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USGS2016

Aqueous exposure to the progestin, levonorgestrel, alters anal fin development and reproductive behavior in the eastern mosquitofish (Gambusia holbrooki)

Endogenous progestogens are important regulators of vertebrate reproduction. Synthetic progestins are components of human contraceptive and hormone replacement pharmaceuticals. Both progestogens and progestins enter the environment through a number of sources, and have been shown to cause profound effects on reproductive health in various aquatic vertebrates. Progestins are designed to bind human progesterone receptors, but they also have been shown to strongly activate androgen receptors in fish. Levonorgestrel (LNG) activates fish androgen receptors and induces development of male secondary sex characteristics in females of other species. Although behavior has been postulated to be a sensitive early indicator of exposure to certain environmental contaminants, no such research on the reproductive behavior of gestagen-exposed fish has been conducted to date. The goal of our study was to examine the exposure effects of a human contraceptive progestin, LNG, on the reproductive development and behavior of the viviparous eastern mosquitofish ( Gambusia holbrooki ). Internal fertilization is a requisite characteristic of viviparous species, and is enabled by an androgen driven elongation of the anal fin into the male gonopodium (i.e., phallus). In this study, we exposed adult mosquitofish to ethanol (EtOH control), 10 ng/L, and 100 ng/L LNG for 8 d using a static replacement exposure design. After 8 d, a subset of males and females from each treatment were examined for differences in the 4:6 anal fin ratio. In addition, paired social interaction trials were performed using individual control males and control females or females treated 10 ng/L or 100 ng/L LNG. Female mosquitofish exposed to LNG were masculinized as evidenced by the elongation of the anal fin rays, a feature normal to males and abnormal to females. LNG caused significant increases in the 4:6 anal fin ratios of female mosquitofish in both the 10 ng/L and 100 ng/L treatments, although these differences were not significant between the two treatments. LNG caused significant increases in the 4:6 anal fin ratio of males exposed to 100 ng/L, with no effects observed in the 10 ng/L treatment. In addition, the reproductive behavior of control males paired with female mosquitofish exposed to 100 ng/L LNG was also altered, for these males spent more time exhibiting no reproductive behavior, had decreased attending behavior, and a lower number of gonopodial thrusts compared to control males paired to control female mosquitofish. Given the rapid effects on both anal fin morphology and behavior observed in this study, the mosquitofish is an excellent sentinel species for the detection of exposure to LNG and likely other 19-nortestosterone derived contraceptive progestins in the environment.

General and Comparative Endocrinology↗

USGS2001

Neuroendocrine and behavioral implications of endocrine disrupting chemicals in quail

Studies in our laboratory have focused on endocrine, neuroendocrine, and behavioral components of reproduction in the Japanese quail. These studies considered various stages in the life cycle, including embryonic development, sexual maturation, adult reproductive function, and aging. A major focus of our research has been the role of neuroendocrine systems that appear to synchronize both endocrine and behavioral responses. These studies provide the basis for our more recent research on the impact of endocrine disrupting chemicals (EDCs) on reproductive function in the Japanese quail. These endocrine active chemicals include pesticides, herbicides, industrial products, and plant phytoestrogens. Many of these chemicals appear to mimic vertebrate steroids, often by interacting with steroid receptors. However, most EDCs have relatively weak biological activity compared to native steroid hormones. Therefore, it becomes important to understand the mode and mechanism of action of classes of these chemicals and sensitive stages in the life history of various species. Precocial birds, such as the Japanese quail, are likely to be sensitive to EDC effects during embryonic development, because sexual differentiation occurs during this period. Accordingly, adult quail may be less impacted by EDC exposure. Because there are a great many data available on normal development and reproductive function in this species, the Japanese quail provides an excellent model for examining the effects of EDCs. Thus, we have begun studies using a Japanese quail model system to study the effects of EDCs on reproductive endocrine and behavioral responses. In this review, we have two goals: first, to provide a summary of reproductive development and sexual differentiation in intact Japanese quail embryos, including ontogenetic patterns in steroid hormones in the embryonic and maturing quail. Second, we discuss some recent data from experiments in our laboratory in which EDCs have been tested in Japanese quail. The Japanese quail provides an excellent avian model for testing EDCs because this species has well-characterized reproductive endocrine and behavioral responses. Considerable research has been conducted in quail in which the effects of embryonic steroid exposure have been studied relative to reproductive behavior. Moreover, developmental processes have been studied extensively and include investigations of the reproductive axis, thyroid system, and stress and immune responses. We have conducted a number of studies, which have considered long-term neuroendocrine consequences as well as behavioral responses to steroids. Some of these studies have specifically tested the effects of embryonic steroid exposure on later reproductive function in a multigenerational context. A multigenerational exposure provides a basis for understanding potential exposure scenarios in the field. In addition, potential routes of exposure to EDCs for avian species are being considered, as well as differential effects due to stage of the life cycle at exposure to an EDC. The studies in our laboratory have used both diet and egg injection as modes of exposure for Japanese quail. In this way, birds were exposed to a specific dose of an EDC at a selected stage in development by injection. Alternatively, dietary exposure appears to be a primary route of exposure; therefore experimental exposure through the diet mimics potential field situations. Thus, experiments should consider a number of aspects of exposure when attempting to replicate field exposures to EDCs.

Hormones and Behavior↗

USGS2009

Ecological Requirements for Pallid Sturgeon Reproduction and Recruitment in the Lower Missouri River: A Research Synthesis 2005-08

This report provides a synthesis of results obtained between 2005 and 2008 from the Comprehensive Sturgeon Research Program, an interagency collaboration between the U.S. Geological Survey, Nebraska Game and Parks Commission, U.S. Fish and Wildlife Service, and the U.S. Army Corps of Engineers' Missouri River Recovery - Integrated Science Program. The goal of the Comprehensive Sturgeon Research Program is to improve fundamental understanding of reproductive ecology of the pallid sturgeon with the intent that improved understanding will inform river and species management decisions. Specific objectives include: *Determining movement, habitat-use, and reproductive behavior of pallid sturgeon; *Understanding reproductive physiology of pallid sturgeon and relations to environmental conditions; *Determining origin, transport, and fate of drifting pallid sturgeon larvae, and evaluating bottlenecks for recruitment of early life stages; *Quantifying availability and dynamics of aquatic habitats needed by pallid sturgeon for all life stages; and *Managing databases, integrating understanding, and publishing relevant information into the public domain. Management actions to increase reproductive success and survival of pallid sturgeon in the Lower Missouri River have been focused on flow regime, channel morphology, and propagation. Integration of 2005-08 Comprehensive Sturgeon Research Program research provides insight into linkages among flow regime, re-engineered channel morphology, and pallid sturgeon reproduction and survival. The research approach of the Comprehensive Sturgeon Research Program integrates opportunistic field studies, field-based experiments, and controlled laboratory studies. The field study plan is designed to explore the role of flow regime and associated environmental cues using two complementary approaches. An upstream-downstream approach compares sturgeon reproductive behavior between an upstream section of the Lower Missouri River with highly altered flow regime to a downstream section that maintains much of its pre-regulation flow variability. The upstream section also has the potential for an experimental approach to compare reproductive behavior in years with pulsed flow modifications ('spring rises') to years without. The reproductive cycle of the female sturgeon requires several years to progress through gonadal development, oocyte maturation, and spawning. Converging lines of evidence support the hypothesis that maturation and readiness to spawn in female sturgeon is cued many months before spawning. Information on reproductive readiness of shovelnose sturgeon indicates that sturgeon at different locations along the Lower Missouri River between St. Louis and Gavins Point Dam are all responding to the same early cue. Although not a perfect surrogate, the more abundant shovelnose sturgeon is morphologically, physiologically, and genetically similar to pallid sturgeon, and thereby provides a useful comparative model for the rarer species. Day length is the likely candidate to define a temporal spawning window. Within the spawning window, one or more additional, short-term, and specific cues may serve to signal ovulation and release of gametes. Of three potential spawning cues - water temperature, water discharge, and day of year - water temperature is the most likely proximate cue because of the fundamental physiological role temperature plays in sturgeon embryo development and survival, and the sensitivity of many fish hormones to temperature change. It also is possible that neither temperature nor discharge is cueing spawning; instead, reproductive behavior may result from the biological clock advancing an individual fish's readiness to spawn day after day through the spawning period until the right moment, independent of local environmental conditions. Separation of the individual effects of discharge events, water temperature, and other possible factors, such as proximity to male

Scientific Investigations Report↗

USGS2006

Reticulate melanism in western painted turtles (Chrysemys picta bellii): Exploring linkages with habitat and heating rates

In western painted turtles (Chrysemys picta bellii), males often exhibit one of two morphs: (1) a reticulated form, characterized by an intricate network of dark markings on the carapace or (2) a non-reticulated form. Although several hypotheses have been proposed to explain the adaptive significance of reticulate melanism (RM) on western painted turtles, no attempts have been made to document whether RM is linked to habitat conditions or if the presence of melanism affects heating rates. To evaluate these questions, we compared the frequency of adult male turtles with RM across three different habitats: riverine (rivers), lacustrine (lakes) and palustrine (wetland) habitats. Using manipulative experiments, we also tested the hypothesis that body heating rates are higher in turtles with RM. Reticulate melanism occurred on 99 (31%) of 320 male turtles captured in South Dakota from 2002 to 2003. Turtles with reticulate melanism were significantly larger than non-reticulated turtles; RM was not observed on male turtles with carapace lengths <15 cm or weights <360 g. The mean proportion of adult male turtles (>15 cm carapace length) with RM was similar among river (0.54), lake (0.50) and wetland (0.64) habitats, implying that RM is not a habitat-linked trait. Heating rates for turtles with RM were similar to those measured for non-reticulated individuals. Body size, however, influenced heating rates; larger-bodied turtles with lower surface area-to-volume ratio heated more slowly than smaller turtles. Whether RM is a by-product of hormonal regulation or serves an adaptive purpose remains unclear. However, other hypotheses, especially those involving communication (e.g., courtship behavior) and/or gamete protection remain untested for western painted turtles and warrant further investigation.

American Midland Naturalist↗

USGS2007

Use of behavioral and physiological indicators to evaluate Scaphirhynchus sturgeon spawning success

Thirty gravid, female shovelnose sturgeon ( Scaphirhynchus platorynchus ) were captured in the Lower Missouri River in March 2004 to evaluate the effectiveness of physiology, telemetry and remote sensor technology coupled with change point analysis in identifying when and where Scaphirhynchus sturgeon spawn. Captured sturgeons were instrumented with ultrasonic transmitters and with archival data storage tags (DST) that recorded temperature and pressure. Female sturgeon were tracked through the suspected spawning period. Thereafter, attempts were made to recapture fish to evaluate spawning success. At the time of transmitter implantation, blood and an ovarian biopsy were taken. Reproductive hormones and cortisol were measured in blood. Polarization indices and germinal vesicle breakdown were assessed on the biopsied oocytes to determine readiness to spawn. Behavioral data collected using telemetry and DST sensors were used to determine the direction and magnitude of possible spawning-related movements and to identify the timing of potential spawning events. Upon recapture observations of the ovaries and blood chemistry provided measures of spawning success and comparative indicators to explain differences in observed behavior. Behavioral and physiological indicators of spawning interpreted along with environmental measures may assist in the determination of variables that may cue sturgeon reproduction and the conditions under which sturgeon successfully spawn.

Journal of Applied Ichthyology↗

USGS2019

In vivo effects of 17α-ethinylestradiol, 17B-estradiol and 4-nonylphenol on insulin-like growth-factor binding proteins (igfbps) in Atlantic salmon

Feminizing endocrine disrupting compounds (EDCs) affect the growth and development of teleost fishes. The major regulator of growth performance, the growth hormone (Gh)/insulin-like growth-factor (Igf) system, is sensitive to estrogenic compounds and mediates certain physiological and potentially behavioral consequences of EDC exposure. Igf binding proteins (Igfbps) are key modulators of Igf activity, but their alteration by EDCs has not been examined. We investigated two life-stages (fry and smolts) of Atlantic salmon ( Salmo salar ), and characterized how the Gh/Igf/Igfbp system responded to waterborne 17α-ethinylestradiol (EE 2 ), 17β-estradiol (E 2 ) and 4-nonylphenol (NP). Fry exposed to EE 2 and NP for 21 days had increased hepatic vitellogenin ( vtg ) mRNA levels while hepatic estrogen receptor α ( erα ), gh receptor (ghr) , igf1 and igf2 mRNA levels were decreased. NP-exposed fry had reduced body mass and total length compared to controls. EE 2 and NP reduced hepatic igfbp1b1 , -2a , -2b1 , -4 , -5b2 and -6b1 , and stimulated igfbp5a . In smolts, hepatic vtg mRNA levels were induced following 4-day exposures to all three EDCs, while erα only responded to EE 2 and E 2 . EDC exposures did not affect body mass or fork length; however, EE 2 diminished plasma Gh and Igf1 levels in parallel with reductions in hepatic ghr and igf1 . In smolts, EE 2 and E 2 diminished hepatic igfbp1b1 , -4 and -6b1 , and stimulated igfbp5a . There were no signs of compromised ionoregulation in smolts, as indicated by unchanged branchial ion pump/transporter mRNA levels. We conclude that hepatic igfbps respond (directly and/or indirectly) to environmental estrogens during two key life-stages of Atlantic salmon, and thus may modulate the growth and development of exposed individuals.

Aquatic Toxicology↗

USGS2012

Does translocation influence physiological stress in the desert tortoise?

Wildlife translocation is increasingly used to mitigate disturbances to animals or habitat due to human activities, yet little is known about the extent to which translocating animals causes stress. To understand the relationship between physiological stress and translocation, we conducted a multiyear study (2007–2009) using a population of desert tortoises (Gopherus agassizii) near Fort Irwin, California. Blood samples were collected from adult tortoises in three treatment groups (resident, translocated and control) for 1 year prior to and 2 years after translocation. Samples were analyzed by radioimmunoassay for plasma total corticosterone (CORT), a glucocorticoid hormone commonly associated with stress responses in reptiles. CORT values were analyzed in relation to potential covariates (animal sex, date, behavior, treatment, handling time, air temperature, home-range size, precipitation and annual plant production) among seasons and years. CORT values in males were higher than in females, and values for both varied monthly throughout the activity season and among years. Year and sex were strong predictors of CORT, and translocation explained little in terms of CORT. Based on these results, we conclude that translocation does not elicit a physiological stress response in desert tortoises.

California↗

USGS2014

Spatiotemporal variations in estrogenicity, hormones, and endocrine-disrupting compounds in influents and effluents of selected wastewater-treatment plants and receiving streams in New York, 2008-09

Endocrine-disrupting compounds (EDCs) in wastewater effluents have been linked to changes in sex ratios, intersex (in males), behavioral modifications, and developmental abnormalities in aquatic organisms. Yet efforts to identify and regulate specific EDCs in complex mixtures are problematic because little is known about the estrogen activity (estrogenicity) levels of many common and emerging contaminants. The potential effects of EDCs on the water quality and health of biota in streams of the New York City water supply is especially worrisome because more than 150 wastewater-treatment plants (WWTPs) are permitted to discharge effluents into surface waters and groundwaters of watersheds that provide potable water to more than 9 million people. In 2008, the U.S. Geological Survey (USGS), the New York State Department of Environmental Conservation (NYSDEC), New York State Department of Health (NYSDOH), and New York City Department of Environmental Protection (NYCDEP) began a pilot study to increase the understanding of estrogenicity and EDCs in effluents and receiving streams mainly in southeastern New York. The primary goals of this study were to document and assess the spatial and temporal variability of estrogenicity levels; the effectiveness of various treatment-plant types to remove estrogenicity; the concentrations of hormones, EDCs, and pharmaceuticals, personal care products (PPCPs); and the relations between estrogenicity and concentrations of hormones, EDCs, and PPCPs. The levels of estrogenicity and selected hormones, non-hormone EDCs, and PPCPs were characterized in samples collected seasonally in effluents from 7 WWTPs, once or twice in effluents from 34 WWTPs, and once in influents to 6 WWTPs. Estrogenicity was quantified, as estradiol equivalents, using both the biological e-screen assay and a chemical model. Results generally show that (1) estrogenicity levels in effluents varied spatially and seasonally, (2) a wide range of known and unknown EDCs were present in both WWTP effluents and receiving streams, (3) some effluents may be important sources of estrogenicity in weakly diluted streams, (4) measured levels of biological estrogenicity were often higher than estimated levels of chemical estrogenicity, and (5) the type of treatment had a large effect on the removal efficacy, and consequently, the estrogenicity levels observed in treated effluents.

New York↗

USGS2010

Endocrine active chemicals and endocrine disruption in Minnesota streams and lakes: Implications for aquatic resources, 1994-2008

The U.S. Geological Survey, in cooperation with St. Cloud State University, Minnesota Department of Health, Minnesota Pollution Control Agency, Minnesota Department of Natural Resources, Metropolitan Council Environmental Services, and the University of Minnesota, has conducted field monitoring studies and laboratory research to determine the presence of endocrine active chemicals and the incidence of endocrine disruption in Minnesota streams and lakes during 1994–2008. Endocrine active chemicals are chemicals that interfere with the natural regulation of endocrine systems, and may mimic or block the function of natural hormones in fish or other organisms. This interference commonly is referred to as endocrine disruption. Indicators of endocrine disruption in fish include vitellogenin (female egg yolk protein normally expressed in female fish) in male fish, oocytes present in male fish testes, reduced reproductive success, and changes in reproductive behavior. The results from a series of studies during 1994–2008 demonstrate that endocrine active chemicals are present in Minnesota surface waters, indicating that aquatic organism exposure is likely. Endocrine active chemicals have been identified in wastewater-treatment plant effluent and surface waters downstream from discharge of wastewater-treatment plant effluent throughout Minnesota at low concentrations. Biological indicators of endocrine disruption have been detected in wild fish throughout Minnesota at sites directly downstream from wastewater-treatment plant effluent, indicating that endocrine active chemicals in effluent contribute to endocrine disruption in fish. This finding was confirmed in a controlled study exposing fathead minnows to wastewater-treatment plant effluent at an onsite fish exposure laboratory. During this controlled study, changes in biological responses coincided with changes in wastewater-treatment plant effluent composition demonstrating that effluent effects on fish endocrine systems are temporally variable. Although chemicals contributing to endocrine disruption in fish are complex, several laboratory studies have further confirmed that certain classes of chemicals, such as hormones and alkylphenols, which are components of wastewater-treatment plant effluent, affect the endocrine systems of fish through biochemical, structural, and behavioral disruption. Although these studies indicate that wastewater-treatment plant effluent is a conduit for endocrine active chemicals to surface waters, endocrine active chemicals also were present in surface waters with no obvious wastewater-treatment plant effluent sources. Endocrine active chemicals were detected and indicators of endocrine disruption in fish were measured at numerous sites upstream from discharge of wastewater-treatment plant effluent. These observations indicate that other unidentified sources of endocrine active chemicals exist, such as runoff from land surfaces, atmospheric deposition, inputs from onsite septic systems, or other groundwater sources. Alternatively, some endocrine active chemicals may not yet have been identified or measured. The presence of biological indicators of endocrine disruption in male fish indicates that the fish are exposed to endocrine active chemicals. However indicators of endocrine disruption in male fish does not indicate an effect on fish reproduction or changes in fish populations.

Minnesota↗

USGS2007

Reproductive physiology of Missouri River gravid pallid sturgeon and shovelnose sturgeon during the 2005 and 2006 spawning seasons: Chapter C in Factors affecting the reproduction, recruitment, habitat, and population dynamics of pallid sturgeon and shovelnose sturgeon in the Missouri River

In a natural, unaltered river, the location and timing of sturgeon spawning will be dictated by the prevailing environmental conditions to which the sturgeon have adapted. A goal of the Comprehensive Sturgeon Research Program (CSRP; see chap. A) at the U.S. Geological Survey Columbia Environmental Research Center is to identify where, when, and under what conditions shovelnose sturgeon ( Scaphirhynchus platorynchus ) and pallid sturgeon ( S. albus ) spawn in the altered Missouri River so that those conditions necessary for spawning success can be defined. One approach to achieving this goal is to exploit what is known about fish reproductive physiology to develop and apply a suite of diagnostic indicators of readiness to spawn. In 2005 and 2006, gravid shovelnose sturgeon and a limited number of pallid sturgeon were fitted with transmitters and tracked on their spawning migration. A suite of physiological indicators of reproductive state such as reproductive hormones and oocyte development were measured. These same measurements were made on tissues collected from additional fish, presumably migrating to spawn, that were not tagged or tracked. The data presented here indicating the sturgeons’ readiness to spawn are to be evaluated together with their behavior and the environmental conditions. The U.S. Army Corps of Engineers (ACOE) Sturgeon Response to Flow Modification (SRFM; see chap. A) study, initiated in 2006, provides additional opportunities to experimentally evaluate the sturgeon reproductive response indicators relative to changes in flow. In this chapter, we report progress made on identifying and developing the physiological indicators and summarize 2 years’ worth of indicator data collected thus far.

Missouri River, Yellowstone River↗

USGS2011

Effects of biologically-active chemical mixtures on fish in a wastewater-impacted urban stream

Stream flow in urban aquatic ecosystems often is maintained by water-reclamation plant (WRP) effluents that contain mixtures of natural and anthropogenic chemicals that persist through the treatment processes. In effluent-impactedstreams, aquatic organisms such as fish are continuously exposed to biologically-activechemicals throughout their life cycles. The North Shore Channel of the Chicago River (Chicago, Illinois) is part of an urban ecosystem in which > 80% of the annual flow consists of effluent from the North Side WRP. In this study, multiple samplings of the effluent and stream water were conducted and fish (largemouth bass and carp) were collected on 2 occasions from the North Shore Channel. Fish also were collected once from the Outer Chicago Harbor in Lake Michigan, a reference site not impacted by WRP discharges. Over 100 organic chemicals with differing behaviors and biological effects were measured, and 23 compounds were detected in all of the water samples analyzed. The most frequently detected and highest concentration (> 100 μg/L) compounds were ethylenediaminetetraacetic acid and 4-nonylphenolmono-to-tetraethoxycarboxylic acids. Other biologically-activechemicals including bisphenol A, 4-nonylphenol, 4-nonylphenolmono-to-tetraethoxylates, 4- tert -octylphenol, and 4- tert -octylphenolmono-to-tetraethoxylates were detected at lower concentrations (< 5 μg/L). The biogenic steroidal hormones 17β-estradiol, estrone, testosterone, 4-androstene-3,17-dione, and cis -androsterone were detected at even lower concentrations (< 0.005 μg/L). There were slight differences in concentrations between the North Side WRP effluent and the North Shore Channel, indicating minimal in-stream attenuation. Fish populations are continuously exposed to mixtures of biologically-activechemicals because of the relative persistency of the chemicals with respect to stream hydraulic residence time, and the lack of a fresh water source for dilution. The majority of male fish exhibited vitellogenin induction, a physiological response consistent with exposure to estrogenic compounds. Tissue-level signs of reproductive disruption, such as ovatestis, were not observed.

Illinois↗

USGS2003

Pharmaceuticals, hormones, personal-care products, and other organic wastewater contaminants in water resources: Recent research activities of the U.S. Geological Survey's toxic substances hydrology program

Recent decades have brought increasing concerns for potential contamination of water resources that could inadvertently result during production, use, and disposal of the numerous chemicals offering improvements in industry, agriculture, medical treatment, and even common household products. Increasing knowledge of the environmental occurrence or toxicological behavior of these contaminants from various studies in Europe, United States, and elsewhere has resulted in increased concern for potential adverse environmental and human health effects (Daughton and Ternes, 1999). Ecologists and public health experts often have incomplete understandings of the toxicological significance of many of these contaminants, particularly long-term, low-level exposure and when they occur in mixtures with other contaminants (Daughton and Ternes, 1999; Kümmerer, 2001). In addition, these ‘emerging contaminants’ are not typically monitored or assessed in ambient water resources. The need to understand the processes controlling the transport and fate of these contaminants in the environment, and the lack of knowledge of the significance of long-term exposures have increased the need to study environmental occurrence down to trace (nanogram per liter) levels. Furthermore, the possibility that mixtures of environmental contaminants may interact synergistically or antagonistically has increased the need to characterize the types of mixtures that are found in our waters. The U.S. Geological Survey’s Toxic Substances Hydrology Program (Toxics Program) is developing information and tools on emerging water-quality issues that will be used to design and improve water-quality monitoring and assessment programs of the USGS and others, and for proactive decision-making by industry, regulators, the research community, and the public (http://toxics.usgs.gov/regional/emc.html). This research on emerging water-quality issues includes a combination of laboratory work to develop new analytical capabilities as well as field work on the occurrence, fate, and effects of these contaminants.

Geohealth News↗

USGS2011

Isotopic tracing of perchlorate in the environment

Isotopic measurements can be used for tracing the sources and behavior of environmental contaminants. Perchlorate (ClO 4 − ) has been detected widely in groundwater, soils, fertilizers, plants, milk, and human urine since 1997, when improved analytical methods for analyzing ClO 4 − concentration became available for routine use. Perchlorate ingestion poses a risk to human health because of its interference with thyroidal hormone production. Consequently, methods for isotopic analysis of ClO 4 − have been developed and applied to assist evaluation of the origin and migration of this common contaminant. Isotopic data are now available for stable isotopes of oxygen and chlorine, as well as 36 Cl isotopic abundances, in ClO 4 − samples from a variety of natural and synthetic sources. These isotopic data provide a basis for distinguishing sources of ClO 4 − found in the environment, and for understanding the origin of natural ClO 4 − . In addition, the isotope effects of microbial ClO 4 − reduction have been measured in laboratory and field experiments, providing a tool for assessing ClO 4 − attenuation in the environment. Isotopic data have been used successfully in some areas for identifying major sources of ClO 4 − contamination in drinking water supplies. Questions about the origin and global biogeochemical cycle of natural ClO 4 − remain to be addressed; such work would benefit from the development of methods for preparation and isotopic analysis of ClO 4 − in samples with low concentrations and complex matrices.

Book chapter↗

USGS2001

Effects of food stress on survival and reproductive performance of seabirds

Traditional field methods of assessing effects of fluctuations in food supply on the survival and reproductive performance of seabirds may give equivocal results. In this project we applied an additional tool: The measure of stress hormones in free-ranging seabirds. Food stress can be quantified by measuring base levels of stress hormones such as corticosterone in the blood of seabirds, or the rise in blood levels of corticosterone in response to a standardized stressor: capture, handling and restraint. We applied these techniques to seabirds breeding in Lower Cook Inlet and also used captive birds for controlled experiments. This study provided a unique opportunity for a concurrent field and captive study of the behavioral and physiological consequences of stress in seabirds. Moreover, this study provides the basis for management of seabird populations in the areas affected by the Exxon Valdez oil spill, which will have broader applications for seabird monitoring programs. This year represents production of a synthesis of the project.

Alaska↗

USGS2013

Smolt physiology and endocrinology

Hormones play a critical role in maintaining body fluid balance in euryhaline fishes during changes in environmental salinity. The neuroendocrine axis senses osmotic and ionic changes, then signals and coordinates tissue-specific responses to regulate water and ion fluxes. Rapid-acting hormones, e.g. angiotensins, cope with immediate challenges by controlling drinking rate and the activity of ion transporters in the gill, gut, and kidney. Slow-acting hormones, e.g. prolactin and growth hormone/insulin-like growth factor-1, reorganize the body for long-term acclimation by altering the abundance of ion transporters and through cell proliferation and differentiation of ionocytes and other osmoregulatory cells. Euryhaline species exist in all groups of fish, including cyclostomes, and cartilaginous and teleost fishes. The diverse strategies for responding to changes in salinity have led to differential regulation and tissue-specific effects of hormones. Combining traditional physiological approaches with genomic, transcriptomic, and proteomic analyses will elucidate the patterns and diversity of the endocrine control of euryhalinity.

Book chapter↗

USGS2013

Pheromonal bile acid 3-ketopetromyzonol sulfate primes the neuroendocrine system in sea lamprey

Background Vertebrate pheromones are known to prime the endocrine system, especially the hypothalamic-pituitary-gonadal (HPG) axis. However, no known pheromone molecule has been shown to modulate directly the synthesis or release of gonadotropin releasing hormone (GnRH), the main regulator of the HPG axis. We selected sea lamprey (Petromyzon marinus) as a model system to determine whether a single pheromone component alters the output of GnRH. Sea lamprey male sex pheromones contain a main component, 7α, 12α, 24-trihydroxy-5α-cholan-3-one 24-sulfate (3 keto-petromyzonol sulfate or 3kPZS), which has been shown to modulate behaviors of mature females. Through a series of experiments, we tested the hypothesis that 3kPZS modulates both synthesis and release of GnRH, and subsequently, HPG output in immature sea lamprey. Results The results showed that natural male pheromone mixtures induced differential steroid responses but facilitated sexual maturation in both sexes of immature animals (χ2 = 5.042, dF = 1, p < 0.05). Exposure to 3kPZS increased plasma 15α-hydroxyprogesterone (15α-P) concentrations (one-way ANOVA, p < 0.05) and brain gene expressions (genes examined: three lamprey (l) GnRH-I transcripts, lGnRH-III, Jun and Jun N-terminal kinase (JNK); one-way ANOVA, p < 0.05), but did not alter the number of GnRH neurons in the hypothalamus in immature animals. In addition, 3kPZS treatments increased lGnRH peptide concentrations in the forebrain and modulated their levels in plasma. Overall, 3kPZS modulation of HPG axis is more pronounced in immature males than in females. Conclusions We conclude that a single male pheromone component primes the HPG axis in immature sea lamprey in a sexually dimorphic manner.

BMC Neuroscience↗

USGS2019

Migratory strategy explains differences in timing of female reproductive development in seasonally sympatric songbirds.

1. Divergent migratory strategies among populations can result in population-level differences in timing of reproduction (allochrony) and local adaptation. However, the mechanisms underlying among-population variation in timing are insufficiently understood, particularly in females. 2. We studied differences in reproductive development and its related mechanisms along the hypothalamic-pituitary-gonadal axis (HPG) in closely related migratory and sedentary (i.e., resident) female dark-eyed juncos (Junco hyemalis) living together in sympatry during early spring. We predicted that residents would be more reproductively developed than migrants. Alternatively, we predicted that females might exhibit similar reproductive development in response to the same environmental cue despite differences in migratory behavior. To compare their degree of reproductive development during seasonal sympatry and the underlying mechanisms of these differences, we collected ovarian and liver tissue in early spring prior to migration, and compared abundance of gene transcripts associated with reproduction using quantitative PCR. We also used stable hydrogen isotopes to infer relative breeding and wintering latitude of migrants. 3. We found more abundant transcripts for luteinizing hormone receptor and aromatase in addition to significantly heavier ovaries in residents than in migrants. Together these results suggest greater sensitivity and response to upstream endocrine stimulation in resident females. Transcript abundance for other receptors associated with reproduction, however, did not differ between populations. When comparing ovarian development within migrants, females with lower hydrogen isotopes (indicating higher breeding latitudes) had smaller ovaries, suggesting that longer-distance migrations may further delay reproductive development. 4. Based on differences in ovary mass and transcript abundance, we conclude that females that differ in migratory strategy also differ in timing of reproductive development. Differences in timing of reproductive development between migrant and resident females could drive reproductive isolation between populations. This study also reveals mechanisms that could drive allochrony and enable us to predict how seasonal organisms will respond to a changing world.

Functional Ecology↗

USGS2011

Effects of supplemental feeding and aggregation on fecal glucocorticoid metabolite concentrations in elk

Habitat modifications and supplemental feeding artificially aggregate some wildlife populations, with potential impacts upon contact and parasite transmission rates. Less well recognized, however, is how increased aggregation may affect wildlife physiology. Crowding has been shown to induce stress responses, and increased glucocorticoid (GC) concentrations can reduce immune function and increase disease susceptibility. We investigated the effects of supplemental feeding and the aggregation that it induces on behavior and fecal glucocorticoid metabolite concentrations (fGCM) in elk (Cervus elaphus) using observational and experimental approaches. We first compared fGCM levels of elk on supplemental feedgrounds to neighboring elk populations wintering in native habitats using data from 2003 to 2008. We then experimentally manipulated the distribution of supplemental food on feedgrounds to investigate whether more widely distributed food would result in lower rates of aggression and stress hormone levels. Contrary to some expectations that fed elk may be less stressed than unfed elk during the winter, we found that elk on feedgrounds had fecal GC levels at least 31% higher than non-feedground populations. Within feedgrounds, fGCM levels were strongly correlated with local measures of elk density (r 2 = 0.81). Dispersing feed more broadly, however, did not have a detectable effect on fGCM levels or aggression rates. Our results suggest that increases in aggregation associated with winter feedgrounds affects elk physiology, and the resulting increases in fGCM levels are not likely to be mitigated by management efforts that distribute the feed more widely. Additional research is needed to assess whether these increases in fGCMs directly alter parasite transmission and disease dynamics.

Journal of Wildlife Management↗