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Prymnesium parvum is a toxin-producing harmful alga that has caused ecological and economic damage worldwide. Effective methods to control blooms of this species in the field, however, are unavailable. This study examined five natural compounds present in the invasive plant Arundo donax and one synthetic derivative (5,6-dichlorogramine) for their effect on P. parvum growth. All compounds except one inhibited growth in the following order of potency: ellipticine > > 5,6-dichlorogramine > 1 H-indole = 2,4,6-trimethyl-benzonitrile > gramine. Ellipticine was by far the most potent inhibitor, with full algicidal activity at concentrations as low as 0.04 mg L−1 and 3- and 9-day IC50 values of 0.012 and 0.007 mg L−1, respectively. A reduction in chlorophyll content and swimming activity and an increase in length and volume (swelling) were documented in algal cells exposed to 0.01–0.02 mg ellipticine L−1. These results show that ellipticine is among the most potent natural algicides identified to date. The sixth compound tested, oleamide, unexpectedly stimulated algal growth above control levels. Overall, these observations confirm the existence of highly potent anti-P. parvum allelochemicals in giant reed and demonstrate potential for using products derived from this plant in the development of natural, environmentally friendly methods to control harmful algal blooms.

Prymnesium parvum is a euryhaline, toxin-producing microalga. Although its abundance in inland waters and growth potential in the laboratory is reduced at high salinity (>20), the ability of inland strains to adjust their growth after long-term residence in high salinity is uncertain. An inland strain of P. parvum maintained at salinity of 5 in modified artificial seawater medium (ASM-5) was subjected to the following treatments over five sequential batch culture rounds: ASM-5 (control); modified ASM at salinity of 30, raised with NaCl; modified ASM at salinity incrementally increased to 30 with NaCl; and Instant Ocean® at salinity of 30 (IO-30). Exponential growth rate (r) was reduced when salinity was increased from 5 to 30 in ASM but returned to control values during the second round. When salinity was incrementally increased, a reduction in r still occurred when salinity reached 25-30. Maximum density was reduced at salinity of 30 in ASM upon abrupt transfer or incremental increase, and compensation did not occur. Growth performance in IO-30 was comparable to control values. In conclusion, (i) long-term compensation for acute inhibitory effects of high salinity occurred for r but not maximum density, (ii) incremental increases in salinity did not prevent growth inhibition, suggesting the existence of a salinity threshold of 25-30 for onset of salinity stress, and (iii) the presence of a seawater-like salt mixture prevented growth inhibition by high salinity. These findings provide new insights on P. parvum's long-term ability to adjust its growth in environments of different salinity and ionic composition.

This study examined the association of air, land, and water variables with the first historical occurrence and current distribution of toxic Prymnesium parvum blooms in reservoirs of the Brazos River and Colorado River, Texas (USA). One impacted and one reference reservoir were selected per basin. Land cover and use variables were estimated for the whole watershed (WW) and a 0.5-km zone on either side of streams (near field, NF). Variables were expressed in annual values. Principal component and trend analyses were used to determine (1) differences in environmental conditions before and after the 2001 onset of toxic blooms in impacted reservoirs (study period, 1992-2017), and (2) traits that uniquely discriminate impacted from reference reservoirs (2001-2017). Of thirty-three variables examined, two positively aligned with the reoccurring appearance of blooms in impacted reservoirs (air CO

Copyright: © Barnes and Patino This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International - CC BY 4.0).

The Jim Bertram Lake System consists of several stream impoundments within the City of Lubbock, Texas (USA). Baseflow in the upstream reach is dominated by nitrogen-rich-treated wastewater. While toxic blooms of Prymnesium parvum have occurred in this system for ∼2 decades during fall or winter-spring, little is known about water quality variables that facilitate blooms or the alga’s spatiotemporal distribution. Water quality associations were examined monthly over a 1-year period. Total phosphorus was largely below the detection limit, suggesting that the system is phosphorus limited. Algal abundance was low during the assessment period and associations were determined using multiple logistic regression. Algal incidence was negatively associated with temperature and positively with organic nitrogen and calcium hardness. These findings conform with earlier reports but positive associations with the latter two variables are noteworthy because they have not been widely confirmed. Spatiotemporal distribution was evaluated in fall and winter-spring of three consecutive years. Prymnesium parvum incidence was higher in the upper than in the lower reach, and detections in the lower reach occurred only after a dense bloom developed in the upper reach contemporaneously with stormwater runoff-associated flooding. Thus, the upstream reach is a major source of propagules for downstream sites. Because urban runoff is a source of phosphorus and its nitrogen: phosphorus ratio is lower than prevailing ratios in the upper reach, what triggered the bloom was likely relief from phosphorus limitation. This study provided water quality, geographic and hydrological indices that may inform prevention and control methods for harmful algae in nitrogen-enriched urban systems.

This study examined the ability of acidic and neutral/alkaline fractions of a methanolic extract from giant reed (Arundo donax) and of two of its constituents, gramine and skatole, to inhibit growth of the ichthyotoxic golden alga (Prymnesium parvum) in batch culture. For this study, growth suppression was defined as inhibition of maximum cell density, algicidal activity as early occurrence of negative growth, and algistatic activity as lack of net growth. The acidic fraction did not affect algal growth. The neutral/alkaline fraction showed growth-suppressing and algicidal activities but no signs of algistatic activity - namely, cells in cultures surviving a partial-algicidal exposure concentration (causing transient negative growth) were later able to initiate positive growth but at higher concentrations, algicidal activity was full and irreversible. Gramine suppressed growth more effectively than skatole and at the highest concentration tested, gramine also showed partial-algicidal and algistatic activity. While the partial-algicidal activities of the neutral/alkaline fraction and of gramine were short-lived (≤6days) and thus may share similar mechanisms, algistatic activity was unique to gramine and persisted for >3 weeks. Given gramine's reported concentration in the neutral/alkaline fraction, its corresponding level of algicidal activity is much lower than the fraction's suggesting the latter contains additional potent algicides. Inhibition of maximum cell density by all test compounds was associated with reductions in exponential growth rate, and in the case of the neutral/alkaline fraction and gramine also reductions in early (pre-exponential) growth. These results indicate that giant reed is a potential source of natural products to control golden alga blooms. Giant reed is an invasive species in North America, thus also providing incentive for research into strategies to couple management efforts for both species.

Sex determination is the first step toward the establishment of phenotypic sex in most vertebrates. Aquatic poikilotherms such as teleost fishes exhibit a high diversity of sex-determination mechanisms and gonadal phenotypes that are remarkably plastic and responsive to a variety of environmental factors (e.g., water temperature, pH, salinity, photoperiod, population density). This chapter reviews current knowledge of genotypic and environmental sex determination systems in fishes with special reference to Atheriniformes-one of the best-characterized taxa in this field-and offers perspectives to guide and stimulate further research.

Introductions of dreissenid mussels in North America have been a significant concern over the last few decades. This study assessed the distribution of synthetic organic compounds (SOCs) in the food web of Lake Mead, Nevada/Arizona, USA and how this distribution was influenced by the introduction of invasive quagga mussels. A clear spatial gradient of SOC concentrations in water was observed between lake basins downstream of populated areas and more rural areas. Within the food web, trophic magnification factors (TMF) indicated statistically significant biomagnification for nine, and biodilution for two, of 22 SOCs examined. The highest value recorded was for PCB 118 (TMF, 5.14), and biomagnification of methyl triclosan (TMF, 3.85) was also apparent. Biodilution was observed for Tonalide® (0.06) and Galaxolide® (0.38). Total SOC concentration in quagga mussels was higher than in three pelagic fishes. Also, 19 of 20 SOC examined in Largemouth Bass (Micropterus salmoides) had substantially lower concentrations in 2013, when quagga mussels had become well established, than in 2007/08, soon after quagga mussels were introduced. Estimates of SOC concentrations in the water column and quagga mussels suggest that a considerable portion (~10.5%) of the SOC mass in the lake has shifted from the pelagic to the benthic environments due to quagga mussel growth. These observations suggest that benthic species, such as the endangered Razorback Sucker, may be experiencing increased risk of SOC exposure. In addition, stable isotope analysis (carbon and nitrogen) indicated a decrease in the nutritional value of zooplankton to consumers (e.g., Razorback Sucker larvae) since quagga mussels became established. These changes could affect Razorback Sucker larval survival and recruitment. Results from this study strongly suggest that the introduction of quagga mussels has greatly altered the dynamics of SOCs and other processes in the food web of Lake Mead.

Intermittent rivers are becoming more ecologically stressed worldwide. Flow cessation occurs naturally and spatiotemporally in these systems and anthropogenic activities such as wastewater discharges can have considerable impacts. Public entities mostly monitor water quality in permanent streams, leading to insufficient monitoring of intermittent streams and consequently to their potentially inadequate management.. This study analyzed spatiotemporal patterns of water quality and associated ecological risk through the quantification of physicochemical and microbiological pollutants in the intermittent river system of El Novillo and San Marcos in Northeast Mexico. Results showed that water quality varied geographically and seasonally. Based on national and international criteria, annual averages of water quality parameters analyzed suggested that streamflow in these river systems is of poor quality and poses high ecological risk to aquatic life. In the urban area, annual mean concentrations of Cd and Pb (0.14 and 0.4 mg/L) were 77- and 10-fold higher than their respective water quality criteria (

Sex determination is the first step toward the establishment of phenotypic sex in most vertebrates. Aquatic poikilotherms such as teleost fishes exhibit a high diversity of sex-determination mechanisms and gonadal phenotypes that are remarkably plastic and responsive to a variety of environmental factors (e.g., water temperature, pH, salinity, photoperiod, population density). This chapter reviews current knowledge of genotypic and environmental sex determination systems in fishes with special reference to Atheriniformes-one of the best-characterized taxa in this field-and offers perspectives to guide and stimulate further research.

Golden alga Prymnesium parvum Carter is a euryhaline, ichthyotoxic haptophyte (Chromista). Because of its presumed coastal/marine origin where SO42- levels are high, the relatively high SO42- concentration of its brackish inland habitats, and the sensitivity of marine chromists to sulfur deficiency, this study examined whether golden alga growth is sensitive to SO42- concentration. Fluoride is a ubiquitous ion that has been reported at higher levels in golden alga habitat; thus, the influence of F- on growth also was examined. In low-salinity (5 psu) artificial seawater medium, overall growth was SO42—dependent up to 1000 mg l-1 using MgSO4 or Na2SO4 as source; the influence on growth rate, however, was more evident with MgSO4. Transfer from 5 to 30 psu inhibited growth when salinity was raised with NaCl but in the presence of seawater levels of SO42-, these effects were fully reversed with MgSO4 as source and only partially reversed with Na2SO4. Growth inhibition was not observed after acute transfer to 30 psu in a commercial sea salt mixture. In 5-psu medium, F- inhibited growth at all concentrations tested. These observations support the hypothesis that spatial differences in SO42- –but not F-–concentration help drive the inland distribution and growth of golden alga and also provide physiological relevance to reports of relatively high Mg2+ concentrations in golden alga habitat. At high salinity, however, the ability of sulfate to maintain growth under osmotic stress was weak and overshadowed by the importance of Mg2+. A mechanistic understanding of growth responses of golden alga to SO42-, Mg2+ and other ions at environmentally relevant levels and under different salinity scenarios will be necessary to clarify their ecophysiological and evolutionary relevance.

Glyphosate-based herbicides (GBH) are widely used around the globe. While generally toxic to phototrophs, organic phosphorus in glyphosate can become available to glyphosate-resistant phytoplankton and contribute to algal bloom development. Few studies have examined the effects of GBH on growth of eukaryotic microalgae and information for the toxic bloom-forming haptophyte, Prymnesium parvum, is limited. Using a batch-culture system, this study examined the effects on P. parvum growth of a single application of Roundup Weed and Grass Killer Super Concentrate Plus® (Roundup SC), Roundup Weed and Grass Killer Ready-to-Use III® (Roundup RtU), and technical-grade glyphosate at low concentrations [0–1000 μg glyphosate acid equivalent (ae) l−1]. Roundup formulations differ in the percent of glyphosate as active ingredient (Roundup SC, ∼50%; Roundup RtU, 2%), allowing indirect evaluation of the influence of inactive ingredients. Roundup SC enhanced exponential growth rate at 10–1000 μg glyphosate ae l−1, and a positive monotonic association was noted between Roundup SC concentration and early (pre-exponential growth) but not maximum cell density. Glyphosate and both Roundup formulations enhanced growth rate at 100 μg glyphosate l−1, but only Roundup SC and glyphosate significantly stimulated early and maximum density. This observation suggests the higher concentration of inactive ingredients and other compounds in Roundup RtU partially counteracts glyphosate-dependent growth stimulation. When phosphate concentration was varied while maintaining other conditions constant, addition of Roundup SC and glyphosate at 100 μg l−1 influenced growth more strongly than equivalent changes in phosphate-associated phosphorus. It appears, therefore, that low doses of glyphosate stimulate growth by mechanisms unrelated to the associated small increases in total phosphorus. In conclusion, glyphosate and GBH stimulate P. parvum growth at low, environmentally relevant concentrations. This finding raises concerns about the potential contribution to P. parvum blooms by glyphosate-contaminated runoff or by direct application of GBH to aquatic environments.

This study examined the effects thyroid-active compounds during early development on body morphology of Zebrafish (Danio rerio). Three-day postfertilization (dpf) larvae were exposed to goitrogen [methimazole (MZ, 0.15mM)], combination of MZ (0.15mM) and thyroxine (T4, 2nM), T4 (2nM), or control (reconstituted water) treatments until 33dpf and subsequently maintained in reconstituted water until 45dpf. Samples were taken at 33 and 45dpf for multivariate analysis of geometric distances between selected homologous landmarks placed on digital images of fish, and for histological assessment of thyrocytes. Body mass, standard length, and pectoral fin length were separately measured on remaining fish at 45dpf. Histological analysis confirmed the hypothyroid effect (increased thyrocyte height) of MZ and rescue effect of T4 co-administration. Geometric distance analysis showed that pectoral and pelvic fins shifted backward along the rostrocaudal axis under hypothyroid conditions at 45dpf and that T4 co-treatment prevented this shift. Pectoral fin length at 45dpf was reduced by exposure to MZ and rescued by co-administration of T4, but it was not associated with standard length. Methimazole caused a reduction in body mass and length at 45dpf that could not be rescued by T4 co-administration, and non-thyroidal effects of MZ on body shape were also recognized at 33 and 45dpf. Alterations in the length and position of paired fins caused by exposure to thyroid-disrupting chemicals during early development, as shown here for Zebrafish, could affect physical aspects of locomotion and consequently other important organismal functions such as foraging, predator avoidance, and ultimately survival and recruitment into the adult population. Results of this study also suggest the need to include rescue treatments in endocrine disruption studies that rely on goitrogens as reference for thyroid-mediated effects.

Adult male Common Carp were sampled in 2007/08 over a full reproductive cycle at Lake Mead National Recreation Area. Sites sampled included a stream dominated by treated wastewater effluent, a lake basin receiving the streamflow, an upstream lake basin (reference), and a site below Hoover Dam. Individual body burdens for 252 contaminants were measured, and biological variables assessed included physiological [plasma vitellogenin (VTG), estradiol-17β (E2), 11-ketotestosterone (11KT)] and organ [gonadosomatic index (GSI)] endpoints. Patterns in contaminant composition and biological condition were determined by Principal Component Analysis, and their associations modeled by Principal Component Regression. Three spatially distinct but temporally stable gradients of contaminant distribution were recognized: a contaminant mixture typical of wastewaters (PBDEs, methyl triclosan, galaxolide), PCBs, and DDTs. Two spatiotemporally variable patterns of biological condition were recognized: a primary pattern consisting of reproductive condition variables (11KT, E2, GSI), and a secondary pattern including general condition traits (condition factor, hematocrit, fork length). VTG was low in all fish, indicating low estrogenic activity of water at all sites. Wastewater contaminants associated negatively with GSI, 11KT and E2; PCBs associated negatively with GSI and 11KT; and DDTs associated positively with GSI and 11KT. Regression of GSI on sex steroids revealed a novel, nonlinear association between these variables. Inclusion of sex steroids in the GSI regression on contaminants rendered wastewater contaminants nonsignificant in the model and reduced the influence of PCBs and DDTs. Thus, the influence of contaminants on GSI may have been partially driven by organismal modes-of-action that include changes in sex steroid production. The positive association of DDTs with 11KT and GSI suggests that lifetime, sub-lethal exposures to DDTs have effects on male carp opposite of those reported by studies where exposure concentrations were relatively high. Lastly, this study highlighted advantages of multivariate/multiple regression approaches for exploring associations between complex contaminant mixtures and gradients and reproductive condition in wild fishes.

Water temperature, dissolved oxygen, and concentrations of salts in surface water bodies can be affected by the natural environment and local human activities such as surface and ground water withdrawals, land use and energy extraction, and variability and long-term trends in atmospheric conditions including temperature and precipitation. Here, we quantify the relationship between 121 indicators of mean and extreme temperature and precipitation and 24 water quality parameters in 57 Texas reservoirs using observational data records covering the period 1960 to 2010. Over time scales ranging from 1 week to 2 years, we find that water temperature, dissolved oxygen, pH, specific conductance, chloride, sulfate, and phosphorus all show consistent correlations with atmospheric predictors, including high and low temperature extremes, dry days, heavy precipitation events, and mean temperature and precipitation. Based on these relationships combined with regional climate projections, we expect climate change to increase water temperatures, decrease dissolved oxygen levels, decrease pH, increase specific conductance, and increase levels of sulfate and chloride in Texas reservoirs. Over decadal time scales, this may affect aquatic ecosystems in the reservoirs, including altering the risk of conditions conducive to algae occurrence, as well as affecting the quality of water available for human consumption and recreation. Copyright © 2015 John Wiley & Sons, Ltd.

Golden alga (Prymnesium parvum) is a toxic haptophyte that has caused considerable ecological damage to marine and inland aquatic ecosystems worldwide. Studies focused primarily on laboratory cultures have indicated that toxicity is poorly correlated with the abundance of golden alga cells. This relationship, however, has not been rigorously evaluated in the field where environmental conditions are much different. The ability to predict toxicity using readily measured environmental variables and golden alga abundance would allow managers rapid assessments of ichthyotoxicity potential without laboratory bioassay confirmation, which requires additional resources to accomplish. To assess the potential utility of these relationships, several a priori models relating lethal levels of golden alga ichthyotoxicity to golden alga abundance and environmental covariates were constructed. Model parameters were estimated using archived data from four river basins in Texas and New Mexico (Colorado, Brazos, Red, Pecos). Model predictive ability was quantified using cross-validation, sensitivity, and specificity, and the relative ranking of environmental covariate models was determined by Akaike Information Criterion values and Akaike weights. Overall, abundance was a generally good predictor of ichthyotoxicity as cross validation of golden alga abundance-only models ranged from ∼ 80% to ∼ 90% (leave-one-out cross-validation). Environmental covariates improved predictions, especially the ability to predict lethally toxic events (i.e., increased sensitivity), and top-ranked environmental covariate models differed among the four basins. These associations may be useful for monitoring as well as understanding the abiotic factors that influence toxicity during blooms.

Summary We assessed reproductive status, fecundity, egg size, and spawning dynamics of shovelnose sturgeon Scaphirhynchus platorynchus in the lower Platte River. Shovelnose sturgeon were captured throughout each year during 2011 and 2012 using a multi-gear approach designed to collect a variety of fish of varying sizes and ages. Fish were collected monthly for a laboratory assessment of reproductive condition. Female shovelnose sturgeon reached fork length at 50% maturity (FL50) at 547 mm and at a minimum length of 449 mm. The average female spawning cycle was 3–5 years. Mean egg count for adult females was 16 098 ± 1103 (SE), and mean egg size was 2.401 ± 0.051 (SE) mm. Total fecundity was positively correlated with length (r2 = 0.728; P 0.05). Male shovelnose sturgeon reached FL50 at 579 mm and at a minimum length of 453 mm. The average male spawning cycle was 1–2 years. Reproductively viable male and female sturgeon occurred during the spring (March–May) and autumn (September–October) in both years, indicating spring and potential autumn spawning events. Shovelnose sturgeon in the lower Platte River are maturing at a shorter length and younger age compared to populations elsewhere. Although it is unknown if the change is plastic or evolutionary, unfavorable environmental conditions or over-harvest may lead to hastened declines compared to other systems.

Since the mid-1980s, fish-killing blooms of Prymnesium parvum spread throughout the USA. In the south central USA, P. parvum blooms have commonly spanned hundreds of kilometers. There is much evidence that physiological stress brought on by inorganic nutrient limitation enhances toxicity. Other factors influence toxin production as well, such as stress experienced at low salinity and temperature. A better understanding of toxin production by P. parvum remains elusive and the identities and functions of chemicals produced are unclear. This limits our understanding of factors that facilitated the spread of P. parvum blooms. In the south central USA, not only is there evidence that the spread of blooms was controlled, in part, by migration limitation. But there are also observations that suggest changed environmental conditions, primarily salinity, facilitated the spread of blooms. Other factors that might have played a role include altered hydrology and nutrient loading. Changes in water hardness, herbicide use, system pH, and the presence of toxin-resistant and/or P. parvum-inhibiting plankton may also have played a role. Management of P. parvum in natural systems has yet to be attempted, but may be guided by successes achieved in small impoundments and mesocosm experiments that employed various chemical and hydraulic control approaches.

This study determined long-term changes in fish assemblages, river discharge, salinity, and local precipitation, and examined hydrological drivers of biotic homogenization in a dryland river ecosystem, the Trans-Pecos region of the Rio Grande/Rio Bravo del Norte (USA/Mexico). Historical (1977–1989) and current (2010–2011) fish assemblages were analyzed by rarefaction analysis (species richness), nonmetric multidimensional scaling (composition/variability), multiresponse permutation procedures (composition), and paired t -test (variability). Trends in hydrological conditions (1970s–2010s) were examined by Kendall tau and quantile regression, and associations between streamflow and specific conductance (salinity) by generalized linear models. Since the 1970s, species richness and variability of fish assemblages decreased in the Rio Grande below the confluence with the Rio Conchos (Mexico), a major tributary, but not above it. There was increased representation of lower-flow/higher-salinity tolerant species, thus making fish communities below the confluence taxonomically and functionally more homogeneous to those above it. Unlike findings elsewhere, this biotic homogenization was due primarily to changes in the relative abundances of native species. While Rio Conchos discharge was > 2-fold higher than Rio Grande discharge above their confluence, Rio Conchos discharge decreased during the study period causing Rio Grande discharge below the confluence to also decrease. Rio Conchos salinity is lower than Rio Grande salinity above their confluence and, as Rio Conchos discharge decreased, it caused Rio Grande salinity below the confluence to increase (reduced dilution). Trends in discharge did not correspond to trends in precipitation except at extreme-high (90th quantile) levels. In conclusion, decreasing discharge from the Rio Conchos has led to decreasing flow and increasing salinity in the Rio Grande below the confluence. This spatially uneven desertification and salinization of the Rio Grande has in turn led to a region-wide homogenization of hydrological conditions and of taxonomic and functional attributes of fish assemblages.

Golden alga (Prymnesium parvum) is a harmful alga that has caused ecological and economic harm in freshwater and marine systems worldwide. In inland systems of North America, toxic blooms have nearly eliminated fish populations in some systems. Modifying nutrient profiles through alterations to land or water use may be a viable alternative for golden alga control in reservoirs. The main objective of this study was to improve our understanding of the nutrient dynamics that influence golden alga bloom formation and toxicity in west Texas reservoirs. We examined eight sites in the Upper Colorado River basin, Texas: three impacted reservoirs that have experienced repeated golden alga blooms; two reference reservoirs where golden alga is present but nontoxic; and three confluence sites downstream of the impacted and reference sites. Total, inorganic, and organic nitrogen and phosphorus and their ratios were quantified monthly along with golden alga abundance and ichthyotoxicity between December 2010 and July 2011. Blooms persisted for several months at the impacted sites, which were characterized by high organic nitrogen and low inorganic nitrogen. At impacted sites, abundance was positively associated with inorganic phosphorus and bloom termination coincided with increases in inorganic nitrogen and decreases in inorganic phosphorus in late spring. Management of both inorganic and organic forms of nutrients may create conditions in reservoirs unfavorable to golden alga.

Estrogens are recognized as modulators of immune responses in mammals and teleosts. While it is known that the effects of estrogens are mediated via leukocyte-specific estrogen receptors (ERs) in humans and mice, leucocyte-specific estrogen receptor expression and the effects of estrogens on this cell population is less explored and poorly understood in teleosts. Here in, we verify that channel catfish (Ictalurus punctaus) leukocytes express ERα and ERβ2. Transcripts of these isoforms were detected in tissue-associated leukocyte populations by PCR, but ERβ2 was rarely detected in PBLs. Expression of these receptors was temporally regulated in PBLs following polyclonal activation by concanavalin A, lipopolysaccharide or alloantigen based on evaluation by quantitative and end-point PCR. Examination of long-term leukocyte cell lines demonstrated that these receptors are differentially expressed depending on leukocyte lineage and phenotype. Expression of ERs was also temporally dynamic in some leukocyte lineages and may reflect stage of cell maturity. Estrogens affect the responsiveness of channel catfish peripheral blood leukocytes (PBLs) to mitogens in vitro. Similarly, bactericidal activity and phorbol 12-myristate 13-acetate induced respiratory burst was modulated by 17β-estradiol. These actions were blocked by the pure ER antagonist ICI 182780 indicating that response is, in part, mediated via ERα. In summary, estrogen receptors are expressed in channel catfish leukocytes and participate in the regulation of the immune response. This is the first time leukocyte lineage expression has been reported in teleost cell lines.

Persistent organochlorine pollutants such as polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethylene (p,p'-DDE), and polybrominated diphenyl ethers (PBDEs) are stable, bioaccumulative, and widely found in the environment, wildlife, and the human population. To explore the hypothesis that reproduction in male fish is associated with environmental exposures in the lower Columbia River (LCR), reproductive and endocrine parameters were studied in male resident, non-anadromous largescale sucker (Catostomus macrocheilus) (LSS) in the same habitats as anadromous salmonids having conservation status. Testes, thyroid tissue and plasma collected in 2010 from Longview (LV), Columbia City (CC), and Skamania (SK; reference) were studied. Sperm morphologies and thyrocyte heights were measured by light microscopy, sperm motilities by computer-assisted sperm motion analysis, sperm adenosine triphosphate (ATP) with luciferase, and plasma vitellogenin (VTG), thyroxine (T4), and triiodothyronine (T3) by immunoassay. Sperm apoptosis, viability, mitochondrial membrane potential, nuclear DNA fragmentation, and reproductive stage were measured by flow cytometry. Sperm quality parameters (except counts) and VTG were significantly different among sites, with correlations between VTG and 7 sperm parameters. Thyrocyte heights, T4, T3, gonadosomatic index and Fulton's condition factor differed among sites, but not significantly. Sperm quality was significantly lower and VTG higher where liver contaminants and water estrogen equivalents were highest (LV site). Total PCBs (specifically PCB-138, -146, -151, -170, -174, -177, -180, -183, -187, -194, and -206) and total PBDEs (specifically BDE-47, -100, -153, and -154) were negatively correlated with sperm motility. PCB-206 and BDE-154 were positively correlated with DNA fragmentation, and pentachloroanisole and VTG were positively correlated with sperm apoptosis and negatively correlated with ATP. BDE-99 was positively correlated with sperm counts and motility; T4 was negatively correlated with counts and positively correlated with motility, thus indicating possible androgenic mechanisms and thyroid endocrine disruption. Male LSS proved to be an informative model for studying reproductive and endocrine biomarkers in the LCR.

The health of Largescale Sucker (Catostomus macrocheilus) in the lower Columbia River (USA) was evaluated using morphometric and histopathological approaches, and its association with organic contaminants accumulated in liver was evaluated in males. Fish were sampled from three sites along a contaminant gradient. In 2009, body length and mass, condition factor, gonadosomatic index, and hematocrit were measured in males and females; liver and gonad tissue were collected from males for histological analyses; and organ composites were analyzed for contaminant content in males. In 2010, additional data were collected for males and females, including external fish condition assessment, histopathologies of spleen, kidney and gill and, for males, liver contaminant content. Multivariate analysis of variance indicated that biological traits in males, but not females, differed among sites in 2009 and 2010. Discriminant function analysis indicated that site-related differences among male populations were relatively small in 2009, but in 2010, when more variables were analyzed, males differed among sites in regards to kidney, spleen, and liver histopathologies and gill parasites. Kidney tubular hyperplasia, liver and spleen macrophage aggregations, and gill parasites were generally more severe in the downstream sites compared to the reference location. The contaminant content of male livers was also generally higher downstream, and the legacy pesticide hexachlorobenzene and flame retardants BDE-47 and BDE-154 were the primary drivers for site discrimination. However, bivariate correlations between biological variables and liver contaminants retained in the discriminant models failed to reveal associations between the two variable sets. In conclusion, whereas certain non-reproductive biological traits and liver contaminant contents of male Largescale Sucker differed according to an upstream-downstream gradient in the lower Columbia River, results from this study did not reveal the specific environmental factors responsible for the differences in health status among fish populations.

Toxic blooms of golden alga (GA, Prymnesium parvum) in Texas typically occur in winter or early spring. In North America, they were first reported in Texas in the 1980s, and a marked range expansion occurred in 2001. Although there is concern about the influence of climate change on the future distribution of GA, factors responsible for past dispersals remain uncertain. To better understand the factors that influence toxic bloom dispersal in reservoirs, this study characterized reservoir water quality associated with toxic GA blooms since 2001, and examined trends in water quality during a 20-year period bracketing the 2001 expansion. Archived data were analyzed for six impacted and six nonimpacted reservoirs from two major Texas basins: Brazos River and Colorado River. Data were simplified for analysis by pooling spatially (across sampling stations) and temporally (winter, December–February) within reservoirs and generating depth-corrected (1 m) monthly values. Classification tree analysis [period of record (POR), 2001–2010] using salinity-associated variables (specific conductance, chloride, sulfate), dissolved oxygen (DO), pH, temperature, total hardness, potassium, nitrate+nitrite, and total phosphorus indicated that salinity best predicts the toxic bloom occurrence. Minimum estimated salinities for toxic bloom formation were 0.59 and 1.02 psu in Brazos and Colorado River reservoirs, respectively. Principal component analysis (POR, 2001–2010) indicated that GA habitat is best defined by higher salinity relative to nonimpacted reservoirs, with winter DO and pH also being slightly higher and winter temperature slightly lower in impacted reservoirs. Trend analysis, however, did not reveal monotonic changes in winter water quality of GA-impacted reservoirs during the 20-year period (1991–2010) bracketing the 2001 dispersal. Therefore, whereas minimum levels of salinity are required for GA establishment and toxic blooms in Texas reservoirs, the lack of trends in water quality suggests that conditions favorable for toxic blooms pre-date the 2001 expansion. These observations are consistent with a climate change-independent scenario of past GA dispersals in Texas reservoirs driven by novel introductions into pre-existing favorable habitat. Reports of latent GA populations in certain nonimpacted reservoirs, however, provide a plausible scenario of future dispersals characterized by prolonged periods between colonization and toxic bloom development and driven by changes in water quality, natural, or anthropogenic.

Common Snook Centropomus undecimalis were once abundant off the Texas coast, but these populations are now characterized by low abundance and erratic recruitment. Most research concerning Common Snook in North America has been conducted in Florida and very little is known about the specific biology and habitat needs of Common Snook in Texas. The primary objective of this study was to describe the habitat use patterns of juvenile Common Snook and their role in the fish assemblage in the lower portion of the Rio Grande, Texas. Secondarily, we documented the relationship between age and juvenile reproductive development. Fish were collected during January-March 2006 from the lower 51.5 km of the Rio Grande using a bottom trawl and boat-mounted electrofisher. Measurements of water quality and other habitat traits were recorded at each sampling site. We captured 225 Common Snook exclusively in freshwater habitats above river kilometer 12.9. The distribution of juvenile Common Snook was not random, but influenced primarily by turbidity and dissolved oxygen. Sex differentiation and gonadal development based on histological examination of gonads established that age-1 and age-2 Common Snook were juvenile, prepubertal males. There was no difference between the age groups in their overall distribution in the river. However, age-2 Common Snook were associated with deeper areas with faster currents, higher conductivity, and steeper banks. Overall, Common Snook in the lower Rio Grande show substantial differences in habitat use than their counterparts in other parts of the range of the species, but it is unclear whether this is due to differences in habitat availability, behavioral plasticity, or some combination thereof.

We examined associations between thyroid condition, gonadal sex and pubertal development in zebrafish. Seventy-two-hour postfertilization larvae were reared in untreated medium or in the presence of goitrogens (sodium perchlorate, 0.82 mM; methimazole, 0.15 and 0.3 mM) or thyroxine (1 and 10 nM) for 30 days. Thyrocyte height, gonadal sex and gonadal development were histologically determined at 45 and 60 days postfertilization (dpf). Thyrocyte hypertrophy, an index of hypothyroidism, was observed at 45 and 60 dpf in perchlorate-treated but only at 45 dpf in methimazole-treated fish. Similarly, gonadal sex ratios were biased toward ovaries relative to control animals at 45 and 60 dpf in perchlorate-treated fish but only at 45 dpf in methimazole-treated fish. Gonadal sex ratios were biased toward testes at 45 and 60 dpf in thyroxine-treated fish. Spermatogenesis was delayed in testes from goitrogen-treated fish at 60 dpf relative to control values, but was unaffected in testes from thyroxine-treated individuals. Oogenesis seemed to be nonspecifically delayed in all treatments relative to control at 60 dpf. This study confirmed the previously reported association between hypothyroid condition and ovarian-skewed ratios, and hyperthyroid condition and testicular-skewed ratios, and also showed that male pubertal development is specifically delayed by experimental hypothyroidism. The simultaneous recovery from the hypothyroid and ovary-inducing effects of methimazole by 60 dpf (27 days post-treatment) suggests that the ovary-skewing effect of goitrogens is reversible when thyroid conditions return to basal levels before developmental commitment of gonadal sex. Conversely, the masculinizing effect of hyperthyroidism seems to be stable and perhaps permanent.

During a 2008 study to assess endocrine and reproductive health of common carp (Cyprinus carpio) in Lake Mead, Nevada (U.S.A.) we identified two fish, one male and one female, as hybrids with goldfish (Carassius auratus) based on morphology, lateral line scale count, and lack of anterior barbels. Gross examination of the female hybrid ovaries indicated presence of vitellogenic ovarian follicles; whereas histological evaluation of the male hybrid testes showed lobule-like structures with open lumens but without germ cells, suggesting it was sterile. Because common carp/goldfish hybrids are more susceptible to gonadal tumors and may have different endocrine profiles than common carp, researchers using common carp as a model for endocrine/reproductive studies should be aware of the possible presence of hybrids.

Several reservoirs in the upper Colorado River and Brazos River basins in Texas have experienced toxic blooms of golden alga Prymnesium parvum and associated fish kills since 2001. There is a paucity of information, however, regarding the population-level effects of such kills in large reservoirs, species-specific resistance to or recovery from kills, or potential differences in the patterns of impacts among basins. We used multiple before–after, control–impact analysis to determine whether repeated golden alga blooms have led to declines in the relative abundance and size structure of fish populations. Sustained declines were noted for 9 of 12 fish species surveyed in the upper Colorado River, whereas only one of eight species was impacted by golden alga in the Brazos River. In the upper Colorado River, White Bass Morone chrysops, White Crappie Pomoxis annularis, Largemouth Bass Micropterus salmoides, Bluegill Lepomis macrochirus, River Carpsucker Carpiodes carpio, Freshwater Drum Aplodinotus gr...

Salinity (5–30) effects on golden alga growth were determined at a standard laboratory temperature (22 °C) and one associated with natural blooms (13 °C). Inoculum-size effects were determined over a wide size range (100–100,000 cells ml −1 ). A strain widely distributed in the USA, UTEX-2797 was the primary study subject but another of limited distribution, UTEX-995 was used to evaluate growth responses in relation to genetic background. Variables examined were exponential growth rate ( r ), maximum cell density (max-D) and, when inoculum size was held constant (100 cells ml −1 ), density at onset of exponential growth (early-D). In UTEX-2797, max-D increased as salinity increased from 5 to ∼10–15 and declined thereafter regardless of temperature but r remained generally stable and only declined at salinity of 25–30. In addition, max-D correlated positively with r and early-D, the latter also being numerically highest at salinity of 15. In UTEX-995, max-D and r responded similarly to changes in salinity − they remained stable at salinity of 5–10 and 5–15, respectively, and declined at higher salinity. Also, max-D correlated with r but not early-D. Inoculum size positively and negatively influenced max-D and r , respectively, in both strains and these effects were significant even when the absolute size difference was small (100 versus 1000 cells ml −1 ). When cultured under similar conditions, UTEX-2797 grew faster and to much higher density than UTEX-995. In conclusion, (1) UTEX-2797’s superior growth performance may explain its relatively wide distribution in the USA, (2) the biphasic growth response of UTEX-2797 to salinity variation, with peak abundance at salinity of 10–15, generally mirrors golden alga abundance-salinity associations in US inland waters, and (3) early cell density – whether artificially manipulated or naturally attained – can influence UTEX-2797 bloom potential.

Channel catfish have an XX female-XY male sex determination system. Although genetic males of this species are readily feminized with exogenous steroid, the pattern of gonadal sex differentiation and the effect of temperature on genetic sex determination are unknown. To document the pattern of gonadal sex differentiation, catfish from a mixed-sex progeny (XX female-XY made cross), known XY males (XX female-YY male cross), and steroid sex-reversed (XY) females were reared at 28°C and collected for histological analysis from Day 7 to Day 90 postfertilization. No signs of gonadal sex differentiation were detected from Day 7 to Day 16. On Day 19, about half the fish from the mixed-sex population as well as the sex-reversed females had gonads with proximal and distal tissue outgrowths. These outgrowths eventually fused to form an ovarian cavity. Germ cell meiosis in these gonads was first detected on Day 22 and growing ovarian follicles were seen at later stages. Thus, ovarian differentiation began about Day 19. Although presumptive testes lose their sensitivity to feminizing steroids about Day 19, the gonads of known males and of putative males from the mixed-sex population showed no clear signs of sex differentiation up to Day 90. Thus, the prolonged temporal dissociation between testicular commitment and differentiation indicates that these two events may be mediated by separate stimuli. To determine the effects of temperature on sex determination, fish of a mixed-sex population were reared at 27°C until Day 10; at 20°, 27°, or 34°C from Day 10 to Day 24; and then at 27°C until Day 102 when sex ratios and gonadal histology were determined. The sex ratio was significantly skewed toward females in fish treated at 34°C (1 male:1.68 female; P < 0.01) but no effects were seen at 20° or 27°C. This observation suggests that genetic and temperature-dependent mechanisms of sex determination are both functional in channel catfish and that this phenomenon is more widespread in fishes than previously believed. The testes of the Day 102 males examined had developed positive histological signs of sex differentiation, suggesting that testicular formation in channel catfish normally starts between Day 90 and 102. © 1996 Wiley-Liss, Inc.

Freshwater salinization is a global concern partly attributable to anthropogenic salt contamination. The authors examined the effects of increased salinity (as NaCl, 250–4,000 µS/cm, specific conductance) on two sympatric freshwater gastropods (Helisoma trivolvis and Physa pomillia). Life stage sensitivities were determined by exposing naive eggs or naive juveniles (through adulthood and reproduction). Additionally, progeny eggs from the juvenile-adult exposures were maintained at their respective parental salinities to examine transgenerational effects. Naive H. trivolvis eggs experienced delayed development at specific conductance > 250 µS/cm; reduced survivorship and reproduction were also seen in juvenile H. trivolvis at 4,000 µS/cm. Survival and growth of P. pomilia were not affected by increased salinity following egg or juvenile exposures. Interestingly, the progeny of H. trivolvis exposed to higher salinity may have gained tolerance to increased salinity whereas P. pomilia progeny may have experienced negative transgenerational effects. The present study demonstrates that freshwater snail species vary in their tolerance to salinization and also highlights the importance of multigenerational studies, as stressor impacts may not be readily apparent from shorter term exposures. Environ. Toxicol. Chem. 2012; 31: 2517–2524. © 2012 SETAC

Campbell, M. D., Patino, R., Tolan, J., Strauss, R., and Diamond, S. L. 2010. Sublethal effects of catch-and-release fishing: measuring capture stress, fish impairment, and predation risk using a condition index. – ICES Journal of Marine Science, 67: 513–521. The sublethal effects of simulated capture of red snapper (Lutjanus campechanus) were analysed using physiological responses, condition indexing, and performance variables. Simulated catch-and-release fishing included combinations of depth of capture and thermocline exposure reflective of environmental conditions experienced in the Gulf of Mexico. Frequency of occurrence of barotrauma and lack of reflex response exhibited considerable individual variation. When combined into a single condition or impairment index, individual variation was reduced, and impairment showed significant increases as depth increased and with the addition of thermocline exposure. Performance variables, such as burst swimming speed (BSS) and simulated predator approach distance (AD), were also significantly different by depth. BSSs and predator ADs decreased with increasing depth, were lowest immediately after release, and were affected for up to 15 min, with longer recovery times required as depth increased. The impairment score developed was positively correlated with cortisol concentration and negatively correlated with both BSS and simulated predator AD. The impairment index proved to be an efficient method to estimate the overall impairment of red snapper in the laboratory simulations of capture and shows promise for use in field conditions, to estimate release mortality and vulnerability to predation.

Xenopus laevis larvae were exposed to cadmium (Cd) at 0, 1, 8, 85 or 860 μg L −1 in FETAX medium from 0 to 86 d postfertilization. Premetamorphic tadpoles were sampled on day 31; pre and prometamorphic tadpoles on day 49; and frogs (NF stage 66) between days 50 and 86. Survival, snout-vent length (SVL), tail length, total length, hindlimb length (HLL), initiation of metamorphic climax, size at and completion of metamorphosis, and gonadal condition and sex ratio (assessed histologically) were determined. Survival was unaffected by Cd until day 49, but increased mortality was observed after day 49 at 860 μg Cd L −1 . On day 31, when tadpoles were in early premetamorphosis, inhibitory effects on tadpole growth were observed only at 860 μg Cd L −1 . On day 49, when most tadpoles where in late premetamorphosis/early prometamorphosis, reductions in SVL, HLL and total length were observed at 8 and 860 but not 85 μg L −1 , thus creating a U-shaped size distribution at 0–85 μg Cd L −1 . However, this U-shaped size pattern was not evident in postmetamorphic individuals. In fact, frog size at completion of metamorphosis was slightly smaller at 85 μg Cd L −1 relative to control animals. These observations confirmed a recent report of a Cd concentration-dependent bimodal growth pattern in late-premetamorphic Xenopus tadpoles, but also showed that growth responses to varying Cd concentrations change with development. The fraction of animals initiating or completing metamorphosis during days 50–86 was reduced in a Cd concentration-dependent manner. Testicular histology and population sex ratios were unaffected by Cd suggesting that, unlike mammals, Cd is not strongly estrogenic in Xenopus tadpoles.

International audience; Although estrogens exert many functions on vertebrate brains, there is little information on the relationship between brain aromatase and estrogen receptors. Here, we report the cloning and characterization of two estrogen receptors, alpha and beta, in pejerrey. Both receptors' mRNAs largely overlap and were predominantly expressed in the brain, pituitary, liver, and gonads. Also brain aromatase and estrogen receptors were up-regulated in the brain of estradiol-treated males. In situ hybridization was performed to study in more detail, the distribution of the two receptors in comparison with brain aromatase mRNA in the brain of adult pejerrey. The estrogen receptors' mRNAs exhibited distinct but partially overlapping patterns of expression in the preoptic area and the mediobasal hypothalamus, as well as in the pituitary gland. Moreover, the estrogen receptor alpha, but not beta, were found to be expressed in cells lining the preoptic recess, similarly as observed for brain aromatase. Finally, it was shown that the onset expression of brain aromatase and both estrogen receptors in the head of larvae preceded the morphological differentiation of the gonads. Because pejerrey sex differentiation is strongly influenced by temperature, brain aromatase expression was measured during the temperature-sensitive window and was found to be significantly higher at male-promoting temperature. Taken together these results suggest close neuroanatomical and functional relationships between brain aromatase and estrogen receptors, probably involved in the sexual differentiation of the brain and raising interesting questions on the origin (central or peripheral) of the brain aromatase substrate.

The post-vitellogenic oocytes of teleost fish are generally unresponsive to maturation-inducing hormone (MIH) until a luteinizing hormone (LH) surge stimulates sensitivity via the acquisition of oocyte-maturational competence (OMC). Heterologous gap junctions (GJs) between granulosa cells and the oocyte have been previously implicated in the regulation of oocyte maturation in various vertebrate species. Although heterologous GJ are present in ovarian follicles of ayu (Plecoglossus altivelis), their role in maturation remains unclear. In the present study, we cloned and characterized complementary DNAs for GJ protein connexin (Cx), and examined the expression pattern of Cx messenger RNAs in the ayu ovary. Four Cx cDNAs with predicted molecular masses of 32.1 (Cx32.1), 34.9 (Cx34.9), 44.1 (Cx44.1), and 44.2 (Cx44.2) kDa, respectively, were cloned. Northern blot analysis revealed that the levels of Cx44.1 and Cx44.2 transcripts were similar during the vitellogenic and ovulatory stages. Cx32.1 transcripts were more abundant during the vitellogenic stage, but their levels declined thereafter. Cx34.9 transcript levels increased during the vitellogenic stage and remained high during the acquisition of OMC. In situ hybridization revealed that Cx44.1 and Cx44.2 signals were restricted to the oocyte, whereas the Cx32.1 and Cx34.9 signals were detected in both cellular fractions. Furthermore, a dye-transfer assay revealed the presence of functional GJs between the oocytes and follicle cells. These results suggest that Cx34.9 contributes to the formation of heterologous GJs between oocytes and granulosa cells. Moreover, GJs formed by Cx34.9 may function during the LH-dependent acquisition of OMC and the MIH-dependent resumption of meiosis in ayu. Mol. Reprod. Dev. 74: 1113–1123, 2007. © 2007 Wiley-Liss, Inc.

The objectives of this study were to determine the effects of prolonged exposure to perchlorate on (1) thyroid status and reproductive performance of adult zebrafish (Danio rerio) and (2) F1 embryo survival and early larval development. Using a static-renewal procedure, mixed sex populations of adult zebrafish were exposed to 0, 10, and 100 mg/l nominal concentrations of waterborne perchlorate for 10 weeks. Thyroid histology was qualitatively assessed, and females and males were separated and further exposed to their respective treatments for six additional weeks. Eight females in each tank replicate (n = 3) were paired weekly with four males from the same respective treatment, and packed-egg (spawn) volume (PEV) was measured each of the last five weeks. At least once during weeks 14-16 of exposure, other end points measured included fertilization rate, fertilized egg diameter, hatching rate, standard length, and craniofacial development of 4-day-postfertilization larvae and thyroid hormone content of 3.5-h embryos and of exposed mothers. At 10 weeks of exposure, perchlorate at both concentrations caused thyroidal hypertrophy and colloid depletion. A marked reduction in PEV was observed toward the end of the 6-week spawning period, but fertilization and embryo hatching rates were unaffected. Fertilized egg diameter and larval length were increased by parental exposure to perchlorate. Larval head depth was unaffected but the forward protrusion of the lower jaw-associated cartilage complexes, Meckel's and ceratohyal, was decreased. Exposure to both concentrations of perchlorate inhibited whole-body thyroxine content in mothers and embryos, but triiodothyronine content was unchanged. In conclusion, prolonged exposure of adult zebrafish to perchlorate not only disrupts their thyroid endocrine system but also impairs reproduction and influences early F1 development.

Hexahydro-1,3,5-trinitro-1,3,5-triazine, a cyclonitramine commonly known as RDX, is used in the production of military munitions. Contamination of soil, sediment, and ground and surface waters with RDX has been reported in different places around the world. Acute and subacute toxicities of RDX have been relatively well documented in terrestrial vertebrates, but among aquatic vertebrates the information available is limited. The objective of this study was to characterize the acute toxicity of RDX to larval zebrafish. Mortality (LC50) and incidence of vertebral column deformities (EC50) were two of the end points measured in this study. The 96-h LC50 was estimated at 22.98 and 25.64 mg l −1 in two different tests. The estimated no-observed-effective-concentration (NOEC) values of RDX on lethality were 13.27 ± 0.05 and 15.32 ± 0.30 mg l −1 ; and the lowest-observed-effective-concentration (LOEC) values were 16.52 ± 0.05 and 19.09 ± 0.23 mg l −1 in these two tests, respectively. The 96-h EC50 for vertebral deformities on survivors from one of the acute lethality tests was estimated at 20.84 mg l −1 , with NOEC and LOEC of 9.75 ± 0.34 and 12.84 ± 0.34 mg l −1 , respectively. Behavioral aberrations were also noted in this acute toxicity study, including the occurrence of whirling movement and lethargic behavior. The acute effects of RDX on survival, incidence of deformities, and behavior of larval zebrafish occurred at the high end of the most frequently reported concentrations of RDX in aquatic environments. The chronic effects of RDX in aquatic vertebrates need to be determined for an adequate assessment of the ecological risk of environmental RDX.

Short-term exposures to elevated temperatures reduce germ cell numbers in the gonads of some fishes, suggesting that heat treatment may be a viable method for delaying or suppressing reproductive development in cultured fishes. The objective of this study was to determine the effects of high temperatures on early gonadal development in channel catfish Ictalurus punctatus. Twenty-three-day-old (after hatching) channel catfish were exposed to temperatures of 27°C (control), 34°C, and 36°C for 4 weeks. Groups of 250 fry were placed in individual 152-L glass aquaria with external biofilters, and four replicated aquaria were used per temperature treatment. Water temperature was regulated with submersible heaters. At the end of treatment, body weight and standard length were measured in 30 individuals per aquarium and gonads were examined histologically in 10 individuals of each sex per aquarium. For statistical analysis, the unit of replication was the aquarium, and differences between treatments were...

Environmental contamination of ground and surface waters by perchlorate, derived from ammonium perchlorate (AP) and other perchlorate salts, is of increasing concern. Exposure to perchlorate can impair the thyroid endocrine system, which is thought to modulate renal and immune function in vertebrates. This study with zebrafish Danio rerio and eastern mosquitofish Gambusia holbrooki examined the histological effects of perchlorate on the trunk kidney, which in teleosts serves excretory and hemopoietic functions and therefore may be a target of perchlorate effects. Adult zebrafish of both sexes were exposed in the laboratory to waterborne, AP-derived perchlorate at measured concentrations of 18 mg/L for 8 weeks. Adult male mosquitofish were exposed to waterborne sodium perchlorate at measured perchlorate concentrations of 1–92 mg/L for 8 weeks. Control fish were kept in untreated water. The region of the body cavity containing the trunk kidney was processed from each fish for histological analysis....

Estrogens are important regulators of physiological functions. Although environmental contaminants (xenoestrogens) which interfere with estrogen signaling are of increasing concern, there is only limited information about their ability to interact with estrogen-binding proteins (SHBG) or receptors (ER). Recombinant ERalpha and beta were obtained after transient transfection of COS-7 cells with channel catfish ER cDNA. Plasma from adult female channel catfish was the source of SHBG. Tritiated estradiol (3H-E2) was used in standard radioligand-binding assays to characterize the binding properties of channel catfish SHBG (ccfSHBG) and to estimate the inhibition constants for various estrogenic compounds. Binding of 3H-E2 to ccfSHBG was saturable and of high affinity with a Kd (+/-SE) of 1.9+/-0.14 nM and a Bmax of 14.3+/-2.4 pmol/mg protein ( n = 3 assays). Additionally, ccfSHBG displayed binding specificity for androgens and estrogens. Endosulfan, 4-nonylphenol, and 4-octylphenol displaced 3H-E2 binding to ccfSHBG albeit only at very high concentrations, whereas dieldrin and atrazine showed little displacement activity even at the highest concentrations used. The synthetic estrogen ethynylestradiol had higher affinity than E2 for ccfSHBG. This finding differs from results with human and rainbow trout SHBG. The alkylphenolic compounds (4-octylphenol and 4-nonylphenol) displayed some ability to displace 3H-E2 binding from ERalpha and beta at high concentrations, but dieldrin and atrazine had little binding activity for both ER subtypes and endosulfan for ERbeta. The xenobiotics tested generally showed equivalent or greater affinity for ERalpha than ERbeta, whereas natural estrogens had much greater affinity for ERbeta than ERalpha. These observations suggest that results of studies using fish tissue ER extracts must be interpreted with caution, since both ER subtypes may be present, and that the binding of xenoestrogens to SHBG must be taken into account for proper assessment of endocrine disruption caused by environmental contaminants.

Adult zebrafish were reared up to eight weeks in control water or in water containing ammonium perchlorate (AP) at measured perchlorate concentrations of 18 (environmentally relevant, high) and 677 ppm. Groups of eight females were paired with four males on a weekly basis to assess AP effects on spawned egg volume, an index of reproductive performance. All treatments were applied to four to five spawning replicates. At 677 ppm, spawn volume was reduced within one week and became negligible after four weeks. At 18 ppm, spawn volume was unaffected even after eight weeks. Also, perchlorate at 18 ppm did not affect percentage egg fertilization. Fish were collected at the end of the exposures (677 ppm, four weeks; control and 18 ppm, eight weeks) for whole-body perchlorate content and thyroid histopathological analysis. Fish perchlorate levels were about one-hundredth of those of treatment water levels, indicating that waterborne perchlorate does not accumulate in whole fish. At 677 ppm for four weeks, perchlorate caused thyroid follicle cell (nuclear) hypertrophy and angiogenesis, whereas at 18 ppm for eight weeks, its effects were more pronounced and included hypertrophy, angiogenesis, hyperplasia, and colloid depletion. In conclusion, an eight-week exposure of adult zebrafish to 18 ppm perchlorate (high environmentally relevant concentrations) affected the histological condition of their thyroid follicles but not their reproductive performance. The effect of 677 ppm perchlorate on reproduction may be due to extrathyroidal toxicity. Further research is needed to determine if AP at lower environmentally relevant concentrations also affects the thyroid follicles of zebrafish.

This study examined the hypothesis that exposure to sublethal concentrations of contaminants alters the gonadal condition of feral common carp Cyprinus carpio. Adult common carp in Lake Mead, Nevada, were collected from a contaminated site (Las Vegas Bay) that receives municipal and industrial effluent and from a reference site (Overton Arm) with a relatively low level of contamination. Fish were sampled seven times over a 1-year period extending over two separate spawning seasons. Morphometric and histopathological parameters of gonadal and germ cell development were determined. In males, the pattern of seasonal changes in the gonadosomatic index (GSI) was similar between the sites and showed no clear association with site-specific seasonal temperature profiles. However, Las Vegas Bay males had consistently lower GSI values and, on one of the sampling dates, a lower proportion of sperm relative to other germ cell stages (determined histologically). Further, Las Vegas Bay males had a higher incid...

The roles of arachidonic acid (AA) and protein kinase C (PKC) during in vitro maturation-inducing hormone (MIH)-dependent meiotic resumption (maturation) and ovulation were studied in ovarian follicles of Atlantic croaker (Micropogonias undulatus). The requirement for cyclooxygenase (COX) metabolites of AA was examined using a nonspecific COX inhibitor, indomethacin (IM), as well as two COX products, prostaglandin (PG) F2α and PGE2, whereas the role of lipoxygenase (LOX) was investigated using a specific LOX inhibitor, nordihydroguaiaretic acid (NDGA). The involvement of PKC was examined using phorbol 12-myristate 13-acetate (PMA), a PKC activator, as well as GF109203X (GF), a specific inhibitor of PKC and 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7), nonspecific inhibitor of protein kinases. Genomic mechanisms were examined with the transcription-inhibitor actinomycin D (ActD) and the functionality of heterologous (oocyte-granulosa) gap junctions (GJ) with a dye transfer assay. The AA (100...

Numerous studies with teleosts have addressed the regulation and mechanisms of oocyte maturation, but largely at the exclusion of ovulation. A smaller but still considerable number of studies have focused on ovulation, and ignored maturation. Consequently, little is known about the mechanistic linkages between these two events. New information is presented here indicating that luteinizing hormone regulates the acquisition not only of oocyte maturational competence, but also ovulatory competence. The thesis is presented that maturation and ovulation are closely integrated and overlapping events that are best viewed conceptually and experimentally as parts of a functional whole.