Diana M. Papoulias

Atrazine reduces reproduction in fathead minnow (Pimephales promelas)

Atrazine, the widely used herbicide, has shown to affect the hypothalamus-pituitary-gonad axis in certain vertebrate species, but few studies have examined reproductive effects of this chemical on fish. Our study was designed to evaluate a population endpoint (egg production) in conjunction with histological (e.g., gonad development) and biochemical (e.g., hormone production) phenotypes associated with atrazine exposure in fathead minnows. Adult virgin breeding groups of 1 male and 2 females were exposed to nominal concentrations of 0, 0.5, 5.0, and 50 microg/L of atrazine in a flow-through diluter for 14 or 30 days. Total egg production was lower (19-39%) in all atrazine-exposed groups as compared to the controls. The decreases in cumulative egg production of atrazine treated fish were significant by 17-20 days of exposure. Reductions in egg production in atrazine treatment groups were most attributable to reduced numbers of spawning events with increased atrazine exposure concentrations. Gonad abnormalities were observed in both male and female fish of atrazine-exposed fish. Our results also indicate that atrazine reduces egg production through alteration of final maturation of oocytes. The reproductive effects observed in this study warrant further investigation and evaluation of the potential risks posed by atrazine, particularly feral populations of fish from streams in agricultural areas with high use of this herbicide.

An in vivo model fish system to test chemical effects on sexual differentiation and development : exposure to ethinyl estradiol

Abstract A model system was characterized which may be used as an in vivo screen for effects of chemicals or environmental mixtures on sexual differentiation and development of reproductive organs and gametes. We evaluated the effects of a model environmental estrogen, ethinyl estradiol (EE 2 ), on the d-rR strain of medaka, Oryzias latipes , using a nano-injection exposure. Gonad histopathology indicated that a single injection of 0.5–2.5 ng EE 2 /egg can cause phenotypic sex-reversal of genetic males to females. Sex-reversals could be detected as early as 7 days post-hatch. Sex-reversed males had female-typical duct development and the secondary sex characteristics we measured were generally consistent with phenotype, with the exception of a few EE 2 -exposed XX and XY females which possessed ambiguous anal fins. Using discriminant analysis, we determined that the presence or absence of the secondary sex characteristic, a dorsal fin notch, was a very reliable indicator of gonadal sex. No instances of gonadal intersexes were observed. Ethinyl estradiol also appeared to reduce growth but not condition (weight-at-length) and exposed XX females appeared to have a higher incidence of atretic follicles relative to controls. Our results suggest that estrogenic chemicals may influence sexual differentiation and development and that the medaka model is well suited to assessing these effects.

Age Estimation for Shovelnose Sturgeon: A Cautionary Note Based on Annulus Formation in Pectoral Fin Rays

Abstract Numerous studies have examined the age and growth of shovelnose sturgeon Scaphirhynchus platorynchus, but only one study attempted to validate age estimation techniques. Therefore, our objective was to use marginal increment analysis to validate annulus formation in pectoral fin rays of shovelnose sturgeon collected from the Missouri River. We also compared the precision of age estimates between two different readers. Marginal increment distance indicated that for most of the populations an opaque band was laid down in pectoral fin rays during the summer. However, opaque bands were formed throughout the year in some individuals, which could be problematic when using fin rays for age estimation. The agreement of age estimates by two readers for shovelnose sturgeon was only 18%, and differences in ages between the two readers increased for older fish. The presence of split annuli, false annuli, spawning bands, imbedded rays, and deteriorating sections made individual growth rings difficult to separ...

A Conceptual Life-History Model for Pallid and Shovelnose Sturgeon

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Atrazine reduces reproduction in Japanese medaka (Oryzias latipes)

Atrazine is an effective broadleaf herbicide and the second most heavily used herbicide in the United States. Effects along the hypothalamus-pituitary-gonad axis in a number of vertebrate taxa have been demonstrated. Seasonally elevated concentrations of atrazine in surface waters may adversely affect fishes, but only a few studies have examined reproductive effects of this chemical. The present study was designed to evaluate a population endpoint (egg production) in conjunction with histological (reproductive stage, gonad pathology) and biochemical (aromatase activity, sex hormone production) phenotypes associated with atrazine exposure in Japanese medaka. Adult virgin breeding groups of one male and four females were exposed to nominal concentrations of 0, 0.5, 5.0, and 50 μg/L (0, 2.3, 23.2, 231 nM) of atrazine in a flow-through diluter for 14 or 38 days. Total egg production was lower (36-42%) in all atrazine-exposed groups as compared to the controls. The decreases in cumulative egg production of atrazine-treated fish were significant by exposure day 24. Reductions in total egg production in atrazine treatment groups were most attributable to a reduced number of eggs ovulated by females in atrazine-treated tanks. Additionally, males exposed to atrazine had a greater number of abnormal germ cells. There was no effect of atrazine on gonadosomatic index, aromatase protein, or whole body 17 β-estradiol or testosterone. Our results suggest that atrazine reduces egg production through alteration of final maturation of oocytes. The reduced egg production observed in this study was very similar to our previously reported results for fathead minnow. This study provides further information with which to evaluate atrazines risk to fish populations.

Environmental contaminants in prey and tissues of the peregrine falcon in the Big Bend Region, Texas, USA

Peregrine falcons (Falco peregrinus) have been recorded nesting in Big Bend National Park, Texas, USA and other areas of the Chihuahuan Desert since the early 1900s. From 1993 to 1996, peregrine falcon productivity rates were very low and coincided with periods of low rainfall. However, low productivity also was suspected to be caused by environmental contaminants. To evaluate potential impacts of contaminants on peregrine falcon populations, likely avian and bat prey species were collected during 1994 and 1997 breeding seasons in selected regions of western Texas, primarily in Big Bend National Park. Tissues of three peregrine falcons found injured or dead and feathers of one live fledgling also were analyzed. Overall, mean concentrations of DDE [1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene], a metabolite of DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane], were low in all prey species except for northern rough-winged swallows (Stelgidopteryx serripennis, mean = 5.1 microg/g ww). Concentrations of mercury and selenium were elevated in some species, up to 2.5 microg/g dw, and 15 microg/g dw, respectively, which upon consumption could seriously affect reproduction of top predators. DDE levels near 5 microg/g ww were detected in carcass of one peregrine falcon found dead but the cause of death was unknown. Mercury, selenium, and DDE to some extent, may be contributing to low reproductive rates of peregrine falcons in the Big Bend region.

A comparative assessment of contaminants in fish from four resacas of the Texas, USA–Tamaulipas, Mexico border region

A recent survey of contaminant information for the Lower Rio Grande Valley (LRGV), Texas, has shown that little is known about contaminants and their impacts on biota of resacas (oxbows) along the US-Mexico border. In 1996, fish were collected from four resacas in the Texas-Tamaulipas border region to assess contaminant loadings and their impacts on fish and birds. Tissue residue concentrations in fish were analyzed and also compared to two histopathological bioindicators of unhealthy environmental conditions. Of the organochlorine insecticides measured, DDE was the most common and was present at relatively high concentrations (10 microg/g w/w) at some sites. DDE concentrations were nearly 20 times greater in fish from resacas in Texas than from resacas in Tamaulipas, although the limited sample sizes obtained precluded statistical comparisons. DDE concentrations in fish from the two Texas resacas were also greater than those reported in fish from nearby areas during the 1980s and 1990s. Most trace element concentrations were similar among resacas from Texas and Tamaulipas. Arsenic, however, was two to six times greater in fish from a downtown resaca in Matamoros than in fish from other resacas in Tamaulipas and Texas. The bioindicators, pigment accumulation, and macrophage aggregates (MAs), in general, reflected the contamination indicated by the tissue residues for each site. Overall, it appears that some resacas of the US-Mexico border region are contaminant sinks and could pose potential health or reproductive problems for fish and wildlife, and humans that consume fish from those sites.

Spawning Habitat Associations and Selection by Fishes in a Flow-Regulated Prairie River

Abstract We used histological features to identify the spawning chronologies of river-dwelling populations of slenderhead darter Percina phoxocephala, suckermouth minnow Phenacobius mirabilis, stonecat Noturus flavus, and red shiner Cyprinella lutrensis and to relate their reproductive status to microhabitat associations. We identified spawning and nonspawning differences in habitat associations resulting from 1 year of field data via logistic regression modeling and identified shifts in microhabitat selection via frequency-of-use and availability histograms. Each species demonstrated different habitat associations between spawning and nonspawning periods. The peak spawning period for slenderhead darters was April to May in high-velocity microhabitats containing cobble. Individuals were associated with similar microhabitats during the postspawn summer and began migrating to deeper habitats in the fall. Most suckermouth minnow spawned from late March through early May in shallow microhabitats. The probabil...

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

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. Environ Toxicol Chem 2016;35:2230-2238. 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.

Optimization of the Ames/Salmonella Mutagenicity Assay for Use with Extracts of Aquatic Sediments

Abstract Non-mutagenic components interfered with the ability of the standard Ames/salmonella assay to detect mutagenicity in extracts of contaminated Great Lakes sediments. The use of gel permeation chromatography (GPC) to remove these macromolecules from methylene chloride extracts prior to Ames testing enhanced the likelihood of transfer of mutagenic components into dimethyl sulf oxide (the assay solvent). Therefore, to optimize the assays sensitivity we pre-treated sediment extracts using GPC and increased metabolic activity through the use of a 30% S9 mix. Increasing the level of Aroclor 1254-induced rat liver S9, typically used to metabolically activate promutagens, had the additional beneficial effect of reducing the cytotoxicity of the extracts. As applied in this study, the Ames assay can serve as a sensitive test for screening the mutagenic potential of large numbers of uncharacterized sediment extracts.