMENT AND FUNCTION. (R827405)

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Vinclozolin is a systemic dicarboximide fungicide that is used on fruits, vegetables, ornamental plants, and turf grass. Vinclozolin and its metabolites are known to be endocrine disruptors and act as androgen receptor antagonists. The hypothesis tested in the current study is...

Is the Factor-of-2 Rule Broadly Applicable for Evaluating the Prediction Accuracy of Metal-Toxicity Models?

Science.gov (United States)

Meyer, Joseph S; Traudt, Elizabeth M; Ranville, James F

2018-01-01

In aquatic toxicology, a toxicity-prediction model is generally deemed acceptable if its predicted median lethal concentrations (LC50 values) or median effect concentrations (EC50 values) are within a factor of 2 of their paired, observed LC50 or EC50 values. However, that rule of thumb is based on results from only two studies: multiple LC50 values for the fathead minnow (Pimephales promelas) exposed to Cu in one type of exposure water, and multiple EC50 values for Daphnia magna exposed to Zn in another type of exposure water. We tested whether the factor-of-2 rule of thumb also is supported in a different dataset in which D. magna were exposed separately to Cd, Cu, Ni, or Zn. Overall, the factor-of-2 rule of thumb appeared to be a good guide to evaluating the acceptability of a toxicity model's underprediction or overprediction of observed LC50 or EC50 values in these acute toxicity tests.

Development of a bioassay using walleye (Sander vitreus) to assess the toxicity of oil sands sediments

Energy Technology Data Exchange (ETDEWEB)

Turcotte, D.; Yuan, H.; Tumber, V.; Parrott, J. [Environment Canada, Ottawa, ON (Canada); Raine, J. [Saskatchewan Univ., Saskatoon, SK (Canada)

2010-07-01

This study examined the effects of sediments from the Athabasca oil sands area on fish development and survival. Walleye (Sander vitreus) which inhabit the Athabasca River are exposed to natural sources of bitumen eroding from the McMurray formation. This study described the design and implementation of a daily-renewal bioassay to evaluate the potential effects of toxicants on walleye development. Eggs were collected and fertilized with milt from spawning wild walleye captured from Lake Diefenbaker in Saskatchewan. The fertilized eggs were exposed to different concentrations of sediments or culture water only (negative controls) until complete yolk absorption of control fish. The walleye embryos were fed brine shrimp daily after hatching and the developing fish were examined for morphological deformities, survival, hatching success, and changes in weight and length between treatments. Organics concentrations in fish tissues and water were measured when possible. Fathead minnows and northern pikes will also be exposed to the same sediments in order to compare the relative sensitivity of the three species.

Toxicogenomic analysis of the hepatic effects of perfluorooctanoic acid on rare minnows (Gobiocypris rarus)

International Nuclear Information System (INIS)

Wei Yanhong; Liu Yang; Wang Jianshe; Tao Yi; Dai Jiayin

2008-01-01

Perfluorooctanoic acid (PFOA) is a ubiquitous environmental contaminant that has been detected in a variety of terrestrial and aquatic organisms. To assess the effects of PFOA in fish and predict its potential mode of action, a toxicogenomic approach was applied to hepatic gene expression profile analysis in male and female rare minnows (Gobiocypris rarus) using a custom cDNA microarray containing 1773 unique genes. Rare minnows were treated with continuous flow-through exposure to PFOA at concentrations of 3, 10, and 30 mg/L for 28 days. Based on the observed histopathological changes, the livers from fish exposed to 10 mg/L PFOA were selected for further hepatic gene expression analysis. While 124 and 171 genes were significantly altered by PFOA in males and females, respectively, of which 43 genes were commonly regulated in both sexes. The affected genes are involved in multiple biological processes, including lipid metabolism and transport, hormone action, immune responses, and mitochondrial functions. PFOA exposure significantly suppressed genes involved in fatty acid biosynthesis and transport but induced genes associated with intracellular trafficking of cholesterol. Alterations in expression of genes associated with mitochondrial fatty acid β-oxidation were only observed in female rare minnows. In addition, PFOA inhibited genes responsible for thyroid hormone biosynthesis and significantly induced estrogen-responsive genes. These findings implicate PFOA in endocrine disruption. This work contributes not only to the elucidation of the potential mode of toxicity of PFOA to aquatic organisms but also to the use of toxicogenomic approaches to address issues in environmental toxicology

Vinclozolin--the lack of a transgenerational effect after oral maternal exposure during organogenesis.

Science.gov (United States)

Schneider, Steffen; Kaufmann, Wolfgang; Buesen, Roland; van Ravenzwaay, Bennard

2008-04-01

The purpose of the study was to investigate a possible transgenerational effect of the fungicide vinclozolin on the male reproductive system following oral exposure since this effect was reported by Anway et al. [Anway MD, Cupp AS, Uzumcu M, Skinner MK. Epigenetic transgenerational actions of endocrine disruptors and male fertility. Science 2005;308(5727 (June 3)):1466-9] after intraperitoneal administration. Pregnant Wistar rats were dosed by oral gavage with vinclozolin 0, 4 or 100mg/(kg bw day) on days 6-15 post coitum (p.c.). F1 male offspring was mated with untreated females to produce F2, which were then similarly mated to produce F3 offspring. F0 maternal treatment had no effect on mating and fertility indices or male offspring sexual development, mean sperm parameters, or histopathology of the sexual organs in F1, F2 or F3 males (at age 127-134 days). Apoptotic germ cell counts were statistically significantly lower in F1, F2 and F3 generations, however, control values showed a pronounced variance over time. Also, as anti-androgenic compounds are more likely to induce the opposite effect (increased apoptosis), this observation is not considered to be treatment related. Consequently, spermatogenesis was not affected by vinclozolin exposure in utero. As vinclozolin has been shown to induce clear anti-androgenic effects in offspring following treatment with 100mg/(kg bw day) during entire gestation, the lack of effects in this study indicates that the window of sensitivity for anti-androgenic effects is from days 16-20 p.c. No transgenerational effect on the male reproductive system was found. The NOAEL was >100mg/(kg bw day) for fertility and reproductive performance, for systemic parental and developmental toxicity in F1, F2 and F3 males.

Bisphenol A, Dichlorodiphenyltrichloroethane (DDT) and Vinclozolin Affect ex-vivo Uterine Contraction in Rats via Uterotonin (Prostaglandin F2α, Acetylcholine and Oxytocin) Related Pathways.

Science.gov (United States)

Salleh, Naguib; Giribabu, Nelli; Feng, Angeline Oh Mei; Myint, Kyaimon

2015-01-01

Bisphenol-A (BPA), dichrolodiphenyltrichloroethane (DDT) and vinclozolin were found able to induce abnormal uterine contraction. The mechanisms involved remains unclear. We hypothesized that the effect of these compounds were mediated via the uterotonin pathways. Therefore, in this study, effects of BPA, vinclozolin and DDT-only and in combination with uterotonins (PGF-2α, acetylcholine and oxytocin) on the force and pattern of uterine contraction were observed. Uteri were harvested from intact adult female rats 24 hours after a single injection (1 mg/kg/b.w) of estrogen to synchronize their oestrous cycle. The uterine horns were subjected for ex-vivo contraction studies in an organ bath connected to Powerlab data acquisition system. Different doses of BPA, vinclozolin and DDT were added into the bathing solution and changes in the pattern and strength of uterine contraction were recorded. Further, increasing doses of uterotonins were concomitantly administered with these compounds and changes in the force and pattern of contraction were observed. In the absence of uterotonins, uterine contractile force decreased with increasing doses of BPA and DDT. However, vinclozolin induced sharp increase in the contractile forces which then gradually decrease. Administration of BPA, DDT and vinclozolin alone reduced the force of uterine contraction following stimulation of contraction by uterotonins. However, BPA, vinclozolin or DDT effects were relieved upon co-administration with uterotonins at increasing doses. The antagonizing effect of uterotonins on BPA, vinclozolin and DDT actions could explain the mechanism underlying the adverse effect of these compounds on uterine contraction.

Additive and synergistic antiandrogenic activities of mixtures of azol fungicides and vinclozolin

Energy Technology Data Exchange (ETDEWEB)

Christen, Verena [University of Applied Sciences and Arts Northwestern Switzerland, School of Life Sciences, Gründenstrasse 40, CH-4132 Muttenz (Switzerland); Crettaz, Pierre [Federal Office of Public Health, Division Chemical Products, 3003 Bern (Switzerland); Fent, Karl, E-mail: karl.fent@fhnw.ch [University of Applied Sciences and Arts Northwestern Switzerland, School of Life Sciences, Gründenstrasse 40, CH-4132 Muttenz (Switzerland); ETH Zürich, Department of Environmental System Sciences, Institute of Biogeochemistry and Pollution Dynamics, Universitätsstrasse 16, CH-8092 Zürich (Switzerland)

2014-09-15

Objective: Many pesticides including pyrethroids and azole fungicides are suspected to have an endocrine disrupting property. At present, the joint activity of compound mixtures is only marginally known. Here we tested the hypothesis that the antiandrogenic activity of mixtures of azole fungicides can be predicted by the concentration addition (CA) model. Methods: The antiandrogenic activity was assessed in MDA-kb2 cells. Following assessing single compounds activities mixtures of azole fungicides and vinclozolin were investigated. Interactions were analyzed by direct comparison between experimental and estimated dose–response curves assuming CA, followed by an analysis by the isobole method and the toxic unit approach. Results: The antiandrogenic activity of pyrethroids deltamethrin, cypermethrin, fenvalerate and permethrin was weak, while the azole fungicides tebuconazole, propiconazole, epoxiconazole, econazole and vinclozolin exhibited strong antiandrogenic activity. Ten binary and one ternary mixture combinations of five antiandrogenic fungicides were assessed at equi-effective concentrations of EC{sub 25} and EC{sub 50}. Isoboles indicated that about 50% of the binary mixtures were additive and 50% synergistic. Synergism was even more frequently indicated by the toxic unit approach. Conclusion: Our data lead to the conclusion that interactions in mixtures follow the CA model. However, a surprisingly high percentage of synergistic interactions occurred. Therefore, the mixture activity of antiandrogenic azole fungicides is at least additive. Practice: Mixtures should also be considered for additive antiandrogenic activity in hazard and risk assessment. Implications: Our evaluation provides an appropriate “proof of concept”, but whether it equally translates to in vivo effects should further be investigated. - Highlights: • Humans are exposed to pesticide mixtures such as pyrethroids and azole fungicides. • We assessed the antiandrogenicity of

Additive and synergistic antiandrogenic activities of mixtures of azol fungicides and vinclozolin

International Nuclear Information System (INIS)

Christen, Verena; Crettaz, Pierre; Fent, Karl

2014-01-01

Objective: Many pesticides including pyrethroids and azole fungicides are suspected to have an endocrine disrupting property. At present, the joint activity of compound mixtures is only marginally known. Here we tested the hypothesis that the antiandrogenic activity of mixtures of azole fungicides can be predicted by the concentration addition (CA) model. Methods: The antiandrogenic activity was assessed in MDA-kb2 cells. Following assessing single compounds activities mixtures of azole fungicides and vinclozolin were investigated. Interactions were analyzed by direct comparison between experimental and estimated dose–response curves assuming CA, followed by an analysis by the isobole method and the toxic unit approach. Results: The antiandrogenic activity of pyrethroids deltamethrin, cypermethrin, fenvalerate and permethrin was weak, while the azole fungicides tebuconazole, propiconazole, epoxiconazole, econazole and vinclozolin exhibited strong antiandrogenic activity. Ten binary and one ternary mixture combinations of five antiandrogenic fungicides were assessed at equi-effective concentrations of EC 25 and EC 50 . Isoboles indicated that about 50% of the binary mixtures were additive and 50% synergistic. Synergism was even more frequently indicated by the toxic unit approach. Conclusion: Our data lead to the conclusion that interactions in mixtures follow the CA model. However, a surprisingly high percentage of synergistic interactions occurred. Therefore, the mixture activity of antiandrogenic azole fungicides is at least additive. Practice: Mixtures should also be considered for additive antiandrogenic activity in hazard and risk assessment. Implications: Our evaluation provides an appropriate “proof of concept”, but whether it equally translates to in vivo effects should further be investigated. - Highlights: • Humans are exposed to pesticide mixtures such as pyrethroids and azole fungicides. • We assessed the antiandrogenicity of pyrethroids and

Prenatal exposure to vinclozolin disrupts selective aspects of the gonadotropin-releasing hormone neuronal system of the rabbit

OpenAIRE

Wadas, B.C.; Hartshorn, C.A.; Aurand, E.R.; Palmer, J.S.; Roselli, C.E.; Noel, M.L.; Gore, A.C.; Veeramachaneni, D.N.R.; Tobet, S.A.

2010-01-01

Developmental exposure to the agricultural fungicide vinclozolin can impair reproductive function in male rabbits and was previously found to decrease the number of immunoreactive-gonadotropin-releasing hormone (ir-GnRH) neurons in the region of the organum vasculosum of the lamina terminalis (OVLT) and rostral preoptic area (rPOA) by postnatal week (PNW) 6. To further examine the disruption of GnRH neurons by fetal vinclozolin exposure, in the current study, pregnant rabbits were dosed orall...

Repeated 28-day oral toxicity study of vinclozolin in rats based on the draft protocol for the "Enhanced OECD Test Guideline No. 407" to detect endocrine effects.

Science.gov (United States)

Shin, Jae-Ho; Moon, Hyun Ju; Kim, Tae Sung; Kang, Il Hyun; Ki, Ho Yeon; Choi, Kwang Sik; Han, Soon Young

2006-09-01

We performed a 28-day repeated-dose toxicity study of vinclozolin, a widely used fungicide, based on the draft protocol of the "Enhanced OECD Test Guideline 407" (Enhanced TG407) to investigate whether vinclozolin has endocrine-mediated properties according to this assay. Seven-week-old SD rats were administered with vinclozolin daily by oral gavage at dose rates of 0, 3.125, 12.5, 50 and 200 mg/kg/day for at least 28 days. The vinclozolin-treated male rats showed a reduction of epididymis and accessory sex organ weights and an alteration of hormonal patterns. A slight prolongation of the estrous cycle and changes in the estrogen/testosterone ratio and luteinizing hormone level were observed in vinclozolin-treated female rats. Thyroxin concentrations were decreased and thyroid-stimulating hormone concentrations were increased in both sexes; however, there were no compound-related microscopic lesions in the thyroid gland or changes in the thyroid weight. The endocrine-related effects of vinclozolin could be detected by the parameters examined in the present study based on the OECD protocol, suggesting the Enhanced TG407 protocol should be a suitable screening test for the detection of endocrine-mediated effects of chemicals.

Electrochemical Impedance Study of Reduction Kinetics of the Pesticide Vinclozoline

Czech Academy of Sciences Publication Activity Database

Pospíšil, Lubomír; Sokolová, Romana; Colombini, M. P.; Giannarelli, S.; Fuoco, R.

2000-01-01

Roč. 67, - (2000), s. 305-312 ISSN 0026-265X R&D Projects: GA MŠk OC D15.10; GA ČR GA203/97/1048 Institutional research plan: CEZ:AV0Z4040901 Keywords : electrochemical impedance * pesticide s * vinclozoline Subject RIV: CG - Electrochemistry Impact factor: 0.884, year: 2000

The effects of the environmental antiandrogen vinclozolin on the induction of granulosa cell apoptosis during follicular atresia in pigs.

Science.gov (United States)

Knet, Malgorzata; Tabarowski, Zbigniew; Slomczynska, Maria; Duda, Malgorzata

2014-06-01

The aim of this study was to investigate whether the androgens testosterone and dihydrotestosterone (DHT) and the antiandrogenic fungicide vinclozolin (Vnz) exert proapoptotic effects on porcine granulosa cells (GCs), and to examine the roles of these compounds in follicular atresia. Granulosa cells isolated from pig follicles were cultured for 24 hours, and then exposed to 0.1 μM testosterone, 0.1 μM DHT, 14 μM Vnz, or the equivalent concentrations of testosterone and Vnz or DHT and Vnz for a further 24 hours. Apoptosis and necrosis of the GCs were determined via Hoechst staining and flow cytometry analyses of annexin V-stained cells. Whole porcine follicles were also exposed to the same compounds and combinations of compounds for 24 hours. The sections were stained with hematoxylin and eosin for morphologic assessments, and a Terminal deoxynucleotidyl Transferase Biotyn-dUTP Nick-End Labeling (TUNEL) assay was performed to determine the number of apoptotic cells. The progesterone and estradiol concentrations secreted into the culture media by isolated GCs and follicles were also measured. Exposure to the androgens resulted in an increased number of apoptotic GCs both in vitro and in the organotypic model. Vinclozolin exposure increased and decreased the number of necrotic and apoptotic GCs, respectively. Furthermore, compared with control follicles, those exposed to testosterone, DHT, or Vnz displayed enhanced atresia, and coadministration of Vnz attenuated the promotive effect of these androgens on atresia. Estradiol secretion was stimulated by the combination of testosterone and Vnz, whereas exposure to Vnz alone reduced it. Progesterone production declined after the combined addition of androgens and the antiandrogen. In summary, Vnz caused massive necrosis of GCs in vitro and induced apoptosis of GCs in whole follicles. The androgens testosterone and DHT enhanced these effects. The results presented here suggest that selective destruction of porcine

Chronic effects of low levels of hydrogen sulfide on freshwater fish

Energy Technology Data Exchange (ETDEWEB)

Smith, L.L. Jr.; Oseid, D.M.

1975-01-01

The purpose of this study was to determine acute toxicity of H/sub 2/S to brook trout, bluegill, fathead minnow, goldfish and amphipod. The sensitivity of various life history stages are compared and acute toxicity is related to no-effect levels of H/sub 2/S based on long-term tests. 12 references, 7 tables.

Ecological diversification associated with the benthic-to-pelagic transition by North American minnows.

Science.gov (United States)

Burress, E D; Holcomb, J M; Tan, M; Armbruster, J W

2017-03-01

Ecological opportunity is often regarded as a key factor that explains why diversity is unevenly distributed across life. Colonization of novel environments or adaptive zones may promote diversification. North American minnows exhibit an ancestral benthic-to-pelagic habitat shift that coincided with a burst in diversification. Here, we evaluate the phenotypic and ecological implications of this habitat shift by assessing craniofacial and dietary traits among 34 species and testing for morphology-diet covariation, convergence and adaptive optima. There were several instances of morphology-diet covariation such as correlations between mouth angle and the consumption of terrestrial insects and between relative gut length and the consumption of algae. After accounting for size and phylogenetic nonindependence, benthic species had longer heads, longer snouts, eyes positioned higher on their head, smaller mouth angles and longer digestive tracts than pelagic minnows. Benthic minnows also consumed more algae but less terrestrial insects, by volume, than pelagic minnows. Lastly, there were three distinct evolutionary regimes and more convergence in morphology and dietary characteristics than expected under a Brownian motion model of evolution. These findings indicate that colonization of the pelagic zone by minnows involved myriad phenotypic and dietary changes associated with exploitation of terrestrial subsidies. Thus, minnows exhibit phenotype-dietary covariation, an expansion of ecological roles and a burst in diversification rates in response to the ecological opportunity afforded by the colonization of a novel habitat. © 2016 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2016 European Society For Evolutionary Biology.

Vinclozolin: a case study on the identification of endocrine active substances in the past and a future perspective.

Science.gov (United States)

van Ravenzwaay, Bennard; Kolle, Susanne N; Ramirez, Tzutzuy; Kamp, Hennicke G

2013-12-16

In the late 1980s vinclozolin was tested for prenatal developmental toxicity in rats for registration purposes in USA. At 1000mg/kgbw, 95% of all fetuses were female upon visual inspection (ano-genital distance determination). Anti-androgenic effects (AA) were also noted in a subsequent 2-generation study. These findings triggered mechanistic investigations at BASF and at US-EPA. Results published by the latter were the starting point of the endocrine disruption (ED) discussion in the 1990s. AA effects of vinclozolin are mediated by two metabolites, which have an antagonistic effect on the androgen receptor. Currently, determination of ED has become a major end-point in toxicology testing and the US-EPA has set up an elaborated testing paradigm to fulfill this requirement. Future screening for ED can be improved making use of new technologies. ED modes of action can be determined by three alternative (3R) methods. Steroid synthesis in H295R cells (1), androgen-receptor binding in modified yeast (2) and metabolomics (3). Using vinclozolin as a case study, results indicate: (1) an effect on steroid synthesis in vitro, (2) an antagonistic effect on the androgen receptor and (3) that the metabolome profile of vinclozolin is similar to that of other receptor mediated anti-androgens (e.g. flutamide). Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Impact of vinclozolin on reproductive behavior and endocrinology in Japanese quail (Coturnix coturnix japonica)

Science.gov (United States)

McGary, S.; Henry, P.F.P.; Ottinger, M.A.

2001-01-01

The impact of endocrine-disrupting chemicals (EDCs) has been demonstrated in mammalian models, but less research is available for avian species. The effects of vinclozolin (VIN), an antiandrogenic fungicide, on sexual differentiation and maturation were investigated in Japanese quail (Coturnix coturnix japonica). On day 4 of incubation, embryos were exposed to no treatment, oil, or 25, 50, or 100 ppm of VIN. Endpoints measured included adult male reproductive behavior, hypothalamic gonadotropin-releasing hormone I (GnRH-I) content in hatchlings and adults, plasma steroid levels in hatchlings and adults, proctodeal gland growth during maturation, and relative testicular weight at seven weeks of age. Results showed that exposure to VIN significantly (p < 0.05) altered GnRH-I in male hatchlings, whereas GnRH-I levels in females remained unaffected. Although steroid levels were unaltered by any VIN treatment, the display of male reproductive behavior seemed delayed, with the number of mounts and the number of cloacal contacts being significantly (p < 0.05) lower in the VIN-treated males. This could have an extreme negative impact on wild avian species that are routinely exposed to similar EDCs.

HYDROLOGIC CONDITIONS AFFECTING THE TROPOSPHERIC FLUX OF VINCLOZOLIN AND ITS DEGRADATION PRODUCTS

Science.gov (United States)

A laboratory chamber was used to determine hydrologic conditions that lead to the tropospheric flux of a suspected anti-androgenic dicarboximide fungicide, vinclozolin (3-(3,5-dichlorophenyl)-5-methyl-5-vinyl-oxzoli-dine-2,4-dione) and three degradation products from sterilized...

Developmental toxicity of endocrine disrupters bisphenol A and vinclozolin in a terrestrial isopod.

Science.gov (United States)

Lemos, M F L; van Gestel, C A M; Soares, A M V M

2010-08-01

Studies of the effects of endocrine-disrupting compounds (EDCs) on invertebrates are still largely underrepresented. This work aims to fill this gap by assessing the effects of bisphenol A (BPA) and vinclozolin (Vz) on the terrestrial isopod Porcellio scaber (common rough woodlouse). Male adult and sexually undifferentiated juvenile woodlice were exposed to the toxicants. Effects on molting regime and growth were investigated independently for males and female woodlice after sexual differentiation. Both chemicals elicited developmental toxicity to P. scaber by causing overall decreased growth. Nevertheless, BPA induced molting, whereas Vz delayed it. Although the LC50 values for juvenile and adult survival were fairly similar, juvenile woodlice showed an increased chronic sensitivity to both chemicals, and female woodlice were most the sensitive to BPA. We recommend the use of adults, juveniles, female, and male woodlice, as well as a large range of toxicant concentrations, to provide valuable information regarding differential dose responses, effects, and threshold values for EDCs.

Vinclozolin--no transgenerational inheritance of anti-androgenic effects after maternal exposure during organogenesis via the intraperitoneal route.

Science.gov (United States)

Schneider, Steffen; Marxfeld, Heike; Gröters, Sibylle; Buesen, Roland; van Ravenzwaay, Bennard

2013-06-01

The goal of this study was to examine the potential transgenerational inheritance of anti-androgenic effects induced by Vinclozolin administered intraperitoneally to pregnant Wistar rats (Crl:WI[Han]). Dams were dosed with Vinclozolin at 0, 4 or 100mg/kg bw/d on gestation days 6-15. Male offspring of F1-F3 generations were bred with untreated females to yield F2-F4 offspring. No evident anti-androgenic effects were observed at 4mg/kg bw/d, but a case of hypospadias as well as delayed sexual maturation in F1 male offspring was observed as a sign of anti-androgenicity at 100mg/kg bw/d. However, F1-F3 males developed normally to sexual maturity and were able to mate and to generate healthy progeny. Sperm count, morphology and motility were not affected in F1-F4 generation male offspring. In conclusion, transgenerational inheritance of Vinclozolin's anti-androgenic effects was not evident in outbred Wistar rats. Copyright © 2013 Elsevier Inc. All rights reserved.

CHARACTERIZATION OF THE PERIOD OF SENSITIVITY OF FETAL MALE SEXUAL DEVELOPMENT TO VINCLOZOLIN

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Characterization of the period of sensitivity of fetal male sexual development to vinclozolin.Wolf CJ, LeBlanc GA, Ostby JS, Gray LE Jr.Endocrinology Branch, MD 72, Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, U....

ENVIRONMENTAL ANTIANDROGENS: LOW DOSES OF VINCLOZOLIN ALTER SEXUAL DIFFERENTIATION OF THE MALE RAT

Science.gov (United States)

In humans and rodents, exposure to antiandrogenic chemicals during sexual differentiation can produce malformations of the reproductive tract. Perinatal administration of 100 or 200 mg vinclozolin (V) kg-1 day-1 during sexual differentiation in rats induces female-like anogenital...

Development of a bioassay to assess the toxicity of oil sands sediments to pike (Esox lucius)

Energy Technology Data Exchange (ETDEWEB)

Turcotte, D.; Yuan, H.; Tumber, V.; Parrott, J. [Environment Canada, Ottawa, ON (Canada); Raine, J. [Saskatchewan Univ., Saskatoon, SK (Canada)

2010-07-01

Pike (Esox lucius) are a commercially sought fish species that inhabit the Athabasca River, which flows through the Athabasca oil sands. The fish are exposed to natural sources of bitumen from the McMurray formation. This study was conducted to design and implement a daily-renewal bioassay to assess the toxicity of oil sands to this fish species and to obtain information regarding the development of pike exposed to bitumen. Eggs were collected and fertilized with milt from spawning wild pike captured from Lake Diefenbaker in Saskatchewan. The fertilized eggs were exposed to different concentrations of sediments or culture water only (negative controls) until complete yolk absorption of control fish, approximately 15 days post-hatch. For the rest of the experiment, brine shrimp were fed to the walleye embryos every day after hatching. The developing fish were examined for morphological deformities, survival, hatching success, and changes in weight and length. The research findings indicated that pike is less sensitive than walleye and fathead minnow to the toxicity of oil sands sediments.

Sub-NOAEL amounts of vinclozolin and xenoestrogens target rat chondrogenesis in vivo.

Science.gov (United States)

Auxietre, Thuy-Anh; Dumontier, Marie-France; Balguy, Irene; Frapart, Yves; Canivenc-Lavier, Marie-Chantal; Berges, Raymond; Boudalia, Sofiane; Auger, Jacques; Corvol, Marie-Therese; Savouret, Jean-François

2014-04-01

Several endocrine disrupting compounds (EDC) elicit skeletal dysgenesis at pharmacological doses. We have investigated the impact of doses below the "No Observed Adverse Effect" (NOAEL) for vinclozolin (V), an anti-androgenic fungicide, alone or associated with xenoestrogens (Genistein, G and bisphenol-A, BPA). V, G, BPA and their combinations were administered orally to female Wistar rats during gestation and lactation. F1 and F2 offspring were investigated for skeletal anomalies at post-natal days 30, 110 (d30, d110). Skeletal development was monitored by measuring caudal vertebrae and long bones dimensions by X-ray micro-CT-scan. A significant increase in Inter Transverse Apophysis (ITA) distance at the upper head of caudal vertebrae, associated with a reduction in vertebral body height was observed in treated F1 females, but not males. Histometrical analysis of vertebral body growth plate cartilage was performed on serial sections of caudal vertebrae. F1 females but not males showed a diminution in growth plate thickness, with greater impact on the hypertrophic zone. All effects were maximal at d30. Effects on ITA width persisted until d110 while effects on growth plate disappeared. These effects were essentially vinclozolin or BPA-dependent. F2 animals were not affected. Our data suggest that vinclozolin and xenoestrogens act as cartilage developmental disruptors. We suggest that present NOAEL values for these compounds, and EDC at large, might be reconsidered using gestational exposure models. Finally, micro CT-scan appears a valuable non-invasive technique to detect EDC effects on live fauna. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Transcriptional expression analysis of ABC efflux transporters and xenobiotic-metabolizing enzymes in the Chinese rare minnow.

Science.gov (United States)

Yuan, Lilai; Lv, Biping; Zha, Jinmiao; Wang, Zijian

2014-05-01

In the present study, the cDNA fragments of five ABC transporter genes (ABCB1, ABCB11, ABCC1, ABCC2, and ABCG2) in the rare minnow were cloned, and their tissue-specific expression patterns were evaluated across eight rare minnow tissues (liver, gill, intestine, kidney, spleen, brain, skin, and muscle). Furthermore, the transcriptional effects on these ABC transporter genes and five xenobiotic-metabolizing enzyme genes (CYP1A, GSTm, GSTp1, GCLC, and UGT1a) were determined in the rare minnow liver after 12 days of pyrene exposure. Basal expression analysis showed that the tissues with high expression of the ABC transporters included the liver, kidney, and intestine. Moreover, the most highly expressed of the ABC genes were ABCB1 and ABCC2 in all eight of the tissues tested. The ABCB11 gene was almost exclusively expressed in the liver of the rare minnow, whereas ABCC1 and ABCG2 showed weak expression in all eight tissues compared to ABCB1 and ABCC2. Our results provide the first thorough examination of the expression patterns of toxicologically relevant ABC transporters in the rare minnow and serve as a necessary basis for further studies of these ABC transporters in fish. Furthermore, synergistic up-regulation of CYP1A, GSTp1, GCLC, UGT1a, and ABCC2 was observed in the rare minnow liver following pyrene exposure, while GSTm, ABCB1, ABCB11, ABCC1, and ABCG2 were not significantly affected (p ABC transporters by pyrene suggests a possible involvement and cooperation of these genes in the detoxification process in rare minnows. Copyright © 2014 Elsevier B.V. All rights reserved.

ANTIANDROGENIC EFFECTS OF VINCLOZOLIN ON MALE RATS ARE PARTIALLY ATTENUATED BY TESTOSTERONE PROPIONATE

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ANTIANDROGENIC EFFECTS OF VINCLOZOLIN ON MALE RATS ARE PARTIALLY ATTENUATED BY TESTOSTERONE PROPIONATECynthia Wolf1,2 , Joe Ostby1, Jonathan Furr 1, Gerald A. LeBlanc2, and L. Earl Gray, Jr.11 US Environmental Protection Agency, NHEERL, RTD, RTP, NC 27711, 2 Departmen...

Acute and chronic sensitivity to copper of a promising ecotoxicological model species, the annual killifish Nothobranchius furzeri.

Science.gov (United States)

Philippe, Charlotte; Grégoir, Arnout F; Janssens, Lizanne; Pinceel, Tom; De Boeck, Gudrun; Brendonck, Luc

2017-10-01

Nothobranchius furzeri is a promising model for ecotoxicological research due to the species' short life cycle and the production of drought-resistant eggs. Although the species is an emerging vertebrate fish model for several fundamental as well as applied research domains, its potential for ecotoxicological research has not yet been tested. The aim of this study was to characterise the acute and chronic sensitivity of this species to copper as compared to other model organisms. Effects of both acute and chronic copper exposure were tested on survival, life history and physiological traits. We report a 24h-LC 50 of 53.93µg Cu/L, which is situated within the sensitivity range of other model species such as Brook Trout, Fathead Minnow and Rainbow Trout. Moreover, in the full life cycle exposure, we show that an exposure concentration of 10.27µg/L did not cause acute adverse effects (96h), but did cause mortality after prolonged exposure (3-4 weeks). Also chronic, sublethal effects were observed, such as a reduction in growth rate, delayed maturation and postponed reproduction. Based on our results, we define the NOEC at 6.68µg Cu/L, making N. furzeri more sensitive to copper as compared to Brook Trout and Fathead Minnow. We found stimulatory effects on peak fecundity at subinhibitory levels of copper concentrations (hormesis). Finally, we found indications for detoxifying and copper-excreting mechanisms, demonstrating the ability of the fish to cope with this essential metal, even when exposed to stressful amounts. The successful application of current ecotoxicological protocols on N. furzeri and its sensitivity range comparable to that of other model organisms forms the basis to exploit this species in further ecotoxicological practices. Copyright © 2017 Elsevier Inc. All rights reserved.

Pharmacokinetics and dosimetry of the anti-androgen vinclozolin after oral administration inthe rat

Science.gov (United States)

Vinclozolin (V) is a fungicide with antiandrogenic properties. To determine the pharmacokinetics and dosimetry of V, adult male rats were administered an oral dose of V (100 mg/kg) in corn oil and sacrificed over time after dosing. V and its metabolites were analyzed in serum and...

Determination and validation of a simple high-performance liquid chromatographic method for simultaneous assay of iprodione and vinclozolin in human urine.

Science.gov (United States)

Carlucci, Giuseppe; Pasquale, Dorina Di; Ruggieri, Fabrizio; Mazzeo, Pietro

2005-12-15

A method based on solid-phase extraction (SPE) and high-performance liquid chromatography (HPLC) was developed for the simultaneous determination of 3-(3,5-diclorophenyl)-5-ethenyl-5-methyl-2,4-oxazolidinedione (vinclozolin) and 3-(3,5-diclorophenyl)-N-(1-methylethyl)-2,4-dioxo-1-imidazolidinecarboxamide (iprodione) in human urine. Urine samples containing vinclozolin and iprodione were collected by solid phase extraction using C(18) cartridges. The chromatographic separation was achieved on a Spherisorb ODS2 (250 mm x 4.6 mm, 5 microm) column with an isocratic mobile phase of acetonitrile-water (60:40, v/v). Detection was UV absorbance at 220 nm. The calibration graphs were linear from 30 to 1000 ng/mL for the two fungicides. Intra- and inter-day R.S.D. did not exceed 2.9%. The quantitation limit was 50 ng/mL for vinclozolin and 30 ng/mL for iprodione, respectively.

Bisphenol A, Dichlorodiphenyltrichloroethane (DDT) and Vinclozolin Affect ex-vivo Uterine Contraction in Rats via Uterotonin (Prostaglandin F2?, Acetylcholine and Oxytocin) Related Pathways

OpenAIRE

Salleh, Naguib; Giribabu, Nelli; Feng, Angeline Oh Mei; Myint, Kyaimon

2015-01-01

Bisphenol-A (BPA), dichrolodiphenyltrichloroethane (DDT) and vinclozolin were found able to induce abnormal uterine contraction. The mechanisms involved remains unclear. We hypothesized that the effect of these compounds were mediated via the uterotonin pathways. Therefore, in this study, effects of BPA, vinclozolin and DDT-only and in combination with uterotonins (PGF-2?, acetylcholine and oxytocin) on the force and pattern of uterine contraction were observed. Methods: Uteri were harvested ...

In vitro metabolism of the anti-androgenic fungicide vinclozolin by rat liver microsomes

Science.gov (United States)

Vinclozolin (V) is a fungicide used in agricultural settings. V administered to rats is hydrolyzed to 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3-butenoic acid (Ml) and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2). V, Ml and M2 have antiandrogenic properties by in...

MODE OF ACTION: INHIBITION OF ANDROGEN RECEPTOR FUNCTION--VINCLOZOLIN-INDUCED MALFORMATIONS IN REPRODUCTIVE DEVELOPMENT

Science.gov (United States)

Vinclozolin is a fungicide that has been shown to cause Leydig cell tumors and atrophy of the accessory sex glands in adult rodents. In addition, exposure of rats during pregnancy causes a pattern of malformations in the male urogenital tract. A wealth of standard toxicological s...

LIQUID CHROMATOGRAPHY DETERMINATION OF ANTI-ANDROGEN VINCLOZOLIN AND ITS METABOLITES IN RAT SERUM

Science.gov (United States)

The objective of this study was to develop a chromatographic method for the analysis of the anti-androgen vinclozolin (V) and its butenoic acid (M1) and enanilide (M2) metabolites in rat serum. V, M1, M2 and M3 were resolved using an HPLC gradient program with a mobile phase con...

Embryonic exposure to the fungicide vinclozolin causes virilization of females and alteration of progesterone receptor expression in vivo: an experimental study in mice

OpenAIRE

Buckley, Jill; Willingham, Emily; Agras, Koray; Baskin, Laurence S

2006-01-01

Abstract Background Vinclozolin is a fungicide that has been reported to have anti-androgenic effects in rats. We have found that in utero exposure to natural or synthetic progesterones can induce hypospadias in mice, and that the synthetic progesterone medroxyprogesterone acetate (MPA) feminizes male and virilizes female genital tubercles. In the current work, we selected a relatively low dose of vinclozolin to examine its in utero effects on the development of the genital tubercle, both at ...

Recovery of a wild fish population from whole-lake additions of a synthetic estrogen.

Science.gov (United States)

Blanchfield, Paul J; Kidd, Karen A; Docker, Margaret F; Palace, Vince P; Park, Brad J; Postma, Lianne D

2015-03-03

Despite widespread recognition that municipal wastewaters contain natural and synthetic estrogens, which interfere with development and reproduction of fishes in freshwaters worldwide, there are limited data on the extent to which natural populations of fish can recover from exposure to these compounds. We conducted whole-lake additions of an active component of the birth control pill (17α-ethynylestradiol; EE2) that resulted in the collapse of the fathead minnow (Pimephales promelas) population. Here we quantify physiological, population, and genetic characteristics of this population over the 7 years after EE2 additions stopped to determine if complete recovery was possible. By 3 years post-treatment, whole-body vitellogenin concentrations in male fathead minnow had returned to baseline, and testicular abnormalities were absent. In the spring of the fourth year, adult size-frequency distribution and abundance had returned to pretreatment levels. Microsatellite analyses clearly showed that postrecovery fish were descendants of the original EE2-treated population. Results from this whole-lake experiment demonstrate that fish can recover from EE2 exposure at the biochemical through population levels, although the timelines to do so are long for multigenerational exposures. These results suggest that wastewater treatment facilities that reduce discharges of estrogens and their mimics can improve the health of resident fish populations in their receiving environments.

Investigation of adaptive responses in fathead minnows (Pimephales promelas) exposed to the model aromatase inhibitor fadrozole

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The vertebrate hypothalamic-pituitary-gonadal (HPG) axis is a highly dynamic system, which, through various feedback mechanisms, strives to maintain physiological conditions conducive to reproduction even in potentially stressful situations. The development of useful predictive m...

VITELLOGENIN GENE EXPRESSION IN FATHEAD MINNOWS EXPOSED TO EE2 IN A WHOLE LAKE DOSING EXPERIMENT

Science.gov (United States)

A whole-lake endocrine disruption experiment was conducted by Fisheries and Oceans Canada at the Experimental Lakes Area (ELA), northwestern Ontario in 2001 and 2002. This experiment examined population, organism, biochemical and cellular-level effects in lake trout, white sucke...

Transgenerational effects of the endocrine disruptor vinclozolin on the prostate transcriptome and adult onset disease.

Science.gov (United States)

Anway, Matthew D; Skinner, Michael K

2008-04-01

The ability of an endocrine disruptor exposure during gonadal sex determination to promote a transgenerational prostate disease phenotype was investigated in the current study. Exposure of an F0 gestating female rat to the endocrine disruptor vinclozolin during F1 embryo gonadal sex determination promoted a transgenerational adult onset prostate disease phenotype. The prostate disease phenotype and physiological parameters were determined for males from F1 to F4 generations and the prostate transcriptome was assessed in the F3 generation. Although the prostate in prepubertal animals develops normally, abnormalities involving epithelial cell atrophy, glandular dysgenesis, prostatitis, and hyperplasia of the ventral prostate develop in older animals. The ventral prostate phenotype was transmitted for four generations (F1-F4). Analysis of the ventral prostate transcriptome demonstrated 954 genes had significantly altered expression between control and vinclozolin F3 generation animals. Analysis of isolated ventral prostate epithelial cells identified 259 genes with significantly altered expression between control and vinclozolin F3 generation animals. Characterization of regulated genes demonstrated several cellular pathways were influenced, including calcium and WNT. A number of genes identified have been shown to be associated with prostate disease and cancer, including beta-microseminoprotein (Msp) and tumor necrosis factor receptor superfamily 6 (Fadd). The ability of an endocrine disruptor to promote transgenerational prostate abnormalities appears to involve an epigenetic transgenerational alteration in the prostate transcriptome and male germ-line. Potential epigenetic transgenerational alteration of prostate gene expression by environmental compounds may be important to consider in the etiology of adult onset prostate disease.

Effects of advanced treatments of wastewater effluents on estrogenic and reproductive health impacts in fish.

Science.gov (United States)

Filby, Amy L; Shears, Janice A; Drage, Briane E; Churchley, John H; Tyler, Charles R

2010-06-01

Whether the implementation of additional treatments for the removal of estrogens from wastewater treatment works (WwTWs) effluents will eliminate their feminizing effects in exposed wildlife has yet to be established, and this information is crucial for future decisions on investment into WwTWs. Here, granular activated carbon (GAC), ozone (O(3)), and chlorine dioxide (ClO(2)) were investigated for their effectiveness in reducing steroidal estrogen levels in a WwTW effluent and assessments made on the associated estrogenic and reproductive responses in fathead minnows (Pimephales promelas) exposed for 21 days. All treatments reduced the estrogenicity of the standard-treated (STD) effluent, but with different efficacies; ranging between 70-100% for total estrogenicity and 53-100% for individual steroid estrogens. In fish exposed to the GAC- and ClO(2)- (but not O(3)-) treated effluents, there was no induction of plasma vitellogenin (VTG) or reduction in the weight of the fatpad, a secondary sex character in males, as occurred for fish exposed to STD effluent. This finding suggests likely benefits of employing these treatment processes for the reproductive health in wild fish populations living in rivers receiving WwTW discharges. Exposure of pair-breeding minnows to the GAC-treated effluent, however, resulted in a similar inhibition of egg production to that occurring for exposure to the STD effluent (34-40%). These data, together with a lack of effect on egg production of the estrogen, ethinylestradiol (10 ng/L), alone, suggest that chemical/physical properties of the effluents rather than their estrogenicity were responsible for the reproductive effect and that these factor(s) were not remediated for through GAC treatment. Collectively, our findings illustrate the importance of assessing integrative biological responses, rather than biomarkers alone, in the assessment and improvement of WwTW technologies for the protection of wild fish populations.

In utero and lactational exposure to low-dose genistein-vinclozolin mixture affects the development and growth factor mRNA expression of the submandibular salivary gland in immature female rats.

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Kouidhi, Wided; Desmetz, Catherine; Nahdi, Afef; Bergès, Raymond; Cravedi, Jean-Pierre; Auger, Jacques; El May, Michèle; Canivenc-Lavier, Marie Chantal

2012-06-01

It has been suggested that hormonally controlled submandibular salivary gland (SSG) development and secretions may be affected by endocrine disruptor compounds. We investigated the effects of oral gestation-lactation exposure to 1 mg/kg body weight daily dose of the estrogenic soy-isoflavone genistein and/or the anti-androgenic food contaminant vinclozolin in female rats. The SSGs of female offspring were collected at postnatal day 35 to study gland morphogenesis and mRNA expression of sex-hormone receptors and endocrine growth factors as sex-dependent biomarkers. Because of high expression in neonatal SSG, mRNA expression of transforming growth factor α was also studied. Exposure to genistein, vinclozolin, or a genistein+vinclozolin mixture resulted in significantly lower numbers of striated ducts linked to an increase in their area and lower acinar proliferation (Ki-67-positive nuclei). Exposure to the mixture had the highest significant effects, which were particularly associated with repression of epidermal growth factor, nerve growth factor, and transforming growth factor α expression. In conclusion, early exposure to low doses of genistein and vinclozolin can affect glandular structure and endocrine gene mRNA expression in prepubertal SSG in female rats, and the effects are potentialized by the genistein+vinclozolin mixture. Our study provides the first evidence that SSG are targeted by both estrogenic and anti-androgenic disrupting compounds and are more sensitive to mixtures.

Sensitive trace enrichment of environmental andiandrogen vinclozolin from natural waters and sediment samples using hollow-fiber liquid-phase microextraction.

Science.gov (United States)

Lambropoulou, Dimitra A; Albanis, Triantafyllos A

2004-12-17

The presence of vinclozolin in the environment as far as the endocrine disruption effects in biota are concerned has raised interest in the environmental fate of this compound. In this respect, the present study attempts to investigate the feasibility of applying a novel quantitative method, liquid-phase microextraction (LPME), so as to determine this environmental andiandrogen in environmental samples such as water and sediment samples. The technique involved the use of a small amount (3 microL) of organic solvent impregnated in a hollow fiber membrane, which was attached to the needle of a conventional GC syringe. The extracted samples were analyzed by gas chromatography coupled with electron-capture detection. Experimental LPME conditions such as extraction solvent, stirring rate, content of NaCl and pH were tested. Once LPME was optimized, the performance of the proposed technique was evaluated for the determination of vinclozolin in different types of natural water samples. The recovery of spiked water samples was from 80 to 99%. The procedure was adequate for quantification of vinclozolin in waters at levels of 0.010 to 50 microg/L (r> 0.994) with a detection limit of 0.001 microg/L (S/N= 3). Natural sediment samples from the Aliakmonas River area (Macedonia, Greece) spiked with the target andiandrogen compound were liquid-liquid extracted and analyzed by the methodology developed in this work. No significant interferences from the samples matrix were noticed, indicating that the reported methodology is an innovative tactic for sample preparation in sediment analysis, with a considerable improvement in the achieved detection limits. The results demonstrated that apart from analyte enrichment, the proposed LPME procedure also serves as clean-up method and could be successfully performed to determine trace amounts of vinclozolin in water and sediment samples.

Effects of oil sands sediments on fish

International Nuclear Information System (INIS)

Parrott, J.; Colavecchia, M.; Hewitt, L.; Sherry, J.; Headley, J.; Turcotte, D.; Liber, K.

2010-01-01

This paper described a collaborative project organized by Natural Resources Canada (NRCan) Panel of Energy Research and Development (PERD) with researchers from Environment Canada and the University of Saskatchewan. The 4-year study was conducted to assess the toxicity of oil sands sediments and river waters, and reclamation ponds and sediments on laboratory-raised fish. Three sediments from rivers were evaluated for their potential to cause adverse impacts on fathead minnow eggs and larvae for a period of 18 days. The study monitored hatching, larval survival, development, and growth. Naphthenic acids (NA), polycyclic aromatic hydrocarbons (PAHs) and metals were measured in the sediments to determine if the compounds can be correlated with observed toxicity. The study will also assess walleye eggs exposed to sediments, and in situ fish exposures. Toxicity identification and evaluation (TIE) studies will be conducted to isolate the fractions that may affect fish development and growth.

Fish gill responses to pollutants from oil sands mining-associated waters

International Nuclear Information System (INIS)

Lee, L.E.J.; Willfang, S.; Lamb, M.P.; Nero, V.; Farwell, A.J.; Dixon, D.G.

2002-01-01

The processing of Athabasca Deposit oil sands results in large amounts of liquid wastes associated with oil sand tailings. In addition to containing polycyclic aromatic hydrocarbons (PAHs), these waste waters are high in salinity and naphthenic acids which may be toxic to aquatic biota and their effects must be clarified. This study presents a suite of tests for in-depth and quick analysis of tailings water toxicity and contributes to the assessment of environmental risk. Yellow perch, fathead minnows, and rainbow trout were exposed to reclamation ponds where both in vivo and in vitro evaluation of crude and individual naphthenic acids and salts were conducted to examine their effect on fish gills which are very susceptible to contaminants. The fish exposed to the reclamation ponds showed higher incidence of gill pathological changes than control fish in Mildred Lake, a reservoir lake whose waters are diverted for use in oil sands extraction. Notable gill histopathological changes were observed when fish were exposed in vivo to sulfate/chloride salts and to abietic acid. Changes in membrane integrity, lysosomal activity and general morphology were observed when fished were exposed in vitro to salts, commercial napthenic acids or crude naphthenic extracts from the reclamation ponds

Effects of anti-androgens cyproterone acetate, linuron, vinclozolin, and p,p'-DDE on the reproductive organs of the copepod Acartia tonsa.

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Watermann, Burkard T; Albanis, Triantafyllos A; Galassi, Silvana; Gnass, Katarina; Kusk, Kresten O; Sakkas, Vasilios A; Wollenberger, Leah

2016-11-09

The study was performed to detect the effects of anti-androgenic compounds on the reproduction. In this paper alterations observed in the marine calanoid copepod Acartia tonsa exposed to environmental concentrations of cyproterone acetate (CPA), linuron (LIN), vinclozolin (VIN), and 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (p,p'-DDE) for 21 days covering a full life cycle are described. Histological alterations were studied with a focus on reproductive organs, gonad and accessory sexual glands. Exposure to ≥1.2 µg L(-1) CPA caused degeneration of spermatocytes and deformation of the spermatophore in males. In a single male exposed to 33 µg L(-1) CPA, an ovotestis was observed. In CPA exposed females, enhancement of oogenesis, increase in apoptosis and a decrease in proliferation occurred. Exposure of males to ≥12 µg L(-1) LIN caused degenerative effects in spermatogonia, spermatocytes and spermatids, and at 4.7 µg L(-1) LIN, the spermatophore wall displayed an irregular formation. In LIN exposed females, no such structural alterations were found; however, the proliferation index was reduced at 29 µg L(-1) LIN. At an exposure concentration of ≥100 µg L(-1) VIN, distinct areas in male gonad were stimulated, whereas others displayed a disturbed spermatogenesis and a deformed spermatophore wall. In VIN exposed female A. tonsa, no effects were observed. Male A. tonsa exposed to p,p'-DDE displayed an impairment of spermatogenesis in all stages with increased degrees of apoptosis. In p,p'-DDE-exposed females, a statistical significant increase of the proliferation index and an intensification of oogenesis were observed at 0.0088 µg L(-1).

Effect of low dose of vinclozolin on reproductive tract development and sperm parameters in CD1 outbred mice.

Science.gov (United States)

Elzeinova, Fatima; Novakova, Vendula; Buckiova, Daniela; Kubatova, Alena; Peknicova, Jana

2008-01-01

The effect of a low dose of vinclozolin within the development of the reproductive tract during gestation (VIN-GD 15-22) and puberty (VIN-PND 23-44) in CD1 mice was tested. We found a decrease in the anogenital distance, prostate weight and pathology of testes in both experimental groups. Sperm counts decreased to 46% (VIN-GD) and to 81% (VIN-PND), and also the acrosomal state (evaluated by antiacrosomal antibody) decreased in both groups to 89% in comparison to the control group (100%). Sperm head abnormalities increased by approximately 18% and 13%, respectively. In this connection, the expression of some genes was changed (arosome-related gene (Acr), apoptosis related genes (p53, p21)). In conclusion, a low dose of vinclozolin affected the reproductive tract, sperm parameters and expression of selected genes in both experimental groups.

Development of Cardiovascular and Neurodevelopmental Metrics as Sublethal Endpoints for the Fish Embryo Toxicity Test.

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Krzykwa, Julie C; Olivas, Alexis; Jeffries, Marlo K Sellin

2018-06-19

The fathead minnow fish embryo toxicity (FET) test has been proposed as a more humane alternative to current toxicity testing methods, as younger organisms are thought to experience less distress during toxicant exposure. However, the FET test protocol does not include endpoints that allow for the prediction of sublethal adverse outcomes, limiting its utility relative to other test types. Researchers have proposed the development of sublethal endpoints for the FET test to increase its utility. The present study 1) developed methods for previously unmeasured sublethal metrics in fathead minnows (i.e., spontaneous contraction frequency and heart rate) and 2) investigated the responsiveness of several sublethal endpoints related to growth (wet weight, length, and growth-related gene expression), neurodevelopment (spontaneous contraction frequency, and neurodevelopmental gene expression), and cardiovascular function and development (pericardial area, eye size and cardiovascular related gene expression) as additional FET test metrics using the model toxicant 3,4-dichloroaniline. Of the growth, neurological and cardiovascular endpoints measured, length, eye size and pericardial area were found to more responsive than the other endpoints, respectively. Future studies linking alterations in these endpoints to longer-term adverse impacts are needed to fully evaluate the predictive power of these metrics in chemical and whole effluent toxicity testing. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Cummulative and antagonistic effects of a mixture of the antiandrogrens vinclozolin and iprodione in the pubertal male rat:

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Vinclozolin and iprodione are dicarboximide fungicides that display anti-androgenic effects in the male rat, which suggests co-exposure to these fungicides would lead to cumulative effects on androgen-sensitive endpoints. Iprodione is a steroid synthesis inhibitor, but AR antagon...

Impaired swim bladder inflation in early-life stage fathead minnows exposed to a deiodinase inhibitor, iopanoic acid

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The thyroid axis plays a critical role in teleost fish development. The present study investigated whether inhibition of deiodinase, the enzyme which converts thyroxine (T4), to the more biologically-active form, 3,5,3'-triiodothyronine (T3), would impact inflation of the posteri...

The effect of turbidity on recognition and generalization of predators and non-predators in aquatic ecosystems.

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Chivers, Douglas P; Al-Batati, Fawaz; Brown, Grant E; Ferrari, Maud C O

2013-02-01

Recent anthropogenic activities have caused a considerable change in the turbidity of freshwater and marine ecosystems. Concomitant with such perturbations are changes in community composition. Understanding the mechanisms through which species interactions are influenced by anthropogenic change has come to the forefront of many ecological disciplines. Here, we examine how a change in the availability of visual information influences the behavior of prey fish exposed to potential predators and non-predators. When fathead minnows, Pimephales promelas, were conditioned to recognize predators and non-predators in clear water, they showed a highly sophisticated ability to distinguish predators from non-predators. However, when learning occurred under conditions of increased turbidity, the ability of the prey to learn and generalize recognition of predators and non-predators was severely impaired. Our work highlights that changes at the community level associated with anthropogenic perturbations may be mediated through altered trophic interactions, and highlights the need to closely examine behavioral interactions to understand how species interactions change.

The effect of turbidity on recognition and generalization of predators and non-predators in aquatic ecosystems

Science.gov (United States)

Chivers, Douglas P; Al-Batati, Fawaz; Brown, Grant E; Ferrari, Maud C O

2013-01-01

Recent anthropogenic activities have caused a considerable change in the turbidity of freshwater and marine ecosystems. Concomitant with such perturbations are changes in community composition. Understanding the mechanisms through which species interactions are influenced by anthropogenic change has come to the forefront of many ecological disciplines. Here, we examine how a change in the availability of visual information influences the behavior of prey fish exposed to potential predators and non-predators. When fathead minnows, Pimephales promelas, were conditioned to recognize predators and non-predators in clear water, they showed a highly sophisticated ability to distinguish predators from non-predators. However, when learning occurred under conditions of increased turbidity, the ability of the prey to learn and generalize recognition of predators and non-predators was severely impaired. Our work highlights that changes at the community level associated with anthropogenic perturbations may be mediated through altered trophic interactions, and highlights the need to closely examine behavioral interactions to understand how species interactions change. PMID:23467451

FIELD VALIDATION OF A SHEEPSHEAD MINNOW ESTROGEN-RESPONSIVE CDNA MACROARRAY

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Hemmer, Michael J., Iris Knoebl, Becky L. Hemmer, Patrick Larkin, Peggy S. Harris and Nancy D. Denslow. In press. Field Validation of a Sheepshead Minnow Estrogen-Responsive cDNA Macroarray (Abstract). To be presented at the SETAC Fourth World Congress, 14-18 November 2004, Portl...

Evaluation of lithium as a toxicant and the modifying effect of sodium

International Nuclear Information System (INIS)

Kszos, L.A.; Stewart, A.J.; O'Reilly, S.E.

1995-01-01

Routine compliance tests conducted for a groundwater treatment facility at the Y-12 Plant on the Department of Energy's (DOE) Oak Ridge Reservation (ORR), TN, showed that the effluent was acutely toxic to Ceriodaphnia dubia and fathead minnow (Pimephales promelas) larvae. An evaluation of suspected contaminants revealed that increased toxicity coincided with increased concentrations of lithium. Lithium is a light, strong metal that is used in DOE operations, including fusion weapons and fission reactors. Little has been published about lithium toxicity. Toxicity tests were conducted with fathead minnows and C. dubia using lithium chloride and lithium tetraborate. Dilute mineral water (DMW) or the receiving stream water (East Fork Poplar Creek) was used as the dilution water in the toxicity evaluation. A concentration of 1 mg Li/L in DMW reduced the survival of both test species; 0.5 mg Li/L in DMW reduced C. dubia reproduction and minnow growth. Sodium appears to influence the toxicity of Li; the metal was six times more toxic in the low-sodium DMW than in stream water containing 30 mg Na/L. Tests with LiCl in combination with NaCl and NA 2 SO 4 demonstrated that the presence of sodium reduced the toxicity of Li to C. dubia. In laboratory tests with a snail (Elimia clavaeformis) common on the ORR, the feeding rate declined in 0.15 mg Li/L. Because Li has also been demonstrated to be toxic to several plant species, tests with LiCi were also conducted using buttercrunch lettuce (Lactuca saliva). The EC 50 for seed growth after 8 d incubation was 37.5 mg Li/L. These findings are significant because of widespread industrial use and potential accumulation of Li in soils

Early development and larval behaviour of a minnow, Barbus ...

African Journals Online (AJOL)

The chubbyhead barb, Barbus anoplus, underwent a population explosion in the early phases of filling of Lake be Roux on the Orange River. This successful colonization was possibly related to the survival strategy of the young stages of this minnow. It is suggested that some of the development traits of B. anoplus enabled ...

EXPRESSION PROFILING OF ESTROGENIC COMPOUNDS USING A SHEEPSHEAD MINNOW CDNA MACROARRAY

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Larkin, Patrick, Leroy C. Folmar, Michael J. Hemmer, Arianna J. Poston and Nancy D. Denslow. 2003. Expression Profiling of Estrogenic Compounds Using a Sheepshead Minnow cDNA Macroarray. Environ. Health Perspect. 111(6):839-846. (ERL,GB 1171). A variety of anthropogenic c...

A mixture of Iprodione and Vinclozolin delays the onset of puberty in the male rat in a cumulative manner.

Science.gov (United States)

Vinclozolin and iprodione are dicarboximide fungicides that display anti-androgenic effects in the male rat, which suggests co-exposure to these fungicides would lead to cumulative effects on androgen-sensitive endpoints. In order to test for cumulative effects by iprodione and v...

ASSOCIATIONS BETWEEN GENETIC DIVERSITY AND ANTHROPOGENIC DISTURBANCE IN MIDWESTERN STREAM-DWELLING MINNOWS

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Anthropogenic disturbances may leave imprints on patterns of intraspecific genetic diversity through their effects on population size, adaptation, migration, and mutation. We examined patterns of genetic diversity for a stream-dwelling minnow (the central stoneroller, Campostoma...

Vinclozolin: a feasibility and sensitivity study of the ILSI-HESI F1-extended one-generation rat reproduction protocol.

Science.gov (United States)

Schneider, Steffen; Kaufmann, Wolfgang; Strauss, Volker; van Ravenzwaay, Bennard

2011-02-01

Feasibility of the ILSI-HESI (ACSA) extended one-generation protocol was tested with vinclozolin (dietary 0, 4, 20, 100mg/kg/day). Parental Wistar rats (n=25/sex/dose) were dosed pre-mating (males 4, females 2 weeks) through F1 offspring weaning (postnatal day PND21); F1 dosing continued through PND70. At PND21, 3 subsets (each 1 pup/sex/litter) were selected for neurotoxicology (functional observational battery, motor activity, neuropathology), clinical pathology (hematology, clinical chemistry, urinalysis, thyroid hormone assay) (subsets 1a, 1b; each n=10/sex/dose), immunotoxicology (IgM) SRBC antibody response and natural killer cell assays (subset 2; n=25/sex/dose), and estrus cycle (subset 3; n=25/dose). Vinclozolin reduced parental and offspring bodyweight and prostate, seminal vesicles and epididymides weight, and increased adrenal weight/induced adrenal cortical hypertrophy at 100mg/kg. Mating, fertility, gestation and lactation were unaffected. At 20 and 100mg/kg, F1 males had reduced anogenital distance and retained areolae; at 100mg/kg only, there was hypospadias, purulent prostatitis and seminal vesicle inflammation with atrophy, and Leydig cell hyperplasia, and in F1 females accelerated vaginal opening. These effects are consistent with vinclozolin's known anti-androgenic developmental effects. Neuro- and immunotoxicology tests were unaffected. F1 Only T4 was reduced at 20 and 100mg/kg. The overall sensitivity of the extended one-generation protocol is comparable to or even greater than the current two-generation study. Thus it reduces animal use while maintaining or enhancing information for risk assessment. Copyright © 2010 Elsevier Inc. All rights reserved.

Fathead minnow whole-mount in situ hybridization (WISH)

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U.S. Environmental Protection Agency — This study demonstrates the potential of whole-mount in situ hybridization (WISH), in conjunction with quantitative real-time polymerase chain reaction (QPCR)...

Effects of synthetic gestagens on fish reproduction.

Science.gov (United States)

Zeilinger, Jana; Steger-Hartmann, Thomas; Maser, Edmund; Goller, Stephan; Vonk, Richardus; Länge, Reinhard

2009-12-01

Although it is well known that estrogenic steroidal hormones are able to affect the sexual development and reproduction of fish at low concentrations, no data on environmental effects of the class of progestogenic hormones are available yet. Synthetic gestagens (progestins) are a component in oral contraceptives. Upon their use, a fraction of the progestins will be excreted via urine into the aquatic environment. On the basis of their pharmacological action in mammals, it is supposed that fish reproduction is the most sensitive endpoint for the progestin treatment. In order to test this assumption, the effects of two progestins currently marketed in contraceptive formulations, levonorgestrel (LNG) and drospirenone (DRSP), were investigated in adult fathead minnows (Pimephales promelas) following an Organization for Economic Cooperation and Development 21-d fish reproduction screening assay draft protocol with additional end points. Levonorgestrel was tested at measured concentrations of 0.8, 3.3, and 29.6 ng/L, and DRSP at concentrations of 0.66, 6.5, and 70 microg/L. Both tested progestins caused an inhibition of reproduction. For LNG, this occurred at concentrations of >or=0.8 ng/L, no no-observed-effect concentration (NOEC) could be defined. Higher concentrations resulted in masculinization of females with de novo synthesis of nuptial tubercles. Drospirenone treatment, however, affected the reproductive success of fathead minnow at concentrations of 6.5 microg/L and higher with a clear dose-response relationship and a NOEC of 0.66 microg/L, which is above environmentally relevant concentrations.

In situ and laboratory toxicity of coalbed natural gas produced waters with elevated sodium bicarbonate

Science.gov (United States)

Farag, Aïda M.; Harper, David D.; Skaar, Don

2014-01-01

Some tributaries in the Powder River Structural Basin, USA, were historically ephemeral, but now contain water year round as a result of discharge of coalbed natural gas (CBNG)-produced waters. This presented the opportunity to study field sites with 100% effluent water with elevated concentrations of sodium bicarbonate. In situ experiments, static renewal experiments performed simultaneously with in situ experiments, and static renewal experiments performed with site water in the laboratory demonstrated that CBNG-produced water reduces survival of fathead minnow (Pimephales promelas) and pallid sturgeon (Scaphirhynchus albus). Age affected survival of fathead minnow, where fish 2 d posthatch (dph) were more sensitive than 6 dph fish, but pallid sturgeon survival was adversely affected at both 4 and 6 dph. This may have implications for acute assays that allow for the use of fish up to 14 dph. The survival of early lifestage fish is reduced significantly in the field when concentrations of NaHCO3 rise to more than 1500 mg/L (also expressed as >1245 mg HCO3 (-) /L). Treatment with the Higgin's Loop technology and dilution of untreated water increased survival in the laboratory. The mixing zones of the 3 outfalls studied ranged from approximately 800 m to 1200 m below the confluence. These experiments addressed the acute toxicity of effluent waters but did not address issues related to the volumes of water that may be added to the watershed.

Genotoxic effects of vinclozolin on the aquatic insect Chironomus riparius (Diptera, Chironomidae).

Science.gov (United States)

Aquilino, Mónica; Sánchez-Argüello, Paloma; Martínez-Guitarte, José-Luis

2018-01-01

Vinclozolin (Vz) is a pollutant found in aquatic environments whose antiandrogenic effects in reproduction are well known in mammals. Although its reproductive effects have been less studied in invertebrates, other effects, including genotoxicity, have been described. Therefore, in this work, we studied the genotoxic effects of Vz in the freshwater benthic invertebrate Chironomus riparius. DNA damage was evaluated with the comet assay (tail area, olive moment, tail moment and % DNA in tail), and the transcriptional levels of different genes involved in DNA repair (ATM, NLK and XRCC1) and apoptosis (DECAY) were measured by RT-PCR. Fourth instar larvae of C. riparius, were exposed to Vz for 24 h at 20 and 200 μg/L. The Vz exposures affected the DNA integrity in this organism, since a dose-response relationship occurred, with DNA strand breaks significantly increased with increased dose for tail area, olive moment and tail moment parameters. Additionally, the lower concentration of Vz produced a significant induction of the transcripts of three genes under study (ATM, NLK and XRCC1) showing the activation of the cellular repair mechanism. In contrast, the expression of these genes with the highest concentration were downregulated, indicating failure of the cellular repair mechanism, which would explain the higher DNA damage. These data report for the first time the alterations of Vz on gene transcription of an insect and confirm the potential genotoxicity of this compound on freshwater invertebrates. Copyright © 2017 Elsevier Ltd. All rights reserved.

Additive and synergistic antiandrogenic activities of mixtures of azol fungicides and vinclozolin.

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Christen, Verena; Crettaz, Pierre; Fent, Karl

2014-09-15

Many pesticides including pyrethroids and azole fungicides are suspected to have an endocrine disrupting property. At present, the joint activity of compound mixtures is only marginally known. Here we tested the hypothesis that the antiandrogenic activity of mixtures of azole fungicides can be predicted by the concentration addition (CA) model. The antiandrogenic activity was assessed in MDA-kb2 cells. Following assessing single compounds activities mixtures of azole fungicides and vinclozolin were investigated. Interactions were analyzed by direct comparison between experimental and estimated dose-response curves assuming CA, followed by an analysis by the isobole method and the toxic unit approach. The antiandrogenic activity of pyrethroids deltamethrin, cypermethrin, fenvalerate and permethrin was weak, while the azole fungicides tebuconazole, propiconazole, epoxiconazole, econazole and vinclozolin exhibited strong antiandrogenic activity. Ten binary and one ternary mixture combinations of five antiandrogenic fungicides were assessed at equi-effective concentrations of EC25 and EC50. Isoboles indicated that about 50% of the binary mixtures were additive and 50% synergistic. Synergism was even more frequently indicated by the toxic unit approach. Our data lead to the conclusion that interactions in mixtures follow the CA model. However, a surprisingly high percentage of synergistic interactions occurred. Therefore, the mixture activity of antiandrogenic azole fungicides is at least additive. Mixtures should also be considered for additive antiandrogenic activity in hazard and risk assessment. Our evaluation provides an appropriate "proof of concept", but whether it equally translates to in vivo effects should further be investigated. Copyright © 2014 Elsevier Inc. All rights reserved.

Evaluation of tributyltin toxicity in Chinese rare minnow larvae by abnormal behavior, energy metabolism and endoplasmic reticulum stress.

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Li, Zhi-Hua; Li, Ping

2015-02-05

Tributyltin (TBT) is a ubiquitous contaminant in aquatic environment, but the detailed mechanisms underlying the toxicity of TBT have not been fully understood. In this study, the effects of TBT on behavior, energy metabolism and endoplasmic reticulum (ER) stress were investigated by using Chinese rare minnow larvae. Fish larvae were exposed at sublethal concentrations of TBT (100, 400 and 800 ng/L) for 7 days. Compared with the control, energy metabolic parameters (RNA/DNA ratio, Na(+)-K(+)-ATPase) were significantly inhibited in fish exposed at highest concentration (800 ng/L), as well as abnormal behaviors observed. Moreover, we found that the PERK (PKR-like ER kinase)-eIF2α (eukaryotic translation initiation factor 2α) pathway, as the main branch was activated by TBT exposure in fish larvae. In short, TBT-induced physiological, biochemical and molecular responses in fish larvae were reflected in parameters measured in this study, which suggest that these biomarkers could be used as potential indicators for monitoring organotin compounds present in aquatic environment. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Tributyltin affects shoaling and anxiety behavior in female rare minnow (Gobiocypris rarus).

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Zhang, Jiliang; Zhang, Chunnuan; Sun, Ping; Shao, Xian

2016-09-01

Effects of tributyltin (TBT) on reproduction are well established in many fish species. However, few studies report the effects of TBT on non-reproductive behaviors, which is a novel aspect of endocrine disruption in fish. Thus, the present study used rare minnow (Gobiocypris rarus) to investigate the effects of TBT, at environmental concentrations of 1, 10 and 100ng/L, on shoaling and anxiety behaviors. The results showed that fish exposed to TBT had less group cohesion during the course of the 10-min observation period as compared with the control fish. Further, TBT altered the shoaling in the Novel tank test, where shoaling is determined as the tendency to leave a shoal of littermates trapped behind a Plexiglas barrier at one end of the test tank. Fish exposed to TBT had shorter latency before leaving shoal mates and spent more time away from shoal than control fish. In addition, we also used Novel tanks to study the anxiety behavior as the tendency to stay at the bottom when introduced into an unfamiliar environment. The fish exposed to TBT showed increased anxiety, manifested as increased latency to enter the upper half and decreased time in upper half when compared with the control fish. TBT exposure increased the levels of dopamine and its metabolite 3,4-dihydroxyphenylacetic acid, and decreased the levels of 5-hydroxytryptamine and its metabolite 5-hydroxy indole acetic acid in the brain. Thus, the hypofunction of the dopaminergic system or of the serotoninergic system or the combination of the two may underlie the observed behavioral change, which might affect the fitness of fish in their natural environment. Copyright © 2016 Elsevier B.V. All rights reserved.

Inter-laboratory comparison of cell lines for susceptibility to three viruses: VHSV, IHNV and IPNV

DEFF Research Database (Denmark)

Lorenzen, Ellen; Carstensen, Bendix; Olesen, Niels Jørgen

1999-01-01

Eleven European National Reference Laboratories participated in an inter-laboratory comparison of the susceptibility of 5 selected cell lines to 3 fish pathogenic viruses. The test included viral hemorrhagic septicaemia virus (VHSV), infectious hematopoietic necrosis virus (IHNV) and infectious...... pancreatic necrosis Virus (IPNV), and the cell lines derived from bluegill fry (BF-2), chinook salmon embryo (CHSE-214), epithelioma papulosum cyprini (EPC), fathead minnow (FHM) and rainbow trout gonad (RTG-2). The results showed that for isolation of VHSV, BF-2 and RTG-2 cells performed equally well...

Comparison of catch per unit effort among four minnow trap models in the three-spined stickleback (Gasterosteus aculeatus) fishery.

Science.gov (United States)

Budria, Alexandre; DeFaveri, Jacquelin; Merilä, Juha

2015-12-21

Minnow traps are commonly used in the stickleback (Gasterostidae) fishery, but the potential differences in catch per unit effort (CPUE) among different minnow trap models are little studied. We compared the CPUE of four different minnow trap models in field experiments conducted with three-spined sticklebacks (Gasterosteus aculeatus). Marked (up to 26 fold) differences in median CPUE among different trap models were observed. Metallic uncoated traps yielded the largest CPUE (2.8 fish/h), followed by metallic black nylon-coated traps (1.3 fish/h). Collapsible canvas traps yielded substantially lower CPUEs (black: 0.7 fish/h; red: 0.1 fish/h) than the metallic traps. Laboratory trials further revealed significant differences in escape probabilities among the different trap models. While the differences in escape probability can explain at least part of the differences in CPUE among the trap models (e.g. high escape rate and low CPUE in red canvas traps), discrepancies between model-specific CPUEs and escape rates suggests that variation in entrance rate also contributes to the differences in CPUE. In general, and in accordance with earlier data on nine-spined stickleback (Pungitius pungitius) trapping, the results suggest that uncoated metallic (Gee-type) traps are superior to the other commonly used minnow trap models in stickleback fisheries.

Epigenetic Transgenerational Actions of Vinclozolin on the Development of Disease and Cancer

Science.gov (United States)

Skinner, Michael K.; Anway, Matthew D.

2018-01-01

Exposure to an environmental endocrine disruptor (e.g., vinclozolin) during embryonic gonadal sex determination appears to alter the male germ line epigenome and subsequently promotes transgenerational adult onset disease. The epigenetic mechanism involves the induction of new imprinted-like genes/DNA sequences in the germ line that appear to transmit disease phenotypes. The disease phenotypes include testis abnormalities, prostate disease, kidney disease, immune abnormalities, and tumor development. This epigenetic transgenerational disease mechanism provides a unique perspective from which to view inheritable adult onset disease states, such as cancer, and ultimately offers new insights into novel diagnostic and therapeutic strategies. PMID:17956218

Impaired swim bladder inflation in early-life stage fathead minnows exposed to a deiodinase inhibitor, iopanoic acid (presentation)

Science.gov (United States)

The present study investigated whether inhibition of deiodinase, the enzyme which converts thyroxine (T4) to the more biologically-active form, 3,5,3'-triiodothyronine (T3), would impact inflation of the posterior and/or anterior chamber of the swim bladder, processes previously ...

Impaired swim bladder inflation in early-life stage fathead minnows exposed to a deiodinase inhibitor, iopanoic acid (article)

Science.gov (United States)

The present study investigated whether inhibition of deiodinase, the enzyme which converts thyroxine (T4) to the more biologically-active form, 3,5,3'-triiodothyronine (T3), would impact inflation of the posterior and/or anterior chamber of the swim bladder, processes previously ...

Morphological alterations in the freshwater rotifer Brachionus calyciflorus Pallas 1766 (Rotifera: Monogononta) caused by vinclozolin chronic exposure.

Science.gov (United States)

Alvarado-Flores, Jesús; Rico-Martínez, Roberto; Adabache-Ortíz, Araceli; Silva-Briano, Marcelo

2015-05-01

Vinclozolin (VZ) is a dicarboximide fungicide widely used on fruits, vegetables and wines, effective against fungi plagues. In this study we characterized the effects of VZ using a 4-day reproductive chronic assay with the freshwater rotifer Brachionus calyciflorus. The assay included observations of several features of asexual and sexual reproduction. Our results indicate that VZ: (a) increased asexual and sexual reproduction, (b) caused severe abnormality in females and (c) these abnormalities were inherited by sexual and asexual reproduction. At 1.2 mg/L three abnormal females were found out of 457 total females (0.66 %). This low percentage is consistent and reproducible according to further analysis, where we increased the number of replicates and total females exposed to 1.2 mg/L of VZ, and found 18 abnormal females out of 2868 total females (0.63 % abnormality). Interestingly, abnormal females found at 5.6 mg/L VZ exposure, were able to show mating behavior. Our results suggest that VZ behaves as a strong endocrine disruptor whose effects show the characteristic inverted-U-shape exposure concentration response curve regarding the intrinsic population increase and the percentage of abnormalities as endpoints.

Transgenerational epigenetic effects of the endocrine disruptor vinclozolin on pregnancies and female adult onset disease

OpenAIRE

Nilsson, Eric E; Anway, Matthew D; Stanfield, Jacob; Skinner, Michael K

2008-01-01

Endocrine disruptor exposure during gonadal sex determination was previously found to induce male rat adult onset transgenerational disease (F1–F4 generation), and this was associated with an alteration in the epigenetic (i.e., DNA methylation) programming of the male germ line. The current study was designed to characterize the transgenerational disease phenotypes of the female adult offspring. Pregnant rats (F0 generation) were treated transiently with vinclozolin (i.e., fungicide with anti...

Physiological Responses in Chinese Rare Minnow Larvae Following Exposure to Low-Dose Tributyltin.

Science.gov (United States)

Li, Ping; Li, Zhi-Hua

2015-11-01

In the present study, the antioxidant response and acetylcholinesterase (AChE) activity were measured in Chinese rare minnow larvae (Gobiocypris rarus) after exposure to tributyltin (TBT) (0, 100, 400 and 800 ngL(-1)) for 7 days, as well as the expression of a series of genes, including cr, aptase and prl genes involved in the ion-regulatory process and igfbp3 and gh related to growth rate. Results shows that oxidative stress was generated in fish exposed to TBT, as evidenced by elevated malondialdehyde levels and the inhibition of antioxidant parameters. The activity of acetylcholinesterase (AChE) was also inhibited in fish under higher TBT stress. Moreover, genes involved in ion regulation and growth were affected, based on the regulated transcription of the cr, atpase, gh, prl and igfbp3 genes in the treated groups. The observed effects of TBT upon antioxidant responses and altered expression of genes provides insight into the use of these molecular biomarkers in evaluating mechanisms of TBT toxicity in fish.

Learning deficits expressed as delayed extinction of a conditioned running response following perinatal exposure to vinclozolin.

Science.gov (United States)

André, Susan M; Markowski, Vincent P

2006-01-01

Vinclozolin (Vz) is one member of a group of fungicides whose metabolites are androgen receptor antagonists. These fungicides have been shown to block androgen-driven development and compromise reproductive function. The current study sought to determine if Vz also affects learning following exposure to low doses during the perinatal period. To test this, an androgen-dependent behavior was examined, the extinction of a previously reinforced running response. Pregnant Long-Evans rats were administered a daily oral dose of 0, 1.5, 3, 6 or 12 mg/kg Vz from the 14th day of gestation through postnatal day 3. After reaching adulthood, male and female offspring were trained to run through a short alleyway for food reinforcement. Acquisition of the response was not affected by Vz exposure. However, males required more trials than females for response extinction once food was no longer available in the apparatus. Males exposed to 6 or 12 mg/kg Vz failed to show any extinction by the end of the procedure, while the lowest dose of Vz appeared to facilitate extinction in both male and female offspring. These results demonstrate that endocrine disrupting antiandrogens can alter nervous system development in addition to the reproductive system.

Quantitative PCR Assays for Detecting Loach Minnow (Rhinichthys cobitis and Spikedace (Meda fulgida in the Southwestern United States.

Directory of Open Access Journals (Sweden)

Joseph C Dysthe

Full Text Available Loach minnow (Rhinichthys cobitis and spikedace (Meda fulgida are legally protected with the status of Endangered under the U.S. Endangered Species Act and are endemic to the Gila River basin of Arizona and New Mexico. Efficient and sensitive methods for monitoring these species' distributions are critical for prioritizing conservation efforts. We developed quantitative PCR assays for detecting loach minnow and spikedace DNA in environmental samples. Each assay reliably detected low concentrations of target DNA without detection of non-target species, including other cyprinid fishes with which they co-occur.

OECD validation of the Hershberger assay in Japan: phase 2 dose response of methyltestosterone, vinclozolin, and p,p'-DDE.

Science.gov (United States)

Yamasaki, Kanji; Sawaki, Masakuni; Ohta, Ryo; Okuda, Hirokazu; Katayama, Seiichi; Yamada, Tomoya; Ohta, Takafumi; Kosaka, Tadashi; Owens, William

2003-01-01

The Organisation for Economic Co-operation and Development has initiated the development of new guidelines for the screening and testing of potential endocrine disruptors. The Hershberger assay is one of the assays selected for validation based on the need for in vivo screening to detect androgen agonists or antagonists by measuring the response of five sex accessory organs and tissues of castrated juvenile male rats: the ventral prostate, the seminal vesicles with coagulating glands, the levator ani and bulbocavernosus muscle complex, the Cowper's glands, and the glans penis. The phase 1 feasibility demonstration stage of the Hershberger validation program has been successfully completed with a single androgen agonist and a single antagonist as reference substances. The phase 2 validation program employs a range of additional androgen agonists and antagonists as well as 5alpha-reductase inhibitors. Seven Japanese laboratories have contributed phase 2 validation studies of the Hershberger assay using methyltestosterone, vinclozolin, and 2,2-bis (4-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE). The methyltestosterone doses were 0, 0.05, 0.5, 5, and 50 mg/kg/day, and the vinclozolin and p,p'-DDE doses were 0, 3, 10, 30, and 100 mg/kg/day. All chemicals were orally administered by gavage for 10 consecutive days. In the antagonist version of the assay using vinclozolin and p,p'-DDE, 0.2 mg/kg/day of testosterone propionate was coadministered by subcutaneous injection. All five accessory sex preproductive organs and tissues consistently responded with statistically significant changes in weight within a narrow window. Therefore, the Japanese studies support the Hershberger assay as a reliable and reproducible screening assay for the detection of androgen agonistic and antagonistic effects. PMID:14644666

Influence of environmentally relevant concentrations of vinclozolin on quality, DNA integrity, and antioxidant responses of sterlet Acipenser ruthenus spermatozoa.

Science.gov (United States)

Gazo, Ievgeniia; Linhartova, Pavla; Shaliutina, Anna; Hulak, Martin

2013-04-25

The effects of vinclozolin (VIN), an anti-androgenic fungicide, on quality, oxidative stress, DNA integrity, and ATP level of sterlet (Acipenser ruthenus) spermatozoa were investigated in vitro. Fish spermatozoa were incubated with different concentrations of vinclozolin (0.5, 2, 10, 15, 20 and 50 μg/l) for 2 h. A dose-dependent reduction in spermatozoa motility and velocity was observed at concentrations of 2-50 μg/l. A dramatic increase in DNA fragmentation was recorded at concentrations 10 μg/l and above. After 2 h exposure at higher test concentrations (10-50 μg/l), oxidative stress was apparent, as reflected by significantly higher levels of protein and lipid oxidation and significantly greater superoxide dismutase activity. Intracellular ATP content of spermatozoa decreased with increasing concentrations of VIN. The results demonstrated that VIN can induce reactive oxygen species stress in fish spermatozoa, which could impair the sperm quality, DNA integrity, ATP content, and the antioxidant defense system. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Effect of low dose of vinclozolin on reproductive tract development and sperm parameters in CD1 outbred mice

Czech Academy of Sciences Publication Activity Database

Elzeinová, Fatima; Nováková, Vendula; Buckiová, Daniela; Kubátová, Alena; Pěknicová, Jana

2008-01-01

Roč. 26, 3-4 (2008), s. 231-238 ISSN 0890-6238 R&D Projects: GA MŠk(CZ) 1M06011; GA MŠk 2B06151 Institutional research plan: CEZ:AV0Z50520701; CEZ:AV0Z50390512 Keywords : Vinclozolin * Intra-acrosomal proteins * Reproductive organs Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 2.957, year: 2008

Acute and chronic toxicity of boron to a variety of freshwater organisms.

Science.gov (United States)

Soucek, David J; Dickinson, Amy; Koch, Brian T

2011-08-01

Boron enters the aquatic environment from various sources, including weathering of borates, sewage effluents, coal combustion, use of cleaning compounds, and agrochemicals. The present study was designed to generate data on acute and chronic boron toxicity in support of an update of water quality standards in Illinois, USA. We examined the acute toxicity of boron to eight different freshwater organisms including a fish, an insect, two crustaceans, and four bivalve mollusks. To our knowledge, this is the first study to present data on the toxicity of boron to freshwater mollusks. We also sought to clarify whether hardness or pH affect boron toxicity to aquatic life, and to quantify chronic effect levels in two freshwater species. Sensitivity among the various species ranged widely, with the fathead minnow (Pimephales promelas) being the most sensitive. Neither pH nor hardness had a consistent effect on acute boron toxicity to two crustaceans (Ceriodaphnia dubia and Hyalella azteca), but we observed evidence that chloride reduces boron toxicity to H. azteca. The fathead minnow, while more acutely sensitive than the other species, had a lower acute to chronic ratio than did H. azteca, which had reduced reproduction at 13 mg/L. While we do not know the extent to which the eight tested species represent the range of sensitivities of native but untested species in Illinois, the current water quality standard for Illinois (1 mg/L) is conservative with regard to the native species tested thus far. Copyright © 2011 SETAC.

Acute toxicity of sodium bicarbonate, a major component of coal bed natural gas produced waters, to 13 aquatic species as defined in the laboratory

Science.gov (United States)

Harper, David D.; Farag, Aïda M.; Skaar, Don

2014-01-01

Water produced during coal bed natural gas (CBNG) extraction in the Powder River Structural Basin of Wyoming and Montana (USA) may contain concentrations of sodium bicarbonate (NaHCO3) of more than 3000 mg/L. The authors evaluated the acute toxicity of NaHCO3, also expressed as bicarbonate (HCO3−), to 13 aquatic organisms. Of the 13 species tested, 7 had a median lethal concentration (LC50) less than 2000 mg/L NaHCO3, or 1300 mg/L HCO3−. The most sensitive species were Ceriodaphnia dubia, freshwater mussels (Lampsilis siliquoidea), pallid sturgeon (Scaphirhynchus albus), and shovelnose sturgeon (Scaphirhynchus platorynchus). The respective LC50s were 989 mg/L, 1120 mg/L, 1249 mg/L, and 1430 mg/L NaHCO3, or 699 mg/L, 844 mg/L, 831 mg/L, and 1038 mg/L HCO3−. Age affected the sensitivity of fathead minnows, even within life stage. Two days posthatch, fathead minnows were more sensitive to NaHCO3 and HCO3− compared with 4-d-old fish, even though fish up to 14 d old are commonly used for toxicity evaluations. The authors recommend that ion toxicity exposures be conducted with organisms less than 24 h posthatch to ensure that experiments document the most sensitive stage of development. The results of the present study, along with historical and current research regarding the toxicity of bicarbonate, may be useful to establish regulatory standards for HCO3−.

75 FR 7625 - Endangered and Threatened Wildlife and Plants; Rio Grande Silvery Minnow (Hybognathus amarus...

Science.gov (United States)

2010-02-22

... modification of its habitat due to dewatering and diversion of water, water impoundment, and modification of... thorough knowledge of the Rio Grande silvery minnow's life history, ecology, and behavior, and the current...

Demasculinization of male fish by wastewater treatment plant effluent

Science.gov (United States)

Vajda, A.M.; Barber, L.B.; Gray, J.L.; Lopez, E.M.; Bolden, A.M.; Schoenfuss, H.L.; Norris, D.O.

2011-01-01

Adult male fathead minnows (Pimephales promelas) were exposed to effluent from the City of Boulder, Colorado wastewater treatment plant (WWTP) under controlled conditions in the field to determine if the effluent induced reproductive disruption in fish. Gonadal intersex and other evidence of reproductive disruption were previously identified in white suckers (Catostomus commersoni) in Boulder Creek downstream from this WWTP effluent outfall. Fish were exposed within a mobile flow-through exposure laboratory in July 2005 and August 2006 to WWTP effluent (EFF), Boulder Creek water (REF), or mixtures of EFF and REF for up to 28 days. Primary (sperm abundance) and secondary (nuptial tubercles and dorsal fat pads) sex characteristics were demasculinized within 14 days of exposure to 50% and 100% EFF. Vitellogenin was maximally elevated in both 50% and 100% EFF treatments within 7 days and significantly elevated by 25% EFF within 14 days. The steroidal estrogens 17??-estradiol, estrone, estriol, and 17??-ethynylestradiol, as well as estrogenic alkylphenols and bisphenol A were identified within the EFF treatments and not in the REF treatment. These results support the hypothesis that the reproductive disruption observed in this watershed is due to endocrine-active chemicals in the WWTP effluent. ?? 2011 Elsevier B.V.

MOLECULAR CHARACTERIZATION OF ENDOCRINE DISRUPTION IN FISH USING CDNA ARRAYS.

Science.gov (United States)

We are developing cDNA macroarrays to measure the induction of gene expression in sheepshead minnows and largemouth bass exposed to anthropogenic chemicals that can mimic the action of endogenous hormones. For sheepshead minnows exposed in aqua, we observed similar genetic profil...

Evaluation of a two-generation reproduction toxicity study adding endpoints to detect endocrine disrupting activity using vinclozolin.

Science.gov (United States)

Matsuura, Ikuo; Saitoh, Tetsuji; Ashina, Michiko; Wako, Yumi; Iwata, Hiroshi; Toyota, Naoto; Ishizuka, Yoshihito; Namiki, Masato; Hoshino, Nobuhito; Tsuchitani, Minoru

2005-12-01

A two-generation reproduction toxicity study in rats adding extra endpoints to detect endocrine disrupting activity was conducted using vinclozolin by dietary administration at 0, 40, 200, and 1000 ppm, for investigation of its utility. The extra endpoints included anogenital distance (AGD), nipple development, sexual maturation (vaginal opening and preputial separation), estrous cycle, spermatogenesis, sex organ weights, and blood hormone concentrations (thyroid and sex hormones). Hepatic drug-metabolizing enzyme activities were also measured. The results revealed changes due to vinclozolin in the AGD, nipple development, sexual maturation, sex organ weights, and blood sex hormone concentrations in males of both parental animals and offspring, even at the lowest dose of 40 ppm, confirmed by results for the classical endpoints of histopathological examination at 200 ppm and mating at 1000 ppm. The effects on parental males included increased pituitary and testis weights, and decreased epididymis weights at 1000 ppm in both generations, and decreased prostate and epididymis weights at 200 and 1000 ppm and seminal vesicle weights at 1000 ppm in F1 males. Histopathological examination revealed hypertrophy of the basophilic cells in the pituitary at these two doses, and diffuse hyperplasia of the testicular interstitial cells and atrophy of the seminal vesicle mucosa at 1000 ppm in F0 and F1 males. In addition, F1 males demonstrated decrease in prostate fluid at 200 and 1000 ppm. Blood hormone analysis revealed increases in LH, FSH, testosterone, and DHT in F0 and F1 males at 1000 ppm. General toxicological effects included suppressed body weight gain in F0 and F1 females and in F1 males, and reduced food consumption in F0 and F1 females at 1000 ppm. Histopathological examination revealed centrilobular hepatocellular hypertrophy in males at 200 and 1000 ppm and in females at 1000 ppm, increased lipid droplets in the adrenal zona fasciculata and zona glomerulosa in

Alternative responses to predation in two headwater stream minnows is reflected in their contrasting diel activity patterns.

Science.gov (United States)

Kadye, Wilbert T; Booth, Anthony J

2014-01-01

Animals exhibit diel periodicity in their activity in part to meet energy requirements whilst evading predation. A competing hypothesis suggests that partitioning of diel activities is less important because animals capitalise on opportunity. To test these hypotheses we examined the diel activity patterns for two cyprinid minnows, chubbyhead barb Barbus anoplus and the Eastern Cape redfin minnow Pseudobarbus afer that both occur within headwater streams in the Eastern Cape, South Africa. Chubbyhead barbs exhibited consistent nocturnal activity based on both field and laboratory observations. Due to the absence of fish predators within its habitat, its nocturnal behaviour suggests a response to the cost associated with diurnal activity, such as predation risk by diving and wading birds. In contrast, redfin minnows showed high diurnal activity and a shoaling behaviour in the wild, whereas, in the laboratory, they showed high refuge use during the diel cycle. Despite their preference for refuge in the laboratory, they were diurnally active, a behaviour that was consistent with observations in the wild. The diurnal activity of this species suggests a response to the cost associated with nocturnal activity. Such a cost could be inferred from the presence of the longfin eel, a native predator that was active at night, whereas the daytime shoaling behaviour suggests an anti-predator mechanism to diurnal visual predators. The implications of these findings relate to the impacts associated with the potential invasions by non-native piscivores that occur in the mainstem sections. Diurnal activity patterns for redfin minnows, that are IUCN-listed as endangered, may, in part, explain their susceptibility to high predation by visual non-native piscivores, such as bass and trout. In contrast, the nocturnal habits of chubbyhead barbs suggest a probable pre-adaptation to visual predation. The likelihood of invasion by nocturnally-active sharptooth catfish Clarias gariepinus

Alternative responses to predation in two headwater stream minnows is reflected in their contrasting diel activity patterns.

Directory of Open Access Journals (Sweden)

Wilbert T Kadye

Full Text Available Animals exhibit diel periodicity in their activity in part to meet energy requirements whilst evading predation. A competing hypothesis suggests that partitioning of diel activities is less important because animals capitalise on opportunity. To test these hypotheses we examined the diel activity patterns for two cyprinid minnows, chubbyhead barb Barbus anoplus and the Eastern Cape redfin minnow Pseudobarbus afer that both occur within headwater streams in the Eastern Cape, South Africa. Chubbyhead barbs exhibited consistent nocturnal activity based on both field and laboratory observations. Due to the absence of fish predators within its habitat, its nocturnal behaviour suggests a response to the cost associated with diurnal activity, such as predation risk by diving and wading birds. In contrast, redfin minnows showed high diurnal activity and a shoaling behaviour in the wild, whereas, in the laboratory, they showed high refuge use during the diel cycle. Despite their preference for refuge in the laboratory, they were diurnally active, a behaviour that was consistent with observations in the wild. The diurnal activity of this species suggests a response to the cost associated with nocturnal activity. Such a cost could be inferred from the presence of the longfin eel, a native predator that was active at night, whereas the daytime shoaling behaviour suggests an anti-predator mechanism to diurnal visual predators. The implications of these findings relate to the impacts associated with the potential invasions by non-native piscivores that occur in the mainstem sections. Diurnal activity patterns for redfin minnows, that are IUCN-listed as endangered, may, in part, explain their susceptibility to high predation by visual non-native piscivores, such as bass and trout. In contrast, the nocturnal habits of chubbyhead barbs suggest a probable pre-adaptation to visual predation. The likelihood of invasion by nocturnally-active sharptooth catfish

Vinclozolin: 3-(3,5-dichlorophenyl-5-ethenyl-5-methyl-1,3-oxazolidine-2,4-dione

Directory of Open Access Journals (Sweden)

Seonghwa Cho

2014-07-01

Full Text Available In the title compound, C12H9Cl2NO3, which is the fungicide vinclozolin, the dihedral angle between the oxazolidine ring mean plane [r.m.s. deviation = 0.029 Å] and the benzene ring is 77.55 (8°. In the crystal, molecules are linked via C—H...O hydrogen bonds, forming chains along [010]. The chains are linked by short Cl...Cl contacts [3.4439 (3 and 3.5798 (3 Å], resulting in a three-dimensional architecture.

Life-cycle toxicity of bis(tributyltin) oxide to the sheepshead minnow (Cyprinodon variegatus).

Science.gov (United States)

Manning, C S; Lytle, T F; Walker, W W; Lytle, J S

1999-08-01

The effects of tributyltin (TBT) to the life cycle of the estuarine fish Cyprinodon variegatus were examined in a 180-day flow-through exposure. The study was initiated with embryos less than 24 h postfertilization and monitored through hatch, maturation, growth, and reproduction under continuous exposure to mean measured TBT concentrations of 5.4, 3.2, 1.3, 0.66, and 0.41 microg TBT/L. Progeny isolated at the onset of reproduction by the parental generation (F0) were monitored for survival as embryos, survival as fry/juveniles, and growth 30 days postisolation. TBT, at a concentration of 5.4 microg/L, significantly reduced embryo survival of the F0 generation sheepshead minnows. By day 145 of the exposure, significant effects to the survival of this generation increased and included all test concentrations >/=0.66 microg/L. Survival of 0.66 microg/L was reduced 59% relative to control survival by termination of the F0 generation. Growth of F0 generation organisms as measured by standard length was significantly reduced only on day 90 at 3.2 microg/L, however no significant reductions of wet or dry weight related to treatment concentration were detected. Due to complete mortality of organisms exposed to 5.4 microg/L by study day 7, effects to fecundity and progeny were monitored at measured concentrations of 3.2, 1.3, 0.66, and 0.41 microg TBT/L. Fecundity, as measured by the production of viable eggs produced per female per day, was unaffected by any of the test treatments. All F1 generation embryos isolated from treatment chambers into 3.2 microg/L died. Survival, standard length, wet and dry weight of the F1 generation at the remaining treatment concentrations were unaffected. The results of this study indicate that exposure to TBT reduced survival of the F0 generation sheepshead minnow and establishes the lowest observed effect concentration (LOEC) as 0.66 microg TBT/L, and the no observed effect concentration (NOEC) as 0.41 microg TBT/L for this species

Assessment of 17α-ethinylestradiol effects and underlying mechanisms in a continuous, multigeneration exposure of the Chinese rare minnow (Gobiocypris rarus)

International Nuclear Information System (INIS)

Zha Jinmiao; Sun Liwei; Zhou Yiqi; Spear, Philip A.; Ma, Mei; Wang Zijian

2008-01-01

17α-Ethinylestradiol (EE 2 ) is a synthetic estrogen used primarily in birth control pills and in hormone replacement therapy. Owing to its occurrence in surface waters at concentrations frequently greater than 1 ng/l and its projected future use, EE 2 is expected to pose a significant risk to aquatic organisms. This study was conducted to obtain long-term exposure data necessary for the establishment of water quality criteria and to investigate mechanisms associated with toxic effects. In a multigeneration experiment, Chinese rare minnows (Gobiocypris rarus) were constantly exposed to environmentally relevant concentrations of the synthetic estrogen EE 2 . Mortality, deformities, reproductive parameters, plasma vitellogenin and histopathology were assessed. The results showed that, in the F 0 generation, all endpoints were significantly affected at concentrations higher than 0.2 ng/l EE 2 . No F 1 phenotypic males developed to maturity at 0.2 ng/l and, when adult females of this exposure group were crossed with unexposed males, no F 2 fertile eggs were produced. Kidney histopathology and ultrastructure suggest anomalies possibly associated with increased vitellogenin accumulation. We concluded that the reproduction of the F 1 minnows was completely inhibited at the lowest concentration tested, 0.2 ng/l EE 2 , a concentration frequently detected in surface waters. Growth effects may be related to increased energy requirements including the energy used in VTG synthesis. Reproductive effects are presumably associated with male feminization and the occurrence of testis-ova in males; however, ovarian degeneration observed in females may also have contributed to reproductive failure

Pathway-based approaches for assessment of real-time exposure to an estrogenic wastewater treatment plant effluent on fathead minnow reproduction

Science.gov (United States)

Wastewater treatment plant (WWTP) effluents are known contributors of chemical mixtures into the environment. Of particular concern are endocrine-disrupting compounds, such as estrogens, that can affect hypothalamic-pituitary-gonadal axis function in exposed organisms. The presen...

Predicting acute aquatic toxicity of structurally diverse chemicals in fish using artificial intelligence approaches.

Science.gov (United States)

Singh, Kunwar P; Gupta, Shikha; Rai, Premanjali

2013-09-01

The research aims to develop global modeling tools capable of categorizing structurally diverse chemicals in various toxicity classes according to the EEC and European Community directives, and to predict their acute toxicity in fathead minnow using set of selected molecular descriptors. Accordingly, artificial intelligence approach based classification and regression models, such as probabilistic neural networks (PNN), generalized regression neural networks (GRNN), multilayer perceptron neural network (MLPN), radial basis function neural network (RBFN), support vector machines (SVM), gene expression programming (GEP), and decision tree (DT) were constructed using the experimental toxicity data. Diversity and non-linearity in the chemicals' data were tested using the Tanimoto similarity index and Brock-Dechert-Scheinkman statistics. Predictive and generalization abilities of various models constructed here were compared using several statistical parameters. PNN and GRNN models performed relatively better than MLPN, RBFN, SVM, GEP, and DT. Both in two and four category classifications, PNN yielded a considerably high accuracy of classification in training (95.85 percent and 90.07 percent) and validation data (91.30 percent and 86.96 percent), respectively. GRNN rendered a high correlation between the measured and model predicted -log LC50 values both for the training (0.929) and validation (0.910) data and low prediction errors (RMSE) of 0.52 and 0.49 for two sets. Efficiency of the selected PNN and GRNN models in predicting acute toxicity of new chemicals was adequately validated using external datasets of different fish species (fathead minnow, bluegill, trout, and guppy). The PNN and GRNN models showed good predictive and generalization abilities and can be used as tools for predicting toxicities of structurally diverse chemical compounds. Copyright © 2013 Elsevier Inc. All rights reserved.

Potential population and assemblage influences of non-native trout on native nongame fish in Nebraska headwater streams

Science.gov (United States)

Turek, Kelly C.; Pegg, Mark A.; Pope, Kevin L.; Schainost, Steve

2014-01-01

Non-native trout are currently stocked to support recreational fisheries in headwater streams throughout Nebraska. The influence of non-native trout introductions on native fish populations and their role in structuring fish assemblages in these systems is unknown. The objectives of this study were to determine (i) if the size structure or relative abundance of native fish differs in the presence and absence of non-native trout, (ii) if native fish-assemblage structure differs in the presence and absence of non-native trout and (iii) if native fish-assemblage structure differs across a gradient in abundances of non-native trout. Longnose dace Rhinichthys cataractae were larger in the presence of brown trout Salmo trutta and smaller in the presence of rainbow trout Oncorhynchus mykiss compared to sites without trout. There was also a greater proportion of larger white suckers Catostomus commersonii in the presence of brown trout. Creek chub Semotilus atromaculatus and fathead minnow Pimephales promelas size structures were similar in the presence and absence of trout. Relative abundances of longnose dace, white sucker, creek chub and fathead minnow were similar in the presence and absence of trout, but there was greater distinction in native fish-assemblage structure between sites with trout compared to sites without trout as trout abundances increased. These results suggest increased risk to native fish assemblages in sites with high abundances of trout. However, more research is needed to determine the role of non-native trout in structuring native fish assemblages in streams, and the mechanisms through which introduced trout may influence native fish populations.

Development of partial life-cycle experiments to assess the effects of endocrine disruptors on the freshwater gastropod Lymnaea stagnalis: a case-study with vinclozolin.

Science.gov (United States)

Ducrot, Virginie; Teixeira-Alves, Mickaël; Lopes, Christelle; Delignette-Muller, Marie-Laure; Charles, Sandrine; Lagadic, Laurent

2010-10-01

Long-term effects of endocrine disruptors (EDs) on aquatic invertebrates remain difficult to assess, mainly due to the lack of appropriate sensitive toxicity test methods and relevant data analysis procedures. This study aimed at identifying windows of sensitivity to EDs along the life-cycle of the freshwater snail Lymnaea stagnalis, a candidate species for the development of forthcoming test guidelines. Juveniles, sub-adults, young adults and adults were exposed for 21 days to the fungicide vinclozolin (VZ). Survival, growth, onset of reproduction, fertility and fecundity were monitored weekly. Data were analyzed using standard statistical analysis procedures and mixed-effect models. No deleterious effect on survival and growth occurred in snails exposed to VZ at environmentally relevant concentrations. A significant impairment of the male function occurred in young adults, leading to infertility at concentrations exceeding 0.025 μg/L. Furthermore, fecundity was impaired in adults exposed to concentrations exceeding 25 μg/L. Biological responses depended on VZ concentration, exposure duration and on their interaction, leading to complex response patterns. The use of a standard statistical approach to analyze those data led to underestimation of VZ effects on reproduction, whereas effects could reliably be analyzed by mixed-effect models. L. stagnalis may be among the most sensitive invertebrate species to VZ, a 21-day reproduction test allowing the detection of deleterious effects at environmentally relevant concentrations of the fungicide. These results thus reinforce the relevance of L. stagnalis as a good candidate species for the development of guidelines devoted to the risk assessment of EDs.

MAXIMIZING GROWTH AND SEXUAL MATURATION OF SHEEPSHEAD MINNOWS IN SUPPORT OF MULTI-GENERATION TEST DEVELOPMENT

Science.gov (United States)

Hemmer, R.L., G.M. Cripe and L.R. Goodman. In press. Maximizing Growth and Sexual Maturation of Sheepshead Minnows (Cyprinodon variegatus) in Support of Multi-Generation Test Development (Abstract). To be presented at the SETAC Fourth World Congress, 14-18 November 2004, Portland...

Vinclozolin modulates hepatic cytochrome P450 isoforms during pregnancy.

Science.gov (United States)

de Oca, Félix Genoveva García-Montes; López-González, Ma de Lourdes; Escobar-Wilches, Derly Constanza; Chavira-Ramírez, Roberto; Sierra-Santoyo, Adolfo

2015-06-01

Vinclozolin (V) is classified as a potent endocrine disruptor. The aim of the present study was to determine the effects of V on rat liver CYP regulation and on serum levels of testosterone and estradiol during pregnancy. Pregnancy decreased the liver total CYP content by 65%, enzyme activities of MROD, PROD, and PNPH, and testosterone hydroxylation activities, as well as the protein content of CYP2A and 3A. V exposure remarkably induced the protein content and enzyme activities of CYP1A, 2A, 2B and 3A subfamilies. Testosterone and estradiol were affected in an opposite manner, provoking a 3.5-fold increase in the estradiol/testosterone ratio. These results suggest that V could regulate the hepatic CYP expression through interaction with receptors and coactivators involved in its expression and may play an important role in hormonal balance during pregnancy. In addition, the results may also contribute to understanding the toxicity of V by in utero exposure. Copyright © 2015 Elsevier Inc. All rights reserved.

Personality differences in two minnow populations that differ in their parasitism and predation risk

Directory of Open Access Journals (Sweden)

Raine eKortet

2015-02-01

Full Text Available Animals are often individually consistent in their behavior, not only over time, but also across different functional contexts. Recent research has focused on phenotypic and evolutionary mechanisms explaining such personality differences through selection. Parasitism and predation induce important mortality and fitness costs, and are thus the main candidates to create and maintain personality differences in the wild. Here, we present data on the behavioral consistency of the Eurasian minnow (Phoxinus phoxinus from two populations that live in different tributaries of the same river, but whose ecological environment differs fundamentally with regard to predation and parasitism. We experimentally demonstrate that minnow in both study populations are consistent in their boldness and activity. However, the two study populations differ notably: in the high predation and parasitism risk population fish show higher mean boldness, but tend to be less active than fish in low predation and parasitism risk population. Parasite (Diplostomum phoxini load was negatively, but not statistically significantly, associated with fish activity level. Our study suggests that parasitism and predation are likely important agents in the ecology and evolution of animal personalities.

The stability of dichlofluanid and vinclozolin and their influence on the quality of strawberries.

Science.gov (United States)

Davídek, J; Hajslová, J; Svobodová, Z

1981-05-01

The content of dichlofluanid and vinclozolin found on strawberries treated with Euoparen and/or Ronilan, respectively, did not exceed the residue tolerance. Further decrease of the amount of these fungicides occurred during heatsterilisation of the fruit and storage of the products. The rate of decomposition of dichlofluanid and/or vincolozolin in model solutions (pH 3.0-6.0) followed pseudo-first-order kinetics. The taste and flavour of untreated fruit was preferred to those of strawberries treated with fungicides. There were differences in the content of reducing sugars, volatile fatty acids and titrable acidity in individual samples. Gas-chromatographic profiles of volatile substances isolated from treated and untreated berries also differed.

Vinclozolin: 3-(3,5-di-chloro-phen-yl)-5-ethenyl-5-methyl-1,3-oxazolidine-2,4-dione.

Science.gov (United States)

Cho, Seonghwa; Kim, Jineun; Lee, Sangjin; Kim, Tae Ho

2014-07-01

In the title compound, C12H9Cl2NO3, which is the fungicide vinclozolin, the dihedral angle between the oxazolidine ring mean plane [r.m.s. deviation = 0.029 Å] and the benzene ring is 77.55 (8)°. In the crystal, mol-ecules are linked via C-H⋯O hydrogen bonds, forming chains along [010]. The chains are linked by short Cl⋯Cl contacts [3.4439 (3) and 3.5798 (3) Å], resulting in a three-dimensional architecture.

Endocrine Disruptor Vinclozolin Induced Epigenetic Transgenerational Adult-Onset Disease

Science.gov (United States)

Anway, Matthew D.; Leathers, Charles; Skinner, Michael K.

2018-01-01

The fetal basis of adult disease is poorly understood on a molecular level and cannot be solely attributed to genetic mutations or a single etiology. Embryonic exposure to environmental compounds has been shown to promote various disease states or lesions in the first generation (F1). The current study used the endocrine disruptor vinclozolin (antiandrogenic compound) in a transient embryonic exposure at the time of gonadal sex determination in rats. Adult animals from the F1 generation and all subsequent generations examined (F1–F4) developed a number of disease states or tissue abnormalities including prostate disease, kidney disease, immune system abnormalities, testis abnormalities, and tumor development (e.g. breast). In addition, a number of blood abnormalities developed including hypercholesterolemia. The incidence or prevalence of these transgenerational disease states was high and consistent across all generations (F1–F4) and, based on data from a previous study, appears to be due in part to epigenetic alterations in the male germ line. The observations demonstrate that an environmental compound, endocrine disruptor, can induce transgenerational disease states or abnormalities, and this suggests a potential epigenetic etiology and molecular basis of adult onset disease. PMID:16973726

Evaluation of whole-mount in situ hybridization as a tool for pathway-based toxicological research with early-life stage fathead minnows

Science.gov (United States)

Early-life stage fish can be more sensitive to chemical exposure than adult fish. Therefore, determining possible adverse outcome pathways (AOPs) for early-life stages is crucial. To determine chemical effects and/or mechanisms of action in exposed fish embryos and larvae, whole-...

Zebra mussels (Dreissena polymorpha) limit food for larval fish (Pimephales promelas) in turbulent systems: A bioenergetics analysis

Science.gov (United States)

Bartsch, L.A.; Richardson, W.B.; Sandheinrich, M.B.

2003-01-01

We conducted a factorial experiment, in outdoor mesocosms, on the effects of zebra mussels and water column mixing (i.e., turbulence) on the diet, growth, and survival of larval fathead minnows (Pimephales promelas). Significant (P zebra mussels, whereas mortality was 37% in treatment with turbulence and 17% and 18% in the zebra mussels treatment, and the control, respectively. The size of individual fish was significantly different among treatments at the end of the experiment and was inversely related to survival. Levels of trophic resources (i.e., phyto and zooplankton) varied among treatments and were treatment specific. Turbulent mixing facilitated removal of phytoplankton by zebra mussels by making the entire water column of the tanks available to these benthic filter feeders. Early in the experiment (Day = 0 to 14) the physical process of turbulent mixing likely caused a reduction in standing stocks of zooplankton. The interactive effect of turbulence and mussels reduced copepod and rotifer stocks, through physical processes and through filtration by zebra mussels, relative to the turbulence treatment. The reductions in the number of total zooplankton in the turbulent mixing mesocosms and the further reduction of rotifer and copepod in the turbulence and mussels treatment coincided with a period of increased reliance of larval fathead minnows on these prey. Estimates of consumption from bioenergetics modeling and measured prey standing stocks indicated caloric resources of suitable prey in turbulence treatments during the early weeks of the experiment were insufficient to prevent starvation. Early mortality in the turbulence and mussels treatment likely released surviving fish from intense intraspecific competition and resulted in higher individual growth rates. A combination of high abundance of zebra mussels in an environment with a well-mixed water column can have significant effects on larval fish survival and growth.

Targeting neurotrophic factors and their receptors, but not cholinesterase or neurotransmitter, in the neurotoxicity of TDCPP in Chinese rare minnow adults (Gobiocypris rarus)

International Nuclear Information System (INIS)

Yuan, Lilai; Li, Jiasu; Zha, Jinmiao; Wang, Zijian

2016-01-01

Organophosphate flame retardants (OPFRs) have been detected at high concentrations in various environmental and biotic samples, but little is known about their toxicity. In this study, the potential neurotoxicity of three OPFRs (TCEP, TDCPP, and TPP) and Chlorpyrifos (CPF, an organophosphate pesticide) were compared in Chinese rare minnow using an acute toxicity test and a 21-day fish assay. The acute test demonstrated significant inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) by CPF. Although significant AChE inhibition at high concentration of TPP was also observed, none of the OPFRs had effects similar to CPF on these enzymes, indicating that their acute toxicities to Chinese rare minnow may be unrelated to cholinesterase inhibition. In addition, the 21-day fish assay with TDCPP demonstrated no significant effects on cholinesterase activities or neurotransmitter levels. Nonetheless, this OPFR exhibited widespread effects on the neurotrophic factors and their receptors (e.g., ntf3, ntrk1, ntrk2, ngfr, and fgf2, fgf11, fgf22, fgfr4), indicating that TDCPP or other OPFRs may elicit neurological effects by targeting neurotrophic factors and their receptors in Chinese rare minnow. - Highlights: • Significant inhibition of AChE and BChE activities by CPF was observed. • None of the OPFRs had similar effects on the cholinesterase like the CPF. • TDCPP showed significant effects on the neurotrophic factor genes in rare minnow. - Although none of the tested OPFRs showed any significant effects on cholinesterase activities and neurotransmitter levels, TDCPP did elicit widespread effects on neurotrophic factor genes.

Changes of hematological and biochemical parameters revealed genotoxicity and immunotoxicity of neonicotinoids on Chinese rare minnows (Gobiocypris rarus).

Science.gov (United States)

Hong, Xiangsheng; Zhao, Xu; Tian, Xue; Li, Jiasu; Zha, Jinmiao

2018-02-01

Adverse impacts of immunity in terrestrial non-target organisms exposed to neonicotinoid insecticides have been reported, but the causal link between insecticide exposure and possible immune alterations in fish remains limited. In the present study, the potential genotoxicity and immunotoxicity of three neonicotinoids (imidacloprid, nitenpyram, and dinotefuran) were assessed in Chinese rare minnows by using a 60-day chronic toxicity test. The hematological and biochemical parameters of juvenile Chinese rare minnows and changes in the transcription of six inflammation-related genes were determined after exposure to neonicotinoids at 0.1, 0.5, or 2.0 mg/L. A clear difference in the frequency of erythrocytes with micronuclei (MN) was observed after treatment with 2.0 mg/L imidacloprid (p < .05). Additionally, exposure to 0.5 or 2.0 mg/L imidacloprid significantly increased the binucleated (BN) erythrocytes and those with notched nuclei (NT) (p < .05). A serum protein electrophoresis (SPE) assay showed significant alterations in the serum protein in all treatments (p < .05), and further analysis indicated decreases in immunoglobulin (Ig) in treatments with 0.5 or 2.0 mg/L imidacloprid or dinotefuran or with 0.1 mg/L nitenpyram (p < .05). Moreover, a biochemical assay confirmed that immunoglobulin M (IgM) levels were indeed significantly decreased upon treatment with imidacloprid or dinotefuran at 0.5 or 2.0 mg/L (p < .05). In addition, the transcriptional levels of the inflammatory cytokines IL-6, INF-α, TNF-α, and IL-1β were markedly down-regulated after all imidacloprid treatments (p < .05), whereas the expression levels of only TNF-α and IL-1β were significantly down-regulated following the 0.5 and 2.0 mg/L dinotefuran treatments (p < .05). Taken together, our results clearly demonstrate that imidacloprid, rather than nitenpyram and dinotefuran, can induce genotoxicity. The responsiveness of these immune indicators

Synthetic Fuels Program

International Nuclear Information System (INIS)

Gehrs, C.W.

1978-01-01

Progress is reported on aquatic transport studies with regard to photolysis of polycyclic compounds in water; volatilization of PAH from water; bioaccumulation of anthracene by fathead minnows; bioaccumulation of polycyclic aromatic hydrocarbons by aquatic invertebrates; bioaccumulation of arylamines by zooplankton; availability of sediment-bound trace metals to bluegill; microbial transformation; transport and transformation of anthracene in natural waters; and microcosm studies. Progress is also reported on acute and chronic aquatic effects; acute and chronic terrestrial effects; leaching and chemical and physical characterization of solid wastes; toxicology of solid wastes; and field site task studies with regard to aquatic transport behavior of trace contaminants in wastewater discharges and airborne contaminants at coking plant field site

Microscopic examination of skin in native and nonnative fish from Lake Tahoe exposed to ultraviolet radiation and fluoranthene

Energy Technology Data Exchange (ETDEWEB)

Gevertz, Amanda K., E-mail: agevertz@geiconsultants.com [Miami University, Department of Zoology, 212 Pearson Hall, Oxford 45056, Ohio (United States); GEI Consultants, Inc. , 4601 DTC Blvd, Suite 900, Denver 80237, Colorado (United States); Oris, James T., E-mail: orisjt@miamioh.edu [Miami University, Department of Zoology, 212 Pearson Hall, Oxford 45056, Ohio (United States)

2014-02-15

Highlights: •PAH cause photo-induced toxicity in aquatic organisms in the natural environment. •Montane lakes like Lake Tahoe receive PAH exposure from recreational watercraft. •These lakes are susceptible to invasion and establishment of non-native species. •Non-natives were less tolerant to photo-toxicity compared to native species. •Sensitivity differences were related to levels of oxidative damage in epidermis. -- Abstract: The presence of nonnative species in Lake Tahoe (CA/NV), USA has been an ongoing concern for many decades, and the management of these species calls for an understanding of their ability to cope with the Lake's stressors and for an understanding of their potential to out-compete and reduce the populations of native species. Decreasing levels of ultraviolet radiation (UVR) due to eutrophication and increasing levels of phototoxic polycyclic aromatic hydrocarbons (PAHs) due to recreational activities may combine to affect the relative ability of native versus nonnative fish species to survive in the lake. Following a series of toxicity tests which exposed larvae of the native Lahontan redside minnow (Richardsonius egregius) and the nonnative warm-water bluegill sunfish (Lepomis macrochirus) to UVR and FLU, the occurrence of skin damage and/or physiologic defense mechanisms were studied using multiple microscopic techniques. The native minnow appeared to exhibit fewer instances of skin damage and increased instances of cellular coping mechanisms. This study supports the results of previous work conducted by the authors, who determined that the native redside minnow is the more tolerant of the two species, and that setting and adhering to a water quality standard for UVR transparency may aid in preventing the spread of the less tolerant nonnative bluegill and similar warm-water species.

Toxicological effects of irregularly shaped and spherical microplastics in a marine teleost, the sheepshead minnow (Cyprinodon variegatus).

Science.gov (United States)

Choi, Jin Soo; Jung, Youn-Joo; Hong, Nam-Hui; Hong, Sang Hee; Park, June-Woo

2018-04-01

The increasing global contamination of plastics in marine environments is raising public concerns about the potential hazards of microplastics to environmental and human health. Microplastics formed by the breakdown of larger plastics are typically irregular in shape. The objective of this study was to compare the effects of spherical or irregular shapes of microplastics on changes in organ distribution, swimming behaviors, gene expression, and enzyme activities in sheepshead minnow (Cyprinodon variegatus). Both types of microplastics accumulated in the digestive system, causing intestinal distention. However, when compared to spherical microplastics, irregular microplastics decreased swimming behavior (i.e., total distance travelled and maximum velocity) of sheepshead minnow. Both microplastics generated cellular reactive oxygen species (ROS), while ROS-related molecular changes (i.e., transcriptional and enzymatic characteristics) differed. This study provides toxicological insights into the impacts of environmentally relevant (fragmented) microplastics on fish and improves our understanding of the environmental effects of microplastics in the ecosystem. Copyright © 2018 Elsevier Ltd. All rights reserved.

Androgen receptor-mediated non-genomic effects of vinclozolin on porcine ovarian follicles and isolated granulosa cells: Vinclozolin and non-genomic effects in porcine ovarian follicles.

Science.gov (United States)

Wartalski, Kamil; Knet-Seweryn, Malgorzata; Hoja-Lukowicz, Dorota; Tabarowski, Zbigniew; Duda, Malgorzata

2016-05-01

The present study investigated the influence of the androgen receptor (AR) agonists testosterone (T) and dihydrotestosterone (DHT), and vinclozolin (Vnz), a fungicide with antiandrogenic activity, on non-genomic signal transduction within ovarian follicles. Porcine granulosa cells (GCs) isolated from mature follicles were cultured for 48h. For the last 24h of culture, they were exposed to T (10(-7)M), DHT (10(-7)M), Vnz (1.4×10(-5)M), T and Vnz (T+Vnz), or DHT and Vnz (DHT+Vnz) at the same concentrations. To better imitate in vivo conditions, whole follicles (4-6mm in diameter) were incubated (24h) in an organ culture system with the same factors. Expression of AR mRNA and protein was determined by real-time PCR and western blot analyses. To demonstrate AR localization in cultured GCs and whole follicles, immunocytochemistry and immunohistochemistry were performed, respectively. To elucidate the possible non-genomic action of Vnz in GCs, protein expression and the activity of ERK1/2 and Akt kinases were determined by western blot and ELISA analyses. The immunocytochemistry and immunohistochemistry results showed that exposure of GCs and follicles to Vnz resulted in cytoplasmic and perinuclear AR localization. Real-time PCR and western blot analysis showed that AR mRNA and protein expression increased (P≤0.001) in GC cultures after combined treatment with an androgen and Vnz. In whole follicles, such treatment also increased AR mRNA with a decrease in the respective protein expression (P≤0.001). Moreover, addition of T or DHT with Vnz increased the activity of ERK1/2 and Akt kinases in cultured GCs (P≤0.001). The results suggest a novel mechanism for Vnz action in porcine ovarian follicles on both AR mRNA and protein levels. Thus, this environmental antiandrogen activates non-genomic signaling pathways, as indicated by the increased activity of both investigated kinases observed within minutes of Vnz addition. Given the widespread presence of Vnz in the

Effects of the UV filter benzophenone-2 on reproduction in fish

International Nuclear Information System (INIS)

Weisbrod, Christin J.; Kunz, Petra Y.; Zenker, Armin K.; Fent, Karl

2007-01-01

The UV filter benzophenone-2 (BP-2) is largely used in personal care products such as cosmetics and in numerous other materials for UV protection. Like other UV filters, BP-2 has been found to be estrogenic in vitro and in vivo, but potential effects on reproduction of fish are unknown. In this study, we evaluate whether BP-2 affects important reproductive parameters such as fecundity, gametogenesis and secondary sex characteristics. After a pre-exposure period of 19 days, reproductively mature fathead minnows (Pimephales promelas) were exposed to 0.002, 0.1, 1.2, 5.0 and 9.7 mg/L BP-2 for 15 days. BP-2 was accumulated in fish up to 3.1 μg/g body weight. In males, a dose-dependent vitellogenin induction and decrease in the number of nuptial tubercles occurred. Moreover, significant dose-related effects on gonads of male and female fish were observed. At concentrations of 1.2 mg/L and higher, spermatocyte and oocyte development was significantly inhibited in male and female fish, respectively. Testes of exposed males had much fewer spermatocytes and ovaries of exposed females had much fewer mature and more atretic follicles. Reproduction was negatively affected in a dose-dependent manner with a decrease in egg production at 5.0 mg/L and a complete cessation of spawning activity at 9.7 mg/L BP-2. Our findings show significant estrogenic effects of the common UV filter BP-2 on vitellogenin induction, secondary sex characteristics, gonadal development, and reproduction in fish

Identification and treatment of lithium as the primary toxicant in a groundwater treatment facility effluent

International Nuclear Information System (INIS)

Kszos, L.A.; Crow, K.R.

1996-01-01

6 Li is used in manufacturing nuclear weapons, shielding, and reactor control rods. Li compounds have been used at DOE facilities and Li-contaminated waste has historically been land disposed. Seep water from burial grounds near Y-12 contain small amounts of chlorinated hydrocarbons, traces of PCBs, and 10-19 mg/L Li. Seep treatment consists of oil-water separation, filtration, air stripping, and carbon adsorption. Routine biomonitoring tests using fathead minnows and Ceriodaphniadubia are conducted. Evaluation of suspected contaminants revealed that toxicity was most likely due to Li. Laboratory tests showed that 1 mg Li/L reduced the survival of both species; 0.5 mg Li/L reduced Ceriodaphnia reproduction and minnow growth. However, the toxicity was greatly reduced in presence of sodium (up to 4 mg Li/L, Na can fully negate the toxic effect of Li). Because of the low Na level discharged from the treatment facility, Li removal from the ground water was desired. SuperLig reg-sign columns were used (Li-selective organic macrocycle bonded to silica gel). Bench-scale tests showed that the material was very effective for removing Li from the effluent, reducing the toxicity

Predicting fecundity of fathead minnows (Pimephales promelas) exposed toeEndocrine-disrupting chemicals using a MATLAB®-based model of oocyte growth dynamics

Science.gov (United States)

Fish spawning is often used as an integrated measure of reproductive toxicity, and an indicator of aquatic ecosystem health in the context of forecasting potential population-level effects considered important for ecological risk assessment. Consequently, there is a need for fle...

Selected endocrine disrupting compounds (vinclozolin, flutamide, ketoconazole and dicofol): effects on survival, occurrence of males, growth, molting and reproduction of Daphnia magna.

Science.gov (United States)

Haeba, Maher H; Hilscherová, Klára; Mazurová, Edita; Bláha, Ludek

2008-05-01

Pollution-induced endocrine disruption in vertebrates and invertebrates is a worldwide environmental problem, but relatively little is known about effects of endocrine disrupting compounds (EDCs) in planktonic crustaceans (including Daphnia magna). Aims of the present study were to investigate acute 48 h toxicity and sub-chronic (4-6 days) and chronic (21 days) effects of selected EDCs in D. magna. We have investigated both traditional endpoints as well as other parameters such as sex determination, maturation, molting or embryogenesis in order to evaluate the sensitivity and possible use of these endpoints in ecological risk assessment. We have studied effects of four model EDCs (vinclozolin, flutamide, ketoconazole and dicofol) on D. magna using (i) an acute 48 h immobilization assay, (ii) a sub-chronic, 4-6 day assay evaluating development and the sex ratio of neonates, and (iii) a chronic, 21 day assay studying number of neonates, sex of neonates, molting frequency, day of maturation and the growth of maternal organisms. Acute EC50 values in the 48 h immobilization test were as follows (mg/L): dicofol 0.2, ketoconazole 1.5, flutamide 2.7, vinclozolin >3. Short-term, 4-6 day assays with sublethal concentrations showed that the sex ratio in Daphnia was modulated by vinclozolin (decreased number of neonate males at 1 mg/L) and dicofol (increase in males at 0.1 mg/L). Flutamide (up to 1 mg/L) had no effect on the sex of neonates, but inhibited embryonic development at certain stages during chronic assay, resulting in abortions. Ketoconazole had no significant effects on the studied processes up to 1 mg/L. Sex ratio modulations by some chemicals (vinclozolin and dicofol) corresponded to the known action of these compounds in vertebrates (i.e. anti-androgenicity and anti-oestrogenicity, respectively). Our study revealed that some chemicals known to affect steroid-regulated processes in vertebrates can also affect sublethal endpoints (e.g. embryonic sex determination

Juvenile exposure to vinclozolin shifts sex ratios and impairs reproductive capacity of zebrafish.

Science.gov (United States)

Lor, Yer; Revak, Andrew; Weigand, Jenna; Hicks, Elisabeth; Howard, David R; King-Heiden, Tisha C

2015-12-01

Exposure to endocrine disruptors during critical periods of development can impact the sustainability of wild fish populations. Anti-androgenic compounds have received less attention, but are capable of modulating gonad differentiation and maturation, and impairing reproduction in fish. The fungicide vinclozolin (VZ) has been shown to impair reproduction in adult fish, but less is known about its effects following exposure earlier in development. Here we show that waterborne exposure to 400μg VZ/L during critical periods of sex differentiation (21-35 days post fertilization) permanently shifts sex ratios towards females, and alters the maturation of the gonad. Both fecundity and fertility were reduced, even when oogenesis and spermatogenesis recover and sperm motility is not altered. These results demonstrate the need to better understand the impacts of early exposure to anti-androgenic compounds on fish. Copyright © 2015 Elsevier Inc. All rights reserved.

Classifying chemical mode of action using gene networks and machine learning: a case study with the herbicide linuron.

Science.gov (United States)

Ornostay, Anna; Cowie, Andrew M; Hindle, Matthew; Baker, Christopher J O; Martyniuk, Christopher J

2013-12-01

The herbicide linuron (LIN) is an endocrine disruptor with an anti-androgenic mode of action. The objectives of this study were to (1) improve knowledge of androgen and anti-androgen signaling in the teleostean ovary and to (2) assess the ability of gene networks and machine learning to classify LIN as an anti-androgen using transcriptomic data. Ovarian explants from vitellogenic fathead minnows (FHMs) were exposed to three concentrations of either 5α-dihydrotestosterone (DHT), flutamide (FLUT), or LIN for 12h. Ovaries exposed to DHT showed a significant increase in 17β-estradiol (E2) production while FLUT and LIN had no effect on E2. To improve understanding of androgen receptor signaling in the ovary, a reciprocal gene expression network was constructed for DHT and FLUT using pathway analysis and these data suggested that steroid metabolism, translation, and DNA replication are processes regulated through AR signaling in the ovary. Sub-network enrichment analysis revealed that FLUT and LIN shared more regulated gene networks in common compared to DHT. Using transcriptomic datasets from different fish species, machine learning algorithms classified LIN successfully with other anti-androgens. This study advances knowledge regarding molecular signaling cascades in the ovary that are responsive to androgens and anti-androgens and provides proof of concept that gene network analysis and machine learning can classify priority chemicals using experimental transcriptomic data collected from different fish species. © 2013.

Development of an operational tool for biomonitoring using constant pressure respirometry

International Nuclear Information System (INIS)

Zachritz, W.H. II; Morrow, J.

1992-01-01

The United States Environmental Protection Agency (US EPA) policy statement (FR 49, 9016, 9 March 1984) for the development of water quality based permit limitations includes toxicity testing of effluents as an important part of a water quality approach to controlling toxics (Pickering, 1988). To assure that state waters are free of toxics, both chemical and biological methods were recommended for assessing effluent quality. The US EPA validated bioassay procedures for toxicity testing of wastewater discharges use three distinctive organisms groups: vertebrates, invertebrates, and algae (Weber, 1989). The specific species for these three groups are fathead minnow, Pimephales promelas; water tea, Ceriodaphnia, dubia; and the green algae, Selenastrum capricornum, respectively. Definitive testing estimates the concentration atwhich a certain percentage of organisms exhibit a certain response. The definitive test exposes several replicate groups of organisms to the target substrate for a predetermined time period effluent concentration. The overall goal of this study is to develop an acceptable protocol for operational biomonitoring based on constant pressure respirometry for LANL. The specific objectives include: Development an appropriate toxicity testing protocol based on constant pressure respirometry for whole effluent toxicity testing, and evaluate the protocol based on factors such as sensitivity, response time, cost of analysis, and simplicity of operation

Species specific anaesthetics for fish anaesthesia and euthanasia.

Science.gov (United States)

Readman, Gareth D; Owen, Stewart F; Knowles, Toby G; Murrell, Joanna C

2017-08-02

There is a need to ensure that the care and welfare for fish maintained in the laboratory are to the highest standards. This extends to the use of anaesthetics for both scientific study, humane killing and euthanasia at end of life. An anaesthetic should not induce negative behaviours and fish should not seek to avoid the anaesthetic. Surprisingly little information is available to facilitate a humane choice of anaesthetic agent for fish despite over 100 years of use and the millions of fish currently held in thousands of laboratories worldwide. Using a chemotaxic choice chamber we found different species specific behavioural responses among four closely related fish species commonly held in the laboratory, exposed to three widely used anaesthetic agents. As previously found for zebrafish (Danio rerio), the use of MS-222 and benzocaine also appears to induce avoidance behaviours in medaka (Oryzias latipes); but etomidate could provide an alternative choice. Carp (Cyprinus carpio), although closely related to zebrafish showed avoidance behaviours to etomidate, but not benzocaine or MS-222; and rainbow trout (Oncorhynchus mykiss) showed no avoidance to the three agents tested. We were unable to ascertain avoidance responses in fathead minnows (Pimephales promelas) and suggest different test paradigms are required for that species.

Transcriptomic effects-based monitoring for endocrine active chemicals: Assessing relative contribution of treated wastewater to downstream pollution

Science.gov (United States)

Martinovic-Weigelt, Dalma; Mehinto, Alvine C.; Ankley, Gerald T.; Denslow, Nancy D.; Barber, Larry B.; Lee, Kathy E.; King, Ryan J.; Schoenfuss, Heiko L.; Schroeder, Anthony L.; Villeneuve, Daniel L.

2014-01-01

The present study investigated whether a combination of targeted analytical chemistry information with unsupervised, data-rich biological methodology (i.e., transcriptomics) could be utilized to evaluate relative contributions of wastewater treatment plant (WWTP) effluents to biological effects. The effects of WWTP effluents on fish exposed to ambient, receiving waters were studied at three locations with distinct WWTP and watershed characteristics. At each location, 4 d exposures of male fathead minnows to the WWTP effluent and upstream and downstream ambient waters were conducted. Transcriptomic analyses were performed on livers using 15 000 feature microarrays, followed by a canonical pathway and gene set enrichment analyses. Enrichment of gene sets indicative of teleost brain–pituitary–gonadal–hepatic (BPGH) axis function indicated that WWTPs serve as an important source of endocrine active chemicals (EACs) that affect the BPGH axis (e.g., cholesterol and steroid metabolism were altered). The results indicated that transcriptomics may even pinpoint pertinent adverse outcomes (i.e., liver vacuolization) and groups of chemicals that preselected chemical analytes may miss. Transcriptomic Effects-Based monitoring was capable of distinguishing sites, and it reflected chemical pollution gradients, thus holding promise for assessment of relative contributions of point sources to pollution and the efficacy of pollution remediation.

Contaminants of emerging concern presence and adverse effects in fish: A case study in the Laurentian Great Lakes

Science.gov (United States)

Jorgenson, Zachary G.; Thomas, Linnea M.; Elliott, Sarah M.; Cavallin, Jenna E.; Randolph, Eric C.; Choy, Steven J.; Alvarez, David; Banda, Jo A.; Gefell, Daniel J.; Lee, Kathy E.; Furlong, Edward T.; Schoenfuss, Heiko L.

2018-01-01

The Laurentian Great Lakes are a valuable natural resource that is affected by contaminants of emerging concern (CECs), including sex steroid hormones, personal care products, pharmaceuticals, industrial chemicals, and new generation pesticides. However, little is known about the fate and biological effects of CECs in tributaries to the Great Lakes. In the current study, 16 sites on three rivers in the Great Lakes basin (Fox, Cuyahoga, and Raquette Rivers) were assessed for CEC presence using polar organic chemical integrative samplers (POCIS) and grab water samplers. Biological activity was assessed through a combination of in vitro bioassays (focused on estrogenic activity) and in vivo assays with larval fathead minnows. In addition, resident sunfish, largemouth bass, and white suckers were assessed for changes in

Transcriptome analyses of sex differential gene expression in brains of rare minnow (Gobiocypris rarus and effects of tributyltin exposure

Directory of Open Access Journals (Sweden)

Ji-liang Zhang

2018-06-01

Full Text Available RNA-sequencing was used to identify sex-biased gene expression in brains of rare minnow (Gobiocypris rarus by comparing transcriptomic profiles between females and males. Furthermore, transcriptomic responses to 10 ng/L tributyltin (TBT in both male and female brains were also investigated to understand whether TBT affects the identified sex-biased genes. Differentially expressed genes (DEGs were identified using the IDEG6 web tool. In this article, we presented male- and female-biased DEGs, and up-regulated and down-regulated DEGs after TBT exposure. The raw reads data supporting the present analyses has been deposited in NCBI Sequence Read Archive (SRA, http://www.ncbi.nlm.nih.gov/Traces/sra with accession number PRJNA376634. The data presented in this article are related to the research article entitled “Transcriptomic analyses of sexual dimorphism of rare minnow (G. rarus brains and effects of tributyltin exposure” (doi: 10.1016/j.ecoenv.2018.02.049.

Epigenetic effects of methoxychlor and vinclozolin on male gametes.

Science.gov (United States)

Paoloni-Giacobino, Ariane

2014-01-01

Imprinting is an epigenetic form of gene regulation that mediates a parent-of-origin-dependent expression of the alleles of a number of genes. Imprinting, which occurs at specific sites within or surrounding the gene, called differentially methylated domains, consists in a methylation of CpGs. The appropriate transmission of genomics imprints is essential for the control of embryonic development and fetal growth. A number of endocrine disruptors (EDs) affect male reproductive tract development and spermatogenesis. It was postulated that the genetic effects of EDs might be induced by alterations in gene imprinting. We tested two EDs: methoxychlor and vinclozolin. Their administration during gestation induced in the offspring a decrease in sperm counts and significant modifications in the methylation pattern of a selection of paternally and maternally expressed canonical imprinted genes. The observation that imprinting was largely untouched in somatic cells suggests that EDs exert their damaging effects via the process of reprogramming that is unique to gamete development. Interestingly, the effects were transgenerational, although disappearing gradually from F1 to F3. A systematic analysis showed a heterogeneity in the CpG sensitivity to EDs. We propose that the deleterious effects of EDs on the male reproductive system are mediated by imprinting defects in the sperm. The reported effects of EDs on human male spermatogenesis might be mediated by analogous imprinting alterations. © 2014 Elsevier Inc. All rights reserved.

Alternations in neuroendocrine and endocrine regulation of reproduction in male goldfish (Carassius auratus) following an acute and chronic exposure to vinclozolin, in vivo.

Science.gov (United States)

Golshan, Mahdi; Hatef, Azadeh; Zare, Ava; Socha, Magdalena; Milla, Sylvain; Gosiewski, Grzegorz; Fontaine, Pascal; Sokołowska-Mikołajczyk, Mirosława; Habibi, Hamid R; Alavi, Sayyed Mohammad Hadi

2014-10-01

The fungicide vinclozolin (VZ) is in use globally and known to disrupt reproductive function in male. The present study tested the hypothesis that VZ disrupts testicular function in goldfish (Carassius auratus) by affecting brain-pituitary-testis axis. Goldfish were exposed to 100, 400 and 800 μg/L VZ and 5 μg/L 17β-estradiol (E2) for comparison. In VZ treated goldfish, 11-ketotesteosterone (11-KT) secretion was changed depending on dose and duration period of treatment. Following 7 days of exposure, 11-KT was decreased in goldfish exposed to 800 μg/L VZ, while it was increased in goldfish exposed to 100 μg/L VZ after 30 days of exposure. Circulating E2 level was unchanged in VZ treated goldfish, however the E2/11-KT ratio was increased in a concentration-related manner. In E2 treated goldfish, circulatory 11-KT and E2 levels were decreased and increased, respectively, which resulted in an increase in the E2/11-KT ratio. Exposure to VZ at 100 μg/L caused a significant increase in the circulatory luteinizing hormone (LH) after 30 days. In E2 treated fish circulatory LH was decreased, significantly. Transcripts of genes encoding gonadotropin-releasing hormone and androgen receptor in the brain, and those of genes encoding LH and follicle-stimulating hormone receptors, StAR, CYP17, and 3β-HSD in the testis changed in VZ-treated goldfish depending on concentration and period of treatment. mRNA of genes encoding vitellogenin and estrogen receptor in the liver and cytochrome P450 aromatase in the brain were increased in E2-treated goldfish. The results suggest that VZ-induced changes in 11-KT were due to disruption in brain-pituitary-testis axis and provide integrated characterization of VZ-related reproductive disorders in male fish. Copyright © 2014 Elsevier B.V. All rights reserved.

An intergeneric hybrid of a native minnow, the golden shiner, and an exotic minnow, the rudd

Science.gov (United States)

Burkhead, N.M.; Williams, J.D.

1991-01-01

The hybrid golden shiner Notemigonus crysoleucas × rudd Scardinius erythrophthalmus is the first known nonsalmonid, intergeneric hybrid of an exotic species and a North American native species. The cross is also the first valid record of a viable hybrid involving the native golden shiner. Meristic and mensural characters of 30 artificially produced hybrids of male golden shiners and female rudds were analyzed. Forty-seven percent of the meristic traits exhibited character states intermediate between those of parents. Twenty-seven percent of the meristic characters were supernumerary, suggesting developmental instability of the hybrid genome. Mensural hybrid characters were significantly skewed to the golden shiner phenotype. The skewed mensural inheritance and other skewed patterns of morphological inheritance also suggest problems in canalization of the hybrid phenome or atypical patterns of dominance. All hybrids were identifiable by intermediate squamation of the cultrate abdomen: the keel was mostly scaled but exhibited a small fleshy ridge posteriorly. This minnow hybrid allows general inferences to be made about the phylogenetic affinity of the golden shiner to other cultrate cyprinids of Eurasia. The hybrid cross has important management and conservation implications for fishes in North America. The hybrid is an example of how an exotic species may negatively affect a native species.

In utero and lactational exposure to vinclozolin and genistein induces genomic changes in the rat mammary gland.

Science.gov (United States)

El Sheikh Saad, H; Toullec, A; Vacher, S; Pocard, M; Bieche, I; Perrot-Applanat, M

2013-02-01

Exposure to low doses of environmental estrogens such as bisphenol A and genistein (G) alters mammary gland development. The effects of environmental anti-androgens, such as the fungicide vinclozolin (V), on mammary gland morphogenesis are unknown. We previously reported that perinatal exposure to G, V, and the GV combination causes histological changes in the mammary gland during the peripubertal period, suggesting alterations to the peripubertal hormone response. We now investigate whether perinatal exposure to these compounds alters the gene expression profiles of the developing glands to identify the dysregulated signaling pathways and the underlying mechanisms. G, V, or GV (1 mg/kg body weight per day) was added to diet of Wistar rats, from conception to weaning; female offspring mammary glands were collected at postnatal days (PNDs) 35 and 50. Genes displaying differential expression and belonging to different functional categories were validated by quantitative PCR and immunocytochemistry. At PND35, G had little effect; the slight changes noted were in genes related to morphogenesis. The changes following exposure to V concerned the functional categories associated with development (Cldn1, Krt17, and Sprr1a), carbohydrate metabolism, and steroidogenesis. The GV mixture upregulated genes (Krt17, Pvalb, and Tnni2) involved in muscle development, indicating effects on myoepithelial cells during mammary gland morphogenesis. Importantly, at PND50, cycling females exposed to GV showed an increase in the expression of genes (Csn2, Wap, and Elf5) related to differentiation, consistent with the previously reported abnormal lobuloalveolar development previously described. Thus, perinatal exposure to GV alters the mammary gland hormone response differently at PND35 (puberty) and in animals with established cycles.

Quantitative PCR assays for detecting loach minnow (Rhinichthys cobitis) and spikedace (Meda fulgida) in the southwestern United States

Science.gov (United States)

Joseph C. Dysthe; Kellie J. Carim; Yvette M. Paroz; Kevin S. McKelvey; Michael K. Young; Michael K. Schwartz

2016-01-01

Loach minnow (Rhinichthys cobitis) and spikedace (Meda fulgida) are legally protected with the status of Endangered under the U.S. Endangered Species Act and are endemic to the Gila River basin of Arizona and New Mexico. Efficient and sensitive methods for monitoring these species’ distributions are critical for prioritizing conservation efforts. We developed...

Dissolved organic carbon ameliorates the effects of UV radiation on a freshwater fish

Energy Technology Data Exchange (ETDEWEB)

Manek, Aditya K., E-mail: aditya.manek@usask.ca [Department of Biology, University of Saskatchewan, Saskatoon, S7N 5E2 SK (Canada); Ferrari, Maud C.O. [Department of Biomedical Sciences, WCVM, University of Saskatchewan, Saskatoon, S7N 5B4 SK (Canada); Chivers, Douglas P.; Niyogi, Som [Department of Biology, University of Saskatchewan, Saskatoon, S7N 5E2 SK (Canada)

2014-08-15

Anthropogenic activities over the past several decades have depleted stratospheric ozone, resulting in a global increase in ultraviolet radiation (UVR). Much of the negative effects of UVR in aquatic systems is minimized by dissolved organic carbon (DOC) which is known to attenuate UVR across the water column. The skin of many fishes contains large epidermal club cells (ECCs) that are known to play a role in innate immune responses and also release chemical alarm cues that warn other fishes of danger. This study investigated the effects of in vivo UVR exposure to fathead minnows (Pimephales promelas), under the influence of two sources of DOC: Sigma Aldrich humic acid, a coal based commercial source of DOC and Luther Marsh natural organic matter, a terrigenous source of DOC. Specifically, we examined ECC investment and physiological stress responses and found that fish exposed to high UVR, in the presence of either source of DOC, had higher ECC investment than fish exposed to high UVR only. Similarly, exposure to high UVR under either source of DOC, reduced cortisol levels relative to that in the high UVR only treatment. This indicates that DOC protects fish from physiological stress associated with UVR exposure and helps maintain production of ECC under conditions of UVR exposure. - Highlights: • We examined the combined effect of UV radiation and Dissolved Organic Carbon on fish. • Physiological stress response and epidermal club cell investment were measured. • Fish exposed to high UVR and DOC had higher ECC investment and reduced cortisol levels. • DOC plays a role in protecting fish from physiological stress and maintains ECC production.

Dissolved organic carbon ameliorates the effects of UV radiation on a freshwater fish

International Nuclear Information System (INIS)

Manek, Aditya K.; Ferrari, Maud C.O.; Chivers, Douglas P.; Niyogi, Som

2014-01-01

Anthropogenic activities over the past several decades have depleted stratospheric ozone, resulting in a global increase in ultraviolet radiation (UVR). Much of the negative effects of UVR in aquatic systems is minimized by dissolved organic carbon (DOC) which is known to attenuate UVR across the water column. The skin of many fishes contains large epidermal club cells (ECCs) that are known to play a role in innate immune responses and also release chemical alarm cues that warn other fishes of danger. This study investigated the effects of in vivo UVR exposure to fathead minnows (Pimephales promelas), under the influence of two sources of DOC: Sigma Aldrich humic acid, a coal based commercial source of DOC and Luther Marsh natural organic matter, a terrigenous source of DOC. Specifically, we examined ECC investment and physiological stress responses and found that fish exposed to high UVR, in the presence of either source of DOC, had higher ECC investment than fish exposed to high UVR only. Similarly, exposure to high UVR under either source of DOC, reduced cortisol levels relative to that in the high UVR only treatment. This indicates that DOC protects fish from physiological stress associated with UVR exposure and helps maintain production of ECC under conditions of UVR exposure. - Highlights: • We examined the combined effect of UV radiation and Dissolved Organic Carbon on fish. • Physiological stress response and epidermal club cell investment were measured. • Fish exposed to high UVR and DOC had higher ECC investment and reduced cortisol levels. • DOC plays a role in protecting fish from physiological stress and maintains ECC production

Water- and sediment-quality effects on Pimephales promelas spawning vary along an agriculture-to-urban land-use gradient.

Science.gov (United States)

Corsi, Steven R; Klaper, Rebecca D; Weber, Daniel N; Bannerman, Roger T

2011-10-15

Many streams in the U.S. are "impaired" due to anthropogenic influence. For watershed managers to achieve practical understanding of these impairments, a multitude of factors must be considered, including point and nonpoint-source influence on water quality. A spawning assay was developed in this study to evaluate water- and sediment-quality effects that influenced Pimephales promelas (fathead minnow) egg production over a gradient of urban and agricultural land use in 27 small watersheds in Eastern Wisconsin. Six pairs of reproducing fathead minnows were contained in separate mesh cartridges within one larger flow-through chamber. Water- and sediment quality were sampled for an array of parameters. Egg production was monitored for each pair providing an assessment of spawning success throughout the 21-day test periods. Incidences of low dissolved oxygen (DO) in many of these streams negatively impacted spawning success. Nine of 27 streams experienced DO less than 3.1mg/L and 15 streams experienced DO less than 4.8mg/L. Low DO was observed in urban and agricultural watersheds, but the upper threshold of minimum DO decreased with increasing urban development. An increase in specific conductance was related to a decrease in spawning success. In previous studies for streams in this region, specific conductance had a linear relation with chloride, suggesting the possibility that chloride could be a factor in egg production. Egg production was lower at sites with substantial urban development, but sites with low egg production were not limited to urban sites. Degradation of water- and sediment-quality parameters with increasing urban development is indicated for multiple parameters while patterns were not detected for others. Results from this study indicate that DO must be a high priority watershed management consideration for this region, specific conductance should be investigated further to determine the mechanism of the relation with egg production, and water- and

Toxicity of uranium and plutonium to the developing embryos of fish

International Nuclear Information System (INIS)

Till, J.E.; Kaye, S.V.; Trabalka, J.R.

1976-07-01

The radiological and chemical toxicity of plutonium and uranium to the developing embryos of fish was investigated using eggs from carp, Cyprinus carpio, and fathead minnows, Pimephales promelas. Freshly fertilized eggs were developed in solutions containing high specific activity 238 Pu or 232 U or low specific activity 244 Pu, 235 U, or 238 U. Quantitative tests to determine the penetration of these elements through the chorion indicated that plutonium accumulated in the contents of carp eggs reaching a maximum concentration factor of approximately 3.0 at hatching. Autoradiographs of 16 μ egg sections showed that plutonium was uniformly distributed in the egg volume. Uranium localized in the yolk material, and the concentration factor in the yolk sac remained constant during development at approximately 3.3. Doses from 238 Pu which affected hatchability of the eggs were estimated to be 1.6 x 10 4 rads and 9.7 x 10 3 rads for C. carpio and P. promelas, respectively; doses from 232 U were 1.3 x 10 4 rads for C. carpio and 2.7 x 10 3 rads for P. promelas. A greater number of abnormal larvae than in control groups was produced by 238 Pu doses of 4.3 x 10 3 rads to carp and 5.7 x 10 2 rads to fathead minnows; 3.2 x 10 3 rads and 2.7 x 10 2 rads were estimated from 232 U. Eggs that were incubated in 20 ppM 244 Pu did not hatch. This mortality may have been the result of chemical toxicity of plutonium. Concentrations of 60 ppM of 235 U and 238 U did not affect egg hatching. Based on these data, concentrations in fish eggs were calculated for representative concentrations of uranium and plutonium in natural waters and the corresponding dose levels are below those levels at which observable effects begin to occur

Toxicity of uranium and plutonium to the developing embryos of fish

International Nuclear Information System (INIS)

Till, J.E.

1976-01-01

The radiological and chemical toxicity of plutonium and uranium to the developing embryos of fish was investigated using eggs from carp, Cyprinus carpio, and fathead minnows, Pimephales promelas. Freshly fertilized eggs were developed in solutions containing high specific activity 238 Pu or 232 U or low specific activity 244 Pu, 235 U, or 238 U. Quantitative tests to determine the penetration of these elements through the chorion indicated that plutonium accumulated in the contents of carp eggs reached a maximum concentration factor of approximately 3.0 at hatching. Autoradiographs of 16 μ egg sections showed that plutonium was uniformly distributed in the egg volume. Uranium localized in the yolk material, and the concentration factor in the yolk sac remained constant during development at approximately 3.3. Doses from 238 Pu which affected hatchability of the eggs were estimated to be 1.6 x 10 4 rads and 9.7 x 10 3 rads for C. carpio and P. promelas, respectively; doses from 232 U were 1.3 x 10 4 rads for C. carpio and 2.7 x 10 3 rads for P. promelas. A greater number of abnormal larvae than in control groups was produced by 238 Pu doses of 4.3 x 10 3 rads to carp and 5.7 x 10 2 rads to fathead minnows; 3.2 x 10 3 rads and 2.7 x 10 2 rads were estimated from 232 U. Eggs that were incubated in 20 ppM 244 Pu did not hatch. This mortality may have been the result of chemical toxicity of plutonium. Concentrations of 60 ppM of 235 U and 238 U did not affect egg hatching. Based on these data, concentrations in fish eggs were calculated for representative concentrations of uranium and plutonium in waste waters and the corresponding dose levels are below those levels at which observable effects begin to occur

Method for aquatic multiple species toxicant testing: acute toxicity of 10 chemicals to 5 vertebrates and 2 invertebrates. [Pimephales promelas; Carassius auratus; Ictalurus punctatus; Lepomis macrochirus; Salmo gairdneri; Orconectes immunis; Aplexa hypnorum

Energy Technology Data Exchange (ETDEWEB)

Phipps, G.L.; Holcombe, G.W.

1985-01-01

A method was developed to simultaneously ascertain 96 h LC/sub 50/ values for seven freshwater species in a single flow through test with measured concentrations. It allows interspecific comparisons, easy determination of the most sensitive species, and cuts cost of labor, materials and chemical analysis for measured concentration tests. Species tested included fathead minnows Pimephales promelas, goldfish Carassius auratus, channel catfish Ictalurus punctatus, bluegill Lepomis macrochirus, rainbow trout Salmo gairdneri, crayfish Orconectes immunis and snails Aplexa hypnorum. Compounds tested were pentachlorophenol, 2-chloroethanol, 2,4-pentanedione, hexachloroethane, ..cap alpha..-bromo-2',5'-dimethoxyacetophenone, benzaldehyde, 1,3-dichloro-4,6-dinitro-benzene, dursban, sevin and cadmium chloride. The LC/sub 50/ values from these multiple species tests compared favourably with those determined using single species tests at this laboratory, usually within 20%.

Effects of aqueous effluents from in situ fossil fuel processing technologies on aquatic systems. Annual progress report, January 1-December 31, 1979

Energy Technology Data Exchange (ETDEWEB)

Bergman, H.L.

1980-01-04

This is the third annual progress report for a continuing EPA-DOE jointly funded project to evaluate the effects of aqueous effluents from in situ fossil-fuel processing technologies on aquatic biota. The project is organized into four project tasks: (1) literature review; (2) process water screening; (3) methods development; and (4) recommendations. Our Bibliography of aquatic ecosystem effects, analytical methods and treatment technologies for organic compounds in advanced fossil-fuel processing effluents was submitted to the EPA for publication. The bibliography contains 1314 citations indexed by chemicals, keywords, taxa and authors. We estimate that the second bibliography volume will contain approximately 1500 citations and be completed in February. We compiled results from several laboratories of inorganic characterizations of 19 process waters: 55 simulated in situ oil-shale retort waters; and Hanna-3, Hanna-4B 01W and Lawrence Livermore Hoe Creek underground coal gasification condenser waters. These process waters were then compared to a published summary of the analyses from 18 simulated in situ oil-shale retort waters. We completed this year 96-h flow-through toxicity bioassays with fathead minnows and rainbow trout and 48-h flow-through bioassays with Daphnia pulicaria exposed to 5 oil-shale process waters, 1 tar-sand process water, 2 underground coal gasification condenser waters, 1 post-gasification backflood condenser water, as well as 2 bioassays with fossil-fuel process water constituents. The LC/sub 50/ toxicity values for these respective species when exposed to these waters are given in detail. (LTN)

Analysis of 3,5-dichloroaniline as a biomarker of vinclozolin and iprodione in human urine using liquid chromatography/triple quadrupole mass spectrometry.

Science.gov (United States)

Lindh, Christian H; Littorin, Margareta; Amilon, Asa; Jönsson, Bo A G

2007-01-01

The fungicides vinclozolin and iprodione are widely used in agriculture. These pesticides are dicarboximide fungicides containing the common moiety 3,5-dichloroaniline (3,5-DCA). It has been suggested that low-level exposures to such compounds may be associated with adverse health effects such as endocrine disruption. In this study a method using liquid chromatography/triple quadrupole mass spectrometry (LC/MS/MS) was developed for the analysis of 3,5-DCA as a biomarker of exposure to these fungicides in human urine. The urine samples were treated by basic hydrolysis to degrade the fungicides, their metabolites and conjugates to 3,5-DCA. The 3,5-DCA was then extracted using toluene and derivatized using pentafluoropropionic anhydride (PFPA). Analysis of the derivative was carried out using selected reaction monitoring (SRM) in the negative ion mode. Quantification of the derivative was performed using [(13)C(6)]-labeled 3,4-DCA as an internal standard with good precision and linearity in the range 0.1-200 ng/mL urine. The limit of detection was determined to be 0.1 ng/mL. The metabolites in urine were found to be stable during storage at -20 degrees C. To validate 3,5-DCA as a biomarker the method was applied in a human experimental exposure to iprodione and vinclozolin. Two healthy volunteers received 200 microg single oral doses of each pesticide followed by urine sampling during 72-120 h post-exposure. Between 78-107% of the dose was recovered as 3,5-DCA in the urine after exposure. Copyright (c) 2007 John Wiley & Sons, Ltd.

Development and application of a density dependent matrix ...

Science.gov (United States)

Ranging along the Atlantic coast from US Florida to the Maritime Provinces of Canada, the Atlantic killifish (Fundulus heteroclitus) is an important and well-studied model organism for understanding the effects of pollutants and other stressors in estuarine and marine ecosystems. Matrix population models are useful tools for ecological risk assessment because they integrate effects across the life cycle, provide a linkage between endpoints observed in the individual and ecological risk to the population as a whole, and project outcomes for many generations in the future. We developed a density dependent matrix population model for Atlantic killifish by modifying a model developed for fathead minnow (Pimephales promelas) that has proved to be extremely useful, e.g. to incorporate data from laboratory studies and project effects of endocrine disrupting chemicals. We developed a size-structured model (as opposed to one that is based upon developmental stages or age class structure) so that we could readily incorporate output from a Dynamic Energy Budget (DEB) model, currently under development. Due to a lack of sufficient data to accurately define killifish responses to density dependence, we tested a number of scenarios realistic for other fish species in order to demonstrate the outcome of including this ecologically important factor. We applied the model using published data for killifish exposed to dioxin-like compounds, and compared our results to those using

Chemical-gene interaction networks and causal reasoning for ...

Science.gov (United States)

Evaluating the potential human health and ecological risks associated with exposures to complex chemical mixtures in the environment is one of the main challenges of chemical safety assessment and environmental protection. There is a need for approaches that can help to integrate chemical monitoring and biological effects data to evaluate risks associated with chemicals present in the environment. Here, we used prior knowledge about chemical-gene interactions to develop a knowledge assembly model for detected chemicals at five locations near the North Branch and Chisago wastewater treatment plants (WWTP) in the St. Croix River Basin, MN and WI. The assembly model was used to generate hypotheses about the biological impacts of the chemicals at each location. The hypotheses were tested using empirical hepatic gene expression data from fathead minnows exposed for 12 d at each location. Empirical gene expression data were also mapped to the assembly models to evaluate the likelihood of a chemical contributing to the observed biological responses using richness and concordance statistics. The prior knowledge approach was able predict the observed biological pathways impacted at one site but not the other. Atrazine was identified as a potential contributor to the observed gene expression responses at a location upstream of the North Branch WTTP. Four chemicals were identified as contributors to the observed biological responses at the effluent and downstream o

Body ion loss as a bioindicator of water quality impaired by coal mining

International Nuclear Information System (INIS)

Grippo, R.S.; Dunson, W.A.

1994-01-01

Protection of surface waters receiving discharges from coal mines is currently based on performance standards set by the EPA after passage of the Clean Water Act. These standards were technology-driven and reflect the Best Achievable Control Technology (BAT) available at the time of promulgation. Changes proposed as part of the upcoming reauthorization of the US Clean Water Act suggest that such technology-based standards may be reevaluated in light of more recent information on the toxicological effect of mine discharges on aquatic biota. The authors present here a physiological-based method for evaluating the site-specific toxicity of mine-derived discharges into receiving waters. They tested the usefulness of the body ion loss rate bioassay by exposing fathead minnows, brook charr and stoneflies to coal mine-impacted waters (elevated acidity and trace metals) in the field and to artificial mine water (AMW) in the laboratory. Body ion loss rate was significantly correlated with levels of mine pollution in the field. Body ion loss measured in AMW revealed strong interactions between metals and acid. Because the test animals exhibited differing levels of sensitivity to mine discharge, the selection of an appropriate organism for the body ion loss bioassay may vary depending on the (1) physical characteristics, (2) chemical characteristics and (3) pre-existing level of mine impact of the receiving waters

Quantitative Adverse Outcome Pathways and Their ...

Science.gov (United States)

A quantitative adverse outcome pathway (qAOP) consists of one or more biologically based, computational models describing key event relationships linking a molecular initiating event (MIE) to an adverse outcome. A qAOP provides quantitative, dose–response, and time-course predictions that can support regulatory decision-making. Herein we describe several facets of qAOPs, including (a) motivation for development, (b) technical considerations, (c) evaluation of confidence, and (d) potential applications. The qAOP used as an illustrative example for these points describes the linkage between inhibition of cytochrome P450 19A aromatase (the MIE) and population-level decreases in the fathead minnow (FHM; Pimephales promelas). The qAOP consists of three linked computational models for the following: (a) the hypothalamic-pitutitary-gonadal axis in female FHMs, where aromatase inhibition decreases the conversion of testosterone to 17β-estradiol (E2), thereby reducing E2-dependent vitellogenin (VTG; egg yolk protein precursor) synthesis, (b) VTG-dependent egg development and spawning (fecundity), and (c) fecundity-dependent population trajectory. While development of the example qAOP was based on experiments with FHMs exposed to the aromatase inhibitor fadrozole, we also show how a toxic equivalence (TEQ) calculation allows use of the qAOP to predict effects of another, untested aromatase inhibitor, iprodione. While qAOP development can be resource-intensive, the quan

Effects of the Antipsychotic Drug, Haloperidol, on Reproduction in the Fathead Minnow

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Haloperidol is a butyrophenone antipsychotic drug used for the treatment of human hyperactive and manic disorders, agitation, and schizophrenia. The drug is thought to act through antagonism of dopaminergic receptors. We have studied a variety of endocrine-disrupting chemicals wi...

Effects of the insecticide fipronil on reproductive endocrinology in the fathead minnow

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Gamma aminobutyric acid (GABA) and GABA receptors play an important role in neuroendocrine regulation in fish. Disruption of the GABAergic system by environmental contaminants could interfere with normal regulation of the hypothalamic pituitary gonadal (HPG) axis, leading to imp...

A method for CRISPR/Cas9 mutation of genes in fathead minnow

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Product Description: CRISPR/Cas9 is a system that can be used to disrupt a gene of interest in any animal. It allows us to study each gene’s role by observing changes in the animal when the gene isn’t functional. We worked out a method to use this technology in the f...

Enantioselective Toxicity and Biotransformation of Fipronil in the Fathead Minnow (Pimephales promelas)

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Fipronil is a relatively new chiral phenylpyrazole insecticide used to control both agricultural and household invertebrate pests. Fipronil is applied as a racemate, or equal mixture, of its two enantiomers. As regulations on older pesticides increase, production and applicatio...

Effects of Environmentally Relevant Concentrations of Bisphenol A on the Fathead Minnow

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Bisphenol A (BPA) is a wide-spread environmental contaminant of concern due, in part, to possible effects on the vertebrate hypothalamic-pituitary-gonadal (HPG) axis, including activation of the estrogen receptor (ER). There is a reasonable amount of toxicological information fo...

On-site evaluation of the suitability of a wetted instream habitat in the Middle Rio Grande, New Mexico, for the Rio Grande silvery minnow (Hybognathus amarus)

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2011-01-01

Two in-situ exposure studies were conducted with the federally-listed endangered Rio Grande silvery minnow (Hybognathus amarus). One-year-old adults were exposed in cages deployed at three sites in the Middle Rio Grande, N. Mex., for 4 days to assess survival and for 26 days to evaluate survival, growth, overall health, and whole-body elemental composition. The test sites were located on the Pueblo of Isleta in the (1) main channel of the Middle Rio Grande, (2) 240-Wasteway irrigation return drain, and (3) wetted instream habitat created below the outfall of the 240-Wasteway irrigation return drain. During the cage exposures, temperature, dissolved oxygen, pH, conductivity, and turbidity were monitored continuously (15-minute intervals) and common constituents, nutrients, carbons, metals, and pesticides were measured at discrete intervals. In both studies, there were statistical differences in several water-quality parameters among sites; and except for turbidity, these differences were small and were not considered to be biologically significant. The cages used in the 4-day exposure study were ineffective at preventing access to the fish by predators, and survival was highly variable (20 percent to 90 percent) across sites. In the 26-day chronic exposure study, weight and condition factor of caged-exposed fish at all sites were significantly lower than those at test initiation. After 26 days of exposure, there were no significant differences in survival, total length, weight, or condition factor of fish across sites, but absolute weight loss and relative reduction in condition factor were significantly greater in fish at the wetted instream habitat site compared to those at the Middle Rio Grande site. There were no statistical differences in health assessment indices, mesenteric fat indices, or prevalence of abnormalities in cage-exposed fish among sites. Cage-exposed fish had higher health assessment indices and prevalence of fin anomalies and a lower mesenteric

Effects of endocrine disruptors on prosobranch snails (Mollusca: Gastropoda) in the laboratory. Part III: Cyproterone acetate and vinclozolin as antiandrogens.

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Tillmann, M; Schulte-Oehlmann, U; Duft, M; Markert, B; Oehlmann, J

2001-12-01

The effects of suspected endocrine disrupting chemicals on freshwater and marine prosobranch species were analysed in laboratory experiments. In this last of three publications, the responses of the fresh water snail Marisa cornuarietis and of two marine prosobranchs (Nucella lapillus, Nassarius (Hinia) reticulatus) to the antiandrogenic model compounds cyproterone acetate (CPA) and vinclozolin (VZ) are presented. The snails were exposed to nominal CPA concentrations of 1.25 mg/L alone and simultaneously to a potent synthetic estrogen (ethinylestradiol), androgen (methyltestosterone) or an indirectly acting xeno-androgen (tributyltin) in experiments with adult specimens and in a life cycle test for 12 months. Marisa and Nucella were furthermore exposed to nominal concentrations of 0.03-1.0 microgram VZ/L for up to 5 months. The antiandrogens induced a number of biological responses in all three species. The length of the penis and of accessory male sex organs (e.g., penis sheath, prostate) were significantly reduced. For Marisa, this effect occurred only in sexually immature specimens and was reversible as the males attained puberty. Typical androgen-mediated responses (imposex development, delayed spermatogenesis, tubulus necrosis of the testis with orchitis and Leydig cell hyperplasia) were partially or totally suppressed by a simultaneous administration of CPA. In the two marine species even adult, sexually mature males responded to antiandrogens with a reduction of the male sex organs and an advancement of the sexual repose phase. The results for CPA and VZ are compared with the effects of an exposure to xeno-estrogens (bisphenol A, octylphenol) and xeno-androgens (triphenyltin, tributyltin) in the same species. Each group of endocrine disruptors induces a characteristic set of toxicological effects in prosobranch snails which can be used as endpoints in an organismic invertebrate test for the identification of endocrine mimetic test compounds. Estrogens cause

PAH related effects on fish in sedimentation ponds for road runoff and potential transfer of PAHs from sediment to biota

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Grung, Merete; Petersen, Karina; Fjeld, Eirik

2016-01-01

of the sedimentation ponds were high (1900-4200ngg(-1)), and even higher levels were observed in plants. Principal component analysis of selected ion chromatograms of PAHs showed a clear separation of plants vs. sediments. The plants preferentially accumulated the high molecular PAHs, both from sedimentation ponds......) the transfer of PAHs from road runoff material to aquatic organisms was substantial. Minnow from a sedimentation pond (Skullerud) near Oslo (Norway) had higher levels of CYP1A enzyme and DNA stand breaks than minnow from the nearby river, but high concentrations of PAH-metabolites in bile revealed that both...... populations were highly exposed. Principal component analysis revealed that CYP1A and age of fish were correlated, while levels of PAH-metabolites were not correlated to CYP1A or DNA damage. Minnow from a lake un-affected by traffic had much lower levels of PAH-metabolites than the exposed fish, and also...

Comparison of anti-androgenic activity of flutamide, vinclozolin, procymidone, linuron, and p, p'-DDE in rodent 10-day Hershberger assay.

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Kang, Il Hyun; Kim, Hyung Sik; Shin, Jae-Ho; Kim, Tae Sung; Moon, Hyun Ju; Kim, In Young; Choi, Kwang Sik; Kil, Kwang Sup; Park, Young In; Dong, Mi Sook; Han, Soon Young

2004-07-01

The rodent Hershberger assay proposed by the Organization for Economic Co-operation and Development (OECD) is in the process of the validating a test method to detecting the androgenic or anti-androgenic compounds. The aim of this study was to compare the anti-androgenic properties of flutamide, vinclozolin, procymidone, linuron, and p,p'-DDE in a 10-day Hershberger assay. In the present study, we used immature Sprague-Dawley male rats castrated at 6 weeks of age. Testosterone propionate (TP) was subcutaneously injected for 10 consecutive days at doses of 0.1, 0.2, 0.4, 0.8, or 1.6 mg/kg per day. To compare the anti-androgenic activity of test compounds, flutamide (1, 5, 10, or 20 mg/kg per day), a pure androgen antagonist was used as a positive control, and administered by oral gavage after TP (0.4 mg/kg per day) treatment. In addition, vinclozolin (25, 50, or 100 mg/kg per day), procymidone (25, 50, or 100 mg/kg per day), linuron (25, 50, or 100 mg/kg per day), and p,p '-DDE (25, 50, or 100 mg/kg per day) were also administered by oral gavage after TP (0.4 mg/kg per day) treatment. As expected, TP dose-dependently increased accessory sex organ weights, and statistically significant effects were observed at doses of 0.1 (only seminal vesicles) or 0.2mg/kg per day and above. Serum testosterone levels increased significantly at 0.4 mg/kg per day and above, while serum LH levels were decreased in a dose-dependent manner. Flutamide significantly inhibited the TP-induced re-growth of seminal vesicles, ventral prostate, and Levator ani plus bulbocavernosus muscles (LABC) at 1mg/kg per day and above, and Cowper's glands and glans penis at 5mg/kg per day and above. In contrast to accessory sex organ weights, flutamide did not affect the serum testosterone levels compared to the control at any concentration, but serum LH levels were significantly increased at doses of 10 and 20 mg/kg per day. Similar to flutamide, vinclozolin caused a statistically significant decrease in

Intergenerational response to the endocrine disruptor vinclozolin is influenced by maternal genotype and crossing scheme.

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Pietryk, Edward W; Clement, Kiristin; Elnagheeb, Marwa; Kuster, Ryan; Kilpatrick, Kayla; Love, Michael I; Ideraabdullah, Folami Y

2018-03-10

In utero exposure to vinclozolin (VIN), an antiandrogenic fungicide, is linked to multigenerational phenotypic and epigenetic effects. Mechanisms remain unclear. We assessed the role of antiandrogenic activity and DNA sequence context by comparing effects of VIN vs. M2 (metabolite with greater antiandrogenic activity) and wild-type C57BL/6 (B6) mice vs. mice carrying mutations at the previously reported VIN-responsive H19/Igf2 locus. First generation offspring from VIN-treated 8nrCG mutant dams exhibited increased body weight and decreased sperm ICR methylation. Second generation pups sired by affected males exhibited decreased neonatal body weight but only when dam was unexposed. Offspring from M2 treatments, B6 dams, 8nrCG sires or additional mutant lines were not similarly affected. Therefore, pup response to VIN over two generations detected here was an 8nrCG-specific maternal effect, independent of antiandrogenic activity. These findings demonstrate that maternal effects and crossing scheme play a major role in multigenerational response to in utero exposures. Copyright © 2018 Elsevier Inc. All rights reserved.

Behavioral observations of the endangered Rio Grande silvery minnow in a conservation aquaculture facility

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Tave Douglas

2018-03-01

Full Text Available A major reason why conservation aquaculture is needed to improve the success of aquaculture-assisted fisheries is that traditional production aquaculture produces fish with mal-adaptive behaviors. These behaviors can be produced via domestication and culture techniques, and preventing these mal-adaptive behaviors requires integrating improvements in genetic management and culture protocols. The genetic protocols needed to minimize hatchery-induced genetic changes have received considerable attention, but changing the way fish are raised has received less effort. Conservation aquaculture cultures fish in environments that resemble their native habitats so that when stocked, they behave like wild fish rather than hatchery fish. A purpose built-conservation aquaculture facility can also be used to learn about a species’ behavior and how it reacts to changes in the environment, something which can be difficult or expensive to study in the wild. These observations can then be used to help direct both propagation and recovery management. This paper provides the rationale for why genetic management, culture systems, and management practices need to be altered to produce fish that are behaviorally similar to wild fish for aquaculture-assisted fisheries programs. It then provides a description of some of the behaviors of the endangered Rio Grande silvery minnow Hybognathus amarus that were observed at the Los Lunas Silvery Minnow Refugium, a purpose-built conservation aquaculture facility, and explains how some of these behaviors can be used in culture and recovery management. Behaviors described are: schooling; predator avoidance; feeding behavior; use of vegetation for cover and predator avoidance; habitat use by bottom substrate; location in the water column; upstream movement via a fish ladder; movement upstream in a high-velocity channel; response to changes in water level; spawning behavior; seine avoidance; and Kaah-chee-nyee Srkaash, a behavior

A Computational Model of the Hypothalamic Pituitary Gonadal Axis in Female Fathead Minnows (Pimephales promelas) exposed to 17á-ethynylestradiol and 17â-trenbolone

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Endocrine disrupting chemicals (e.g., estrogens, androgens and their mimics) are known to affect reproduction in fish. 17a-ethynylestradiol is a synthetic estrogen used in birth control pills. 17a-trenbolone is a relatively stable metabolite of trenbolone acetate, a synthetic and...

Social learning in a high-risk environment: incomplete disregard for the 'minnow that cried pike' results in culturally transmitted neophobia.

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Crane, Adam L; Mathiron, Anthony G E; Ferrari, Maud C O

2015-08-07

Many prey species rely on conspecifics to gather information about unknown predation threats, but little is known about the role of varying environmental conditions on the efficacy of social learning. We examined predator-naive minnows that had the opportunity to learn about predators from experienced models that were raised in either a low- or high-risk environment. There were striking differences in behaviour among models; high-risk models showed a weaker response to the predator cue and became neophobic in response to the control cue (a novel odour, NO). Observers that were previously paired with low-risk models acquired a strong antipredator response only to the predator cue. However, observers that interacted with high-risk models, displayed a much weaker response to the predator odour and a weak neophobic response to the NO. This is the first study reporting such different outcomes of social learning under different environmental conditions, and suggests high-risk environments promote the cultural transmission of neophobia more so than social learning. If such a transfer can be considered similar to secondary traumatization in humans, culturally transmitted neophobia in minnows may provide a good model system for understanding more about the social ecology of fear disorders. © 2015 The Author(s).

Induction of gene expression in sheepshead minnows (Cyprinodon variegatus) treated with 17beta-estradiol, diethylstilbestrol, or ethinylestradiol: the use of mRNA fingerprints as an indicator of gene regulation.

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Denslow, N D; Bowman, C J; Ferguson, R J; Lee, H S; Hemmer, M J; Folmar, L C

2001-03-01

The recent interest in hormonally active environmental contaminants has sparked a drive to find sensitive methods to measure their effects on wildlife. A molecular-based assay has been developed to measure the induction of gene expression in sheepshead minnows (Cyprinodon variegatus) exposed in vivo to the natural and pharmaceutical estrogens 17beta-estradiol, ethinylestradiol, and diethylstilbestrol. This method used differential display reverse transcriptase polymerase chain reaction assays to compare the expression of individual mRNAs from control and estrogen-exposed fish. Forty-eight differentially expressed cDNAs were isolated by this method, including cDNAs for vitelline envelope proteins and vitellogenin. The mRNA expression patterns for fish injected with a pharmacological dose of estradiol (5 mg/kg) were identical to those obtained in fish receiving constant aqueous exposure to 212 ng estradiol/liter. Further, the cDNA "fingerprint" pattern observed in the estradiol-treated fish also matched that obtained in fish receiving continuous-flow aqueous exposures to 192 ng ethinyl estradiol/liter and a nominal concentration of 200 ng diethylstilbestrol/liter. The results demonstrate a characteristic expression pattern for genes upregulated by exposure to a variety of natural and anthropogenic estrogens and suggest this approach may be valuable to examine the potential effects of environmental contaminants on other endocrine-mediated pathways of reproduction, growth, and development. Copyright 2001 Academic Press.

Variability and condition of common minnow (Phoxinus phoxinus, Cyprinidae, Cypriniformes) on the Semipalatinsk range and out of its influence

International Nuclear Information System (INIS)

Mitrofanov, I.V.; Matmuratov, S.A.

2003-01-01

Populations of common minnow (Phoxinus phoxinus) from five rivers of Irtysh and Balkhash basins were investigated. Population from the Shagan river situated inside area of Semipalatinsk Range influence. This population has normal sexual structure and high growth rate. Condition factor of all fishes is high. Population from Semipalatinsk Range has increased variability of characteristics due to increasing of stochastic part of general variability. Abnormalities of external fish morphology are not observed in this area. Asymmetry of bilateral characteristics is low. (author)

Tertiary Epimutations – A Novel Aspect of Epigenetic Transgenerational Inheritance Promoting Genome Instability

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McCarrey, John R.; Lehle, Jake D.; Raju, Seetha S.; Wang, Yufeng; Nilsson, Eric E.; Skinner, Michael K.

2016-01-01

Exposure to environmental factors can induce the epigenetic transgenerational inheritance of disease. Alterations to the epigenome termed “epimutations” include “primary epimutations” which are epigenetic alterations in the absence of genetic change and “secondary epimutations” which form following an initial genetic change. To determine if secondary epimutations contribute to transgenerational transmission of disease following in utero exposure to the endocrine disruptor vinclozolin, we exposed pregnant female rats carrying the lacI mutation-reporter transgene to vinclozolin and assessed the frequency of mutations in kidney tissue and sperm recovered from F1 and F3 generation progeny. Our results confirm that vinclozolin induces primary epimutations rather than secondary epimutations, but also suggest that some primary epimutations can predispose a subsequent accelerated accumulation of genetic mutations in F3 generation descendants that have the potential to contribute to transgenerational phenotypes. We therefore propose the existence of “tertiary epimutations” which are initial primary epimutations that promote genome instability leading to an accelerated accumulation of genetic mutations. PMID:27992467

Effects of Xenoestrogen and Androgen Mixtures on Ovarian Transcriptome of the Fathead Minnow

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Endocrine disrupting chemicals (EDCs), such as the estrogens ethinylestradiol (EE2) and bisphenol A (BPA), and androgens like 17â-trenbolone (TRB) can occur as mixtures in aquatic environments. To date, however, most studies with EDCs in fish have focused on their effects as indi...

Effectiveness of thiophanate-methyl, trifloxystrobin and vinclozolin on canker caused by Phoma exigua Desm. on ash tree seedlings

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Cuvelier M.

2006-01-01

Full Text Available In recent years several cases of cankers caused by Phoma exigua on ash tree seedlings have been reported in Belgian nurseries, leading to a total loss of the affected crops. Similar symptoms have been observed on ash tree seedlings elsewhere in Europe, notably in France and in Great Britain, but the pathogenicity was never established. Inoculation and re-isolation tests were therefore undertaken and demonstrated the phytopathogenic character of P. exigua on ash. Moreover the effectiveness of three fungicides (thiophanate-methyl, trifloxystrobin, vinclozolin against stem canker of ash tree seedlings was studied. In vitro tests were conducted to evaluate the ability of these fungicides to inhibit mycelium growth and spore germination. The extent to which they reduced the symptoms was estimated in a field trial. The results of this study allowed to get by the Belgian proper authorities the use extension of thiophanate-methyl for the control of canker caused by P. exigua in forest nurseries.

Effects of the antimicrobial contaminant triclocarban and co-exposure with the androgen 17â-trenbolone, on reproductive function and ovarian transcriptome of the fathead minnow (Pimephales promelas)

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Triclocarban (TCC) is a widely used antimicrobial agent that is routinely detected in surface waters. The present study was designed to examine TCC’s efficacy and mode of action as a reproductive toxicant in fish. Reproductively mature Pimephales promelas were continuously expose...

Atrazine affects kidney and adrenal hormones (AHs) related genes expressions of rare minnow (Gobiocypris rarus).

Science.gov (United States)

Yang, Lihua; Zha, Jinmiao; Li, Wei; Li, Zhaoli; Wang, Zijian

2010-05-05

Atrazine, one of the most widely used herbicides, has been proved to interfere with sexual hormones. However few studies have considered the effects of atrazine on adrenal hormones (AH). In this study, rare minnow (Gobiocypris rarus) was exposed to 0, 3, 10, 33, 100 and 333microg/l atrazine for 28 days. The histopathology of kidney and gill was examined and the expressions of AHs-related genes including Na(+),K(+)-ATPase, glucocorticoid receptor (gr), heat shock protein 70 (hsp70), and heat shock protein 90 (hsp90) in kidney and gill were quantitatively determined. Histopathological observation revealed obvious lesions in gill including hyperplasia, necrosis in epithelium region, aneurysm and lamellar fusion at concentrations as low as 10microg/l. The observed lesions in kidney included extensive expansion in the lumen, degenerative and necrotic changes of the tubular epithelia, shrinkage of the glomerulus as well as increase of the Bowman's space at concentrations as low as 10microg/l. The expressions of Na(+),K(+)-ATPase, gr, hsp70 and hsp90 in the kidney of females were significantly decreased at all concentrations. For males, the expressions of hsp90 in the kidney of all treated groups were significantly down-regulated, while gr at all concentrations and hsp70 at 10, 33, 100microg/l were significantly up-regulated. However in the gill, the expressions of these genes were not significantly different from the control. These results indicated that exposure to atrazine caused impairments of kidney and gill of fish at environmental related concentrations. Histopathological lesions could partly attribute to the changes of the expressions of AHs-related genes in kidney. We concluded also that atrazine is a potential AHs-disruptor and AHs-related genes in kidney of fish could be used as sensitive molecular biomarkers.

Atrazine affects kidney and adrenal hormones (AHs) related genes expressions of rare minnow (Gobiocypris rarus)

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Yang Lihua; Zha Jinmiao; Li Wei; Li Zhaoli [State Key Laboratory of Environmental Aquatic Chemistry, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Shuangqing Road 18, P.O. Box 2871, Beijing 100085 (China); Wang Zijian, E-mail: wangzj@rcees.ac.cn [State Key Laboratory of Environmental Aquatic Chemistry, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Shuangqing Road 18, P.O. Box 2871, Beijing 100085 (China)

2010-05-05

Atrazine, one of the most widely used herbicides, has been proved to interfere with sexual hormones. However few studies have considered the effects of atrazine on adrenal hormones (AH). In this study, rare minnow (Gobiocypris rarus) was exposed to 0, 3, 10, 33, 100 and 333 {mu}g/l atrazine for 28 days. The histopathology of kidney and gill was examined and the expressions of AHs-related genes including Na{sup +},K{sup +}-ATPase, glucocorticoid receptor (gr), heat shock protein 70 (hsp70), and heat shock protein 90 (hsp90) in kidney and gill were quantitatively determined. Histopathological observation revealed obvious lesions in gill including hyperplasia, necrosis in epithelium region, aneurysm and lamellar fusion at concentrations as low as 10 {mu}g/l. The observed lesions in kidney included extensive expansion in the lumen, degenerative and necrotic changes of the tubular epithelia, shrinkage of the glomerulus as well as increase of the Bowman's space at concentrations as low as 10 {mu}g/l. The expressions of Na{sup +},K{sup +}-ATPase, gr, hsp70 and hsp90 in the kidney of females were significantly decreased at all concentrations. For males, the expressions of hsp90 in the kidney of all treated groups were significantly down-regulated, while gr at all concentrations and hsp70 at 10, 33, 100 {mu}g/l were significantly up-regulated. However in the gill, the expressions of these genes were not significantly different from the control. These results indicated that exposure to atrazine caused impairments of kidney and gill of fish at environmental related concentrations. Histopathological lesions could partly attribute to the changes of the expressions of AHs-related genes in kidney. We concluded also that atrazine is a potential AHs-disruptor and AHs-related genes in kidney of fish could be used as sensitive molecular biomarkers.

Atrazine affects kidney and adrenal hormones (AHs) related genes expressions of rare minnow (Gobiocypris rarus)

International Nuclear Information System (INIS)

Yang Lihua; Zha Jinmiao; Li Wei; Li Zhaoli; Wang Zijian

2010-01-01

Atrazine, one of the most widely used herbicides, has been proved to interfere with sexual hormones. However few studies have considered the effects of atrazine on adrenal hormones (AH). In this study, rare minnow (Gobiocypris rarus) was exposed to 0, 3, 10, 33, 100 and 333 μg/l atrazine for 28 days. The histopathology of kidney and gill was examined and the expressions of AHs-related genes including Na + ,K + -ATPase, glucocorticoid receptor (gr), heat shock protein 70 (hsp70), and heat shock protein 90 (hsp90) in kidney and gill were quantitatively determined. Histopathological observation revealed obvious lesions in gill including hyperplasia, necrosis in epithelium region, aneurysm and lamellar fusion at concentrations as low as 10 μg/l. The observed lesions in kidney included extensive expansion in the lumen, degenerative and necrotic changes of the tubular epithelia, shrinkage of the glomerulus as well as increase of the Bowman's space at concentrations as low as 10 μg/l. The expressions of Na + ,K + -ATPase, gr, hsp70 and hsp90 in the kidney of females were significantly decreased at all concentrations. For males, the expressions of hsp90 in the kidney of all treated groups were significantly down-regulated, while gr at all concentrations and hsp70 at 10, 33, 100 μg/l were significantly up-regulated. However in the gill, the expressions of these genes were not significantly different from the control. These results indicated that exposure to atrazine caused impairments of kidney and gill of fish at environmental related concentrations. Histopathological lesions could partly attribute to the changes of the expressions of AHs-related genes in kidney. We concluded also that atrazine is a potential AHs-disruptor and AHs-related genes in kidney of fish could be used as sensitive molecular biomarkers.

Development of a methodology for successful multigeneration life-cycle testing of the estuarine sheepshead minnow, Cyprinodon variegatus.

Science.gov (United States)

Cripe, G M; Hemmer, B L; Goodman, L R; Vennari, J C

2009-04-01

Evaluation of effects on fish reproduction and development during chemical exposures lasting for multiple generations is sometimes limited by variable reproductive responses and the time required for the exposure. Established testing methods and the short life cycle of the sheepshead minnow, Cyprinodon variegatus, make this species particularly suitable for use in identifying potential impacts of contaminants in estuarine and marine environments. This study describes the refinement of life-cycle exposure methods that increased the reliability of reproduction in sheepshead minnows and reduced the time to maturation for larvae and juvenile fishes. A test of three spawning chamber designs, three sex ratios, and two photoperiods identified conditions that reduced the coefficient of variation in egg production from >100% to as little as 32%. The most reliable results were produced with groups of three female and two male fishes (all of similar size) when they were placed in a rectangular chamber and acclimated for 12 days. A test water temperature of 26.5 +/- 2 degrees C and a 14L:10D photoperiod resulted in fish producing a mean of 74 embryos per female per day, with a coefficient of variation of 31.8%. Egg fertility exceeded 90%, with a hatch rate of 95% for normal embryos (>or=80% yolk) and a hatch rate of size (>or=2.7 cm standard length) was critical for spawning readiness. Adult fish were prepared for the spawning assessment by adding frozen brine shrimp to their diet. Results of these experiments provide methods that are of particular interest in assessment of endocrine-disrupting chemicals that are known to affect reproduction.

Distribution of Eurasian minnows (Phoxinus: Cypriniformes in the Western Balkans

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Vucić Matej

2018-01-01

Full Text Available The Eurasian minnows of the genus Phoxinus are small cyprinid species, widely distributed across Europe and Asia. Currently, there are at least 15 species in Phoxinus, with preliminary data suggesting more to be described. Despite the discernible increase in research integrating molecular phylogenetic approaches with traditional taxonomy and systematics, inter- and intraspecific relationships in the genus Phoxinus are still poorly known. The aims of this research were to: (i compile data on the distribution of Phoxinus species in Croatia and Bosnia and Herzegovina from the sampling conducted by authors from 2006 to 2016 and from literature sources, and (ii provide a detailed insight on distribution ranges of genetic clades and species of Phoxinus in the western Balkans. Additional 118 localities in Croatia and 8 locations in Bosnia and Herzegovina were added to already known 160 localities from the literature data. Molecular analyses of mitochondrial DNA indicate: (i the presence of Phoxinus marsilii in Croatia (Drava drainage, the Papuk Mountain, around 260 km south of its known distribution range in Hungary, and (ii an exceptional genetic variability of P. lumaireul in the Western Balkans.

Propagation and isolation of ranaviruses in cell culture

DEFF Research Database (Denmark)

Ariel, Ellen; Nicolajsen, Nicole; Christophersen, Maj-Britt

2009-01-01

The optimal in vitro propagation procedure for a panel of ranavirus isolates and the best method for isolation of Epizootic haematopoietic necrosis virus (EHNV) from organ material in cell-culture were investigated. The panel of ranavirus isolates included: Frog virus 3 (FV3), Bohle iridovirus (BIV......), Pike-perch iridovirus (PPIV), European catfish virus (ECV), European sheatfish virus (ESV), EHNV, Doctor fish virus (DFV), Guppy virus 6 (GF6), short-finned eel virus (SERV) and Rana esculenta virus Italy 282/102 (REV 282/102). Each isolate was titrated in five cell lines: bluegill fry (BF-2......), epithelioma papulosum cyprini (EPC), chinook salmon embryo (CHSE-214) rainbow trout gonad (RTG-2) and fathead minnow (FHM), and incubated at 10, 15, 20, 24 and 28 °C for two weeks. BF-2, EPC and CHSE-214 cells performed well and titers obtained in the three cell lines were similar, whereas FHM and RTG-2 cells...

Gene expression profiling of the androgen receptor antagonists flutamide and vinclozolin in zebrafish (Danio rerio) gonads

International Nuclear Information System (INIS)

Martinovic-Weigelt, Dalma; Wang Ronglin; Villeneuve, Daniel L.; Bencic, David C.; Lazorchak, Jim; Ankley, Gerald T.

2011-01-01

The studies presented in this manuscript focus on characterization of transcriptomic responses to anti-androgens in zebrafish (Danio rerio). Research on the effects of anti-androgens in fish has been characterized by a heavy reliance on apical endpoints, and molecular mechanisms of action (MOA) of anti-androgens remain poorly elucidated. In the present study, we examined effects of a short term exposure (24-96 h) to the androgen receptor antagonists flutamide (FLU) and vinclozolin (VZ) on gene expression in gonads of sexually mature zebrafish, using commercially available zebrafish oligonucleotide microarrays (4 x 44 K platform). We found that VZ and FLU potentially impact reproductive processes via multiple pathways related to steroidogenesis, spermatogenesis, and fertilization. Observed changes in gene expression often were shared by VZ and FLU, as demonstrated by overlap in differentially-expressed genes and enrichment of several common key pathways including: (1) integrin and actin signaling, (2) nuclear receptor 5A1 signaling, (3) fibroblast growth factor receptor signaling, (4) polyamine synthesis, and (5) androgen synthesis. This information should prove useful to elucidating specific mechanisms of reproductive effects of anti-androgens in fish, as well as developing biomarkers for this important class of endocrine-active chemicals.

Gene expression profiling of the androgen receptor antagonists flutamide and vinclozolin in zebrafish (Danio rerio) gonads

Energy Technology Data Exchange (ETDEWEB)

Martinovic-Weigelt, Dalma, E-mail: dalma@stthomas.edu [US Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Mid-Continent Ecology Division, 6201 Congdon Blvd., Duluth, MN 55804 (United States); University of St. Thomas, 2115 Summit Ave, Saint Paul, MN 55105 (United States); Wang Ronglin [US Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory, Ecological Exposure Research Division, 26W. Martin Luther King Dr., Cincinnati, OH 45268 (United States); Villeneuve, Daniel L. [US Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Mid-Continent Ecology Division, 6201 Congdon Blvd., Duluth, MN 55804 (United States); Bencic, David C.; Lazorchak, Jim [US Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory, Ecological Exposure Research Division, 26W. Martin Luther King Dr., Cincinnati, OH 45268 (United States); Ankley, Gerald T. [US Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Mid-Continent Ecology Division, 6201 Congdon Blvd., Duluth, MN 55804 (United States)

2011-01-25

The studies presented in this manuscript focus on characterization of transcriptomic responses to anti-androgens in zebrafish (Danio rerio). Research on the effects of anti-androgens in fish has been characterized by a heavy reliance on apical endpoints, and molecular mechanisms of action (MOA) of anti-androgens remain poorly elucidated. In the present study, we examined effects of a short term exposure (24-96 h) to the androgen receptor antagonists flutamide (FLU) and vinclozolin (VZ) on gene expression in gonads of sexually mature zebrafish, using commercially available zebrafish oligonucleotide microarrays (4 x 44 K platform). We found that VZ and FLU potentially impact reproductive processes via multiple pathways related to steroidogenesis, spermatogenesis, and fertilization. Observed changes in gene expression often were shared by VZ and FLU, as demonstrated by overlap in differentially-expressed genes and enrichment of several common key pathways including: (1) integrin and actin signaling, (2) nuclear receptor 5A1 signaling, (3) fibroblast growth factor receptor signaling, (4) polyamine synthesis, and (5) androgen synthesis. This information should prove useful to elucidating specific mechanisms of reproductive effects of anti-androgens in fish, as well as developing biomarkers for this important class of endocrine-active chemicals.

A computational approach to evaluate the androgenic affinity of iprodione, procymidone, vinclozolin and their metabolites.

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Corrado Lodovico Galli

Full Text Available Our research is aimed at devising and assessing a computational approach to evaluate the affinity of endocrine active substances (EASs and their metabolites towards the ligand binding domain (LBD of the androgen receptor (AR in three distantly related species: human, rat, and zebrafish. We computed the affinity for all the selected molecules following a computational approach based on molecular modelling and docking. Three different classes of molecules with well-known endocrine activity (iprodione, procymidone, vinclozolin, and a selection of their metabolites were evaluated. Our approach was demonstrated useful as the first step of chemical safety evaluation since ligand-target interaction is a necessary condition for exerting any biological effect. Moreover, a different sensitivity concerning AR LBD was computed for the tested species (rat being the least sensitive of the three. This evidence suggests that, in order not to over-/under-estimate the risks connected with the use of a chemical entity, further in vitro and/or in vivo tests should be carried out only after an accurate evaluation of the most suitable cellular system or animal species. The introduction of in silico approaches to evaluate hazard can accelerate discovery and innovation with a lower economic effort than with a fully wet strategy.

A computational approach to evaluate the androgenic affinity of iprodione, procymidone, vinclozolin and their metabolites.

Science.gov (United States)

Galli, Corrado Lodovico; Sensi, Cristina; Fumagalli, Amos; Parravicini, Chiara; Marinovich, Marina; Eberini, Ivano

2014-01-01

Our research is aimed at devising and assessing a computational approach to evaluate the affinity of endocrine active substances (EASs) and their metabolites towards the ligand binding domain (LBD) of the androgen receptor (AR) in three distantly related species: human, rat, and zebrafish. We computed the affinity for all the selected molecules following a computational approach based on molecular modelling and docking. Three different classes of molecules with well-known endocrine activity (iprodione, procymidone, vinclozolin, and a selection of their metabolites) were evaluated. Our approach was demonstrated useful as the first step of chemical safety evaluation since ligand-target interaction is a necessary condition for exerting any biological effect. Moreover, a different sensitivity concerning AR LBD was computed for the tested species (rat being the least sensitive of the three). This evidence suggests that, in order not to over-/under-estimate the risks connected with the use of a chemical entity, further in vitro and/or in vivo tests should be carried out only after an accurate evaluation of the most suitable cellular system or animal species. The introduction of in silico approaches to evaluate hazard can accelerate discovery and innovation with a lower economic effort than with a fully wet strategy.

Investigations of putative reproductive toxicity of low-dose exposures to vinclozolin in Wistar rats.

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Flick, Burkhard; Schneider, Steffen; Melching-Kollmuss, Stephanie; Fussell, Karma C; Gröters, Sibylle; Buesen, Roland; Strauss, Volker; van Ravenzwaay, Bennard

2017-04-01

The current investigation examines whether the fungicide vinclozolin, which has an anti-androgenic mode of action, is capable of disrupting endocrine homeostasis at very low doses. The data generated clarify whether a non-monotonic dose-response relationship exists to enhance the current debate about the regulation of endocrine disruptors. Moreover, it is part of a series of investigations assessing the dose-response relationship of single and combined administration of anti-androgenic substances. A pre-postnatal in vivo study design was chosen which was compliant with regulatory testing protocols. The test design was improved by additional endpoints addressing hormone levels, morphology and histopathological examinations. Doses were chosen to represent an effect level (20 mg/kg bw/d), the current NOAEL (4 mg/kg bw/d), and a dose close to the "ADI" (0.005 mg/kg bw/d) for the detection of a possible non-monotonic dose-response curve. Anti-androgenic changes were observable at the effect level but not at lower exposures. Nipple/areola counts appeared to be the most sensitive measure of effect, followed by male sex organ weights at sexual maturation, and finally gross and histopathological findings. The results indicate the absence of evidence for effects at low or very low dose levels. A non-monotonic dose-response relationship was not evident.

Free-to-play: About addicted Whales, at risk Dolphins and healthy Minnows. Monetarization design and Internet Gaming Disorder.

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Dreier, M; Wölfling, K; Duven, E; Giralt, S; Beutel, M E; Müller, K W

2017-01-01

Video games are not only changing due to technical innovation, but also because of new game design and monetization approaches. Moreover, elite gamer groups with financial in-game-investments co-finance all users of free-to-play-games. Besides questions on youth protection, the growing popularity of free-to-play games has fostered discussions on supposed associations to Internet Gaming Disorder (IGD). Children and adolescents using free-to-play browser games were examined in a German school-based representative study (N=3967; age range 12 to 18). Based on a clinical self-report AICA-S (Wölfling et al., 2011), students were categorized into non-problematic, risky, and addicted users. Psycho-social problems (SDQ; Goodman, 1997), perceived stress (PSS; Cohen, Kamarck & Mermelstein, 1983), coping strategies (BriefCOPE; Carver, 1997), and Average Revenue per (Paying) User (ARPU) were investigated as dependent variables. Furthermore, an industry classification (Freeloaders, Minnows, Dolphins, and Whales) for free-to-play gamers was used for additional relations regarding IGD, SDQ, PSS, BriefCOPE, and ARPU. Among free-to-play gamers the prevalence of IGD amounted to 5.2%. Subjects classified with IGD displayed higher psycho-social symptoms than non-problematic users, reported higher degrees of perceived stress, and applied dysfunctional coping strategies more frequently. Additionally, we found a higher ARPU among subjects with IGD. ARPU is significantly associated with IGD. Whales share significant characteristics with addicted video gamers; Dolphins might be classified as risky consumers; Minnows and Freeloaders are rather non-pathological gamers. Vulnerability for stress, dysfunctional coping, and free-to-play gaming represent an unhealthy combination. Copyright © 2016 Elsevier Ltd. All rights reserved.

The potential effects of sodium bicarbonate, a major constituent from coalbed natural gas production, on aquatic life

Science.gov (United States)

Farag, Aïda M.; Harper, David D.

2012-01-01

,181 milligrams calcium carbonate per liter (mg CaCO3/L)) that varied across species and lifestage within a species. The age at which fish were exposed to NaHCO3 significantly affected the severity of toxic responses for some organisms. The chronic toxicity of NaHCO3 was defined in experiments that lasted from 7—60 days post-hatch. For these experiments, sublethal effects such as growth and reproduction, in addition to significant reductions in survival were included in the final determination of effects. Chronic toxicity was observed at concentrations that ranged from 450 to 800mg NaHCO3/L (also defined as 430 to 657 mg HCO3-/L or total alkalinity expressed as 354 to 539 mg CaCO3/L) and the specific concentration depended on the sensitivity of the four species of invertebrates and fish exposed. Sublethal investigations during chronic studies revealed percent decrease in the activity of sodium-potassium adenosine triphosphatase (Na/K ATPase, an enzyme involved in ionoregulation) and the age of the fish at the onset of the decrease may affect the ability of fathead minnow to survive exposures to NaHCO3. A database of toxicity evaluations of NaHCO3 on aquatic life has been constructed. Using these data, sample acute and chronic criteria of 459 and 381 mg NaHCO3/L, respectively, can be calculated for the protection of aquatic life. The final derivation and implementation of such criteria is, of course, left to the discretion of the concerned management agencies. A combination of in situ experiments, static-renewal experiments performed simultaneously with in situ experiments, and static renewal experiments performed with site water in the laboratory, demonstrated that untreated coalbed natural gas (CBNG) product water from the Tongue and Powder River Basins reduces survival of fathead minnow and pallid sturgeon. More precisely, the survival of early-lifestage fathead minnow, especially those less than 6-days post hatch (dph), likely is reduced significantly in the field when

Effects of Gemfibrozil on Cholesterol Metabolism and Steroidogenesis in the Fathead Minnow (Pimephales promelas)

Science.gov (United States)

Fibrates are a class of pharmaceuticals that indirectly modulate cholesterol biosynthesis through effects on peroxisome proliferator-activated receptors (PPAR), which are transcriptional cofactors that regulate expression of genes related to lipid metabolism. Gemfibrozil is a fib...

Analysis of porcine granulosa cell death signaling pathways induced by vinclozolin.

Science.gov (United States)

Knet, Malgorzata; Wartalski, Kamil; Hoja-Lukowicz, Dorota; Tabarowski, Zbigniew; Slomczynska, Maria; Duda, Malgorzata

2015-10-01

Recent studies suggest that disturbing androgen-signaling pathways in porcine ovarian follicles may cause granulosa cell (GC) death. For this reason, we investigated which apoptotic pathway is initiated after GC exposure to an environmental antiandrogen, vinclozolin (Vnz), in vitro. Immunocytochemistry, Western blots, and fluorometric assays were used to quantify caspase-3 and -9 expression and activity. To elucidate the specific mechanism of Vnz action and toxicity, GCs were assessed for viability, cytotoxicity, and apoptotic activity using the ApoTox-Glo Triplex Assay. To further determine the mechanism of GC death induced by Vnz, we used the Apoptosis Antibody Array Kit. In response to Vnz stimulus, we found an increased level of caspase-3 protein expression (P ≤ 0.001) and an increase in caspase-3 proteolytic activity (P ≤ 0.001), confirming that Vnz is a potent proapoptotic factor. The strong immunoreaction of caspase-9 after Vnz treatment (P ≤ 0.001) suggests that intrinsic mitochondrial apoptosis pathway was activated during GC death. On the other hand, caspase-8, being a part of the extrinsic receptor pathway, was also activated (P ≤ 0.001). Therefore, it is possible that Vnz induces porcine granulosal apoptosis also through a parallel pathway. Activation of these two pathways was confirmed by the Apoptosis Antibody Array Kit. In conclusion, it is possible that the intrinsic signaling pathway may not act as an initial trigger for GC apoptosis but might contribute to the amplification and propagation of apoptotic cell death in the granulosa layer after treatment with this antiandrogen. Moreover, Vnz disturbs the physiological process of programmed cell death. Consequently, this could explain why atretic follicles are rapidly removed and suggests that normal function of the ovarian follicle may be destroyed. Copyright © 2015 Elsevier Inc. All rights reserved.

MSPD procedure for determining buprofezin, tetradifon, vinclozolin, and bifenthrin residues in propolis by gas chromatography-mass spectrometry.

Science.gov (United States)

dos Santos, Thaíse Fernanda Santana; Aquino, Adriano; Dórea, Haroldo Silveira; Navickiene, Sandro

2008-03-01

A simple and effective extraction method based on matrix solid-phase dispersion (MSPD) was developed to determine bifenthrin, buprofezin, tetradifon, and vinclozolin in propolis using gas chromatography-mass spectrometry in selected ion monitoring mode (GC-MS, SIM). Different method conditions were evaluated, for example type of solid phase (C(18), alumina, silica, and Florisil), the amount of solid phase and eluent (n-hexane, dichloromethane, dichloromethane-n-hexane (8:2 and 1:1, v/v) and dichloromethane-ethyl acetate (9:1, 8:2 and 7:3, v/v)). The best results were obtained using 0.5 g propolis, 1.0 g silica as dispersant sorbent, 1.0 g Florisil as clean-up sorbent, and dichloromethane-ethyl acetate (9:1, v/v) as eluting solvent. The method was validated by analysis of propolis samples fortified at different concentration levels (0.25 to 1.0 mg kg(-1)). Average recoveries (four replicates) ranged from 67% to 175% with relative standard deviation between 5.6% and 12.1%. Detection and quantification limits ranged from 0.05 to 0.10 mg kg(-1) and 0.15 to 0.25 mg kg(-1) propolis, respectively.

Distinct actions of ancestral vinclozolin and juvenile stress on neural gene expression in the male rat

Directory of Open Access Journals (Sweden)

Ross eGillette

2015-03-01

Full Text Available Exposure to the endocrine disrupting chemical vinclozolin during gestation of an F0 generation and/or chronic restraint stress during adolescence of the F3 descendants affects behavior, physiology, and gene expression in the brain. Genes related to the networks of growth factors, signaling peptides and receptors, steroid hormone receptors and enzymes, and epigenetic related factors were measured using quantitative polymerase chain reaction via Taqman low density arrays targeting 48 genes in the central amygdaloid nucleus, medial amygdaloid nucleus, medial preoptic area, lateral hypothalamus, and the ventromedial nucleus of the hypothalamus. We found that growth factors are particularly vulnerable to ancestral exposure in the central and medial amygdala; restraint stress during adolescence affected neural growth factors in the medial amygdala. Signaling peptides were affected by both ancestral exposure and stress during adolescence primarily in hypothalamic nuclei. Steroid hormone receptors and enzymes were strongly affected by restraint stress in the medial preoptic area. Epigenetic related genes were affected by stress in the ventromedial hypothalamus and by both ancestral exposure and stress during adolescence independently in the central amygdala. It is noteworthy that the lateral hypothalamus showed no effects of either manipulation. Gene expression is discussed in the context of behavioral and physiological measures previously published.

Distinct actions of ancestral vinclozolin and juvenile stress on neural gene expression in the male rat.

Science.gov (United States)

Gillette, Ross; Miller-Crews, Isaac; Skinner, Michael K; Crews, David

2015-01-01

Exposure to the endocrine disrupting chemical vinclozolin during gestation of an F0 generation and/or chronic restraint stress during adolescence of the F3 descendants affects behavior, physiology, and gene expression in the brain. Genes related to the networks of growth factors, signaling peptides, and receptors, steroid hormone receptors and enzymes, and epigenetic related factors were measured using quantitative polymerase chain reaction via Taqman low density arrays targeting 48 genes in the central amygdaloid nucleus, medial amygdaloid nucleus, medial preoptic area (mPOA), lateral hypothalamus (LH), and the ventromedial nucleus of the hypothalamus. We found that growth factors are particularly vulnerable to ancestral exposure in the central and medial amygdala; restraint stress during adolescence affected neural growth factors in the medial amygdala. Signaling peptides were affected by both ancestral exposure and stress during adolescence primarily in hypothalamic nuclei. Steroid hormone receptors and enzymes were strongly affected by restraint stress in the mPOA. Epigenetic related genes were affected by stress in the ventromedial nucleus and by both ancestral exposure and stress during adolescence independently in the central amygdala. It is noteworthy that the LH showed no effects of either manipulation. Gene expression is discussed in the context of behavioral and physiological measures previously published.

Gene expression profiling of the androgen receptor antagonists flutamide and vinclozolin in zebrafish (Danio rerio) gonads.

Science.gov (United States)

Martinović-Weigelt, Dalma; Wang, Rong-Lin; Villeneuve, Daniel L; Bencic, David C; Lazorchak, Jim; Ankley, Gerald T

2011-01-25

The studies presented in this manuscript focus on characterization of transcriptomic responses to anti-androgens in zebrafish (Danio rerio). Research on the effects of anti-androgens in fish has been characterized by a heavy reliance on apical endpoints, and molecular mechanisms of action (MOA) of anti-androgens remain poorly elucidated. In the present study, we examined effects of a short term exposure (24-96h) to the androgen receptor antagonists flutamide (FLU) and vinclozolin (VZ) on gene expression in gonads of sexually mature zebrafish, using commercially available zebrafish oligonucleotide microarrays (4×44K platform). We found that VZ and FLU potentially impact reproductive processes via multiple pathways related to steroidogenesis, spermatogenesis, and fertilization. Observed changes in gene expression often were shared by VZ and FLU, as demonstrated by overlap in differentially-expressed genes and enrichment of several common key pathways including: (1) integrin and actin signaling, (2) nuclear receptor 5A1 signaling, (3) fibroblast growth factor receptor signaling, (4) polyamine synthesis, and (5) androgen synthesis. This information should prove useful to elucidating specific mechanisms of reproductive effects of anti-androgens in fish, as well as developing biomarkers for this important class of endocrine-active chemicals. 2010 Elsevier B.V. All rights reserved.

Cannibalistic-morph Tiger Salamanders in unexpected ecological contexts

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McLean, Kyle I.; Stockwell, Craig A.; Mushet, David M.

2016-01-01

Barred tiger salamanders [Ambystoma mavortium (Baird, 1850)] exhibit two trophic morphologies; a typical and a cannibalistic morph. Cannibalistic morphs, distinguished by enlarged vomerine teeth, wide heads, slender bodies, and cannibalistic tendencies, are often found where conspecifics occur at high density. During 2012 and 2013, 162 North Dakota wetlands and lakes were sampled for salamanders. Fifty-one contained A. mavortium populations; four of these contained cannibalistic morph individuals. Two populations with cannibalistic morphs occurred at sites with high abundances of conspecifics. However, the other two populations occurred at sites with unexpectedly low conspecific but high fathead minnow [Pimephales promelas (Rafinesque, 1820)] abundances. Further, no typical morphs were observed in either of these later two populations, contrasting with earlier research suggesting cannibalistic morphs only occur at low frequencies in salamander populations. Another anomaly of all four populations was the occurrence of cannibalistic morphs in permanent water sites, suggesting their presence was due to factors other than faster growth allowing them to occupy ephemeral habitats. Therefore, our findings suggest environmental factors inducing the cannibalistic morphism may be more complex than previously thought.

Oxidation of danofloxacin by free chlorine-kinetic study, structural identification of by-products by LC-MS/MS and potential toxicity of by-products using in silico test.

Science.gov (United States)

Yassine, Montaha; Rifai, Ahmad; Doumyati, Samah; Trivella, Aurélien; Mazellier, Patrick; Budzinski, Hélène; Al Iskandarani, Mohamad

2017-03-01

In this study, we aimed to investigate the kinetics and the mechanism of reaction of the fluoroquinolone antibacterial danofloxacin (DANO) by free available chlorine (FAC) during water chlorination process. Kinetic study was thus performed at pH 7.2, 20 °C in the presence of an excess of total chlorine. Under these experimental conditions, a second-order reaction rate constant (first-order relative to DANO concentration and first-order relative to FAC concentration) was evaluated to k~1446 M -1  s -1 . Five degradation products were identified at different reaction times. Their structures were investigated by using fragmentations obtained at different CID collision energies in MS/MS experiments. Moreover, the toxicity of the proposed structures was predicted by using T.E.S.T. The results indicated that all by-products may have a developmental toxicity. The oral rat LD 50 concentration was predicted to be lower than that of DANO. Furthermore, two degradation compounds presented a concentration level for fathead minnow LC 50 (96 h) lower than that of DANO and presented toxicity for the marine animals.

Identification of the cause of weak acute toxicity to rainbow trout at a petroleum refinery

International Nuclear Information System (INIS)

Arnold, W.R.; Zaleski, R.T.; Biddinger, G.R.

1995-01-01

The refinery in question performs flow through acute toxicity tests on its effluent four times per month using three fish species: fathead minnows (Pimephales promelas), threespine sticklebacks (Gasterosteus oculeatus) and rainbow trout (Oncorhynchus mykiss). Several months of monitoring data indicated a transient low level acute toxicity to rainbow trout. In most cases, several days were required for mortality to occur in the flow through tests and numerous attempts to reproduce toxicity in static and static renewal tests were unsuccessful. A decision was made to manipulate the effluent in an attempt to enhance the toxic effect in the static mode so that conventional methods could be used to identify the cause. these tests indicated that toxicity was pH dependent. Additional testing, using EPA's Phase 1 Toxicity Identification Evaluation methods suggested that the cause of toxicity was probably an organic acid. Experiments were subsequently begun to identify the specific cause and source of toxicity. This paper reviews the problems confronted during the various phases of the study and the decisions that were made that eventually led to an understanding of the basis of toxicity

Interaction of chemical cues from fish tissues and organophosphorous pesticides on Ceriodaphnia dubia survival

International Nuclear Information System (INIS)

Maul, Jonathan D.; Farris, Jerry L.; Lydy, Michael J.

2006-01-01

Cladocera are frequently used as test organisms for assessing chemical and effluent toxicity and have been shown to respond to stimuli and cues from potential predators. In this study, the interactive effects of visual and chemical cues of fish and two organophosphorous pesticides on survival of Ceriodaphnia dubia were examined. A significant chemical cue (homogenized Pimephales promelas) and malathion interaction was observed on C. dubia survival (P = 0.006). Chemical cue and 2.82 μg/L malathion resulted in a 76.0% reduction in survival compared to malathion alone (P < 0.01). Furthermore, potentiation of malathion toxicity varied based on the source of chemical cues (i.e., epithelial or whole body). It is unclear in this study whether these chemical cues elicited a predation-related stress in C. dubia. Future research should examine the mechanism of this interaction and determine what role, if any, stress responses by C. dubia might play in the interaction. - Potentiation of organophosphorous pesticide toxicity to Ceriodaphnia dubia by fathead minnow (Pimephales promelas) chemical cues was observed

Development of an ecotoxicity QSAR model for the KAshinhou Tool for Ecotoxicity (KATE) system, March 2009 version.

Science.gov (United States)

Furuhama, A; Toida, T; Nishikawa, N; Aoki, Y; Yoshioka, Y; Shiraishi, H

2010-07-01

The KAshinhou Tool for Ecotoxicity (KATE) system, including ecotoxicity quantitative structure-activity relationship (QSAR) models, was developed by the Japanese National Institute for Environmental Studies (NIES) using the database of aquatic toxicity results gathered by the Japanese Ministry of the Environment and the US EPA fathead minnow database. In this system chemicals can be entered according to their one-dimensional structures and classified by substructure. The QSAR equations for predicting the toxicity of a chemical compound assume a linear correlation between its log P value and its aquatic toxicity. KATE uses a structural domain called C-judgement, defined by the substructures of specified functional groups in the QSAR models. Internal validation by the leave-one-out method confirms that the QSAR equations, with r(2 )> 0.7, RMSE 5, give acceptable q(2) values. Such external validation indicates that a group of chemicals with an in-domain of KATE C-judgements exhibits a lower root mean square error (RMSE). These findings demonstrate that the KATE system has the potential to enable chemicals to be categorised as potential hazards.

Gene transcription ontogeny of thyroid-axis development in early-life stage fathead minnows (Pimephales promelas)

Science.gov (United States)

Disruption of thyroid hormone signaling is a form of endocrine disruption that is of concern to both human health and ecosystems. Research is being conducted to define the biological targets chemicals may interact with to disrupt thyroid hormone signaling and the stages in develo...

Hypothesis testing with computational modeling: linking aromatase inhibition with plasma vitellogenin dynamics in fathead minnows

Science.gov (United States)

Fadrozole inhibits aromatase (CYP19A), a key enzyme that converts testosterone to estradiol (E2). In fish, E2 concentrations control hepatic synthesis ofthe glycolipoprotein vitellogenin (VTG), an egg yolk precursor protein essential to oocyte development and larval survival. Whe...

Effects of Gemfibrozil on Cholesterol Metabolism, Steroidogenesis, and Reproduction in the Fathead Minnow (Pimephales promelas)

Science.gov (United States)

Fibrates are a class of pharmaceuticals that indirectly modulate cholesterol biosynthesis through effects on peroxisome proliferator-activated receptors, which are transcriptional cofactors that regulate expression of genes related to lipid metabolism. Gemfibrozil is a fibrate th...

Effects of gemfibrozil on lipid metabolism, steroidogenesis and reproduction in the fathead minnow (Pimephales promelas)

Science.gov (United States)

Fibrates are a class of pharmaceuticals that indirectly modulate cholesterol biosynthesis through effects on peroxisome proliferator-activated receptors (PPARs), which are transcriptional cofactors that regulate expression of genes related to lipid metabolism. Gemfibrozil is a fi...

Prior knowledge-based approach for associating contaminants with biological effects: A case study in the St. Croix River basin, MN, WI, USA

Science.gov (United States)

Schroeder, Anthony L.; Martinovic-Weigelt, Dalma; Ankley, Gerald T.; Lee, Kathy E.; Garcia-Reyero, Natalia; Perkins, Edward J.; Schoenfuss, Heiko L.; Villeneuve, Daniel L.

2017-01-01

Evaluating potential adverse effects of complex chemical mixtures in the environment is challenging. One way to address that challenge is through more integrated analysis of chemical monitoring and biological effects data. In the present study, water samples from five locations near two municipal wastewater treatment plants in the St. Croix River basin, on the border of MN and WI, USA, were analyzed for 127 organic contaminants. Known chemical-gene interactions were used to develop site-specific knowledge assembly models (KAMs) and formulate hypotheses concerning possible biological effects associated with chemicals detected in water samples from each location. Additionally, hepatic gene expression data were collected for fathead minnows (Pimephales promelas) exposed in situ, for 12 d, at each location. Expression data from oligonucleotide microarrays were analyzed to identify functional annotation terms enriched among the differentially-expressed probes. The general nature of many of the terms made hypothesis formulation on the basis of the transcriptome-level response alone difficult. However, integrated analysis of the transcriptome data in the context of the site-specific KAMs allowed for evaluation of the likelihood of specific chemicals contributing to observed biological responses. Thirteen chemicals (atrazine, carbamazepine, metformin, thiabendazole, diazepam, cholesterol, p-cresol, phenytoin, omeprazole, ethyromycin, 17β-estradiol, cimetidine, and estrone), for which there was statistically significant concordance between occurrence at a site and expected biological response as represented in the KAM, were identified. While not definitive, the approach provides a line of evidence for evaluating potential cause-effect relationships between components of a complex mixture of contaminants and biological effects data, which can inform subsequent monitoring and investigation.

In vitro metabolism of the anti-androgenic fungicide vinclozolin by rat liver microsomes.

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Sierra-Santoyo, Adolfo; Angeles-Soto, Esperanza; de Lourdes López-González, Ma; Harrison, Randy A; Hughes, Michael F

2012-03-01

Vinclozolin (V) is a fungicide used in agricultural settings. V administered to rats is hydrolyzed to 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3-butenoic acid (M1) and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2). V, M1 and M2 have antiandrogenic properties by interacting with the androgen receptor. Data on V, M1 and M2 biotransformation are limited. Our objective was to characterize V metabolism by rat liver microsomes. V was incubated with non-treated adult male Long-Evans rat liver microsomes and NADPH. Several metabolites were detected following the extraction of incubate with acetonitrile and analysis by HPLC/DAD/MSD. One metabolite was identified as [3-(3,5-dichlorophenyl)-5-methyl-5-(1,2-dihydroxyethyl)-1,3-oxazolidine-2,4-dione] (M4), which was gradually converted to 3',5'-dichloro-2,3,4-trihydroxy-2-methylbutylanilide (M5). Both co-eluted in the same HPLC peak. Another metabolite ([M7]) was detected by UV but was unstable for mass spectral analysis. The K(M app) for co-eluted M4/M5 and [M7] was 53.7 and 135.4 μM, the V(max app) was 0.812 and 0.669 nmoles/min/mg protein, and CL(int) was 15.1 and 4.9 ml/min/g protein, respectively. Pilocarpine, orphenadrine and proadifen and anti-rat cytochrome P450 (CYP)2A, 2B and 3A antibodies inhibited M4/M5 and [M7] formation. These results indicate that V is efficiently metabolized by CYP. Determination of the metabolites of V will provide further insight into the relationship between toxicity and tissue dose of V and its metabolites.

Catchment-scale determinants of nonindigenous minnow richness in the eastern United States

Science.gov (United States)

Peoples, Brandon K.; Midway, Stephen R.; DeWeber, Jefferson T.; Wagner, Tyler

2018-01-01

Understanding the drivers of biological invasions is critical for preserving aquatic biodiversity. Stream fishes make excellent model taxa for examining mechanisms driving species introduction success because their distributions are naturally limited by catchment boundaries. In this study, we compared the relative importance of catchment-scale abiotic and biotic predictors of native and nonindigenous minnow (Cyprinidae) richness in 170 catchments throughout the eastern United States. We compared historic and contemporary cyprinid distributional data to determine catchment-wise native/nonindigenous status for 152 species. Catchment-scale model predictor variables described natural (elevation, precipitation, flow accumulation) and anthropogenic (developed land cover, number of dams) abiotic features, as well as native congener richness. Native congener richness may represent either biotic resistance via interspecific competition, or trait preadaptation according to Darwin's naturalisation hypothesis. We used generalised linear mixed models to examine evidence supporting the relative roles of abiotic and biotic predictors of cyprinid introduction success. Native congener richness was positively correlated with nonindigenous cyprinid richness and was the most important variable predicting nonindigenous cyprinid richness. Mean elevation had a weak positive effect, and effects of other abiotic factors were insignificant and less important. Our results suggest that at this spatial scale, trait preadaptation may be more important than intrageneric competition for determining richness of nonindigenous fishes.

An environmentally relevant endocrine-disrupting antiandrogen, vinclozolin, affects calling behavior of male Xenopus laevis.

Science.gov (United States)

Hoffmann, Frauke; Kloas, Werner

2010-09-01

Vinclozolin (VIN) is an antiandrogenic model substance as well as a common fungicide that can affect the endocrine system of vertebrates. The objective of this study was to investigate how VIN affects mate calling behavior of South African clawed frogs (Xenopus laevis) and whether it is effective at environmentally relevant concentrations. Male X. laevis were injected with human chorionic gonadotropin (hCG) to stimulate their androgen-controlled mate calling behavior and were treated with VIN at concentrations of 10(-6), 10(-8) and 10(-10)M. VIN at 10(-6)M reduced calling activity. Furthermore, the vocalization composition of VIN-treated X. laevis was altered. The call types advertisement calls and chirping are uttered by reproductively active males, whereas the call types growling, ticking, and rasping indicate a sexually unaroused state of a male. VIN at any of the tested concentrations led to a decrease in utterance of calls, which indicate a sexually aroused state of the males, and an increase in relative proportions of calls, indicating a sexually unaroused state of the males. Additionally, the mean duration of clicks and the number of accentuated clicks during the advertisement calls decreased at all concentrations of VIN. No significant differences were observed in any other temporal or spectral calling parameters between the treatments. This study illustrates that exposure to the antiandrogen VIN might result in a reduced reproductive success by altering mate calling behavior of X. laevis. Moreover, it suggests that the behavioral parameters examined in this study can be used as sensitive biomarkers for detecting antiandrogenic endocrine disrupting compounds in amphibians. Copyright (c) 2010 Elsevier Inc. All rights reserved.

Tennessee's East Fork Poplar Creek: A biological monitoring and abatement program

International Nuclear Information System (INIS)

Halbrook, R.S.; Loar, J.M.; Adams, S.M.; Black, M.C.; Boston, H.L.; Greeley, M.S. Jr.; Hill, W.R.; Hinzman, R.L.; McCarthy, J.F.; Peterson, M.J.; Ryon, M.G.; Schilling, E.M.; Smith, J.G.; Southworth, G.R.; Stewart, A.J.; Gatz, A.J.

1991-01-01

On May 1985, a Biological Monitoring Program was developed for East Fork Poplar Creek (EFPC) in eastern Tennessee, United States. This stream originates within the Oak Ridge Y-12 Plant that produces nuclear weapons components for the Department of Energy. Water and sediment in the stream contain metals, organic chemicals, and radionuclides from releases that have occurred over the past 45 years. The creek also receives urban and some agricultural runoff and effluent from the City of Oak Ridge's Wastewater Treatment Facility (WTF). The biological monitoring program includes four major tasks: (1) ambient toxicity testing: (2) bioaccumulation studies; (3) biological indicator studies; and (4) ecological monitoring of stream communities, including periphyton, benthic macroinvertebrates, and fish. Biological conditions are monitored at six sites on EFPC ranging from kilometer 24.4 near the headwaters to kilometer 6.3 near the month. A site on Brushy Fork, A stream just north of Oak Ridge, is used as reference. Ambient (instream) toxicity was monitored through the use of 7-day static-renewal tests that measured the survival and growth of fathead minnow (Pimephales promelas) larvae and the survival and reproduction of a microstrustacean (Ceriodaphnia dubia). Full-strength water from EFPC within the Y-12 Plant boundary was frequently toxic to Ceriodaphnia, but less frequently toxic to the minnow larvae. Chlorine has been identified as an important toxicant in upper EFPC. Water samples from six sites in EFPC downstream from the Y-12 Plant boundary were tested eight times with both species during a 2-year period (October, 1986 through October, 1988). These sites were ranked by the number of times they were ''best'' or ''worst'' for each species. Water samples collected for use in the ambient toxicity tests were routinely analyzed for conductivity, pH, alkalinity, hardness, total residual and free chlorine, and temperature

Cloning and initial characterization of nuclear and membrane progesterone receptors in the Fathead Minnow, Pimephales promelas

Science.gov (United States)

Both native progestagens and synthetic progestins have important effects on reproduction that are mediated through progesterone receptors (PRs). They regulate gamete maturation and can serve as precursors for other steroid hormones in vertebrates and act as reproductive pheromone...

Effects of Fadrozole, Ketoconazole, and 17β-trenbolone on Ex Vivo Steroidogenesis in the Fathead Minnow

Science.gov (United States)

A variety of endocrine-disrupting chemicals have the ability to disrupt steroidogenesis through interaction with the hypothalamic-pituitary-gonadal (HPG) axis. We examined the effects of the competitive aromatase inhibitor fadrozole (0, 3, and 30 g/L), the cytochrome P450 enzyme...

Short-term effects of propiconazole on hypothalamic-pituitary-gonadal function in the fathead minnows (Pimephales promelas)

Science.gov (United States)

Propiconazole is an ergosterol inhibitor commonly used in agriculture and has been detected in aquatic environments. Ergosterol inhibitors decrease fungal growth through effects on 14á-demethylase, a cytochrome P450 (CYP), isoform important for ergosterol biosynthesis. In higher ...

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.

Science.gov (United States)

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

Pharmacokinetics and dosimetry of the antiandrogen vinclozolin after oral administration in the rat.

Science.gov (United States)

Sierra-Santoyo, Adolfo; Castañeda-Hernández, Gilberto; Harrison, Randy A; Barton, Hugh A; Hughes, Michael F

2008-11-01

Vinclozolin (V) is a fungicide with antiandrogenic properties. To determine the pharmacokinetics and dosimetry of V, adult male rats were administered an oral dose of V (100 mg/kg) in corn oil and sacrificed over time after dosing. V and its metabolites were analyzed in serum and tissues by high performance liquid chromatography/diode array detector/mass spectrometer. V, 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3-butenoic acid (M1), and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2), and five other metabolites were detected in serum and tissues. One metabolite was identified as 3',5'-dichloro-2,3,4-trihydroxy-2-methylbutylanilide (M5). The mean serum concentration data for V were fitted to a one-compartment model for kinetic analysis. At 2 h, V serum concentration peaked; whereas only trace levels were detected at 24 h (t(1/2 elim) = 3.6 h). V was detected in all tissues and preferentially accumulated in fat. M1 serum levels increased until 8 h, being at least 2-fold higher than those of V at this time, and then declined with a t(1/2) = 3.3 h. M5 was the main metabolite in serum and tissues. Serum M5 levels were 5-fold higher than V and 2-fold greater than M1 at all times. At 48 h, M5 remained the main metabolite (t(1/2 elim) = 13.1 h). Liver and kidney exhibited the highest levels of M5, V, and M1. M2 and 3,5-dichloroaniline had the lowest levels of V metabolites in serum and tissues. V is well absorbed, extensively metabolized and widely distributed. M5, the most abundant V metabolite, may be used as an exposure biomarker for pharmacokinetic modeling. These results may clarify the relationship between toxicity and tissue dose of V and its metabolites.

Bioavailability study for the Paducah Gaseous Diffusion Plant

Energy Technology Data Exchange (ETDEWEB)

Phipps, T.L.; Kszos, L.A.

1996-08-01

The overall purpose of this plan is to assess the bioavailability of metals in the continuous and intermittent outfalls. The results may be used to determine alternative metal limits that more appropriately measure the portion of metal present necessary for toxicity to aquatic life. These limits must remain protective of in-stream aquatic life; thus, the highest concentration of metal in the water will be determined concurrently with an assessment of acute or chronic toxicity on laboratory tests. Using the method developed by the Kentucky Division of Water (KDOW), biomonitoring results and chemical data will be used to recommend alternative metal limits for the outfalls of concern. The data will be used to meet the objectives of the study: (1) evaluate the toxicity of continuous outfalls and intermittent outfalls at Paducah Gaseous Diffusion Plant; (2) determine the mean ratio of dissolved to Total Recoverable metal for Cd, Cr, Cu, Pb, Ni, and Zn in the continuous and intermittent outfalls; (3) determine whether the concentration of total recoverable metal discharged causes toxicity to fathead minnows and /or Ceriodaphnia; and (4) determine alternative metal limits for each metal of concern (Cd, Cr, Cu, Pb, Ni, and Zn).

Geochemical pathways and biological uptake of radium in small Canadian Shield lakes

International Nuclear Information System (INIS)

Hesslein, R.H.; Slavicek, E.

1984-01-01

The sediment-water interactions and biological uptake of 226 Ra are described for four small Canadian Shield lakes at the Experimental Lakes Area, Kenora, Ontario. A single addition of 226 Ra was made to each lake between 1970 and 1976. Approximately 90 percent of the added 226 Ra initially sorbed to the sediments. Outflow from the lakes showed losses of only 5-11 percent 226 Ra per year. Models are proposed for adsorption and outflow of 226 Ra from lakes. Biological uptake and long-term 226 Ra concentrations were measured in three species of macrophytes, crayfish, and five species of fish. Bioaccumulation ranged from 1100 to 5000 in macrophytes, 705 in crayfish, from 30 to 80 in large trout (Salvelinus namaycush), white sucker (Catostomus commersoni), and lake whitefish (Coregonus clupeaformis), and from 230 to 1200 in fathead minnows (Pimephales promelas), pearl dace (Semotilus margarita), and northern redbelly dace (Chrosomus eos). The concept of Ra/Ca ratio in organisms versus water and food is used to explain the differences in bioaccumulation. 226 Ra is discriminated against versus calcium by fish but favoured by macrophytes and crayfish

Fate of the pyrethroid insecticide deltamethrin in small ponds: a mass balance study

International Nuclear Information System (INIS)

Muir, D.C.G.; Rawn, G.P.; Grift, N.P.

1985-01-01

The fate and distribution of 14 C-radiolabeled deltamethrin (1(R)[la(S),3a]-cyano-(3-phenoxyphenyl)methyl 3-(2,2-dibromoethenyl)-2,2-dimethylcyclopropanecarboxylate) were monitored for 306 days, following a single application at 10 g/ha to two small outdoor ponds (17 m 2 surface area). Initial concentrations of the insecticide in filtered water ranged from 1.28 to 2.50 μg/L. Deltamethrin ([ 14 C]cyclopropyl acid or benzyl alcohol labeled) rapidly partitioned into suspended solids, plants, sediment, and air, with a half-life of 2-4 h in water. Duckweed (Lemna sp.) and a submerged pondweed (Potamogeton berchtoldi) accumulated deltamethrin concentrations ranging from 253 to 1021 ng/g, respectively, at 24 h posttreatment. Sediments were the major sink for radioactivity at 306 days posttreatment, and intact deltamethrin was present at concentrations ranging from 3 to 5 ng/g. Deltamethrin levels in air above the water ranged from 10-100 ng/m 3 during a 48-h monitoring period following application. Fathead minnows (Pimephales promelas) accumulated levels of extractable radioactivity 248-907-fold higher than concentrations in water 24 h posttreatment, but no fish mortality was observed

Small nonnative fishes as predators of larval razorback suckers

Science.gov (United States)

Carpenter, J.; Mueller, G.A.

2008-01-01

The razorback sucker (Xyrauchen texanus), an endangered big-river fish of the Colorado River basin, has demonstrated no sustainable recruitment in 4 decades, despite presence of spawning adults and larvae. Lack of adequate recruitment has been attributed to several factors, including predation by nonnative fishes. Substantial funding and effort has been expended on mechanically removing nonnative game fishes, typically targeting large predators. As a result, abundance of larger predators has declined, but the abundance of small nonnative fishes has increased in some areas. We conducted laboratory experiments to determine if small nonnative fishes would consume larval razorback suckers. We tested adults of three small species (threadfin shad, Dorosoma petenense; red shiner, Cyprinella lutrensis; fathead minnow, Pimephales promelas) and juveniles of six larger species (common carp, Cyprinus carpio; yellow bullhead, Ameiurus natalis; channel catfish, Ictalurus punctatus; rainbow trout, Oncorhynchus mykiss; green sunfish, Lepomis cyanellus; bluegill, L. macrochirus). These nonnative fishes span a broad ecological range and are abundant within the historical range of the razorback sucker. All nine species fed on larval razorback suckers (total length, 9-16 mm). Our results suggest that predation by small nonnative fishes could be responsible for limiting recovery of this endangered species.

Hazard evaluation of soil contaminants from an abandoned oil refinery site with chemical and biological assays

International Nuclear Information System (INIS)

Ramanathan, A.; Yates, C.W.; Burks, S.L.

1993-01-01

The phytotoxic characteristics of soil and leachates of soil from an abandoned oil refinery site were evaluated with rice (Oryza sativa L.) seed germinations and root elongation assays. Toxicity of soil leachates to aquatic animals was determined with acute and martial chronic toxicity tests with Ceriodaphnia dubia, fathead minnows, and Microtox reg-sign. Soil samples from uncontaminated (control) and selected contaminated areas within the old refinery were extracted with Toxic Characteristics Leachate Procedure (TCLP), an aqueous procedure and a supercritical carbon dioxide method. Aqueous extracts of soil from the oil leaded gasoline storage area exhibited greatest effects in all tests. Aqueous extracts from this site also caused a significant reduction in rice root development. Supercritical carbon dioxide extraction proved to be a quick and non-toxic procedure for isolating non-polar organics for assay with aquatic toxicity tests. Subsequent supercritical extracts collected in solvent can help characterize the class of toxicants through HPLC and Gas Chromatography. The toxic constituents were characterized with a Toxicity Identification/Toxicity Reduction Evaluation protocol to fractionate the contaminants into conventional non-polar organics, weak acids, base-neutrals, or heavy metals for subsequent analysis

Methoxychlor and Vinclozolin Induce Rapid Changes in Intercellular and Intracellular Signaling in Liver Progenitor Cells.

Science.gov (United States)

Babica, Pavel; Zurabian, Rimma; Kumar, Esha R; Chopra, Rajus; Mianecki, Maxwell J; Park, Joon-Suk; Jaša, Libor; Trosko, James E; Upham, Brad L

2016-09-01

Methoxychlor (MXC) and vinclozolin (VIN) are well-recognized endocrine disrupting chemicals known to alter epigenetic regulations and transgenerational inheritance; however, non-endocrine disruption endpoints are also important. Thus, we determined the effects of MXC and VIN on the dysregulation of gap junctional intercellular communication (GJIC) and activation of mitogen-activated protein kinases (MAPKs) in WB-F344 rat liver epithelial cells. Both chemicals induced a rapid dysregulation of GJIC at non-cytotoxic doses, with 30 min EC50 values for GJIC inhibition being 10 µM for MXC and 126 µM for VIN. MXC inhibited GJIC for at least 24 h, while VIN effects were transient and GJIC recovered after 4 h. VIN induced rapid hyperphosphorylation and internalization of gap junction protein connexin43, and both chemicals also activated MAPK ERK1/2 and p38. Effects on GJIC were not prevented by MEK1/2 inhibitor, but by an inhibitor of phosphatidylcholine-specific phospholipase C (PC-PLC), resveratrol, and in the case of VIN, also, by a p38 inhibitor. Estrogen (ER) and androgen receptor (AR) modulators (estradiol, ICI 182,780, HPTE, testosterone, flutamide, VIN M2) did not attenuate MXC or VIN effects on GJIC. Our data also indicate that the effects were elicited by the parental compounds of MXC and VIN. Our study provides new evidence that MXC and VIN dysregulate GJIC via mechanisms involving rapid activation of PC-PLC occurring independently of ER- or AR-dependent genomic signaling. Such alterations of rapid intercellular and intracellular signaling events involved in regulations of gene expression, tissue development, function and homeostasis, could also contribute to transgenerational epigenetic effects of endocrine disruptors. © The Author 2016. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Unsupervised Analysis of the Effects of a Wastewater Treatment Plant Effluent on the Fathead Minnow Ovarian Transcriptome

Science.gov (United States)

Wastewater treatment plant (WWTP) effluents contain complex mixtures of chemicals, potentially including endocrine active chemicals (EACs), pharmaceuticals, and other contaminants of emerging concern (CECs). Due to the complex and variable nature of effluents, biological monitori...

Gene transcription ontogeny of hypothalamic-pituitary-thyroid-axis development in early-life stage fathead minnow and zebrafish

Science.gov (United States)

Disruption of thyroid hormone signaling is a form of endocrine disruption that is of concern to both human health and ecosystems. Research is being conducted to define the biological targets chemicals may interact with to disrupt thyroid hormone signaling and the stages in develo...

Cloning and initial characterization of nuclear and four membrane progesterone receptors in the fathead minnow(Pimephales promelas)

Science.gov (United States)

Both native progestagens and synthetic progestins have important effects on reproduction that are mediated through progesterone receptors (PRs). Progestagens regulate gamete maturation in vertebrates, are critical regulators of placental mammal pregnancy, and act as reproductive ...

Is Low Efficacy of Fungicides always a Consequence of Fungicide Resistance Development in Pathogen Populations?

Directory of Open Access Journals (Sweden)

Brankica Tanović

2011-01-01

Full Text Available Efficacy of four fungicides with different modes of action (vinclozolin, pyrimethanil, benomyland fenhexamid in control of B. cinerea in raspberry, was investigated in the paper.The trials were conducted at two localities in commercial raspberry plantations. In the caseof unsatisfactory fungicide efficacy, qualitative and/or quantitative test of the susceptibilityof the isolates to particular fungicide was performed, to determine whether the low efficacyis a consequence of resistance development in the pathogen population. At both localities,pyrimethanil and fenhexamid demonstrated the highest efficacy (73.2-89.6%, whilethe efficacy of vinclozolin was statistically significantly lower (48.7-63.4% at both localities.However, qualitative and quantitative test of susceptibility to vinclozolin showed that all theisolates were susceptible to vinclozolin and that the reason for unsatisfactory efficacy shouldbe primarily sought in inadequate fungicide application

Effects of Fungicides on Rat’s Neurosteroid Synthetic Enzymes

Directory of Open Access Journals (Sweden)

Xiuwei Shen

2017-01-01

Full Text Available Exposure to environmental endocrine disruptors may interfere with nervous system’s activity. Fungicides such as tebuconazole, triadimefon, and vinclozolin have antifungal activities and are used to prevent fungal infections in agricultural plants. In the present study, we studied effects of tebuconazole, triadimefon, and vinclozolin on rat’s neurosteroidogenic 5α-reductase 1 (5α-Red1, 3α-hydroxysteroid dehydrogenase (3α-HSD, and retinol dehydrogenase 2 (RDH2. Rat’s 5α-Red1, 3α-HSD, and RDH2 were cloned and expressed in COS-1 cells, and effects of these fungicides on them were measured. Tebuconazole and triadimefon competitively inhibited 5α-Red1, with IC50 values of 8.670 ± 0.771 × 10−6 M and 17.390 ± 0.079 × 10−6 M, respectively, while vinclozolin did not inhibit the enzyme at 100 × 10−6 M. Triadimefon competitively inhibited 3α-HSD, with IC50 value of 26.493 ± 0.076 × 10−6 M. Tebuconazole and vinclozolin weakly inhibited 3α-HSD, with IC50 values about 100 × 10−6 M, while vinclozolin did not inhibit the enzyme even at 100 × 10−6 M. Tebuconazole and triadimefon weakly inhibited RDH2 with IC50 values over 100 × 10−6 M and vinclozolin did not inhibit this enzyme at 100 × 10−6 M. Docking study showed that tebuconazole, triadimefon, and vinclozolin bound to the steroid-binding pocket of 3α-HSD. In conclusion, triadimefon potently inhibited rat’s neurosteroidogenic enzymes, 5α-Red1 and 3α-HSD.

PAH related effects on fish in sedimentation ponds for road runoff and potential transfer of PAHs from sediment to biota.

Science.gov (United States)

Grung, Merete; Petersen, Karina; Fjeld, Eirik; Allan, Ian; Christensen, Jan H; Malmqvist, Linus M V; Meland, Sondre; Ranneklev, Sissel

2016-10-01

Road runoff is an important source of pollution to the aquatic environment, and sedimentation ponds have been installed to mitigate effects on the aquatic environment. The purpose of this study was to investigate if a) fish from sedimentation ponds were affected by road pollution and; b) the transfer of PAHs from road runoff material to aquatic organisms was substantial. Minnow from a sedimentation pond (Skullerud) near Oslo (Norway) had higher levels of CYP1A enzyme and DNA stand breaks than minnow from the nearby river, but high concentrations of PAH-metabolites in bile revealed that both populations were highly exposed. Principal component analysis revealed that CYP1A and age of fish were correlated, while levels of PAH-metabolites were not correlated to CYP1A or DNA damage. Minnow from a lake un-affected by traffic had much lower levels of PAH-metabolites than the exposed fish, and also an improved condition. The latter results indicate that fish health was affected by road runoff. A closer investigation of PAH levels of the ecosystems of two sedimentation ponds (Skullerud and Vassum) and nearby environments were conducted. The concentration of the 16 EPA PAHs in sediments of the sedimentation ponds were high (1900-4200ngg(-1)), and even higher levels were observed in plants. Principal component analysis of selected ion chromatograms of PAHs showed a clear separation of plants vs. sediments. The plants preferentially accumulated the high molecular PAHs, both from sedimentation ponds with a petrogenic PAH isomer ratio in sediments; and from a lake with pyrogenic PAH isomer ratio in sediments. Copyright © 2016 Elsevier B.V. All rights reserved.

IPRODIONE DELAYS MALE RAT PUBERTAL DEVELOPMENT, REDUCING SERUM TESTOSTERONE AND EX VIVO TESTOSTERONE PRODUCTION

Science.gov (United States)

Iprodione (IPRO) is a dichlorophenyl dicarboximide fungicide similar to the androgen receptor (AR) antagonist vinclozolin. The current studies were designed to determine if IPRO would delay male rat pubertal development like vinclozolin and to identify the mechanism(s) of action...

Development of a site-specific water quality criterion for hexavalent chromium

International Nuclear Information System (INIS)

McIntyre, D.O.; Sticko, J.P.; Reash, R.J.

1995-01-01

The effluent of treated fly ash from a coal-fired power plant located on the Ohio River periodically exceeds its NPDES acute permit limit for hexavalent chromium of 15 microg/L. The increased levels of hexavalent chromium in the effluent are a recent occurrence which are likely due to changes in coal blends burned in the generating units. Ohio EPA determined the use designation of the receiving stream (Limited Resource Water) was being attained and a one-year biomonitoring program of the effluent detected no acute toxicity to Ceriodaphnia dubia or Daphnia magna. The water-effect ratio (WER) procedure was selected to develop a site-specific criterion maximum concentration for hexavalent chromium for the effluent's receiving stream. WER procedures followed those described in EPA's ''Interim Guidance on Determination and Use of Water-Effect Ratios for Metals'' (1994). Site water used in the WER determinations was undiluted effluent since the receiving stream originates at the discharge point of the outfall. 48-hour acute D. magna and 96-hour acute fathead minnow toxicity tests were selected as the primary and secondary tests, respectively for use in three seasonal WER determinations. The results of the three WER determinations and the status of the regulatory process will be presented

Correlation of BTEX levels and toxicity of condensate contaminated groundwater

International Nuclear Information System (INIS)

Headley, J.; Goudey, S.; Birkholz, D.; Hardisty, P.

1995-01-01

The concentration of BTEX was determined for 60 groundwater samples collected from 6 gas plants in Western Canada, using conventional purge-and-trap GC/MS procedures. The gas plants were selected to cover different types of operations with different amine process chemicals employed for the sweetening of the raw sour-gas condensates. Aliquots of the ground water samples were subjected to toxicity screening tests, specifically, (a) bacterial luminescence (microtox); (b) daphnia mortality and (c) fathead minnow mortality. For the toxicity tests, sample handling procedures were developed to minimize the loss of volatile organics during the experiments. To account for possible losses, the levels of BTEX were monitored at the start and upon completion of these tests. The results indicated that the toxicity of the groundwater was in general, well correlated to the concentration of BTEX (primarily xylene). Approximately 5% of the samples, however, were observed to be toxic although the concentration of BTEX were below the method detection limit (1 microg/1). Thiophenic volatile organics were implicated for the latter. Based on the laboratory results, the remediation of BTEX is expected to correlate with the removal of the toxicity of the groundwater. These findings are of direct relevance to present technologies employed for remediation of ground water at the Sourgas plants

Using a risk assessment approach to handle 'hot spot' remediation in an environmentally-sensitive habitat

International Nuclear Information System (INIS)

BenKinney, M.T.; Guiseppi-Eli, A.; Novick, N.J.; Naro, P.A.

1993-01-01

A number of different approaches have been used to address site restoration following an accidental discharge or spill. Removal of all contamination with subsequent site restoration has been a favored approach. The authors' presentation will focus on a case study where a risk assessment approach was used to establish the criteria for is the restoration. Waste oil from a gasoline service station had leaked, over time, into a swale system in an ecologically-sensitive coastal forested habitat. The initial regulatory recommendation was to excavate all soils and restore the area. Their initial investigations revealed that only a limited area was heavily contaminated, predominantly with paraffinic hydrocarbons. They used a risk assessment approach coupled with sediment elutriate testing to demonstrate that leaving the lightly contaminated material in place would not present an environmental hazard from the waste oil, while causing less damage to the swale bed and forested area. Bioassay tests were conducted on sediments collected along the length of the swale from the areas with the highest remaining hydrocarbon concentrations using fathead minnow and Daphnia magna. The bioassay studies confirmed the results of the risk assessment which determined that no environmental toxicity would be expected from allowing lightly contaminated material to remain in-place at this site

From "Duck Factory" to "Fish Factory": Climate induced changes in vertebrate communities of prairie pothole wetlands and small lakes

Science.gov (United States)

McLean, Kyle I.; Mushet, David M.; Stockwell, Craig A.

2016-01-01

The Prairie Pothole Region’s myriad wetlands and small lakes contribute to its stature as the “duck factory” of North America. The fishless nature of the region’s aquatic habitats, a result of frequent drying, freezing, and high salinity, influences its importance to waterfowl. Recent precipitation increases have resulted in higher water levels and wetland/lake freshening. In 2012–13, we sampled chemical characteristics and vertebrates (fish and salamanders) of 162 Prairie Pothole wetlands and small lakes. We used non-metric multidimensional scaling, principal component analysis, and bootstrapping techniques to reveal relationships. We found fish present in a majority of sites (84 %). Fish responses to water chemistry varied by species. Fathead minnows (Pimephales promelas) and brook sticklebacks (Culaea inconstans) occurred across the broadest range of conditions. Yellow perch (Perca flavescens) occurred in a smaller, chemically defined, subset. Iowa darters (Etheostoma exile) were restricted to the narrowest range of conditions. Tiger salamanders (Ambystoma mavortium) rarely occurred in lakes with fish. We also compared our chemical data to similar data collected in 1966–1976 to explore factors contributing to the expansion of fish into previously fishless sites. Our work contributes to a better understanding of relationships between aquatic biota and climate-induced changes in this ecologically important area.

Histopathological changes induced by malathion in the gills of bluegill Lepomis macrochirus

Energy Technology Data Exchange (ETDEWEB)

Richmonds, C.; Dutta, H.M. (Kent State Univ., OH (USA))

1989-07-01

Malathion is a widely used broad spectrum organophosphorus insecticide. Its wide use provides many occasions for its entry into aquatic environments. The presence of this chemical in the aquatic environment would adversely affect many non-target species like fish. About 50 to 90% of the absorbed malathion can be eliminated in one to three days by the fish. About 25% of malathion remained in river water after 2 wk, and 10% remained after 4 wk from the time of its entry. Respiratory distress is one of the early symptoms of pesticide poisoning. These toxicants appear to cause a loss of adhesion between the epithelial cells and the underlying pillar cell system, accompanied by a collapse of the structural integrity of the secondary lamellae. Gills are important in respiration as well as osmoregulation of the fish. Therefore it was decided to study the effects of malathion on the gills of bluegill sunfish, Lepomis macrochirus. Bluegills were selected for this study due to the following reasons: (1) Bluegills are more sensitive to malathion when compared to fathead minnows and goldfish. (2) They are important both as edible and game fish. (3) They are easily available and easy to maintain in the laboratory.

Singapore grouper iridovirus, a large DNA virus, induces nonapoptotic cell death by a cell type dependent fashion and evokes ERK signaling.

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Huang, Xiaohong; Huang, Youhua; Ouyang, Zhengliang; Xu, Lixiao; Yan, Yang; Cui, Huachun; Han, Xin; Qin, Qiwei

2011-08-01

Virus induced cell death, including apoptosis and nonapoptotic cell death, plays a critical role in the pathogenesis of viral diseases. Singapore grouper iridovirus (SGIV), a novel iridovirus of genus Ranavirus, causes high mortality and heavy economic losses in grouper aquaculture. Here, using fluorescence microscopy, electron microscopy and biochemical assays, we found that SGIV infection in host (grouper spleen, EAGS) cells evoked nonapoptotic programmed cell death (PCD), characterized by appearance of cytoplasmic vacuoles and distended endoplasmic reticulum, in the absence of DNA fragmentation, apoptotic bodies and caspase activation. In contrast, SGIV induced typical apoptosis in non-host (fathead minnow, FHM) cells, as evidenced by caspase activation and DNA fragmentation, suggesting that SGIV infection induced nonapoptotic cell death by a cell type dependent fashion. Furthermore, viral replication was essential for SGIV induced nonapoptotic cell death, but not for apoptosis. Notably, the disruption of mitochondrial transmembrane potential (ΔΨm) and externalization of phosphatidylserine (PS) were not detected in EAGS cells but in FHM cells after SGIV infection. Moreover, the extracellular signal-regulated kinase (ERK) signaling was involved in SGIV infection induced nonapoptotic cell death and viral replication. This is a first demonstration of ERK-mediated nonapoptotic cell death induced by a DNA virus. These findings contribute to understanding the mechanisms of iridovirus pathogenesis.

Developing confidence in adverse outcome pathway-based toxicity predictions effects of the fungicide imazalil on fathead minnow reproduction

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An adverse outcome pathway (AOP) description linking inhibition of aromatase (cytochrome P450 [cyp] 19) to reproductive dysfunction was reviewed for scientific and technical quality and endorsed by the OECD (https://aopwiki.org/wiki/index.php/Aop:25). An intended application of t...

Impaired anterior swim bladder inflation following exposure to the thyroid peroxidase inhibitor 2-Mercaptobenzothiazole Part I: Fathead minnow

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Development of adverse outcome pathways linking specific chemical-induced pathway perturbations to adverse outcomes relevant to regulatory decision-making has potential to support the development of alternatives to traditional whole organism toxicity tests, such as the fish early...

Crafting a gene expression timeline for the thyroid in the early-life stages of fathead minnows (Pimephales promelas)

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The hypothalamic-pituitary-thyroid (HPT) axis plays a number of critical roles in chordate physiology including regulation of metabolism, growth, and development. While the role of the HPT axis and thyroid hormone signaling in mammalian and amphibian development is well establis...

Transcriptomic analyses of tributyltin-induced sexual dimorphisms in rare minnow (Gobiocypris rarus) brains.

Science.gov (United States)

Zhang, Ji-Liang; Liu, Min; Zhang, Chun-Nuan; Li, Er-Chao; Fan, Ming-Zhen; Huang, Mao-Xian

2018-07-30

The brain of fish displays sexual dimorphisms and exhibits remarkable sexual plasticity throughout their life span. Although reproductive toxicity of tributyltin (TBT) in fish is well documented in fish, it remains unknown whether TBT interrupts sexual dimorphisms of fish brains. In this work, brain transcriptomic profiles of rare minnow (Gobiocypris rarus) was characterized and sex-biased genes were identified using RNA sequencing. Functional annotation and enrichment analysis were performed to reveal differences of gene products and pathways between the brains of male and female fish. Furthermore, transcriptomic responses of male and female brains to TBT at 10 ng/L were also investigated to understand effects of TBT on brain sexual dimorphisms. Only 345 male-biased and 273 female-biased genes were found in the brains. However, significant female-biased pathways of circadian rhythm and phototransduction were identified in the brains by enrichment analysis. Interestingly, following TBT exposure in the female fish, the circadian rhythm pathway was significantly disrupted based on enrichment analysis, while in the male fish, the phototransduction pathway was significantly disrupted. In the female fish, expression of genes (Per, Cry, Rev-Erb α, Ror, Dec and CK1δ/ε) in the circadian rhythm pathway was down-regulated after TBT exposure; while in the male fish, expression of genes (Rec, GNAT1_2, GNGT1, Rh/opsin, PDE and Arr) in the phototransduction pathway was up-regulated after TBT exposure. Overall, our results not only provide key data on the molecular basis of brain sexual dimorphisms in fish, but also offer valuable resources for investigating molecular mechanisms by which environmental chemicals might influence brain sexual plasticity. Copyright © 2018 Elsevier Inc. All rights reserved.

Transgenerational effects of the endocrine disruptor vinclozolin on the methylation pattern of imprinted genes in the mouse sperm.

Science.gov (United States)

Stouder, Christelle; Paoloni-Giacobino, Ariane

2010-02-01

Endocrine-disrupting chemicals (EDCs), among which is the antiandrogen vinclozolin (VCZ), have been reported to affect the male reproductive system. In this study, VCZ was administered to pregnant mice at the time of embryo sex determination, and its possible effects on the differentially methylated domains (DMDs) of two paternally (H19 and Gtl2) and three maternally (Peg1, Snrpn, and Peg3) imprinted genes were tested in the male offspring. The CpGs methylation status within the five gene DMDs was analyzed in the sperm, tail, liver, and skeletal muscle DNAs by pyrosequencing. In the sperm of controls, the percentages of methylated CpGs were close to the theoretical values of 100 and 0% in paternally or maternally imprinted genes respectively. VCZ decreased the percentages of methylated CpGs of H19 and Gtl2 (respective values 83.1 and 91.5%) and increased those of Peg1, Snrpn, and Peg3 (respective values 11.3, 18.3, and 11.2%). The effects of VCZ were transgenerational, but they disappeared gradually from F1 to F3. The mean sperm concentration of the VCZ-administered female offspring was only 56% of that of the controls in the F1 offspring, and it was back to normal values in the F2 and F3 offspring. In the somatic cells of controls, the percentages of methylated CpGs were close to the theoretical value of 50% and, surprisingly, VCZ altered the methylation of Peg3. We propose that the deleterious effects of VCZ on the male reproductive system are mediated by imprinting defects in the sperm. The reported effects of EDCs on human male spermatogenesis might be mediated by analogous imprinting alterations.

Perinatal Exposure to Low Levels of the Environmental Antiandrogen Vinclozolin Alters Sex-Differentiated Social Play and Sexual Behaviors in the Rat

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Colbert, Nathan K.W.; Pelletier, Nicole C.; Cote, Joyce M.; Concannon, John B.; Jurdak, Nicole A.; Minott, Sara B.; Markowski, Vincent P.

2005-01-01

In this study we examined the effects of exposure to the antiandrogenic fungicide vinclozolin (Vz) on the development of two sex-differentiated behaviors that are organized by the perinatal actions of androgens. Pregnant Long-Evans rats were administered a daily oral dose of 0, 1.5, 3, 6, or 12 mg/kg Vz from the 14th day of gestation through postnatal day (PND)3. The social play behavior of juvenile offspring was examined on PND22 and again on PND34 during play sessions with a same-sex littermate. After they reached adulthood, the male offspring were examined with the ex copula penile reflex procedure to assess erectile function. Vz did not produce any gross maternal or neonatal toxicity, nor did it reduce the anogenital distance in male pups. We observed no effects of Vz on play behavior on PND22. However, the 12-mg/kg Vz dose significantly increased play behavior in the male offspring on PND34 compared with controls. The most dramatic increases were seen with the nape contact and pounce behavior components of play. The Vz effect was more pronounced in male than in female offspring. As adults, male offspring showed a significant reduction of erections at all dose levels during the ex copula penile reflex tests. The 12-mg/kg dose was also associated with an increase in seminal emissions. These effects demonstrate that perinatal Vz disrupts the development of androgen-mediated behavioral functions at exposure levels that do not produce obvious structural changes or weight reductions in androgen-sensitive reproductive organs. PMID:15929892

Toxicity of sediment-associated substituted phenylamine antioxidants on the early life stages of Pimephales promelas and a characterization of effects on freshwater organisms.

Science.gov (United States)

Prosser, Ryan S; Parrott, Joanne L; Galicia, Melissa; Shires, Kallie; Sullivan, Cheryl; Toito, John; Bartlett, Adrienne J; Milani, Danielle; Gillis, Patty L; Balakrishnan, Vimal K

2017-10-01

Substituted phenylamine antioxidants (SPAs) are high production volume chemicals that are incorporated into a variety of commercial products (e.g., polymers, dyes, lubricants). There are few data on chronic toxicity of SPAs to fish and no data on the toxicity of SPAs to the early life stages of fish. The physicochemical properties of SPAs would suggest that if they were to enter an aquatic ecosystem they would partition into sediment. Therefore, the present study focused on investigating the chronic effect of sediment-associated SPAs to the early life stages of the fathead minnow (Pimephales promelas). Eggs and larvae were exposed to sediment spiked with diphenylamine (DPA), N-phenyl-1-napthylamine (PNA), N-(1,3-dimethylbutyl)-N'-phenyl-1,4-phenylenediamine (DPPDA), or 4,4'-methylene-bis[N-sec-butylaniline] (MBA). The most sensitive endpoint for DPA, PNA, and DPPDA was total survival with 21-d median lethal concentrations (LC50s) based on concentration in overlying water of 1920, 74, and 35 μg/L, respectively. The most sensitive endpoint for MBA was growth with a 21-d median effective concentration (EC50) of 71 μg/L. The same endpoints were the most sensitive in terms of concentrations of DPA, PNA, DPPDA, and MBA in sediment (101, 54, 111, and 76 μg/g dry wt, respectively). Species sensitivity distributions (SSDs) were constructed for each SPA based on acute and chronic toxicity data generated in the present study and found in the literature. Overall, P. promelas was in the midrange of chronic sensitivity, with the most sensitive species being Tubifex tubifex. The SSDs indicate that DPA based on concentration in water is the least toxic to aquatic biota of the 4 SPAs investigated. The constructed SSDs indicate that a concentration in water and sediment of 1 μg/L and 1 μg/g dry weight, respectively, would be protective of >95% of the aquatic species tested. Environ Toxicol Chem 2017;36:2730-2738. © 2017 SETAC. © 2017 SETAC.

Quantitative Cross-Species Extrapolation between Humans and Fish: The Case of the Anti-Depressant Fluoxetine

Science.gov (United States)

Margiotta-Casaluci, Luigi; Owen, Stewart F.; Cumming, Rob I.; de Polo, Anna; Winter, Matthew J.; Panter, Grace H.; Rand-Weaver, Mariann; Sumpter, John P.

2014-01-01

Fish are an important model for the pharmacological and toxicological characterization of human pharmaceuticals in drug discovery, drug safety assessment and environmental toxicology. However, do fish respond to pharmaceuticals as humans do? To address this question, we provide a novel quantitative cross-species extrapolation approach (qCSE) based on the hypothesis that similar plasma concentrations of pharmaceuticals cause comparable target-mediated effects in both humans and fish at similar level of biological organization (Read-Across Hypothesis). To validate this hypothesis, the behavioural effects of the anti-depressant drug fluoxetine on the fish model fathead minnow (Pimephales promelas) were used as test case. Fish were exposed for 28 days to a range of measured water concentrations of fluoxetine (0.1, 1.0, 8.0, 16, 32, 64 µg/L) to produce plasma concentrations below, equal and above the range of Human Therapeutic Plasma Concentrations (HTPCs). Fluoxetine and its metabolite, norfluoxetine, were quantified in the plasma of individual fish and linked to behavioural anxiety-related endpoints. The minimum drug plasma concentrations that elicited anxiolytic responses in fish were above the upper value of the HTPC range, whereas no effects were observed at plasma concentrations below the HTPCs. In vivo metabolism of fluoxetine in humans and fish was similar, and displayed bi-phasic concentration-dependent kinetics driven by the auto-inhibitory dynamics and saturation of the enzymes that convert fluoxetine into norfluoxetine. The sensitivity of fish to fluoxetine was not so dissimilar from that of patients affected by general anxiety disorders. These results represent the first direct evidence of measured internal dose response effect of a pharmaceutical in fish, hence validating the Read-Across hypothesis applied to fluoxetine. Overall, this study demonstrates that the qCSE approach, anchored to internal drug concentrations, is a powerful tool to guide the

Quantitative cross-species extrapolation between humans and fish: the case of the anti-depressant fluoxetine.

Directory of Open Access Journals (Sweden)

Luigi Margiotta-Casaluci

Full Text Available Fish are an important model for the pharmacological and toxicological characterization of human pharmaceuticals in drug discovery, drug safety assessment and environmental toxicology. However, do fish respond to pharmaceuticals as humans do? To address this question, we provide a novel quantitative cross-species extrapolation approach (qCSE based on the hypothesis that similar plasma concentrations of pharmaceuticals cause comparable target-mediated effects in both humans and fish at similar level of biological organization (Read-Across Hypothesis. To validate this hypothesis, the behavioural effects of the anti-depressant drug fluoxetine on the fish model fathead minnow (Pimephales promelas were used as test case. Fish were exposed for 28 days to a range of measured water concentrations of fluoxetine (0.1, 1.0, 8.0, 16, 32, 64 µg/L to produce plasma concentrations below, equal and above the range of Human Therapeutic Plasma Concentrations (H(TPCs. Fluoxetine and its metabolite, norfluoxetine, were quantified in the plasma of individual fish and linked to behavioural anxiety-related endpoints. The minimum drug plasma concentrations that elicited anxiolytic responses in fish were above the upper value of the H(TPC range, whereas no effects were observed at plasma concentrations below the H(TPCs. In vivo metabolism of fluoxetine in humans and fish was similar, and displayed bi-phasic concentration-dependent kinetics driven by the auto-inhibitory dynamics and saturation of the enzymes that convert fluoxetine into norfluoxetine. The sensitivity of fish to fluoxetine was not so dissimilar from that of patients affected by general anxiety disorders. These results represent the first direct evidence of measured internal dose response effect of a pharmaceutical in fish, hence validating the Read-Across hypothesis applied to fluoxetine. Overall, this study demonstrates that the qCSE approach, anchored to internal drug concentrations, is a powerful tool

Mutations in the C-terminal region affect subcellular localization of crucian carp herpesvirus (CaHV) GPCR.

Science.gov (United States)

Wang, Jun; Gui, Lang; Chen, Zong-Yan; Zhang, Qi-Ya

2016-08-01

G protein-coupled receptors (GPCRs) are known as seven transmembrane domain receptors and consequently can mediate diverse biological functions via regulation of their subcellular localization. Crucian carp herpesvirus (CaHV) was recently isolated from infected fish with acute gill hemorrhage. CaHV GPCR of 349 amino acids (aa) was identified based on amino acid identity. A series of variants with truncation/deletion/substitution mutation in the C-terminal (aa 315-349) were constructed and expressed in fathead minnow (FHM) cells. The roles of three key C-terminal regions in subcellular localization of CaHV GPCR were determined. Lysine-315 (K-315) directed the aggregation of the protein preferentially at the nuclear side. Predicted N-myristoylation site (GGGWTR, aa 335-340) was responsible for punctate distribution in periplasm or throughout the cytoplasm. Predicted phosphorylation site (SSR, aa 327-329) and GGGWTR together determined the punctate distribution in cytoplasm. Detection of organelles localization by specific markers showed that the protein retaining K-315 colocalized with the Golgi apparatus. These experiments provided first evidence that different mutations of CaHV GPCR C-terminals have different affects on the subcellular localization of fish herpesvirus-encoded GPCRs. The study provided valuable information and new insights into the precise interactions between herpesvirus and fish cells, and could also provide useful targets for antiviral agents in aquaculture.

Use of various acute, sublethal and early life-stage tests to evaluate the toxicity of refinery effluents

International Nuclear Information System (INIS)

Sherry, J.; Scott, B.; Dutka, B.

1997-01-01

The toxicities of effluents from three Ontario, Canada, refineries were assessed with microbes, plants, invertebrates, and fish. Acute toxicity was assessed by the Microtox test, an assay based on electron transport activity in submitochondrial particles, and Daphnia magna (water flea); growth of Selenastrum capricornutum (alga); growth of Lemna minor (aquatic plant); germination of Lactuca sativa (nonaquatic plant); survival, growth, and maturation of Panagrellus redivivus (nematode); and genotoxicity in the SOS-Chromotest. Only the Microtox test and the submitochondrial particle test detected acute toxicity in the effluent samples. Reduced survival and sublethal responses were caused by some effluents, but not all effluents were toxic, and none caused a response in all of the tests applied. The results suggest that the effluent treatment systems used at Ontario refineries have largely eliminated acute toxicity to the organisms in their test battery. Although reduced survival and sublethal effects were detected in some of the effluents, the effects were minor. Some of the tests provided evidence, albeit weak, of variations in the responses of the test organisms to a temporal series of effluent samples. Not unexpectedly, there were also minor differences in the responses of the tests to effluents from the three refineries. The fathead minnow test seems to be a sensitive indicator of the sublethal toxicity of Ontario refinery effluents

Effects-Directed Analysis of Dissolved Organic Compounds in Oil Sands Process-Affected Water.

Science.gov (United States)

Morandi, Garrett D; Wiseman, Steve B; Pereira, Alberto; Mankidy, Rishikesh; Gault, Ian G M; Martin, Jonathan W; Giesy, John P

2015-10-20

Acute toxicity of oil sands process-affected water (OSPW) is caused by its complex mixture of bitumen-derived organics, but the specific chemical classes that are most toxic have not been demonstrated. Here, effects-directed analysis was used to determine the most acutely toxic chemical classes in OSPW collected from the world's first oil sands end-pit lake. Three sequential rounds of fractionation, chemical analysis (ultrahigh resolution mass spectrometry), and acute toxicity testing (96 h fathead minnow embryo lethality and 15 min Microtox bioassay) were conducted. Following primary fractionation, toxicity was primarily attributable to the neutral extractable fraction (F1-NE), containing 27% of original organics mass. In secondary fractionation, F1-NE was subfractionated by alkaline water washing, and toxicity was primarily isolated to the ionizable fraction (F2-NE2), containing 18.5% of the original organic mass. In the final round, chromatographic subfractionation of F2-NE2 resulted in two toxic fractions, with the most potent (F3-NE2a, 11% of original organic mass) containing predominantly naphthenic acids (O2(-)). The less-toxic fraction (F3-NE2b, 8% of original organic mass) contained predominantly nonacid species (O(+), O2(+), SO(+), NO(+)). Evidence supports naphthenic acids as among the most acutely toxic chemical classes in OSPW, but nonacidic species also contribute to acute toxicity of OSPW.

Developing confidence in adverse outcome pathway-based toxicity predictions effects of the fungicide imazalil on fathead minnow reproduction (Poster)

Science.gov (United States)

An adverse outcome pathway (AOP) description linking inhibition of aromatase (cytochrome P450 [cyp] 19) to reproductive dysfunction was reviewed for scientific and technical quality and endorsed by the OECD. An intended application of the AOP framework is to support the use of me...

The Stepwise Behavioral Responses: Behavioral Adjustment of the Chinese Rare Minnow (Gobiocypris rarus in the Exposure of Carbamate Pesticides

Directory of Open Access Journals (Sweden)

Zongming Ren

2013-01-01

Full Text Available In order to illustrate the behavioral regulation in environmental stress, the behavioral responses of the Chinese rare minnow (Gobiocypris rarus to arprocarb, carbofuran, and oxamyl were analyzed with an online monitoring system. The Self-Organizing Map (SOM was used to define the patterns of the behavioral data obtained from treatments at concentrations of 0.1 toxic unit (TU, 1 TU, 2 TU, 5 TU, 10 TU, and 20 TU and a control. In certain cases, differences among the carbamate pesticides (CPs tested were observed. The profiles of behavioral strength (BS in SOM varied according to the concentration used. The time of the first significant decrease of the BS varied inversely with the CP concentrations. The results suggested that the behavioral regulation in the stepwise behavioral responses (SBR was evident. The primary movement behaviors shown by the SBR model included no effect, stimulation, acclimation, adjustment (readjustment, and toxic effect, especially at the lower concentrations. However, higher stress (10 TU and 20 TU might limit the function of the behavioral adjustment produced by the intrinsic response mechanisms. It was concluded that SBR, which were affected by both the concentration and the exposure time, could be used as a suitable indicator in the ecotoxicological risk assessment of CPs.

Maternal exposure to anti-androgenic compounds, vinclozolin, flutamide and procymidone, has no effects on spermatogenesis and DNA methylation in male rats of subsequent generations

International Nuclear Information System (INIS)

Inawaka, Kunifumi; Kawabe, Mayumi; Takahashi, Satoru; Doi, Yuko; Tomigahara, Yoshitaka; Tarui, Hirokazu; Abe, Jun; Kawamura, Satoshi; Shirai, Tomoyuki

2009-01-01

To verify whether anti-androgens cause transgenerational effects on spermatogenesis and DNA methylation in rats, gravid Crl:CD(SD) female rats (4 or 5/group, gestational day (GD) 0 = day sperm detected) were intraperitoneally treated with anti-androgenic compounds, such as vinclozolin (100 mg/kg/day), procymidone (100 mg/kg/day), or flutamide (10 mg/kg/day), from GD 8 to GD 15. Testes were collected from F1 male pups at postnatal day (PND) 6 for DNA methylation analysis of the region (210 bp including 7 CpG sites) within the lysophospholipase gene by bisulfite DNA sequencing method. F0 and F1 males underwent the sperm analysis (count, motility and morphology), followed by DNA methylation analysis of the sperm. Remaining F1 males were cohabited with untreated-females to obtain F2 male pups for subsequent DNA methylation analysis of the testes at PND 6. These analyses showed no effects on spermatogenesis and fertility in F1 males of any treatment group. DNA methylation status in testes (F1 and F2 pups at PND 6) or sperms (F1 males at 13 weeks old) of the treatment groups were comparable to the control at all observation points, although DNA methylation rates in testes were slightly lower than those in sperm. In F0 males, no abnormalities in the spermatogenesis, fertility and DNA methylation status of sperm were observed. No transgenerational abnormalities of spermatogenesis and DNA methylation status caused by anti-androgenic compounds were observed.

Maternal exposure to anti-androgenic compounds, vinclozolin, flutamide and procymidone, has no effects on spermatogenesis and DNA methylation in male rats of subsequent generations.

Science.gov (United States)

Inawaka, Kunifumi; Kawabe, Mayumi; Takahashi, Satoru; Doi, Yuko; Tomigahara, Yoshitaka; Tarui, Hirokazu; Abe, Jun; Kawamura, Satoshi; Shirai, Tomoyuki

2009-06-01

To verify whether anti-androgens cause transgenerational effects on spermatogenesis and DNA methylation in rats, gravid Crl:CD(SD) female rats (4 or 5/group, gestational day (GD) 0=day sperm detected) were intraperitoneally treated with anti-androgenic compounds, such as vinclozolin (100 mg/kg/day), procymidone (100 mg/kg/day), or flutamide (10 mg/kg/day), from GD 8 to GD 15. Testes were collected from F1 male pups at postnatal day (PND) 6 for DNA methylation analysis of the region (210 bp including 7 CpG sites) within the lysophospholipase gene by bisulfite DNA sequencing method. F0 and F1 males underwent the sperm analysis (count, motility and morphology), followed by DNA methylation analysis of the sperm. Remaining F1 males were cohabited with untreated-females to obtain F2 male pups for subsequent DNA methylation analysis of the testes at PND 6. These analyses showed no effects on spermatogenesis and fertility in F1 males of any treatment group. DNA methylation status in testes (F1 and F2 pups at PND 6) or sperms (F1 males at 13 weeks old) of the treatment groups were comparable to the control at all observation points, although DNA methylation rates in testes were slightly lower than those in sperm. In F0 males, no abnormalities in the spermatogenesis, fertility and DNA methylation status of sperm were observed. No transgenerational abnormalities of spermatogenesis and DNA methylation status caused by anti-androgenic compounds were observed.

Marine Mammals and Low-Frequency Sound: Progress Since 1994

Science.gov (United States)

2000-03-03

Band (dB re 1 uPa (%lm) 5-m Zodiac inflatable boat0 6,300 152 Bell 212 helicopter6 16 159 Large tanker 100+125 177 Icebreaker 100 183 ATOC 75 195...coastline during two days (May 12 and 13, 1996) in the Kyparissiakos Gulf in Greece. There was no external sign of injury or disease in any of these...in two fish species, sheepshead minnows (Cyprinodon variegatus) and killifish (Fundulus similis), were significantly lower in aquariums exposed to

Probabilistic cumulative risk assessment of anti-androgenic pesticides in food

DEFF Research Database (Denmark)

Müller, Anne Kirstine; Bosgra, Sieto; Boon, Polly E.

2009-01-01

In this paper, we present a cumulative risk assessment of three anti-androgenic pesticides (vinclozolin, procymidone and prochloraz) using the relative potency factor (RPF) approach and an integrated probabilistic risk assessment (IPRA) model. RPFs for each substance were estimated for three......) and the fraction of individuals with IMoEs vinclozolin, procymidone and prochloraz is not likely to be of concern for the reproductive development of their male foetuses. However...

Generational comparisons (F1 versus F3) of vinclozolin induced epigenetic transgenerational inheritance of sperm differential DNA methylation regions (epimutations) using MeDIP-Seq.

Science.gov (United States)

Beck, Daniel; Sadler-Riggleman, Ingrid; Skinner, Michael K

2017-07-01

Environmentally induced epigenetic transgenerational inheritance of disease and phenotypic variation has been shown to involve DNA methylation alterations in the germline (e.g. sperm). These differential DNA methylation regions (DMRs) are termed epimutations and in part transmit the transgenerational phenotypes. The agricultural fungicide vinclozolin exposure of a gestating female rat has previously been shown to promote transgenerational disease and epimutations in F3 generation (great-grand-offspring) animals. The current study was designed to investigate the actions of direct fetal exposure on the F1 generation rat sperm DMRs compared to the F3 transgenerational sperm DMRs. A protocol involving methylated DNA immunoprecipitation (MeDIP) followed by next-generation sequencing (Seq) was used in the current study. Bioinformatics analysis of the MeDIP-Seq data was developed and several different variations in the bioinformatic analysis were evaluated. Observations indicate needs to be considered. Interestingly, the F1 generation DMRs were found to be fewer in number and for the most part distinct from the F3 generation epimutations. Observations suggest the direct exposure induced F1 generation sperm DMRs appear to promote in subsequent generations alterations in the germ cell developmental programming that leads to the distinct epimutations in the F3 generation. This may help explain the differences in disease and phenotypes between the direct exposure F1 generation and transgenerational F3 generation. Observations demonstrate a distinction between the direct exposure versus transgenerational epigenetic programming induced by environmental exposures and provide insights into the molecular mechanisms involved in the epigenetic transgenerational inheritance phenomenon.

Impacts of WWTP Effluents on the Hepatic Metabolome of Male and Female Fathead Minnows in the South Platte River, Colorado

Science.gov (United States)

Metabolomics is rapidly becoming established as an eefective tool for studying the responses of organisms, such as fish, to various environmentally relevant stressors. While the majority of the work has been laboratory-based, successful application of the technique in recent year...

CLONING, EXPRESSION AND CHARACTERIZATION OF THE ANDROGEN RECEPTOR AND ISOLATION OF ESTROGEN RECEPTOR ALPHA FROM THE FATHEAD MINNOW (PIMEPHALES PROMELAS)

Science.gov (United States)

In vitro screening assays designed to identify hormone mimics or antagonists, including those recommended for use in the EPA's Tier 1 screening battery, typically use mammalian estrogen (ER) and androgen receptors (AR) such as rat or human. Although we know that the amino acid s...

Mixtures of 17â-trenbolone with Ethinylestradiol or Bisphenol A Altered Tubercle Formation and Steroid Production in the Fathead Minnow

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The xenoestrogens, ethinylestradiol (EE2) and bisphenol A (BPA), and the androgen 17â-trenbolone (TRB) are examples of endocrine disrupting compounds (EDCs) commonly detected in the environment. All have been shown to affect fish reproductive endocrinology individually, but littl...

Eryptosis in lead-exposed workers

International Nuclear Information System (INIS)

Aguilar-Dorado, Itzel-Citlalli; Hernández, Gerardo; Quintanar-Escorza, Martha-Angelica; Maldonado-Vega, María; Rosas-Flores, Margarita; Calderón-Salinas, José-Víctor

2014-01-01

Eryptosis is a physiological phenomenon in which old and damaged erythrocytes are removed from circulation. Erythrocytes incubated with lead have exhibited major eryptosis. In the present work we found evidence of high levels of eryptosis in lead exposed workers possibly via oxidation. Blood samples were taken from 40 male workers exposed to lead (mean blood lead concentration 64.8 μg/dl) and non-exposed workers (4.2 μg/dl). The exposure to lead produced an intoxication characterized by 88.3% less δ-aminolevulinic acid dehydratase (δALAD) activity in lead exposed workers with respect to non-lead exposed workers. An increment of oxidation in lead exposed workers was characterized by 2.4 times higher thiobarbituric acid-reactive substance (TBARS) concentration and 32.8% lower reduced/oxidized glutathione (GSH/GSSG) ratio. Oxidative stress in erythrocytes of lead exposed workers is expressed in 192% higher free calcium concentration [Ca 2+ ] i and 1.6 times higher μ-calpain activity with respect to non-lead exposed workers. The adenosine triphosphate (ATP) concentration was not significantly different between the two worker groups. No externalization of phosphatidylserine (PS) was found in non-lead exposed workers (< 0.1%), but lead exposed workers showed 2.82% externalization. Lead intoxication induces eryptosis possibly through a molecular pathway that includes oxidation, depletion of reduced glutathione (GSH), increment of [Ca 2+ ], μ-calpain activation and externalization of PS in erythrocytes. Identifying molecular signals that induce eryptosis in lead intoxication is necessary to understand its physiopathology and chronic complications. - Graphical abstract: Fig. 1. (A) Blood lead concentration (PbB) and (B) phosphatidylserine externalization on erythrocyte membranes of non-lead exposed (□) and lead exposed workers (■). Values are mean ± SD. *Significantly different (P < 0.001). - Highlights: • Erythrocytes of lead exposed workers showed higher PS

Eryptosis in lead-exposed workers

Energy Technology Data Exchange (ETDEWEB)

Aguilar-Dorado, Itzel-Citlalli [Biochemistry Department, Centro de Investigación y Estudios Avanzados IPN, México, DF (Mexico); Hernández, Gerardo [Section of Methodology of Science, Centro de Investigación y Estudios Avanzados IPN, México, DF (Mexico); Quintanar-Escorza, Martha-Angelica [Faculty of Medicine, UJED, Durango, DGO (Mexico); Maldonado-Vega, María [CIATEC, León, GTO (Mexico); Rosas-Flores, Margarita [Biochemistry Department, Centro de Investigación y Estudios Avanzados IPN, México, DF (Mexico); Calderón-Salinas, José-Víctor, E-mail: jcalder@cinvestav.mx [Biochemistry Department, Centro de Investigación y Estudios Avanzados IPN, México, DF (Mexico)

2014-12-01

Eryptosis is a physiological phenomenon in which old and damaged erythrocytes are removed from circulation. Erythrocytes incubated with lead have exhibited major eryptosis. In the present work we found evidence of high levels of eryptosis in lead exposed workers possibly via oxidation. Blood samples were taken from 40 male workers exposed to lead (mean blood lead concentration 64.8 μg/dl) and non-exposed workers (4.2 μg/dl). The exposure to lead produced an intoxication characterized by 88.3% less δ-aminolevulinic acid dehydratase (δALAD) activity in lead exposed workers with respect to non-lead exposed workers. An increment of oxidation in lead exposed workers was characterized by 2.4 times higher thiobarbituric acid-reactive substance (TBARS) concentration and 32.8% lower reduced/oxidized glutathione (GSH/GSSG) ratio. Oxidative stress in erythrocytes of lead exposed workers is expressed in 192% higher free calcium concentration [Ca{sup 2+}]{sub i} and 1.6 times higher μ-calpain activity with respect to non-lead exposed workers. The adenosine triphosphate (ATP) concentration was not significantly different between the two worker groups. No externalization of phosphatidylserine (PS) was found in non-lead exposed workers (< 0.1%), but lead exposed workers showed 2.82% externalization. Lead intoxication induces eryptosis possibly through a molecular pathway that includes oxidation, depletion of reduced glutathione (GSH), increment of [Ca{sup 2+}], μ-calpain activation and externalization of PS in erythrocytes. Identifying molecular signals that induce eryptosis in lead intoxication is necessary to understand its physiopathology and chronic complications. - Graphical abstract: Fig. 1. (A) Blood lead concentration (PbB) and (B) phosphatidylserine externalization on erythrocyte membranes of non-lead exposed (□) and lead exposed workers (■). Values are mean ± SD. *Significantly different (P < 0.001). - Highlights: • Erythrocytes of lead exposed workers

Linking field-based metabolomics and chemical analyses to prioritize contaminants of emerging concern in the Great Lakes basin

Science.gov (United States)

Davis, John M.; Ekman, Drew R.; Teng, Quincy; Ankley, Gerald T.; Berninger, Jason P.; Cavallin, Jenna E.; Jensen, Kathleen M.; Kahl, Michael D.; Schroeder, Anthony L.; Villeneuve, Daniel L.; Jorgenson, Zachary G.; Lee, Kathy E.; Collette, Timothy W.

2016-01-01

The ability to focus on the most biologically relevant contaminants affecting aquatic ecosystems can be challenging because toxicity-assessment programs have not kept pace with the growing number of contaminants requiring testing. Because it has proven effective at assessing the biological impacts of potentially toxic contaminants, profiling of endogenous metabolites (metabolomics) may help screen out contaminants with a lower likelihood of eliciting biological impacts, thereby prioritizing the most biologically important contaminants. The authors present results from a study that utilized cage-deployed fathead minnows (Pimephales promelas) at 18 sites across the Great Lakes basin. They measured water temperature and contaminant concentrations in water samples (132 contaminants targeted, 86 detected) and used 1H-nuclear magnetic resonance spectroscopy to measure endogenous metabolites in polar extracts of livers. They used partial least-squares regression to compare relative abundances of endogenous metabolites with contaminant concentrations and temperature. The results indicated that profiles of endogenous polar metabolites covaried with at most 49 contaminants. The authors identified up to 52% of detected contaminants as not significantly covarying with changes in endogenous metabolites, suggesting they likely were not eliciting measurable impacts at these sites. This represents a first step in screening for the biological relevance of detected contaminants by shortening lists of contaminants potentially affecting these sites. Such information may allow risk assessors to prioritize contaminants and focus toxicity testing on the most biologically relevant contaminants. Environ Toxicol Chem 2016;35:2493–2502.

Sensitivity of screening-level toxicity tests using soils from a former petroleum refinery

International Nuclear Information System (INIS)

Pauwels, S.; Bureau, J.; Roy, Y.; Allen, B.; Robidoux, P.Y.; Soucy, M.

1995-01-01

The authors tested five composite soil samples from a former refinery. The samples included a reference soil (Mineral Oil and Grease, MO and G < 40 ppm), thermally-treated soil, biotreated soil, and two untreated soils. They evaluated toxicity using the earthworm E. foetida, lettuce, cress, barley, Microtox, green algae, fathead minnow, and D. magna. The endpoints measured were lethality, seed germination, root elongation, growth, and bioluminescence. Toxicity, as measured by the number of positive responses, increased as follows: biotreated soil < untreated soil No. 1 < reference soil < thermally-treated soil and untreated soil No. 2. The biotreated soil generated only one positive response, whereas the thermally-treated soil and untreated soil No. 2 generated five positive responses. The most sensitive and discriminant terrestrial endpoint was lettuce root elongation which responded to untreated soil No. 1, thermally-treated soil, and reference soil. The least sensitive was barley seed germination for which no toxicity was detected. The most sensitive and discriminant aquatic endpoint was green algae growth which responded to untreated soil No. 1, thermally-treated soil, and reference soil. The least sensitive was D. magna for which no toxicity was detected. Overall, soil and aqueous extract toxicity was spotty and no consistent patterns emerged to differentiate the five soils. Biotreatment significantly reduced the effects of the contamination. Aqueous toxicity was measured in the reference soil, probably because of the presence of unknown dissolved compounds in the aqueous extract. Finally, clear differences in sensitivity existed among the test species

Acute toxicity of metals and reference toxicants to a freshwater ostracod, Cypris subglobosa Sowerby, 1840 and correlation to EC{sub 50} values of other test models

Energy Technology Data Exchange (ETDEWEB)

Khangarot, B.S., E-mail: bkhangarot@hotmail.com [Ecotoxicology Division, Indian Institute of Toxicology Research (Formerly: Industrial Toxicology Research Centre), Post Box No. 80, Mahatma Gandhi Marg, Lucknow 226001 (India); Das, Sangita [Ecotoxicology Division, Indian Institute of Toxicology Research (Formerly: Industrial Toxicology Research Centre), Post Box No. 80, Mahatma Gandhi Marg, Lucknow 226001 (India)

2009-12-30

The ostracod Cypris subglobosa Sowerby, 1840 static bioassay test on the basis of a 48 h of 50% of immobilization (EC{sub 50}) has been used to measure the toxicity of 36 metals and metalloids and 12 reference toxicants. Among the 36 metals and metalloids, osmium (Os) was found to be the most toxic in the test while boron (B), the least toxic. The EC{sub 50} values of this study revealed positive linear relationship with the established test models of cladoceran (Daphnia magna), sludge worm (Tubifex tubifex), chironomid larvae (Chironomus tentans), protozoan (Tetrahymena pyriformis), fathead minnow (Pimephales promelas), bluegill sunfish (Lepomis macrochirus), and aquatic macrophyte duckweed (Lemna minor). Correlation coefficients (r{sup 2}) for 17 physicochemical properties of metals or metal ions and EC{sub 50}s (as pM) were examined by linear regression analysis. The electronegativity, ionization potential, melting point, solubility product of metal sulfides (pK{sub sp}), softness parameter and some other physicochemical characteristics were significantly correlated with EC{sub 50}s of metals to C. subglobosa. The reproducibility of toxicity test was determined using 12 reference toxicants. The coefficient of variability of the EC{sub 50}s ranged from 6.95% to 55.37% and variability was comparable to that noticed for D. magna and other aquatic test models. The study demonstrated the need to include crustacean ostracods in a battery of biotests to detect the presence of hazardous chemicals in soils, sewage sludges, sediments and aquatic systems.

Acute toxicity of metals and reference toxicants to a freshwater ostracod, Cypris subglobosa Sowerby, 1840 and correlation to EC50 values of other test models

International Nuclear Information System (INIS)

Khangarot, B.S.; Das, Sangita

2009-01-01

The ostracod Cypris subglobosa Sowerby, 1840 static bioassay test on the basis of a 48 h of 50% of immobilization (EC 50 ) has been used to measure the toxicity of 36 metals and metalloids and 12 reference toxicants. Among the 36 metals and metalloids, osmium (Os) was found to be the most toxic in the test while boron (B), the least toxic. The EC 50 values of this study revealed positive linear relationship with the established test models of cladoceran (Daphnia magna), sludge worm (Tubifex tubifex), chironomid larvae (Chironomus tentans), protozoan (Tetrahymena pyriformis), fathead minnow (Pimephales promelas), bluegill sunfish (Lepomis macrochirus), and aquatic macrophyte duckweed (Lemna minor). Correlation coefficients (r 2 ) for 17 physicochemical properties of metals or metal ions and EC 50 s (as pM) were examined by linear regression analysis. The electronegativity, ionization potential, melting point, solubility product of metal sulfides (pK sp ), softness parameter and some other physicochemical characteristics were significantly correlated with EC 50 s of metals to C. subglobosa. The reproducibility of toxicity test was determined using 12 reference toxicants. The coefficient of variability of the EC 50 s ranged from 6.95% to 55.37% and variability was comparable to that noticed for D. magna and other aquatic test models. The study demonstrated the need to include crustacean ostracods in a battery of biotests to detect the presence of hazardous chemicals in soils, sewage sludges, sediments and aquatic systems.

Dysregulation of autism-associated synaptic proteins by psychoactive pharmaceuticals at environmental concentrations.

Science.gov (United States)

Kaushik, Gaurav; Xia, Yu; Pfau, Jean C; Thomas, Michael A

2017-11-20

Autism Spectrum Disorders (ASD) are complex neurological disorders for which the prevalence in the U.S. is currently estimated to be 1 in 50 children. A majority of cases of idiopathic autism in children likely result from unknown environmental triggers in genetically susceptible individuals. These triggers may include maternal exposure of a developing embryo to environmentally relevant minute concentrations of psychoactive pharmaceuticals through ineffectively purified drinking water. Previous studies in our lab examined the extent to which gene sets associated with neuronal development were up- and down-regulated (enriched) in the brains of fathead minnows treated with psychoactive pharmaceuticals at environmental concentrations. The aim of this study was to determine whether similar treatments would alter in vitro expression of ASD-associated synaptic proteins on differentiated human neuronal cells. Human SK-N-SH neuroblastoma cells were differentiated for two weeks with 10μM retinoic acid (RA) and treated with environmentally relevant concentrations of fluoxetine, carbamazepine or venlafaxine, and flow cytometry technique was used to analyze expression of ASD-associated synaptic proteins. Data showed that carbamazepine individually, venlafaxine individually and mixture treatment at environmental concentrations significantly altered the expression of key synaptic proteins (NMDAR1, PSD95, SV2A, HTR1B, HTR2C and OXTR). Data indicated that psychoactive pharmaceuticals at extremely low concentrations altered the in vitro expression of key synaptic proteins that may potentially contribute to neurological disorders like ASD by disrupting neuronal development. Copyright © 2017 Elsevier B.V. All rights reserved.

Eryptosis in lead-exposed workers.

Science.gov (United States)

Aguilar-Dorado, Itzel-Citlalli; Hernández, Gerardo; Quintanar-Escorza, Martha-Angelica; Maldonado-Vega, María; Rosas-Flores, Margarita; Calderón-Salinas, José-Víctor

2014-12-01

Eryptosis is a physiological phenomenon in which old and damaged erythrocytes are removed from circulation. Erythrocytes incubated with lead have exhibited major eryptosis. In the present work we found evidence of high levels of eryptosis in lead exposed workers possibly via oxidation. Blood samples were taken from 40 male workers exposed to lead (mean blood lead concentration 64.8μg/dl) and non-exposed workers (4.2μg/dl). The exposure to lead produced an intoxication characterized by 88.3% less δ-aminolevulinic acid dehydratase (δALAD) activity in lead exposed workers with respect to non-lead exposed workers. An increment of oxidation in lead exposed workers was characterized by 2.4 times higher thiobarbituric acid-reactive substance (TBARS) concentration and 32.8% lower reduced/oxidized glutathione (GSH/GSSG) ratio. Oxidative stress in erythrocytes of lead exposed workers is expressed in 192% higher free calcium concentration [Ca(2+)]i and 1.6 times higher μ-calpain activity with respect to non-lead exposed workers. The adenosine triphosphate (ATP) concentration was not significantly different between the two worker groups. No externalization of phosphatidylserine (PS) was found in non-lead exposed workers (lead exposed workers showed 2.82% externalization. Lead intoxication induces eryptosis possibly through a molecular pathway that includes oxidation, depletion of reduced glutathione (GSH), increment of [Ca(2+)], μ-calpain activation and externalization of PS in erythrocytes. Identifying molecular signals that induce eryptosis in lead intoxication is necessary to understand its physiopathology and chronic complications. Copyright © 2014 Elsevier Inc. All rights reserved.

Analysis of 3',5'-dichloro-2,3,4-trihydroxy-2-methylbutylanilide (DTMBA) as a new potential biomarker of exposure to vinclozolin in urine.

Science.gov (United States)

Cruz-Hurtado, Marycarmen; López-González, Ma de Lourdes; Escobar-Wilches, Derly Constanza; Sierra-Santoyo, Adolfo

2018-05-01

Vinclozolin (V) is a fungicide with anti-androgenic properties whose metabolism is not fully understood, and data on urinary elimination of either V or its metabolites are limited. Therefore the kinetics of urinary elimination of V and its metabolites, after an oral dose in adult male rats were investigated. A single oral dose of V (100 mg/kg) suspended in corn oil was administered to male adult Wistar rats, and urine was collected at different times after dosing. V and its metabolites were extracted from urine, then enzymatically hydrolyzed using β-glucuronidase/sulfatase of H. pomatia, and analyzed by HPLC/DAD. Urinary pharmacokinetic parameters were calculated using the analyte concentrations adjusted by creatinine levels. V and its metabolites 3',5'-dichloro-2,3,4-trihydroxy-2-methylbutylanilide (DTMBA, formerly denoted as M5), 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3-butenoic acid (M1), 3,5-dichloroaniline (M3), and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2) were efficiently detected. The mean urine concentrations of V and M1 metabolite were fitted to a two-compartmental model for pharmacokinetic analysis. DTMBA approximately represented 88% of the total excreted metabolites, it was easily detected up to 168 h after dosing and its half-lives were 21.5 and 74.1 h, respectively. M1 was the second most abundant metabolite and was detected up to 144 h after being void. V and M3 were detected before 48 h, and M2 exhibited the lowest levels during the first 8 h after dosing. DTMBA, the most abundant V metabolite is quickly eliminated by urine, it is chemically stable, specific and could represent a useful alternative to be used as a biomarker of exposure to V. Copyright © 2018 Elsevier Inc. All rights reserved.

Buildings exposed to fire

International Nuclear Information System (INIS)

1987-01-01

The 24 lectures presented to the colloquium cover the following subject fields: (1) Behaviour of structural components exposed to fire; (2) Behaviour of building materials exposed to fire; (3) Thermal processes; (4) Safety related, theoretical studies. (PW) [de

Metabolomics for informing adverse outcome pathways: Androgen receptor activation and the pharmaceutical spironolactone

International Nuclear Information System (INIS)

Davis, J.M.; Ekman, D.R.; Skelton, D.M.; LaLone, C.A.; Ankley, G.T.; Cavallin, J.E.; Villeneuve, D.L.; Collette, T.W.

2017-01-01

Highlights: • Metabolomics identified potential key events in an androgen receptor activation AOP. • Metabolomics indicate spironolactone may elicit effects via multiple nuclear receptors. • Spironolactone exposure may elicit interactive effects in multi-stressor environments. - Abstract: One objective in developing adverse outcome pathways (AOPs) is to connect biological changes that are relevant to risk assessors (i.e., fecundity) to molecular and cellular-level alterations that might be detectable at earlier stages of a chemical exposure. Here, we examined biochemical responses of fathead minnows (Pimephales promelas) to inform an AOP relevant to spironolactone’s activation of the androgen receptor, as well as explore other biological impacts possibly unrelated to this receptor. Liquid chromatography with high resolution mass spectrometry (LC–MS) was used to measure changes in endogenous polar metabolites in livers of male and female fish that were exposed to five water concentrations of spironolactone (0, 0.05, 0.5, 5, or 50 μg L"−"1) for 21 days. Metabolite profiles were affected at the two highest concentrations (5 and 50 μg L"−"1), but not in the lower-level exposures, which agreed with earlier reported results of reduced female fecundity and plasma vitellogenin (VTG) levels. We then applied partial least squares regression to assess whether metabolite alterations covaried with changes in fecundity, VTG gene expression and protein concentrations, and plasma 17β-estradiol and testosterone concentrations. Metabolite profiles significantly covaried with all measured endpoints in females, but only with plasma testosterone in males. Fecundity reductions occurred in parallel with changes in metabolites important in osmoregulation (e.g., betaine), membrane transport (e.g., L-carnitine), and biosynthesis of carnitine (e.g., methionine) and VTG (e.g., glutamate). Based on a network analysis program (i.e., mummichog), spironolactone also affected

Endocrine active chemicals, pharmaceuticals, and other chemicals of concern in surface water, wastewater-treatment plant effluent, and bed sediment, and biological characteristics in selected streams, Minnesota-design, methods, and data, 2009

Science.gov (United States)

Lee, Kathy E.; Langer, Susan K.; Barber, Larry B.; Writer, Jeff H.; Ferrey, Mark L.; Schoenfuss, Heiko L.; Furlong, Edward T.; Foreman, William T.; Gray, James L.; ReVello, Rhiannon C.; Martinovic, Dalma; Woodruff, Olivia R.; Keefe, Steffanie H.; Brown, Greg K.; Taylor, Howard E.; Ferrer, Imma; Thurman, E. Michael

2011-01-01

remaining samples were archived. Biological characteristics were determined by using an in-vitro bioassay to determine total estrogenicity in water samples and a caged fish study to determine characteristics of fish from experiments that exposed fish to wastewater effluent in 2009. St. Cloud State University deployed and processed caged fathead minnows at 13 stream sites during September 2009 for the caged fish study. Measured fish data included length, weight, body condition factor, and vitellogenin concentrations.

An Assessment of the Model of Concentration Addition for Predicting the Estrogenic Activity of Chemical Mixtures in Wastewater Treatment Works Effluents

Science.gov (United States)

Thorpe, Karen L.; Gross-Sorokin, Melanie; Johnson, Ian; Brighty, Geoff; Tyler, Charles R.

2006-01-01

The effects of simple mixtures of chemicals, with similar mechanisms of action, can be predicted using the concentration addition model (CA). The ability of this model to predict the estrogenic effects of more complex mixtures such as effluent discharges, however, has yet to be established. Effluents from 43 U.K. wastewater treatment works were analyzed for the presence of the principal estrogenic chemical contaminants, estradiol, estrone, ethinylestradiol, and nonylphenol. The measured concentrations were used to predict the estrogenic activity of each effluent, employing the model of CA, based on the relative potencies of the individual chemicals in an in vitro recombinant yeast estrogen screen (rYES) and a short-term (14-day) in vivo rainbow trout vitellogenin induction assay. Based on the measured concentrations of the four chemicals in the effluents and their relative potencies in each assay, the calculated in vitro and in vivo responses compared well and ranged between 3.5 and 87 ng/L of estradiol equivalents (E2 EQ) for the different effluents. In the rYES, however, the measured E2 EQ concentrations in the effluents ranged between 0.65 and 43 ng E2 EQ/L, and they varied against those predicted by the CA model. Deviations in the estimation of the estrogenic potency of the effluents by the CA model, compared with the measured responses in the rYES, are likely to have resulted from inaccuracies associated with the measurement of the chemicals in the extracts derived from the complex effluents. Such deviations could also result as a consequence of interactions between chemicals present in the extracts that disrupted the activation of the estrogen response elements in the rYES. E2 EQ concentrations derived from the vitellogenic response in fathead minnows exposed to a series of effluent dilutions were highly comparable with the E2 EQ concentrations derived from assessments of the estrogenic potency of these dilutions in the rYES. Together these data support the

Metabolomics for informing adverse outcome pathways: Androgen receptor activation and the pharmaceutical spironolactone

Energy Technology Data Exchange (ETDEWEB)

Davis, J.M., E-mail: davis.john@epa.gov [U.S. EPA, National Exposure Research Laboratory, 960 College Station Rd., Athens, GA 30605 (United States); Ekman, D.R., E-mail: ekman.drew@epa.gov [U.S. EPA, National Exposure Research Laboratory, 960 College Station Rd., Athens, GA 30605 (United States); Skelton, D.M. [U.S. EPA, National Exposure Research Laboratory, 960 College Station Rd., Athens, GA 30605 (United States); LaLone, C.A.; Ankley, G.T.; Cavallin, J.E.; Villeneuve, D.L. [U.S. EPA, National Health and Environmental Effects Research Laboratory, 6201 Congdon Blvd., Duluth, MN 55804 (United States); Collette, T.W. [U.S. EPA, National Exposure Research Laboratory, 960 College Station Rd., Athens, GA 30605 (United States)

2017-03-15

Highlights: • Metabolomics identified potential key events in an androgen receptor activation AOP. • Metabolomics indicate spironolactone may elicit effects via multiple nuclear receptors. • Spironolactone exposure may elicit interactive effects in multi-stressor environments. - Abstract: One objective in developing adverse outcome pathways (AOPs) is to connect biological changes that are relevant to risk assessors (i.e., fecundity) to molecular and cellular-level alterations that might be detectable at earlier stages of a chemical exposure. Here, we examined biochemical responses of fathead minnows (Pimephales promelas) to inform an AOP relevant to spironolactone’s activation of the androgen receptor, as well as explore other biological impacts possibly unrelated to this receptor. Liquid chromatography with high resolution mass spectrometry (LC–MS) was used to measure changes in endogenous polar metabolites in livers of male and female fish that were exposed to five water concentrations of spironolactone (0, 0.05, 0.5, 5, or 50 μg L{sup −1}) for 21 days. Metabolite profiles were affected at the two highest concentrations (5 and 50 μg L{sup −1}), but not in the lower-level exposures, which agreed with earlier reported results of reduced female fecundity and plasma vitellogenin (VTG) levels. We then applied partial least squares regression to assess whether metabolite alterations covaried with changes in fecundity, VTG gene expression and protein concentrations, and plasma 17β-estradiol and testosterone concentrations. Metabolite profiles significantly covaried with all measured endpoints in females, but only with plasma testosterone in males. Fecundity reductions occurred in parallel with changes in metabolites important in osmoregulation (e.g., betaine), membrane transport (e.g., L-carnitine), and biosynthesis of carnitine (e.g., methionine) and VTG (e.g., glutamate). Based on a network analysis program (i.e., mummichog), spironolactone also

Environmentally induced epigenetic transgenerational inheritance of altered Sertoli cell transcriptome and epigenome: molecular etiology of male infertility.

Directory of Open Access Journals (Sweden)

Carlos Guerrero-Bosagna

Full Text Available Environmental toxicants have been shown to induce the epigenetic transgenerational inheritance of adult onset disease, including testis disease and male infertility. The current study was designed to determine the impact of an altered sperm epigenome on the subsequent development of an adult somatic cell (Sertoli cell that influences the onset of a specific disease (male infertility. A gestating female rat (F0 generation was exposed to the agriculture fungicide vinclozolin during gonadal sex determination and then the subsequent F3 generation progeny used for the isolation of Sertoli cells and assessment of testis disease. As previously observed, enhanced spermatogenic cell apoptosis was observed. The Sertoli cells provide the physical and nutritional support for the spermatogenic cells. Over 400 genes were differentially expressed in the F3 generation control versus vinclozolin lineage Sertoli cells. A number of specific cellular pathways were identified to be transgenerationally altered. One of the key metabolic processes affected was pyruvate/lactate production that is directly linked to spermatogenic cell viability. The Sertoli cell epigenome was also altered with over 100 promoter differential DNA methylation regions (DMR modified. The genomic features and overlap with the sperm DMR were investigated. Observations demonstrate that the transgenerational sperm epigenetic alterations subsequently alters the development of a specific somatic cell (Sertoli cell epigenome and transcriptome that correlates with adult onset disease (male infertility. The environmentally induced epigenetic transgenerational inheritance of testis disease appears to be a component of the molecular etiology of male infertility.

RNA-sequencing and pathway analysis reveal alteration of hepatic steroid biosynthesis and retinol metabolism by tributyltin exposure in male rare minnow (Gobiocypris rarus).

Science.gov (United States)

Zhang, Jiliang; Zhang, Chunnuan; Sun, Ping; Huang, Maoxian; Fan, Mingzhen; Liu, Min

2017-07-01

Tributyltin (TBT) is widely spread in aquatic ecosystems. Although adverse effects of TBT on reproduction and lipogenesis are observed in fishes, the underlying mechanisms, especially in livers, are still scarce and inconclusive. Thus, RNA-sequencing runs were performed on the hepatic libraries of adult male rare minnow (Gobiocypris rarus) after TBT exposure for 60d. After differentially expressed genes were identified, enrichment analysis and validation by quantitative real-time PCR were conducted. The results showed that TBT up-regulated the profile of hepatic genes in the steroid biosynthesis pathway and down-regulated the profile of hepatic genes in the retinol metabolism pathway. In the hepatic steroid biosynthesis pathway, TBT might induce biosynthesis of cholesterol, which could affect the bioavailability of steroid hormones. More important, 3beta-hydroxysteroid 3-dehydrogenase, a key enzyme in the biosynthesis of all active steroid hormones, was up-regulated by TBT exposure. In the hepatic retinol metabolism pathway, TBT impaired retinoic acid homeostasis which plays essential roles in both reproduction and lipogenesis. The results of two pathways offered new mechanisms underlying the toxicology of TBT and represented a starting point from which detailed mechanistic links should be explored. Copyright © 2017 Elsevier B.V. All rights reserved.

Evaluation of whole mount in situ hybridization as a tool for pathway-based toxicological research in early-life stage fathead minnows

Science.gov (United States)

Early-life stage fish can be more sensitive to chemical exposure than mature, adult fish. Therefore, defining adverse outcome pathways (AOPs) relevant to early-life stages is critical for linking perturbations of key events during fish development to potential adverse outcomes of...

The use of adverse outcome pathway-based toxicity predictions: A case study evaluating the effects of imazalil on fathead minnow reproduction

Science.gov (United States)

Product Description: As a means to increase the efficiency of chemical safety assessment, there is an interest in using data from molecular and cellular bioassays, conducted in a highly automated fashion using modern robotics, to predict toxicity in humans and wildlife. The prese...

EXPOSE-R2: The Astrobiological ESA Mission on Board of the International Space Station

Directory of Open Access Journals (Sweden)

Elke Rabbow

2017-08-01

Full Text Available On July 23, 2014, the Progress cargo spacecraft 56P was launched from Baikonur to the International Space Station (ISS, carrying EXPOSE-R2, the third ESA (European Space Agency EXPOSE facility, the second EXPOSE on the outside platform of the Russian Zvezda module, with four international astrobiological experiments into space. More than 600 biological samples of archaea, bacteria (as biofilms and in planktonic form, lichens, fungi, plant seeds, triops eggs, mosses and 150 samples of organic compounds were exposed to the harsh space environment and to parameters similar to those on the Mars surface. Radiation dosimeters distributed over the whole facility complemented the scientific payload. Three extravehicular activities later the chemical samples were returned to Earth on March 2, 2016, with Soyuz 44S, having spent 588 days in space. The biological samples arrived back later, on June 18, 2016, with 45S, after a total duration in space of 531 days. The exposure of the samples to Low Earth Orbit vacuum lasted for 531 days and was divided in two parts: protected against solar irradiation during the first 62 days, followed by exposure to solar radiation during the subsequent 469 days. In parallel to the space mission, a Mission Ground Reference (MGR experiment with a flight identical Hardware and a complete flight identical set of samples was performed at the premises of DLR (German Aerospace Center in Cologne by MUSC (Microgravity User Support Center, according to the mission data either downloaded from the ISS (temperature data, facility status, inner pressure status or provided by RedShift Design and Engineering BVBA, Belgium (calculated ultra violet radiation fluence data. In this paper, the EXPOSE-R2 facility, the experimental samples, mission parameters, environmental parameters, and the overall mission and MGR sequences are described, building the background for the research papers of the individual experiments, their analysis and results.

Identification of aberrant gene expression associated with aberrant promoter methylation in primordial germ cells between E13 and E16 rat F3 generation vinclozolin lineage.

Science.gov (United States)

Taguchi, Y-h

2015-01-01

Transgenerational epigenetics (TGE) are currently considered important in disease, but the mechanisms involved are not yet fully understood. TGE abnormalities expected to cause disease are likely to be initiated during development and to be mediated by aberrant gene expression associated with aberrant promoter methylation that is heritable between generations. However, because methylation is removed and then re-established during development, it is not easy to identify promoter methylation abnormalities by comparing normal lineages with those expected to exhibit TGE abnormalities. This study applied the recently proposed principal component analysis (PCA)-based unsupervised feature extraction to previously reported and publically available gene expression/promoter methylation profiles of rat primordial germ cells, between E13 and E16 of the F3 generation vinclozolin lineage that are expected to exhibit TGE abnormalities, to identify multiple genes that exhibited aberrant gene expression/promoter methylation during development. The biological feasibility of the identified genes were tested via enrichment analyses of various biological concepts including pathway analysis, gene ontology terms and protein-protein interactions. All validations suggested superiority of the proposed method over three conventional and popular supervised methods that employed t test, limma and significance analysis of microarrays, respectively. The identified genes were globally related to tumors, the prostate, kidney, testis and the immune system and were previously reported to be related to various diseases caused by TGE. Among the genes reported by PCA-based unsupervised feature extraction, we propose that chemokine signaling pathways and leucine rich repeat proteins are key factors that initiate transgenerational epigenetic-mediated diseases, because multiple genes included in these two categories were identified in this study.

Evaluation of targeted and untargeted effects-based monitoring tools to assess impacts of contaminants of emerging concern on fish in the South Platte River, CO.

Science.gov (United States)

Ekman, Drew R; Keteles, Kristen; Beihoffer, Jon; Cavallin, Jenna E; Dahlin, Kenneth; Davis, John M; Jastrow, Aaron; Lazorchak, James M; Mills, Marc A; Murphy, Mark; Nguyen, David; Vajda, Alan M; Villeneuve, Daniel L; Winkelman, Dana L; Collette, Timothy W

2018-08-01

Rivers in the arid Western United States face increasing influences from anthropogenic contaminants due to population growth, urbanization, and drought. To better understand and more effectively track the impacts of these contaminants, biologically-based monitoring tools are increasingly being used to complement routine chemical monitoring. This study was initiated to assess the ability of both targeted and untargeted biologically-based monitoring tools to discriminate impacts of two adjacent wastewater treatment plants (WWTPs) on Colorado's South Platte River. A cell-based estrogen assay (in vitro, targeted) determined that water samples collected downstream of the larger of the two WWTPs displayed considerable estrogenic activity in its two separate effluent streams. Hepatic vitellogenin mRNA expression (in vivo, targeted) and NMR-based metabolomic analyses (in vivo, untargeted) from caged male fathead minnows also suggested estrogenic activity downstream of the larger WWTP, but detected significant differences in responses from its two effluent streams. The metabolomic results suggested that these differences were associated with oxidative stress levels. Finally, partial least squares regression was used to explore linkages between the metabolomics responses and the chemical contaminants that were detected at the sites. This analysis, along with univariate statistical approaches, identified significant covariance between the biological endpoints and estrone concentrations, suggesting the importance of this contaminant and recommending increased focus on its presence in the environment. These results underscore the benefits of a combined targeted and untargeted biologically-based monitoring strategy when used alongside contaminant monitoring to more effectively assess ecological impacts of exposures to complex mixtures in surface waters. Published by Elsevier Ltd.

The aquatic toxicity and chemical forms of coke plant effluent cyanide -- Implications for discharge limits

International Nuclear Information System (INIS)

Garibay, R.; Rupnow, M.; Godwin-Saad, E.; Hall, S.

1995-01-01

Cyanide is present in treated cokemaking process waters at concentrations as high as 8.0 mg/L. In assessing options for managing the discharge of a treated effluent, the development and implementation of discharge limits for cyanide became a critical issue. A study was initiated to evaluate possible alternatives to cyanide permit limits at the US Steel Gary Works Facility. The objectives of the study were to: (1) evaluation the forms of cyanide present in coke plant effluent; (2) determine whether these forms of cyanide are toxic to selected aquatic organisms; (3) compare the aquatic toxicity of various chemical forms of cyanide; (4) identify if the receiving water modifies cyanide bioavailability; and (5) confirm, with respect to water quality-based effluent limits, an appropriate analytical method for monitoring cyanide in a coke plant effluent. The results of aquatic toxicity tests and corresponding analytical data are presented. Toxicity tests were conducted with various pure chemical forms of cyanide as well as whole coke plant effluent (generated from a pilot-scale treatment system). Test species included the fathead minnow (Pimephales promelas), rainbow trout (Oncorhynchus mykiss), Ceriodaphnia dubia (C. dubia) and Daphnia magna (D. magna). Analytical measurements for cyanide included total, weak acid dissociable, diffusible cyanide and selected metal species of cyanide. The findings presented by the paper are relevant with respect to the application of cyanide water quality criteria for a coke plant effluent discharge, the translation of these water quality-based effluent limits to permit limits, and methods for compliance monitoring for cyanide

Aspects of the winter predator--prey relationship between sauger and threadfin shad in Watts Bar Reservoir, Tennessee

International Nuclear Information System (INIS)

McGee, M.V.; Griffith, J.S.; McLean, R.B.

1978-01-01

This study sought to determine the impact of cold-induced mortality and impingement of threadfin shad (Dorsoma petenense) on the food consumption and prey selection of sauger (Stizostedion canadense), and to estimate the ability of sauger to digest meals consumed at low temperatures in winter. Prey selection of sauger was monitored from November 1976 through April 1977. Stomach contents of 536 sauger indicated threadfin provided the entire forage base for sauger through January. Food consumption of sauger was reduced and prey selection shifted to other species after January due to the combined effects of predation, impingement, and natural mortality of cold-stressed threadfin. Threadfin shad of a size available to most sauger were virtually eliminated by February. From February through April some sauger utilized alternate prey species. Laboratory digestion rate studies of sauger indicated digestion of force-fed meals of 4 to 7 g fathead minnows (Pimephales promelas) could proceed to 90 percent completion in 54 h at 5 C, 47 hr at 10 C, and 25 hr at 15 C. Conclusions of this study are: (1) that threadfin shad were the most abundant and vulnerable prey species available to and utilized by sauger during the late fall and winter months; (2) extensive mortalities of threadfin due to cold-stress increased sauger predation on four alternate prey species; (3) sauger continued feeding and digesting meals at temperatures between 5 and 15 C every 1 to 3 days; (4) sauger stored excess energy available from threadfin early in the winter as visceral fat which was available later when food consumption was reduced

Aspects of the winter predator--prey relationship between sauger and threadfin shad in Watts Bar Reservoir, Tennessee

Energy Technology Data Exchange (ETDEWEB)

McGee, M.V.; Griffith, J.S.; McLean, R.B.

1978-04-01

This study sought to determine the impact of cold-induced mortality and impingement of threadfin shad (Dorsoma petenense) on the food consumption and prey selection of sauger (Stizostedion canadense), and to estimate the ability of sauger to digest meals consumed at low temperatures in winter. Prey selection of sauger was monitored from November 1976 through April 1977. Stomach contents of 536 sauger indicated threadfin provided the entire forage base for sauger through January. Food consumption of sauger was reduced and prey selection shifted to other species after January due to the combined effects of predation, impingement, and natural mortality of cold-stressed threadfin. Threadfin shad of a size available to most sauger were virtually eliminated by February. From February through April some sauger utilized alternate prey species. Laboratory digestion rate studies of sauger indicated digestion of force-fed meals of 4 to 7 g fathead minnows (Pimephales promelas) could proceed to 90 percent completion in 54 h at 5 C, 47 hr at 10 C, and 25 hr at 15 C. Conclusions of this study are: (1) that threadfin shad were the most abundant and vulnerable prey species available to and utilized by sauger during the late fall and winter months; (2) extensive mortalities of threadfin due to cold-stress increased sauger predation on four alternate prey species; (3) sauger continued feeding and digesting meals at temperatures between 5 and 15 C every 1 to 3 days; (4) sauger stored excess energy available from threadfin early in the winter as visceral fat which was available later when food consumption was reduced.

The chronic toxicity of sodium bicarbonate, a major component of coal bed natural gas produced waters

Science.gov (United States)

Farag, Aïda M.; Harper, David D.

2014-01-01

Sodium bicarbonate (NaHCO3) is the principal salt in coal bed natural gas produced water from the Powder River Structural Basin, Wyoming, USA, and concentrations of up to 3000 mg NaHCO3/L have been documented at some locations. No adequate studies have been performed to assess the chronic effects of NaHCO3 exposure. The present study was initiated to investigate the chronic toxicity and define sublethal effects at the individual organism level to explain the mechanisms of NaHCO3 toxicity. Three chronic experiments were completed with fathead minnows (Pimephales promelas), 1 with white suckers (Catostomus commersoni), 1 with Ceriodaphnia dubia, and 1 with a freshwater mussel, (Lampsilis siliquoidea). The data demonstrated that approximately 500 mg NaHCO3/L to 1000 mg NaHCO3/L affected all species of experimental aquatic animals in chronic exposure conditions. Freshwater mussels were the least sensitive to NaHCO3 exposure, with a 10-d inhibition concentration that affects 20% of the sample population (IC20) of 952 mg NaHCO3/L. The IC20 for C. dubia was the smallest, at 359 mg NaHCO3/L. A significant decrease in sodium–potassium adenosine triphosphatase (Na+/K+ ATPase) together with the lack of growth effects suggests that Na+/K+ ATPase activity was shut down before the onset of death. Several histological anomalies, including increased incidence of necrotic cells, suggested that fish were adversely affected as a result of exposure to >450 mg NaHCO3/L.

Lipid accumulation, oxidative stress and immune-related molecules affected by tributyltin exposure in muscle tissues of rare minnow (Gobiocypris rarus).

Science.gov (United States)

Zhang, Jiliang; Zhang, Chunnuan; Ma, Dongdong; Liu, Min; Huang, Shuntao

2017-12-01

Tributyltin (TBT) is reported to induce adipogenesis in fish, which might affect nutritional qualities and health status. Muscle tissues account for the majority of body mass, and have been described as a major site of fat deposition and an immunologically active organ. Therefore, the present study aims to evaluate whether chronic exposures of TBT, at environmental concentrations of 1, 10 and 100 ng/L, affects lipid accumulation, oxidative stress and immune status in muscle tissues of rare minnow (Gobiocypris rarus). After 60 d of exposure, TBT increased contents of total lipid, total cholesterol, triglyceride and fatty acids in muscle tissues. Interestingly, TBT exposure disrupted fatty acid composition and increased contents of unsaturated fatty acids (such as eicosapentaenoic acid and docosahexaenoic acid) in muscle tissues, which might be a response to preserve membrane functions from TBT exposure. Meanwhile, the concentrations of hepatic fatty acid desaturase 2 (Δ6-desaturase) and stearoyl-CoA desaturase (Δ9-desaturase) were increased after TBT exposure, which might contribute the increase of unsaturated fatty acids. Furthermore, TBT increased muscle lipid peroxidation products, antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase), and the expression of immune-related molecules (tumor necrosis factor alpha, interleukin 1 beta and nuclear factor kappa B) in muscle tissues. The disruption of TBT on the lipid accumulation, oxidative stress and immune-toxic effects in muscle tissues of fish might reduce nutritional qualities, and affect growth and health status, which might pose a constant and serious threat to fish and result in economic loss in aquaculture. Copyright © 2017 Elsevier Ltd. All rights reserved.

Nutritional Status among the Children of Age Group 5-14 Years in Selected Arsenic Exposed and Non-Exposed Areas of Bangladesh.

Science.gov (United States)

Rezaul Karim, Mohammad; Ahmad, Sk Akhtar

2014-12-01

To assess and compare the nutritional status of children aged 5-14 years in arsenic exposed and non- exposed areas. It was a cross sectional study conducted on 600 children of age 5-14 years from arsenic exposed and non-exposed areas in Bangladesh. Designed questionnaire and check list were used for collection of data. To estimate BMI necessary anthropometric measurements of the studied children were done. Dietary intakes of the study children were assessed using 24-hours recall method. The difference of socio-economic conditions between the children of exposed area and non-exposed area was not significant. On an average the body mass index was found to be significantly (p < 0.01) lower among the children of arsenic exposed area (49%) in comparison to that of children in non-exposed area (38%). Stunting (p < 0.01), wasting (p < 0.05) and underweight (p < 0.05) were significantly higher in exposed group in comparison to non-exposed group. No significant difference of nutrition intake was found between exposed and non-exposed children as well as thin and normal children. In this study children exposed to arsenic contaminated water were found to be suffered from lower nutritional status.

Nutritional Status among the Children of Age Group 5-14 Years in Selected Arsenic Exposed and Non-Exposed Areas of Bangladesh.

Directory of Open Access Journals (Sweden)

Mohammad Rezaul Karim

2014-12-01

Full Text Available To assess and compare the nutritional status of children aged 5-14 years in arsenic exposed and non- exposed areas.It was a cross sectional study conducted on 600 children of age 5-14 years from arsenic exposed and non-exposed areas in Bangladesh. Designed questionnaire and check list were used for collection of data. To estimate BMI necessary anthropometric measurements of the studied children were done. Dietary intakes of the study children were assessed using 24-hours recall method.The difference of socio-economic conditions between the children of exposed area and non-exposed area was not significant. On an average the body mass index was found to be significantly (p < 0.01 lower among the children of arsenic exposed area (49% in comparison to that of children in non-exposed area (38%. Stunting (p < 0.01, wasting (p < 0.05 and underweight (p < 0.05 were significantly higher in exposed group in comparison to non-exposed group. No significant difference of nutrition intake was found between exposed and non-exposed children as well as thin and normal children.In this study children exposed to arsenic contaminated water were found to be suffered from lower nutritional status.

Evaluation of whole mount in situ hybridization as a tool for pathway-based toxicological research in early-life stage fathead minnows (poster)

Science.gov (United States)

Early-life stage fish can be more sensitive to chemical exposure than mature, adult fish. Therefore, defining adverse outcome pathways (AOPs) relevant to early-life stages is critical for linking perturbations of key events during fish development to potential adverse outcomes of...

A PHYSIOLOGICALLY BASED COMPUTATIONAL MODEL OF THE BPG AXIS IN FATHEAD MINNOWS: PREDICTING EFFECTS OF ENDOCRINE DISRUPTING CHEMICAL EXPOSURE ON REPRODUCTIVE ENDPOINTS

Science.gov (United States)

This presentation describes development and application of a physiologically-based computational model that simulates the brain-pituitary-gonadal (BPG) axis and other endpoints important in reproduction such as concentrations of sex steroid hormones, 17-estradiol, testosterone, a...

Impact assessment of dredging to remove coal fly ash at the Tennessee Valley Authority Kingston Fossil plant using fathead minnow elutriate exposures.

Science.gov (United States)

Stanley, Jacob K; Kennedy, Alan J; Bednar, Anthony J; Chappell, Mark A; Seiter, Jennifer M; Averett, Daniel E; Steevens, Jeffery A

2013-04-01

On December 22, 2008, failure of an earthen containment structure resulted in the release of approximately 4.1 million m(3) of coal fly ash into the Emory River and the surrounding area from the Tennessee Valley Authority Kingston Fossil Plant near Kingston, Tennessee, USA. The purpose of the present study was to assess the potential of dredging activities performed to remove the fly ash from the river to result in increased risk to pelagic fish, with special consideration of mobilization of metals. Elutriates were created using two sources of fly ash by bubbling with air over 10 d. This elutriate preparation method was designed to represent worst-case conditions for oxidation, metal release, and dissolution. Larval and juvenile Pimephales promelas underwent 10-d exposures to these elutriates. Larval end points included survival and biomass, and juvenile end points included survival, length, biomass, liver somatic index, and bioaccumulation. No significant toxicity was observed. Bioaccumulation of metals in juveniles was found to be primarily attributable to metals associated with particles in the gut. Results suggest little potential for toxicity to related fish species due to fly ash removal dredging activities given the extreme conditions represented by the elutriates in the present study. Copyright © 2013 SETAC.

Disposal of tritium-exposed metal hydrides

International Nuclear Information System (INIS)

Nobile, A.; Motyka, T.

1991-01-01

A plan has been established for disposal of tritium-exposed metal hydrides used in Savannah River Site (SRS) tritium production or Materials Test Facility (MTF) R ampersand D operations. The recommended plan assumes that the first tritium-exposed metal hydrides will be disposed of after startup of the Solid Waste Disposal Facility (SWDF) Expansion Project in 1992, and thus the plan is consistent with the new disposal requiremkents that will be in effect for the SWDF Expansion Project. Process beds containing tritium-exposed metal hydride powder will be disposed of without removal of the powder from the bed; however, disposal of tritium-exposed metal hydride powder that has been removed from its process vessel is also addressed

9 CFR 78.8 - Brucellosis exposed cattle.

Science.gov (United States)

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Brucellosis exposed cattle. 78.8... Restrictions on Interstate Movement of Cattle Because of Brucellosis § 78.8 Brucellosis exposed cattle. Brucellosis exposed cattle may be moved interstate only as follows: (a) Movement to recognized slaughtering...

Leukemias in the progeny of exposed parents

International Nuclear Information System (INIS)

Kosenko, M.M.; Gudkova, N.V.

1996-01-01

The purpose of this study was to assess the incidence of leukemias among the progeny of exposed parents. The parents were exposed as a result of discharge of radioactive waste from the Mayak atomic plant into the Techa river in the Southern Urals. The doses per parents gonads, ranging from 0.035 to 1.27 Sv, were due to external exposure in 1950-1956 and to incorporation of Cs-137. Nine cases with leukemia and four with lympohoma were recorded in 13.500 antenatally exposed subjects and descendants of exposed parents over the period of 1950 to 1988. The leukemia morbidity index for the progeny of exposed parents was 2.51, which virtually not statistically differ from that in control group. Refs. 7, figs. 3, tabs. 3

The properties degradation of exposed GFRP roof

Science.gov (United States)

Zainudin, Mohammad; Diharjo, Kuncoro; Kaavessina, Mujtahid; Setyanto, Djoko

2018-02-01

There is much consideration of roof selection as a protector of a building against the outside weather, such as lightweight, strong stiff, corrosion resistant and guarantee for the availability of products. Based on these considerations, glass fiber reinforced polymer (GFRP) roof is a roof which can fulfill the requirement. The objective of this research is to investigate the degradation of physical and mechanical properties of GFRP roof exposed in outside weather. This GFRP roof composite was produced using a sheet molding compound (SMC) supplied by PT Intec Persada, Tangerang, Indonesia. There are two kinds GFRP roofs evaluated in this research that are GFRP roof exposed within 7 years and new GFRP roof that has not been exposed. The GFRP roofs were cut manually for preparing the specimens for hardness test, tensile test, SEM and FTIR test. The results show that the GFRP roof exposed within 7 years had the degradation of properties compared to the new GFRP roof. The exposed GFRP roof had lower strength and hardness compared to the new GFRP roof. The SEM observation indicates that exposed GFRP roof had the debonding of fiber on the surface, and in contrast, there are no debonding of fiber in the new GFRP roof surface. It can be recommended that the exposed GFRP roof may be repaired to enhance its performance and can re-increase its properties using the coating.

The DNA methylation status alteration of two steroidogenic genes in gonads of rare minnow after bisphenol A exposure.

Science.gov (United States)

Zhang, Ting; Liu, Yan; Chen, Hong; Gao, Jiancao; Zhang, Yingying; Yuan, Cong; Wang, Zaizhao

2017-08-01

Both cytochrome P450c17 (CYP17A1) and P-450 side chain cleavage (CYP11A1) play important roles in steroid biosynthesis. According to our previous studies, bisphenol A (BPA) could regulate the mRNA expression of cyp17a1 and cyp11a1 in rare minnow Gobiocypris rarus. However, the potential mechanism of the regulation is barely understood. In the present study, aiming to explore how BPA affects the mRNA expression of cyp17a1 and cyp11a1 in testes and ovaries of G. rarus, we firstly cloned 340-bp fragment of 5' flanking region of cyp11a1 and then detected the methylation level of CpG loci involved in 5' flanking of cyp11a1 and cyp17a1 and their mRNA expression levels. Results showed that exposure to BPA significantly increased serum estradiol (E2) and 11-ketotesterone (11-KT) concentrations. Ovarian mRNA expression of cyp17a1 and cyp11a1 were significantly decreased after BPA exposure 7- for and 14-days. However, transcriptions of testicular cyp17a1 and cyp11a1 were significantly increased and decreased respectively after BPA treatment for 14days. The DNA methylation levels of cyp17a1 were decreased in ovaries on day 7 and increased in ovaries and decreased in testes respectively on day 14. The methylation levels of cyp11a1 were increased in ovaries on day 7 and both ovaries and testes on day 14. There were a significant correlation between DNA methylation at specific CpG loci and cyp17a1 and cyp11a1 genes transcription levels. In conclusion, the CpG loci methylation in 5' flanking region appears to involve in the regulation of mRNA expression of cyp17a1 and cyp11a1 mediated by BPA. Copyright © 2017 Elsevier Inc. All rights reserved.

Protein A from orange-spotted nervous necrosis virus triggers type I interferon production in fish cell.

Science.gov (United States)

Huang, Runqing; Zhou, Qiong; Shi, Yan; Zhang, Jing; He, Jianguo; Xie, Junfeng

2018-05-04

Family Nodaviridae consists of two genera: Alphanodavirus and Betanodavirus, and the latter is classified into four genotypes, including red-spotted grouper nervous necrosis virus, tiger puffer nervous necrosis virus, striped jack nervous necrosis virus, and barfin flounder nervous necrosis virus. Type I interferons (IFNs) play a central role in the innate immune system and antiviral responses, and the interactions between IFN and NNV have been investigated in this study. We have found that the RNA-dependent RNA polymerase (RdRp) from orange-spotted nervous necrosis virus (OGNNV), named protein A, was capable of activating IFN promoter in fathead minnow (FHM) cells. Transient expression of protein A was found to induce IFN expression and secretion, endowing FHM cells with anti-tiger frog virus ability. Protein A from SJNNV can also induce IFN expression in FHM cells but that from Flock House virus (FHV), a well-studied representative species of genus Alphanodavirus, cannot. RdRp activity and mitochondrial localization were shown to be required for protein A to induce IFN expression by means of activating IRF3 but not NFκB. Furthermore, DsRNA synthesized in vitro transcription and poly I:C activated IFN promoter activity when transfected into FHM cells, and dsRNA were also detected in NNV-infected cells. We postulated that dsRNA, a PAMP, was produced by protein A, leading to activation of innate immune response. These results suggest that protein As from NNV are the agonists of innate immune response. This is the first work to demonstrate the interaction between NNV protein A and innate immune system, and may help to understand pathogenesis of NNV. Copyright © 2018. Published by Elsevier Ltd.

A strain of Siniperca chuatsi rhabdovirus causes high mortality among cultured Largemouth Bass in South China.

Science.gov (United States)

Ma, Dongmei; Deng, Guocheng; Bai, Junjie; Li, Shengjie; Yu, Lingyun; Quan, Yingchun; Yang, Xiaojing; Jiang, Xiaoyan; Zhu, Zemin; Ye, Xing

2013-09-01

In April 2011, 40% mortality of Largemouth Bass Micropterus salmoides juveniles occurred at a farm of Zhongshan City, Guangdong Province, China. Infected fish became lethargic, exhibited corkscrew and irregular swimming, and developed a distended abdomen and crooked body. Fish began to die within 2 d after the appearance of clinical signs. In order to analyze the pathogeny and diagnose the disease earlier, observation of clinical signs, cell infection, titer calculation, electron microscopy, immersion infection assay for fish, and nucleotide sequence analysis were carried out. Fathead minnow (FHM) cell cultures, inoculated with filtrate of liver and spleen homogenates from the diseased fish, developed the obvious cytopathic effect 46 h after inoculation in the primary culture and 24 h at the first passage. Typical rhabdovirus particles, 115-143 nm in length and 62-78 nm in diameter, were observed in infected FHM cells by direct transmission electron microscopy. The isolated virus produced a titer of 10(7.15) TCID50/mL. Immersion-Fish infected with the virus had similar clinical signs and 80% mortality with 10(2.5) LD50/mL. The data indicated that the rhabdovirus was the lethal pathogeny of the current disease. Based on nucleoprotein-gene nucleotide sequence multiple alignment analysis, the newly isolated virus is a strain of Siniperca chuatsi rhabdovirus (SCRV) under family Rhabdoviridae, which was initially isolated from Mandarin Fish Siniperca chuatsi. Up to the present, at least four virus strains have been isolated from diseased Largemouth Bass, which have had different clinical signs. Comparison of the clinical signs can help in an early diagnosis of the disease.

Endocrine disrupting chemicals in fish: developing exposure indicators and predictive models of effects based on mechanism of action.

Science.gov (United States)

Ankley, Gerald T; Bencic, David C; Breen, Michael S; Collette, Timothy W; Conolly, Rory B; Denslow, Nancy D; Edwards, Stephen W; Ekman, Drew R; Garcia-Reyero, Natalia; Jensen, Kathleen M; Lazorchak, James M; Martinović, Dalma; Miller, David H; Perkins, Edward J; Orlando, Edward F; Villeneuve, Daniel L; Wang, Rong-Lin; Watanabe, Karen H

2009-05-05

Knowledge of possible toxic mechanisms (or modes) of action (MOA) of chemicals can provide valuable insights as to appropriate methods for assessing exposure and effects, thereby reducing uncertainties related to extrapolation across species, endpoints and chemical structure. However, MOA-based testing seldom has been used for assessing the ecological risk of chemicals. This is in part because past regulatory mandates have focused more on adverse effects of chemicals (reductions in survival, growth or reproduction) than the pathways through which these effects are elicited. A recent departure from this involves endocrine-disrupting chemicals (EDCs), where there is a need to understand both MOA and adverse outcomes. To achieve this understanding, advances in predictive approaches are required whereby mechanistic changes caused by chemicals at the molecular level can be translated into apical responses meaningful to ecological risk assessment. In this paper we provide an overview and illustrative results from a large, integrated project that assesses the effects of EDCs on two small fish models, the fathead minnow (Pimephales promelas) and zebrafish (Danio rerio). For this work a systems-based approach is being used to delineate toxicity pathways for 12 model EDCs with different known or hypothesized toxic MOA. The studies employ a combination of state-of-the-art genomic (transcriptomic, proteomic, metabolomic), bioinformatic and modeling approaches, in conjunction with whole animal testing, to develop response linkages across biological levels of organization. This understanding forms the basis for predictive approaches for species, endpoint and chemical extrapolation. Although our project is focused specifically on EDCs in fish, we believe that the basic conceptual approach has utility for systematically assessing exposure and effects of chemicals with other MOA across a variety of biological systems.

Site Specific Metal Criteria Developed Using Kentucky Division of Water Procedures

International Nuclear Information System (INIS)

Kszos, L.A.; Phipps, T.L.

1999-01-01

Alternative limits for Cu, Ni, Pb, and Zn were developed for treated wastewater from four outfalls at a Gaseous Diffusion Plant. Guidance from the Kentucky Division of Water (KDOW) was used to (1) estimate the toxicity of the effluents using water fleas (Ceriodaphnia dubia) and fathead minnow (Pimephales promelas) larvae; (2) determine total recoverable and dissolved concentrations of Cu, Pb, Ni, and Zn ; (3) calculate ratios of dissolved metal (DM) to total recoverable metal (TRM); and (4) assess chemical characteristics of the effluents. Three effluent samples from each outfall were collected during each of six test periods; thus, a total of 18 samples from each outfall were evaluated for toxicity, DM and TRM. Subsamples were analyzed for alkalinity, hardness, pH, conductivity, and total suspended solids. Short-term (6 or 7 d), static renewal toxicity tests were conducted according to EPA methodology. Ceriodaphnia reproduction was reduced in one test of effluent from Outfall A , and effluent from Outfall B was acutely toxic to both test species during one test. However, the toxicity was not related to the metals present in the effluents. Of the 18 samples from each outfall, more than 65% of the metal concentrations were estimated quantities. With the exception of two total recoverable Cu values in Outfall C, all metal concentrations were below the permit limits and the federal water quality criteria. Ranges of TR for all outfalls were: Cd, ,0.1-0.4 microg/L; Cr,1.07-3.93 microg/L; Cu, 1.59-7.24 microg/L; Pb, 39%. Thus, the alternative, calculated total recoverable metal concentration provides the discharger with a ''cushion'' for meeting permit limits

Analysis of early mortality rates of survivors exposed within Japanese wooden houses in Hiroshima by exposed distance

International Nuclear Information System (INIS)

Hayakawa, Norihiko; Munaka, Masaki; Kurihara, Minoru; Ohkita, Takeshi.

1986-01-01

Mortality for 3,215 A-bomb survivors who were exposed in Japanese wooden houses at ≤ 1,300 m from the hypocenter on August 6, 1945 was examined. An overall mortality was 51 % (1,640/3,215 survivors) within 61 days after the exposure. According to the distance from the hypocenter, it was 100 % in A-bomb survivors exposed at ≤ 600 m, and 20 % in those exposed between 1,201 m and 1,300 m. The mortality decreased with increasing the distance from the hypocenter. In conjunction with the duration after the exposure and the distance from the hypocenter, the mortality was 100 % 12 days after the exposure in survivors exposed at ≤ 600 m. In survivors exposed at > 800 m, the mortality tended to be higher two weeks after the exposure than immediately after that. The distance from the hypocenter causing 50 per cent mortality was estimated to be 1,026 m from August 6 to October 5; 1,002 m from August 6 to September 10; 887 m from August 7 to September 10; and 867 m from August 20 to September 16. However, these figures were probably lower than the real mortality rates, since no information was available when whole family died. (Namekawa, K.)

Occurrence of organic wastewater and other contaminants in cave streams in northeastern Oklahoma and northwestern Arkansas

Science.gov (United States)

Bidwell, Joseph R.; Becker, C.; Hensley, S.; Stark, R.; Meyer, M.T.

2010-01-01

found in caves and surface-water sites included brominated flame retardants, organochlorine pesticides (chlordane and nonachlor), and polychlorinated biphenyls. The placement of samplers in the caves (near the cave mouth compared to farther in the system) might have influenced the number of halogenated organics detected due to possible aerial transport of residues. Guano from cave-dwelling bats also might have been a source of some of these chlorinated organics. Seven-day survival and growth bioassays with fathead minnows (Pimephales promelas) exposed to samples of cave water indicated initial toxicity in water from two of the caves, but these effects were transient, with no toxicity observed in follow-up tests. ??Springer Science+Business Media, LLC 2009.

The chronic toxicity of molybdate to freshwater organisms. I. Generating reliable effects data

Energy Technology Data Exchange (ETDEWEB)

De Schamphelaere, K.A.C., E-mail: karel.deschamphelaere@Ugent.be [Laboratory of Environmental Toxicology and Aquatic Ecology, Faculty of Bioscience Engineering, Ghent University (Belgium); Stubblefield, W. [Oregon State University, Department of Environmental and Molecular Toxicology, 421 Weniger Hall, Corvallis, OR 97331 (United States); Rodriguez, P. [Centro de Investigacion Minera y Metalurgica (CIMM), Santiago (Chile); Vleminckx, K. [Department for Molecular Biomedical Research, Ghent University (Belgium); Janssen, C.R. [Laboratory of Environmental Toxicology and Aquatic Ecology, Faculty of Bioscience Engineering, Ghent University (Belgium)

2010-10-15

The European Union regulation on Registration, Evaluation, Authorization and Restriction of Chemical substances (REACH) (EC, 2006) requires the characterization of the chronic toxicity of many chemicals in the aquatic environment, including molybdate (MoO{sub 4}{sup 2-}). Our literature review on the ecotoxicity of molybdate revealed that a limited amount of reliable chronic no observed effect concentrations (NOECs) for the derivation of a predicted no-effect concentration (PNEC) existed. This paper presents the results of additional ecotoxicity experiments that were conducted in order to fulfill the requirements for the derivation of a PNEC by means of the scientifically most robust species sensitivity distribution (SSD) approach (also called the statistical extrapolation approach). Ten test species were chronically exposed to molybdate (added as sodium molybdate dihydrate, Na{sub 2}MoO{sub 4}.2H{sub 2}O) according to internationally accepted standard testing guidelines or equivalent. The 10% effective concentrations (EC10, expressed as measured dissolved molybdenum) for the most sensitive endpoint per species were 62.8-105.6 (mg Mo)/L for Daphnia magna (21 day-reproduction), 78.2 (mg Mo)/L for Ceriodaphnia dubia (7 day-reproduction), 61.2-366.2 (mg Mo)/L for the green alga Pseudokirchneriella subcapitata (72 h-growth rate), 193.6 (mg Mo)/L for the rotifer Brachionus calyciflorus (48 h-population growth rate), 121.4 (mg Mo)/L for the midge Chironomus riparius (14 day-growth), 211.3 (mg Mo)/L for the snail Lymnaea stagnalis (28 day-growth rate), 115.9 (mg Mo)/L for the frog Xenopus laevis (4 day-larval development), 241.5 (mg Mo)/L for the higher plant Lemna minor (7 day-growth rate), 39.3 (mg Mo)/L for the fathead minnow Pimephales promelas (34 day-dry weight/biomass), and 43.2 (mg Mo)/L for the rainbow trout Oncorhynchus mykiss (78 day-biomass). These effect concentrations are in line with the few reliable data currently available in the open literature. The data

The chronic toxicity of molybdate to freshwater organisms. I. Generating reliable effects data

International Nuclear Information System (INIS)

De Schamphelaere, K.A.C.; Stubblefield, W.; Rodriguez, P.; Vleminckx, K.; Janssen, C.R.

2010-01-01

The European Union regulation on Registration, Evaluation, Authorization and Restriction of Chemical substances (REACH) (EC, 2006) requires the characterization of the chronic toxicity of many chemicals in the aquatic environment, including molybdate (MoO 4 2- ). Our literature review on the ecotoxicity of molybdate revealed that a limited amount of reliable chronic no observed effect concentrations (NOECs) for the derivation of a predicted no-effect concentration (PNEC) existed. This paper presents the results of additional ecotoxicity experiments that were conducted in order to fulfill the requirements for the derivation of a PNEC by means of the scientifically most robust species sensitivity distribution (SSD) approach (also called the statistical extrapolation approach). Ten test species were chronically exposed to molybdate (added as sodium molybdate dihydrate, Na 2 MoO 4 .2H 2 O) according to internationally accepted standard testing guidelines or equivalent. The 10% effective concentrations (EC10, expressed as measured dissolved molybdenum) for the most sensitive endpoint per species were 62.8-105.6 (mg Mo)/L for Daphnia magna (21 day-reproduction), 78.2 (mg Mo)/L for Ceriodaphnia dubia (7 day-reproduction), 61.2-366.2 (mg Mo)/L for the green alga Pseudokirchneriella subcapitata (72 h-growth rate), 193.6 (mg Mo)/L for the rotifer Brachionus calyciflorus (48 h-population growth rate), 121.4 (mg Mo)/L for the midge Chironomus riparius (14 day-growth), 211.3 (mg Mo)/L for the snail Lymnaea stagnalis (28 day-growth rate), 115.9 (mg Mo)/L for the frog Xenopus laevis (4 day-larval development), 241.5 (mg Mo)/L for the higher plant Lemna minor (7 day-growth rate), 39.3 (mg Mo)/L for the fathead minnow Pimephales promelas (34 day-dry weight/biomass), and 43.2 (mg Mo)/L for the rainbow trout Oncorhynchus mykiss (78 day-biomass). These effect concentrations are in line with the few reliable data currently available in the open literature. The data presented in this

EFEITO DA APLICAÇÃO DE FUNGICIDAS NO SOLO SOBRE A GERMINAÇÃO CARPOGÊNICA E MICELIOGÊNICA DE ESCLERÓDIOS DE Sclerotinia sclerotiorum EFFECT OF FUNGICIDE APPLICATION IN THE SOIL ON THE CARPOGENIC AND MYCELIOGENIC GERMINATION OF Sclerotinia sclerotiorum

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Gesimária Ribeiro Costa

2007-09-01

Full Text Available

Os fungicidas benomyl, procymidone, iprodione, vinclozolin, fluazinan e tiofanato metílico foram utilizados para verificar seus efeitos na germinação carpogênica e miceliogênica de escleródios de Sclerotinia sclerotiorum, no solo. O solo coletado foi distribuído em caixas gerbox. Em cada caixa foram colocados 300 g de solo e, posteriormente, foi efetuado o enterrio de 25 escleródios à profundidade de 2,0 cm. Os fungicidas foram aplicados no solo, na dose de 0,5 kg.ha-1 de i.a., simulando uma lâmina de água de 6,0 mm. Em geral, o vinclozolin foi o melhor tratamento, apresentando uma eficiência de 100% na inibição de estipes e apotécios. O fluazinan permitiu apenas a formação de estipes inviáveis, resultando na ausência de apotécios. Na germinação miceliogênica dos escleródios, observou-se que após quinze dias de incubação, o tiofanato metílico reduziu em 75% a germinação miceliogênica, seguido dos fungicidas procymidone e vinclozolin, que apresentaram, em média, 60% de inibição. Os fungicidas benomyl, fluazinan e iprodione foram os menos eficientes na inibição da germinação miceliogênica dos escleródios, não diferindo estatisticamente do controle. Após trinta dias de incubação dos escleródios no solo, os fungicidas procymidone, tiofanato metílico, vinclozolin, iprodione e fluazinam não diferiram significativamente entre si quanto à inibição da germinação miceliogênica, mas diferiram do fungicida benomyl e do tratamento testemunha.

PALAVRAS-CHAVE: Controle químico; mofo branco; Phaseolus vulgaris.

The effects of benomyl, procymidone, iprodione, vinclozolin, fluazinan and methyl thiophanate on the carpogenic and myceliogenic germination of Sclerotinia sclerotiorum sclerotia, in soil, were

CLONING AND IN VITRO EXPRESSION AND CHARACTERIZATION OF THE ANDROGEN RECEPTOR AND ISOLATION OF ESTROGEN RECEPTOR α FROM THE FATHEAD MINNOW (PIMEPHALES PROMELAS)

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In vitro screening assays designed to identify hormone mimics or antagonists typically use mammalian (rat, human) estrogen (ER) and androgen receptors (AR). Although we know that the amino acid sequences of steroid receptors in nonmammalian vertebrates are not identical to the ma...

EVALUATION OF PILOT TREATMENT EFFLUENTS FROM SUMMITVILLE MINE, CO, USING CERIODAPHNIA DUBIA, FATHEAD MINNOW (PIMEPHALES PROMELAS), AND RAINBOW TROUT (ONCORHYNCHUS MYKISS) TOXICITY TESTS

Science.gov (United States)

As part of a Superfund Innovative Technology Evaluation (SITE) Program, the U.S. Environmental Protection Agency evaluated a remediation technology at the Summitville Mine Superfund site in southern Colorado. The technology evaluated was a successive alkalinity producing system ...

[A survey of occupational health among polyether-exposed workers].

Science.gov (United States)

Fu, Xu-ying; Yu, Bin; Zhang, Chun-ping; Zheng, Guan-hua; Bai, Lan; Zhang, Pan-pan

2013-06-01

To investigate the occupational health of the workers simultaneously exposed to acrylonitrile, epoxyethane, epoxypropane, and styrene. A questionnaire survey was conducted in 70 front-line workers simultaneously exposed to acrylonitrile, epoxyethane, epoxypropane, and styrene (exposure group) and 50 managers (control group) in a polyether manufacturer; in addition, air monitoring at workplace and occupational health examination were also performed. The obtained data were analyzed. The female workers in exposure group and the spouses of male workers in exposure group had significantly higher spontaneous abortion rates than their counterparts in control group (P polyether-exposed working years had significantly higher mean levels of DNA damage than the control group (P polyether-exposed working years and those with not less than 20 polyether-exposed working years had significantly higher mean micronucleus rates than the control group (P polyether-exposed working years (P > 0.05); the workers with not less than 5 and less than 20 polyether-exposed working years and workers with not less than 20 polyether-exposed working years had significantly higher mean micronucleus rates than those with less than 5 polyether-exposed working years (P polyether manufacturer.

Exposure to endocrine disruptor induces transgenerational epigenetic deregulation of microRNAs in primordial germ cells.

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Miguel A Brieño-Enríquez

Full Text Available In mammals, germ cell differentiation is initiated in the Primordial Germ Cells (PGCs during fetal development. Prenatal exposure to environmental toxicants such as endocrine disruptors may alter PGC differentiation, development of the male germline and induce transgenerational epigenetic disorders. The anti-androgenic compound vinclozolin represents a paradigmatic example of molecule causing transgenerational effects on germ cells. We performed prenatal exposure to vinclozolin in mice and analyzed the phenotypic and molecular changes in three successive generations. A reduction in the number of embryonic PGCs and increased rate of apoptotic cells along with decrease of fertility rate in adult males were observed in F1 to F3 generations. Blimp1 is a crucial regulator of PGC differentiation. We show that prenatal exposure to vinclozolin deregulates specific microRNAs in PGCs, such as miR-23b and miR-21, inducing disequilibrium in the Lin28/let-7/Blimp1 pathway in three successive generations of males. As determined by global maps of cytosine methylation, we found no evidence for prominent changes in DNA methylation in PGCs or mature sperm. Our data suggest that embryonic exposure to environmental endocrine disruptors induces transgenerational epigenetic deregulation of expression of microRNAs affecting key regulatory pathways of germ cells differentiation.

Proximally exposed A-bomb survivors. 2

International Nuclear Information System (INIS)

Kamada, Nanao

1992-01-01

Methods for observing chromosomes can be chronologically divided into the era of non-differential staining technique (1962-1975) and the era of differential staining method (since 1976). This paper reviews the literature of chromosomal aberrations in bone marrow cells found in the two eras. Findings during the era of 1962-1975 include the frequency of chromosomal aberrations in bone marrow cells, comparison of chromosomal aberrations in bone marrow cells and T lymphocytes, and annual variation of chromosomal aberrations. The frequency of chromosomal aberrations was high in proximally exposed A-bomb survivors (90.5% and 52.6% in A-bomb survivors exposed within 500 m and at 501-1,000 m, respectively); on the contrary, it was low in those exposed far from 1,000 m (6.2% or less). The frequency of chromosomal aberrations in bone marrow cells was lower than that in T lymphocytes (21.5% vs 27.1% in those exposed within 500 m and 14.1% vs 23% in those exposed at 501-1,000 m). Annual analysis for chromosomal aberrations has shown the somewhat dependence upon medullary hematopoiesis and virus infection. The advent of differential staining technique since 1976 has made it possible to clarify the type of chromosomal aberrations and site of breakage. Of 710 bone marrow cells taken from 13 A-bomb survivors exposed within 1,000 m, 121 cells (from 11 A-bomb survivors) exhibited chromosomal aberrations. In differential staining analysis, all 121 cells but one were found to be of stable type, such as translocation and inversion. Furthermore, the site of breakage was found to be non-randomly distributed. Analysis of chromosomal aberrations in bone marrow cells has advantages of reflecting dynamic condition of these cells and determining gradual progression into leukemia. (N.K.)

Self-reported hearing performance in workers exposed to solvents

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Adrian Fuente

2013-02-01

Full Text Available OBJECTIVE: To compare hearing performance relating to the peripheral and central auditory system between solvent-exposed and non-exposed workers. METHODS: Forty-eight workers exposed to a mixture of solvents and 48 non-exposed control subjects of matched age, gender and educational level were selected to participate in the study. The evaluation procedures included: pure-tone audiometry (500 - 8,000 Hz, to investigate the peripheral auditory system; the Random Gap Detection test, to assess the central auditory system; and the Amsterdam Inventory for Auditory Disability and Handicap, to investigate subjects' self-reported hearing performance in daily-life activities. A Student t test and analyses of covariance (ANCOVA were computed to determine possible significant differences between solvent-exposed and non-exposed subjects for the hearing level, Random Gap Detection test and Amsterdam Inventory for Auditory Disability and Handicap. Pearson correlations among the three measures were also calculated. RESULTS: Solvent-exposed subjects exhibited significantly poorer hearing thresholds for the right ear than non-exposed subjects. Also, solvent-exposed subjects exhibited poorer results for the Random Gap Detection test and self-reported poorer listening performance than non-exposed subjects. Results of the Amsterdam Inventory for Auditory Disability and Handicap were significantly correlated with the binaural average of subject pure-tone thresholds and Random Gap Detection test performance. CONCLUSIONS: Solvent exposure is associated with poorer hearing performance in daily life activities that relate to the function of the peripheral and central auditory system.

Plerocercoids of the cestode Diphyllobothrium ditremum in brown trout Salmo trutta: substantial increase in infection after establishment of European minnow Phoxinus phoxinus.

Science.gov (United States)

Borgstrøm, R; Trømborg, J; Haugen, T O; Rosseland, B O

2017-09-01

This study focuses on plerocercoids of the cestode Diphyllobothrium ditremum in brown trout Salmo trutta from the subalpine lake Øvre Heimdalsvatn in south-central Norway. Salmo trutta was the only fish species in this lake until European minnow Phoxinus phoxinus was registered in 1969. The P. phoxinus population increased substantially in the following years. In contrast with the 1969-1972 period, when plerocercoids of D. ditremum were practically absent in S. trutta, there was a high prevalence and intensity of infection in the 2013 S. trutta samples. Because the life cycle of D. ditremum involves two larval stages, in copepods and salmonids and mature worms in piscivorous birds, such as mergansers and loons, a change in feeding ecology of S. trutta or changes in population densities of copepods, fish or birds might have influenced the infection pattern. No relationships between D. ditremum infection and muscle-tissue δ 15 N signature or Hg concentration were found, indicating that infection is not a result of piscivory or cannibalism. Furthermore, consumption of copepods by S. trutta during summer and autumn was low. On the other hand, the number of piscivorous birds has increased, probably due to the presence of P. phoxinus as a new and numerous prey. An increased number of final D. ditremum hosts may have produced a higher output of cestode eggs, resulting in more infected copepods that in turn are consumed by S. trutta. Indirectly, P. phoxinus may therefore have caused the observed increased infection in S. trutta and thereby imposed further negative effects on S. trutta in high mountain areas. © 2017 The Fisheries Society of the British Isles.

Survival of Spores of Trichoderma longibrachiatum in Space: data from the Space Experiment SPORES on EXPOSE-R

Science.gov (United States)

Neuberger, Katja; Lux-Endrich, Astrid; Panitz, Corinna

2015-01-01

In the space experiment `Spores in artificial meteorites' (SPORES), spores of the fungus Trichoderma longibrachiatum were exposed to low-Earth orbit for nearly 2 years on board the EXPOSE-R facility outside of the International Space Station. The environmental conditions tested in space were: space vacuum at 10-7-10-4 Pa or argon atmosphere at 105 Pa as inert gas atmosphere, solar extraterrestrial ultraviolet (UV) radiation at λ > 110 nm or λ > 200 nm with fluences up to 5.8 × 108 J m-2, cosmic radiation of a total dose range from 225 to 320 mGy, and temperature fluctuations from -25 to +50°C, applied isolated or in combination. Comparable control experiments were performed on ground. After retrieval, viability of spores was analysed by two methods: (i) ethidium bromide staining and (ii) test of germination capability. About 30% of the spores in vacuum survived the space travel, if shielded against insolation. However, in most cases no significant decrease was observed for spores exposed in addition to the full spectrum of solar UV irradiation. As the spores were exposed in clusters, the outer layers of spores may have shielded the inner part. The results give some information about the likelihood of lithopanspermia, the natural transfer of micro-organisms between planets. In addition to the parameters of outer space, sojourn time in space seems to be one of the limiting parameters.

NEW DESIGN FOR AIRLIFT PUMP USED IN FISH CULTURE TANKS WITH THE ENDANGERED RIO GRANDE SILVERY MINNOW (Hybognathus amarus

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Alison M. Hutson

2012-10-01

Full Text Available This article describes an airlift pump used to produce a circular flow in a fish culture tank that does not attach to the tank. The design produces an airlift pump that does not swing back and forth or float upwards while in use. It is easy to build, inexpensive, and can be quickly installed and removed so that it does not interfere with sampling or harvest. The airlift pump was evaluated during a 30-d survival trial with the endangered Rio Grande silvery minnow (Hybognathus amarus in 2.44-m-diameter circular tanks (3,666 l. Because the fish is endangered, all new culture units must be evaluated in a survival trial. To be able to use a new 15-tank system, survival had to be evaluated in a random representation of three tanks. U.S. Fish and Wildlife Service, which regulates all activities with this endangered species, decided that permitted take (maximum permitted mortality was 60% for the survival trial; consequently, survival >40% in each tank would be considered successful. Two airlift pumps were placed in each tank. The two airlift pumps moved a mean±SD of 33.697±5.563 l/min; this produced total tank turnovers through the airlift pumps of 110.65±16.93 min. Water velocities were measured at nine locations in the tanks. Water velocities were 0.0-0.04 m/sec. Dissolved oxygen concentration never went below 6.30 mg/l. The airlift pumps operated flawlessly and required no maintenance. They produced water velocities preferred by the fish and helped keep dissolved oxygen concentration above the permitted minimum (5 mg/l. The airlift pumps will be used in future fish culture activities in these and other tanks. Survival in the three tanks was 78%, 94% and 96%; overall survival was 89.3%. Because take (10.7% was under the permitted level (60%, the trial was successful.

Contaminations of inner surface of magnesium fluoride windows in the `Expose-R' experiment on the International Space Station

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Skurat, V. E.

2017-10-01

A series of experiments was carried out previously on board of the International Space Station in `EXPOSE-R', a multi-user expose facility, provided by European Space Agency attached to the external surface of the Russian Segment. In one experiment, spores of microorganisms and species of higher plant seeds, in heat-sealed polymer bags were irradiated by solar radiation passed through MgF2 windows in a high space vacuum. After sample exposure, it was found that in many cases the inner surfaces of windows were contaminated. Analysis of the contamination revealed the presence of chemical groups CH2, CH3, NH, OH, C═O, Si-CH3 (Demets et al. in 2015). Their presence in deposits was explained by photofixation of gaseous precursors - some of the vapours of glues and additives in polymeric materials in the core facility of `Expose-R'. Carbon-, oxygen- and silicon-containing groups may be deposited from outer intrinsic atmosphere. This atmosphere is connected with sample compartments and core facility. However, the presence of NH groups on inner surfaces of windows was not expected. This paper shows that the process responsible for carbon-, nitrogen- and oxygen-containing group formation can be a photopolymerization of caprolactam, which is released from the outer Nylon 6 layer of polymer bags under Solar vacuum ultraviolet radiation.

9 CFR 78.23 - Brucellosis exposed bison.

Science.gov (United States)

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Brucellosis exposed bison. 78.23... AGRICULTURE INTERSTATE TRANSPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS BRUCELLOSIS Restrictions on Interstate Movement of Bison Because of Brucellosis § 78.23 Brucellosis exposed bison...

9 CFR 78.32 - Brucellosis exposed swine.

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2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Brucellosis exposed swine. 78.32... AGRICULTURE INTERSTATE TRANSPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS BRUCELLOSIS Restrictions on Interstate Movement of Swine Because of Brucellosis § 78.32 Brucellosis exposed swine. (a...

Endocrine Disruption and Human Prostate Cancer

National Research Council Canada - National Science Library

Risbridger, Gail

2008-01-01

.... In order to test the concept that Vinclozolin alters human prostate development and induces disease, we used our model system to study human prostate development and maturation over 8-12 weeks...

UDS in lymphocytes of occupationally radiation exposed persons

International Nuclear Information System (INIS)

Tuschl, H.; Kovac, R.

1982-01-01

To determine a possible effect of low dose radiation on DNA repair processes, peripheral lymphocytes of mine workers exposed to 222 Rn in the thermal gallery of Badgastein (Austria) and employees of the Austrian Research Centre Seibersdorf, exposed to varying doses of gamma radiation, were investigated. The capacity for unscheduled DNA synthesis (UDS) induced by in vitro UV irradiation was measured by autoradiography of isolated lymphocytes of exposed persons and unexposed controls. In all 222 Rn-exposed mine workers a significant increase of UDS above control values could be observed. Gamma irradiation 31 mrad had a significant effect on UDS, indicating a stimulation of DNA repair capability by chronic low dose exposure. (Author)

To be a worker (exposed?) or not to be a worker (exposed?) that is the question

International Nuclear Information System (INIS)

Ammerich, M.

2008-01-01

The notion of personnel is detailed in this article in order to know exactly what personnel is considered as exposed and what radiation doses are under this term. The regulatory texts are studied in different articles of the French law and show that different kind of exposed personnel are considered. The definitions are varying with the notion of risk, of radiation doses and the work itself. This article asks for a better and more precise definition that will help the actors of radiation protection. (N.C.)

Fire exposed aluminium structures

NARCIS (Netherlands)

Maljaars, J.; Fellinger, J.E.J.; Soetens, F.

2005-01-01

Material properties and mechanical response models for fire design of steel structures are based on extensive research and experience. Contrarily, the behaviour of aluminium load bearing structures exposed to fire is relatively unexplored. This article gives an overview of physical and mechanical

Pulmonary function and oxidative stress in workers exposed to styrene in plastic factory: occupational hazards in styrene-exposed plastic factory workers.

Science.gov (United States)

Sati, Prakash Chandra; Khaliq, Farah; Vaney, Neelam; Ahmed, Tanzeel; Tripathi, Ashok K; Banerjee, Basu Dev

2011-11-01

Styrene is a volatile organic compound used in factories for synthesis of plastic products. The pneumotoxicity of styrene in experimental animals is known. The aim of the present study was to study the effect of styrene on lung function and oxidative stress in occupationally exposed workers in plastic factory. Thirty-four male workers, between 18 and 40 years of age, exposed to styrene for atleast 8 hours a day for more than a year were studied, while 30 age- and sex-matched healthy subjects not exposed to styrene served as controls. Assessment of lung functions showed a statistically significant reduction (p volumes, capacities (FVC, FEV(1), VC, ERV, IRV, and IC) and flow rates (PEFR, MEF(75%), and MVV) in the study group (workers) as compared to controls. Malondialdehyde (MDA) was observed to be significantly high (p < 0.05) while ferric-reducing ability of plasma (FRAP) was significantly low (p < 0.05) in styrene-exposed subjects. Reduced glutathione (GSH) level was significantly depleted in exposed subjects as compared to control group. The mean value of serum cytochrome c in styrene-exposed subjects was found to be 1.1 ng/ml (0.89-1.89) while in control its levels were under detection limit (0.05 ng/ml). It shows that styrene inhalation by workers leads to increased level of oxidative stress, which is supposed to be the cause of lung damage.

Response of exposed bark and exposed lichen to an urban area

Energy Technology Data Exchange (ETDEWEB)

Cruz, A.M.J. [Polytechnic Institute of Coimbra, Oliveira do Hospital (Portugal). Oliveira do Hospital College of Technology and Management; Freitas, M.C.; Canha, N. [URSN, Sacavem (Portugal). Inst. Tecnologico e Nuclear (ITN); Verburg, T.G.; Wolterbeek, H.T. [Technical Univ. of Delft (Netherlands). Dept. of Radiation, Radionuclides and Reactors

2011-07-01

The aim of this study is to understand emission sources of chemical elements using biomonitoring as a tool. The selected lichen and bark were respectively Parmotrema bangii and Criptomeria japonica, sampled in the pollution-free atmosphere of Azores (Sao Miguel island), Portugal, and were exposed in the courtyards of 22 basic schools of Lisbon. The exposure was from January to May 2008 and from June to October 2008 (designated through the text as winter and summer respectively). The chemical element concentrations were determined by INAA. Conductivity of the lichen samples was measured. Factor analysis (MCTTFA) was applied to winter/summer bark/lichen exposed datasets. Arsenic emission sources, soil with anthropogenic contamination, a Se source, traffic, industry, and a sea contribution, were identified. In lichens, a physiological source based on the conductivity values was found. The spatial study showed contribution of sources to specific school positioning. Conductivity values were high in summer in locations as international Lisbon airport and downtown. Lisbon is spatially influenced by marine air mass transportation. It is concluded that one air sampler in Lisbon might be enough to define the emission sources under which they are influenced. (orig.)

Serum-thyroxine levels in microwave-exposed rats

International Nuclear Information System (INIS)

Lu, S.T.; Lebda, N.; Michaelson, S.M.; Pettit, S.

1985-01-01

The nature of the response of the thyroid gland in animals exposed to microwave irradiation is controversial. Animal experimentation has contributed to the controversy because both increased and decreased thyroid functions have been reported. The thyroxine concentration in rats as representative of thyroid function in animals exposed to 2.45-GHz, 120-Hz amplitude-modulated microwaves has been studied. These studies covered a long time span; rats from two commercial sources (BS and CR) were used and subjected to different numbers of exposures, and therefore these data were evaluated for their stability. Two factors could influence in the result significantly, i.e., source of animal and number of sham exposures. Rats used in the 2-hr exposures were from two different commercial sources; rats from CR had a higher (but normal) thyroxine concentration than did rats from BS. Therefore the data of these animals were separated by commercial source for reevaluation. Instead of increased thyroxine concentration in rats exposed at 25, 30, and 40 mW/cm 2 , changes were not noted in any microwave-exposed rats. The influence of sham exposure revealed that appropriate concurrent control and specification of animal source are needed in longitudinal studies. Furthermore, statistical procedures used can greatly influence the conclusions. Thus the specificity of changes in thyroxine concentration in rats exposed to microwaves because of its sporadic occurrence and because of inconsistencies among experiments was doubted

Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II

International Nuclear Information System (INIS)

Sample, Robert; Bryan, Locke; Long, Scott; Majji, Sai; Hoskins, Glenn; Sinning, Allan; Olivier, Jake; Chinchar, V. Gregory

2007-01-01

Frog virus 3 (FV3) is a large DNA virus that encodes ∼ 100 proteins. Although the general features of FV3 replication are known, the specific roles that most viral proteins play in the virus life cycle have not yet been elucidated. To address the question of viral gene function, antisense morpholino oligonucleotides (asMOs) were used to transiently knock-down expression of specific viral genes and thus infer their role in virus replication. We designed asMOs directed against the major capsid protein (MCP), an 18 kDa immediate-early protein (18K) that was thought to be a viral regulatory protein, and the viral homologue of the largest subunit of RNA polymerase II (vPol-IIα). All three asMOs successfully inhibited translation of the targeted protein, and two of the three asMOs resulted in marked phenotypic changes. Knock-down of the MCP resulted in a marked reduction in viral titer without a corresponding drop in the synthesis of other late viral proteins. Transmission electron microscopy (TEM) showed that in cells treated with the anti-MCP MO assembly sites were devoid of viral particles and contained numerous aberrant structures. In contrast, inhibition of 18K synthesis did not block virion formation, suggesting that the 18K protein was not essential for replication of FV3 in fathead minnow (FHM) cells. Finally, consistent with the view that late viral gene expression is catalyzed by a virus-encoded or virus-modified Pol-II-like protein, knock-down of vPol-IIα triggered a global decline in late gene expression and virus yields without affecting the synthesis of early viral genes. Collectively, these results demonstrate the utility of using asMOs to elucidate the function of FV3 proteins

Long-term trends of native and non-native fish faunas in the American Southwest

Directory of Open Access Journals (Sweden)

Olden, J. D.

2005-06-01

Full Text Available Environmental degradation and the proliferation of non-native fish species threaten the endemic, and highly unique fish faunas of the American Southwest. The present study examines long-term trends (> 160 years of fish species distributions in the Lower Colorado River Basin and identifies those native species (n = 28 exhibiting the greatest rates of decline and those non-native species (n = 48 exhibiting the highest rates of spread. Among the fastest expanding invaders in the basin are red shiner (Cyprinella lutrensis, fathead minnow (Pimephales promelas, green sunfish (Lepomis cyanellus, largemouth bass (Micropterus salmoides, western mosquitofish (Gambussia affinis and channel catfish (Ictalurus punctatus; species considered to be the most invasive in terms of their negative impacts on native fish communities. Interestingly, non-native species that have been recently introduced (1950+ have generally spread at substantially lower rates as compared to species introduced prior to this time (especially from 1920 to 1950, likely reflecting reductions in human-aided spread of species. We found general agreement between patterns of species decline and extant distribution sizes and official listing status under the U.S. Endangered Species Act. ‘Endangered’ species have generally experienced greater declines and have smaller present-day distributions compared to ‘threatened’ species, which in turn have shown greater declines and smaller distributions than those species not currently listed. A number of notable exceptions did exist, however, and these may provide critical information to help guide the future listing of species (i.e., identification of candidates and the upgrading or downgrading of current listed species that are endemic to the Lower Colorado River Basin. The strong correlation between probability estimates of local extirpation and patterns of native species decline and present-day distributions suggest a possible proactive

Reducing aquatic hazards of industrial chemicals: probabilistic assessment of sustainable molecular design guidelines.

Science.gov (United States)

Connors, Kristin A; Voutchkova-Kostal, Adelina M; Kostal, Jakub; Anastas, Paul; Zimmerman, Julie B; Brooks, Bryan W

2014-08-01

Basic toxicological information is lacking for the majority of industrial chemicals. In addition to increasing empirical toxicity data through additional testing, prospective computational approaches to drug development aim to serve as a rational basis for the design of chemicals with reduced toxicity. Recent work has resulted in the derivation of a "rule of 2," wherein chemicals with an octanol-water partition coefficient (log P) less than 2 and a difference between the lowest unoccupied molecular orbital and the highest occupied molecular orbital (ΔE) greater than 9 (log P9 eV) are predicted to be 4 to 5 times less likely to elicit acute or chronic toxicity to model aquatic organisms. The present study examines potential reduction of aquatic toxicity hazards from industrial chemicals if these 2 molecular design guidelines were employed. Probabilistic hazard assessment approaches were used to model the likelihood of encountering industrial chemicals exceeding toxicological categories of concern both with and without the rule of 2. Modeling predicted that utilization of these molecular design guidelines for log P and ΔE would appreciably decrease the number of chemicals that would be designated to be of "high" and "very high" concern for acute and chronic toxicity to standard model aquatic organisms and end points as defined by the US Environmental Protection Agency. For example, 14.5% of chemicals were categorized as having high and very high acute toxicity to the fathead minnow model, whereas only 3.3% of chemicals conforming to the design guidelines were predicted to be in these categories. Considerations of specific chemical classes (e.g., aldehydes), chemical attributes (e.g., ionization), and adverse outcome pathways in representative species (e.g., receptor-mediated responses) could be used to derive future property guidelines for broader classes of contaminants. © 2014 SETAC.

Effects of depletion sampling by standard three-pass pulsed DC electrofishing on blood chemistry parameters of fishes from Appalachian streams

Science.gov (United States)

Densmore, Christine L.; Panek, Frank M.

2013-01-01

Adverse effects on fishes captured by electrofishing techniques have long been recognized, although the extent of associated physical injury and behavioral alterations are highly variable and dependent on a number of factors. We examined the effects of three-pass pulsed DC (PDC) electrofishing on two salmonid species (Rainbow Trout Oncorhynchus mykiss and Brook Trout Salvelinus fontinalis) and five other genera (Green Sunfish Lepomis cyanellus, Potomac Sculpin Cottus girardi, Fathead Minnow Pimephales promelas, Largemouth Bass Micropterus salmoides, and Channel Catfish Ictalurus punctatus) common to Appalachian streams. We examined the corresponding effects of PDC electroshock on the following physiological indicators of stress and trauma: blood glucose and serum lactate, as well as on other blood chemistry, namely, enzymes, electrolytes, minerals, and proteins. All species demonstrated physiological responses to PDC electroshock, indicated by the biochemical differences in blood parameters in unshocked and shocked groups of fish with or without gross evidence of hemorrhagic trauma. Serum lactate was the most consistent indicator of these effects. Significant differences in whole blood glucose levels were also noted in treatment groups in all species except Green Sunfish, although the patterns observed were not as consistent as for serum lactate. Elevations in the serum enzymes, aspartate aminotransferase and creatine kinase, in the electroshocked fish occurred only in the two salmonid species. In many instances, although blood parameters were elevated in electroshocked fish compared with the unshocked controls for a given species, there were no differences in those levels in electroshocked fish based on the presence of gross hemorrhagic trauma to axial musculature. While some of the blood parameters examined correlated with both the occurrence of electroshock and the resultant tissue injury, there was no apparent link between the altered blood chemistry and

Evaluation of water quality threats to the endangered Okaloosa darter (Etheostoma okaloosae) in East Turkey Creek on Eglin Air Force Base.

Science.gov (United States)

Weil, R E; Spade, D J; Knoebl, I; Hemming, J M; Tongue, M L; Szabo, N J; Kroll, K J; Tate, W B; Denslow, N D

2012-04-01

The threatened Okaloosa darter (Etheostoma okaloosae) is found almost exclusively on the Eglin Air Force Base in the Choctawhatchee Bay watershed of Florida. Portions of this limited habitat are threatened with soil erosion, altered hydrology, and impaired water quality. In the present study, general water quality parameters (i.e., dissolved oxygen, specific conductance, pH, temperature, relative turbidity, and primary productivity) were characterized in East Turkey Creek, which is a body of water potentially impacted by treated wastewater sprayfields, and Long Creek, an adjacent reference stream that does not border the sprayfields. Water quality was assessed during a 30-day exposure using passive samplers for both non-polar and polar effluent parameters. Because the Okaloosa darter was listed as endangered at the time of sampling we chose a closely related species from the same creeks, the sailfin shiner (Pteronotropis hypseleotris) in which to measure metal body burdens. Additionally, fathead minnows (Pimephales promelas) were used for microarray analysis on gonad and liver tissues after 48 h exposures to water collected from the two creeks and brought into the laboratory. Waters from all sites, including reference sites, affected the expression of genes related to various biological processes including transcription and translation, cell cycle control, metabolism, and signaling pathways, suggesting that the sum of anthropogenic compounds in the site waters may cause a generalized stress response in both liver and testis, an effect that could be related to the generally low populations of the Okaloosa darter. Furthermore, effects of site waters on fish gene expression may be related to the impact of human activities other than the wastewater sprayfields, as nearby areas are closed to the public for military testing, training, and administrative activities and due to ordnance contamination. Copyright © 2012 Elsevier B.V. All rights reserved.

Survey on Urinary Levels of Aflatoxins in Professionally Exposed Workers

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Fulvio Ferri

2017-03-01

Full Text Available Feed mill workers may handle or process maize contaminated with aflatoxins (AFs. This condition may lead to an unacceptable intake of toxins deriving from occupational exposure. This study assessed the serological and urinary levels of AFs in workers exposed to potentially contaminated dusts in two mills. From March to April 2014, blood and urine samples were collected, on Monday and Friday morning of the same working week from 29 exposed workers and 30 non-exposed controls. AFs (M1, G2, G1, B1, B2 and aflatoxicol (AFOH A were analyzed. Each subject filled in a questionnaire to evaluate potential food-borne exposures to mycotoxins. AFs contamination in environmental dust was measured in both plants. No serum sample was found to be positive. Seventy four percent of urine samples (73.7% revealed AFM1 presence. AFM1 mean concentration was 0.035 and 0.027 ng/mL in exposed and non-exposed workers, respectively (p = 0.432; the concentration was slightly higher in Friday’s than in Monday’s samples, in exposed workers, 0.040 versus (vs. 0.031 and non-exposed controls (0.030 vs. 0.024, p = 0.437. Environmental AFs contamination ranged from 7.2 to 125.4 µg/kg. The findings of this study reveal the presence of higher AFs concentration in exposed workers than in non-exposed controls, although these differences are to be considered consistent with random fluctuations.

Going beyond the most exposed people in a dose assessment

Energy Technology Data Exchange (ETDEWEB)

Hjerpe, Thomas; Broed, Robert [Facilia AB, Gustavslundsvaegen 151C, SE-167 51 Bromma (Sweden); Ikonen, Ari T.K. [Environmental Research and Assessment, EnviroCase, Ltd., Hallituskatu 1 D 4, FI-28 100 Pori (Finland)

2014-07-01

The dose assessment in a long-term radiation safety assessment often focus on assessing dose of a representative person to be used for determining compliance with a radiation dose constraint. This representative person is often assumed to receive a dose that is representative of the most exposed people, i.e., the more highly exposed individuals in the population. This is not always sufficient, the Finnish regulations for disposal of nuclear waste has radiation dose constraint to the most exposed people as well as for larger groups of exposed people. This work presents the methodology to assessing dose of a representative person for a larger group of exposed people as applied by Posiva in the TURVA-2012 safety case for the spent nuclear fuel disposal at Olkiluoto. In addition, annual doses from the set of biosphere calculation cases analysed in TURVA-2012 are presented and discussed. Special focus is given on explaining the differences in exposure levels and exposure routes between the estimated annual doses to representative persons for most exposed people and a larger exposed group. The results show that the annual doses to a larger group of people ranges from one to three orders of magnitude below the annual doses to the most exposed people. Furthermore, the exposure route related to food ingestion is less significant for the larger group of people compared to the most exposed people and that the exposure route related to water ingestion shows the opposite behaviour. (authors)

Reprocessing of nonoptimally exposed holograms

International Nuclear Information System (INIS)

Phipps, G.S.; Robertson, C.E.; Tamashiro, F.M.

1980-01-01

Two reprocessing techniques have been investigated that are capable of correcting the effects of nonoptimum optical density of photographic amplitude holograms recorded on Agfa-Gevaert type 10E75 plates. In some cases a reprocessed hologram will exhibit a diffraction efficiency even higher than that obtainable from a hologram exposed and processed to the optimum density. The SNR of the reprocessed holograms is much higher than that of the same holograms belached with cupric bromide. In some cases the SNR approaches the optimum value for a properly exposed amplitude hologram. Subjective image quality and resolution of reprocessed hologram reconstructins appear to be no different than for normal single-development holograms. Repeated reprocessing is feasible and in some cases desirable as a means of increasing diffraction efficiency