ABSTRACT
Persistent organic pollutants (POPs) are highly lipophilic compounds that accumulate at increased concentrations in high tropic level organisms like marine mammals. Marine mammals' reliance on blubber makes them susceptible to accumulating POPs at potentially toxic concentrations. In this study, we analyzed POP concentrations, (polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers (PBDEs), and methoxylated-BDE (MeOBDE), in the blubber of 16 subsistence harvested sub-adult, male northern fur seals as well as assessed changes in mRNA gene expression of nine relevant biomarkers including the aryl hydrocarbon receptor, thyroid receptor-α, and adiponectin. PBDE and MeOBDE concentrations were significantly lower than PCB and OCP concentrations. A negative relationship was observed between percent lipid in the blubber and contaminant concentrations, both individual and sum. Expression changes in eight biomarkers were correlated with individual and sum contaminant concentrations. This study shows that contaminant concentrations measured are correlated to changes in expression of genes from different physiological systems, metabolism and endocrine, that are important for the regulation of blubber metabolism. Northern fur seals are reliant on blubber as an energy source during times of low food intake. Potential contaminant induced changes in blubber metabolism pathways could have significant impacts on the health of individuals during critical periods.
Subject(s)
Fur Seals , Polychlorinated Biphenyls , Water Pollutants, Chemical , Animals , Male , Adipose Tissue/chemistry , Adipose Tissue/metabolism , Alaska , Biomarkers/metabolism , Environmental Monitoring , Fur Seals/metabolism , Gene Expression , Halogenated Diphenyl Ethers/toxicity , Halogenated Diphenyl Ethers/metabolism , Persistent Organic Pollutants/metabolism , Polychlorinated Biphenyls/analysis , Water Pollutants, Chemical/analysisABSTRACT
Current evidence suggests thyroid hormones (THs) impact development of the immune system, but few studies have explored the connection between the thyroid and immune systems, especially in fish. This is important as some environmental contaminants disrupt TH homeostasis and may thus have negative impacts on the immune system. To determine the long-term consequences of early life stage (ELS) hypothyroidism on immune function, fathead minnows were exposed to the model thyroid hormone suppressant propylthiouracil (PTU) from < 1 to 30 days post hatch. Fish were transferred to clean water and raised to adulthood (5-7 months post hatch) at which time, several aspects of immune function were evaluated. Ex vivo assessment of immune cell function revealed significant decreases (1.2-fold) in the phagocytic cell activity of PTU-treated fish relative to the controls. Fish were also injected with Yersinia ruckeri to evaluate their in vivo immune responses across a suite of endpoints (i.e., transcriptomic analysis, leukocyte counts, spleen index, hematocrit, bacterial load and pathogen resistance). The transcriptomic response to infection was significantly different between control and PTU-treated fish, though no differences in bacterial load or pathogen resistance were noted. Overall, these results suggest that early life stage TH suppression causes long-term impacts on immune function at the molecular and cellular levels suggesting a key role for TH signaling in normal immune system development. This study lays the foundation for further exploration into thyroid-immune crosstalk in fish. This is noteworthy as disruption of the thyroid system during development, which can occur in response to chemicals present in the environment, may have lasting effects on immune function in adulthood.
Subject(s)
Cyprinidae/immunology , Fish Diseases/immunology , Host-Pathogen Interactions/immunology , Thyroid Gland/physiopathology , Yersinia Infections/veterinary , Animals , Antithyroid Agents/pharmacology , Female , Fish Diseases/microbiology , Fish Proteins/genetics , Gene Expression Regulation , Host-Pathogen Interactions/physiology , Immunity, Cellular , Immunity, Innate , Kidney/microbiology , Kidney/physiology , Propylthiouracil/pharmacology , Thyroid Gland/drug effects , Thyroid Gland/immunology , Thyroid Hormones/metabolism , Yersinia Infections/immunology , Yersinia Infections/microbiology , Yersinia ruckeriABSTRACT
Systemic acquired resistance (SAR) is an inducible defense mechanism that systemically enhances resistance against pathogens in foliar tissues. SAR, which engages salicylic acid (SA) signaling, shares molecular components with the autonomous pathway, which is involved in controlling flowering time in Arabidopsis thaliana. FLOWERING LOCUS D (FLD) is one such autonomous pathway component that is required for flowering time and the systemic accumulation of SA during SAR. Here, we show that CYP720A1, a putative cytochrome P450 monoxygenase, controls FLD expression and is required for the timing of flowering and the manifestation of SAR. The delayed flowering time in the cyp720a1 mutant correlated with the elevated transcript level of the floral repressor FLC, while the SAR deficiency phenotype of the cyp720a1 mutant correlated with the inability to systemically accumulate SA. CYP720A1 transcript abundance in shoots is poor compared with roots. Reciprocal root-shoot grafting confirmed that CYP720A1 function in the roots is critical for flowering time and SAR. We therefore suggest that root to shoot communication involving a CYP720A1-dependent factor contributes to the timing of reproductive development and defense in the foliage.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Flowers/genetics , Flowers/metabolism , Gene Expression Regulation, Plant , Immunity, Innate , MADS Domain Proteins/genetics , Salicylic AcidABSTRACT
Measures of respiratory burst and phagocytic cell activity are frequently utilized to assess cellular immune function in teleosts. Respiratory burst predominately occurs in neutrophils and causes the release of reactive oxygen species to kill pathogens. Phagocytosis is the process by which pathogens are engulfed and destroyed by various immune cells. Though a variety of approaches have been utilized to measure respiratory burst and phagocytic cell activity, assays that rely only on common laboratory equipment (e.g., plate reader) may offer advantages over those that rely on more specialized equipment (e.g., flow cytometer). The goal of the current study was to optimize and validate the use of a colorimetric plate-based respiratory burst and fluorometric plate-based phagocytic cell activity assays for use with kidney cells from the fathead minnow (Pimephales promelas), an emerging immunotoxicity model. In addition, a protocol for the dissection of kidney tissue followed by the extraction of kidney cells, as well as recommendations and resources for future experiments utilizing each of these assays, are provided.â¢All methods are optimized for use with the fathead minnow or similar teleost species.â¢Respiratory burst and phagocytic cell activity are measured using a standard plate reader.
ABSTRACT
Recent studies have utilized the fathead minnow (Pimephales promelas) to explore the immunotoxic effects associated with a variety of environmental contaminants in the absence of immunological stimuli. Though this approach allows for alterations in the resting immune system to be detected, previous evidence suggests that many immunotoxic effects may only manifest in the activated immune system. However, basic immune responses to pathogens have not been well described in this species. To expand the utility of the fathead minnow as a model for immunotoxicity testing, a more comprehensive understanding of the activated immune system is required. As such, the main goal of this study was to characterize the transcriptomic response to pathogen infection in the fathead minnow using RNA sequencing. To achieve this goal, female fathead minnows were intraperitoneally injected with either Hank's Balanced Salt Solution (sham-injected) or Yersinia ruckeri (pathogen-injected). Eight hours following injection, fish were sacrificed for the assessment of general morphological (i.e., mass, length, condition factor, hepatic index) and immunological (i.e., leukocyte counts, spleen index) endpoints. To assess the molecular immune response to Y. ruckeri, kidney tissue was collected for transcriptomic analysis. A comparison of sham- and pathogen-injected fish revealed that >1800 genes and >500 gene networks were differentially expressed.Gene networks associated with inflammation, innate immunity, complement, hemorrhaging and iron absorption are highlighted and their utility within the context of immunotoxicity is discussed. These data reveal pathogen-related molecular endpoints to improve data interpretation of future studies utilizing the fathead minnow as a model for immunotoxicity.
Subject(s)
Cyprinidae , Fish Diseases/immunology , Immunity, Innate , Transcriptome/immunology , Yersinia Infections/veterinary , Animals , Female , Fish Diseases/microbiology , Kidney/immunology , Kidney/microbiology , Models, Animal , Yersinia Infections/immunology , Yersinia Infections/microbiology , Yersinia ruckeri/physiologyABSTRACT
Abietane diterpenoids are tricyclic diterpenes whose biological functions in angiosperms are largely unknown. Here, we show that dehydroabietinal (DA) fosters transition from the vegetative phase to reproductive development in Arabidopsis thaliana by promoting flowering time. DA's promotion of flowering time was mediated through up-regulation of the autonomous pathway genes FLOWERING LOCUS D (FLD), RELATIVE OF EARLY FLOWERING 6 (REF6), and FVE, which repress expression of FLOWERING LOCUS C (FLC), a negative regulator of the key floral integrator FLOWERING LOCUS T (FT). Our results further indicate that FLD, REF6, and FVE are also required for systemic acquired resistance (SAR), an inducible defense mechanism that is also activated by DA. However, unlike flowering time, FT was not required for DA-induced SAR. Conversely, salicylic acid, which is essential for the manifestation of SAR, was not required for the DA-promoted flowering time. Thus, although the autonomous pathway genes FLD, REF6, and FVE are involved in SAR and flowering time, these biological processes are not interdependent. We suggest that SAR and flowering time signaling pathways bifurcate at a step downstream of FLD, REF6, and FVE, with an FLC-dependent arm controlling flowering time, and an FLC-independent pathway controlling SAR.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Abietanes , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Flowers/genetics , Flowers/metabolism , Gene Expression Regulation, Plant , MADS Domain Proteins/genetics , MADS Domain Proteins/metabolism , Mutation , Transcription FactorsABSTRACT
Lignin biosynthesis is evolutionarily conserved among higher plants and features a critical 3-hydroxylation reaction involving phenolic esters. However, increasing evidence questions the involvement of a single pathway to lignin formation in vascular plants. Here we describe an enzyme catalyzing the direct 3-hydroxylation of 4-coumarate to caffeate in lignin biosynthesis as a bifunctional peroxidase that oxidizes both ascorbate and 4-coumarate at comparable rates. A combination of biochemical and genetic evidence in the model plants Brachypodium distachyon and Arabidopsis thaliana supports a role for this coumarate 3-hydroxylase (C3H) in the early steps of lignin biosynthesis. The subsequent efficient O-methylation of caffeate to ferulate in grasses is substantiated by in vivo biochemical assays. Our results identify C3H as the only non-membrane bound hydroxylase in the lignin pathway and revise the currently accepted models of lignin biosynthesis, suggesting new gene targets to improve forage and bioenergy crops.
Subject(s)
Cytosol/enzymology , Lignin/biosynthesis , Arabidopsis/metabolism , Ascorbate Peroxidases , Brachypodium/metabolism , Caffeic Acids/metabolism , Coumaric Acids/metabolism , Mixed Function Oxygenases/genetics , Mixed Function Oxygenases/metabolism , Plant Proteins/genetics , Plant Proteins/metabolismABSTRACT
Synthetic sex steroids, like the synthetic progestin norethindrone (NET), can affect a wide variety of biological processes via highly conserved mechanisms. NET is prevalent in surface waters, yet the sub-lethal effects of NET exposure are not are net yet well characterized in aquatic biota. A few targeted gene expression and behavioral studies have concluded that NET affects the vision of adult fish; however, early life stage (ELS) fish are often more sensitive to contaminants. Furthermore, many species of fish rely heavily on visual perception for survival during development. The goal of the present study was to characterize the effects of developmental exposure to environmentally relevant concentrations of NET on the visual system of ELS zebrafish, using transcriptomics and histological methods. Results indicate that exposure to relatively low levels of NET in aquatic systems may be sufficient to affect the visual function of developing fish.
Subject(s)
Contraceptives, Oral, Synthetic/toxicity , Gene Expression Regulation, Developmental/drug effects , Norethindrone/toxicity , Transcriptome/drug effects , Vision, Ocular/genetics , Water Pollutants, Chemical/toxicity , Zebrafish/genetics , Animals , Embryo, Nonmammalian/drug effects , Eye/anatomy & histology , Eye/drug effectsABSTRACT
Metformin is one of the most prevalent pharmaceuticals in both surface and waste waters, yet little is known about the bioavailability and/or effects of developmental exposure on early life stage (ELS) fish. Here, we demonstrate that embryo-larval stages of medaka are capable of taking up metformin from the aquatic environment, provided exposure occurs prior to chorion hardening (â¼6-hpf). Once transferred to clean water, ELS medaka are able to completely depurate metformin in <24-hours. Furthermore, ELS medaka exposed to a range of relevant concentrations of waterborne metformin (from 6 hpf through 28-days post hatch) had significantly reduced growth metrics, altered metabolomes, and changes in the expression of genes associated with cell growth. The range of concentrations investigated were 1.0, 3.2, 10, 32, and 100 µg·L-1. To examine effects of chronic, low level metformin exposure across the full medaka life-cycle, we exposed newly fertilized embryos to 3.2 µg L-1 waterborne metformin for 165-days. The weight and length of adult fish were examined, as were effects on the production of some steroid hormones, specifically a significant increase (control females: 0.161 ± 0.023 pg/mg; metformin treated females: 3.42 ± 0.543) in the production of 11-ketotestosterone was observed in adult female medaka. Collectively, these results suggest that current environmental exposure scenarios may be sufficient to cause effects on developing fish.
Subject(s)
Embryo, Nonmammalian/drug effects , Environmental Exposure , Metformin/toxicity , Oryzias , Animals , Female , Fish Proteins/genetics , Gene Expression Regulation/drug effects , Water Pollutants, Chemical/toxicityABSTRACT
Mercury is a global contaminant, which may be microbially transformed into methylmercury (MeHg), which bioaccumulates. This results in potentially toxic body burdens in high trophic level organisms in aquatic ecosystems and maternal transfer to offspring. We previously demonstrated effects on developing fish including hyperactivity, altered time-to-hatch, reduced survival, and dysregulation of the dopaminergic system. A link between gut microbiota and central nervous system function in teleosts has been established with implications for behavior. We sequenced gut microbiomes of fathead minnows exposed to dietary MeHg to determine microbiome effects. Dietary exposures were repeated with adult CD-1 mice. Metabolomics was used to screen for metabolome changes in mouse brain and larval fish, and results indicate effects on lipid metabolism and neurotransmission, supported by microbiome data. Findings suggest environmentally relevant exposure scenarios may cause xenobiotic-mediated dysbiosis of the gut microbiome, contributing to neurotoxicity. Furthermore, small-bodied teleosts may be a useful model species for studying certain types of neurodegenerative diseases, in lieu of higher vertebrates.
Subject(s)
Cyprinidae , Gastrointestinal Microbiome , Water Pollutants, Chemical , Animals , Ecosystem , Metabolome , MiceABSTRACT
Exposures to polybrominated diphenyl ethers (PBDEs) have been shown to alter immune function in adult organisms across a variety of taxa. However, few if any studies have investigated the long-term consequences of early life stage PBDE exposures on immune function in fish. This study sought to determine the effects of early life stage BDE-47 exposure on pathogen resistance in the fathead minnow (Pimephales promelas) following an extended depuration period (≥180â¯d). Minnows were exposed to BDE-47 via a combination of maternal transfer and diet through 34 days post fertilization (dpf), raised to adulthood (>215 dpf) on a clean diet, then subjected to pathogen resistance trials. Early life stage exposures to BDE-47 did not affect the ability of females to survive from Yersinia ruckeri infection. However, the survival of BDE-47 exposed males was significantly reduced relative to controls, indicating that developmental exposures to BDE-47 altered male immunity. Because BDE-47 is a known thyroid hormone disruptor and thyroid hormone disruptors have the potential to adversely impact immune development and function, metrics indicative of thyroid disruption were evaluated, as were immune parameters known to be altered in response to thyroid disruption. BDE-47 exposed minnows exhibited signs of thyroid disruption (i.e., reduced growth); however, no alterations were observed in immune parameters known to be influenced by thyroid hormones (i.e., thymus size, expression of genes associated with lymphoid development) suggesting that the observed alterations in immunocompetence may occur through alternative mechanisms. Regardless of the mechanisms responsible, the results of this study demonstrate the potential for early life stage PBDE exposures to adversely impact immunity and illustrate that the immunological consequences of PBDE exposures are sex dependent.
Subject(s)
Cyprinidae/immunology , Disease Resistance/drug effects , Endocrine Disruptors/toxicity , Flame Retardants/toxicity , Halogenated Diphenyl Ethers/toxicity , Animals , Cyprinidae/genetics , Embryo, Nonmammalian , Female , Gene Expression Regulation/drug effects , Larva , Male , Sex Characteristics , Thymus Gland , Thyroid Gland , Yersinia Infections/immunology , Yersinia Infections/veterinary , Yersinia ruckeriABSTRACT
Docosahexaenoic acid (DHA) is an important and abundant fatty acid moiety in vertebrate brains. We measured brain phospholipid composition during development in red-winged blackbirds (Agelaius phoeniceus), an altricial species that breeds in aquatic habitats. We also manipulated diet by feeding nestlings fish oil or sunflower oil. Finally, we assessed selective uptake of yolk by comparing the yolk fatty acid composition of freshly laid eggs and day-old hatchlings. Relative to other altricial species, blackbirds achieved high DHA in brain phospholipids (20% of phospholipid fatty acids in day-old hatchlings). This was not a result of selective uptake from the yolk, but rather a consequence of a high proportion of DHA in the yolk (2.5% of total lipids) at laying. Our dietary study confirmed that nestling brains are sensitive to fatty acid supply. Red-winged blackbirds may be able to advance cognitive development relative to other altricial species owing to their aquatic maternal diet.
Subject(s)
Brain Chemistry , Diet/veterinary , Docosahexaenoic Acids/metabolism , Egg Yolk/chemistry , Songbirds/metabolism , Animal Feed/analysis , Animals , Embryo, Nonmammalian/chemistry , Embryonic Development , Fish Oils/administration & dosage , Fish Oils/metabolism , Random Allocation , Songbirds/embryology , Sunflower Oil/administration & dosage , Sunflower Oil/metabolismABSTRACT
Polybrominated diphenyl ethers (PBDEs) are highly lipophilic components of brominated flame retardants that are environmentally persistent and bioaccumulate. PBDEs are taken up from the gastrointestinal tract and accumulate mainly in fat depots and liver tissues. Seal species inhabiting Arctic and sub-Arctic regions can have upwards of 30% of their body mass composed of blubber. When those blubber stores are mobilized for energy, stored toxicants are also released into circulation. Most studies reporting accumulation of PBDEs in seals have focused on harbor and grey seals with few examining harp and hooded seals. In this study, PBDEs concentrations were analyzed in seal blubber from 21 stranded harp and 9 stranded hooded seals sampled along the northeast coast of the U.S. (1999-2010). A PBDE congener profile was determined for each individual. The results show that both species of seals are accumulating PBDEs with BDE-47 being the dominant congener. Mean Æ©PBDE concentrations in harp seals were 70.55⯱â¯33.59â¯ng/g ww and for hooded seals 94.28⯱â¯42.65â¯ng/g ww. The results of this study are consistent with previous studies reporting a decrease in bioaccumulation with an increase in bromination. For both species, BDE-47 represented the highest percentage of the Æ©PBDEs, composing over 50% of the Æ©PBDEs in harp seals. When compared to stranding condition code, animals found alive had overall higher PBDE concentrations than those found in a state of moderate decomposition. This difference could be due to decreased blubber levels in the decomposed animals or potential degradation of the compounds in the blubber. Almost all seals used in this study were yearlings which is the most likely age class to strand. Yearling seals are at a crucial stage of development, especially of their immune system, which can be impacted by high levels of contaminants like PBDEs and increase the susceptibility to disease.
Subject(s)
Environmental Monitoring , Halogenated Diphenyl Ethers/metabolism , Seals, Earless/metabolism , Water Pollutants, Chemical/metabolism , Animals , Arctic Regions , New EnglandABSTRACT
The 'membrane pacemaker' hypothesis proposes a biochemical explanation for among-species variation in resting metabolism, based on the positive correlation between membrane docosahexaenoic acid (DHA) and metabolic rate. We tested this hypothesis using a novel model, altricial red-winged blackbird nestlings, predicting that the proportion of DHA in muscle and liver membranes should increase with the increasing metabolic rate of the nestling as it develops endothermy. We also used a dietary manipulation, supplementing the natural diet with fish oil (high DHA) or sunflower oil (high linoleic acid) to alter membrane composition and then assessed metabolic rate. In support of the membrane pacemaker hypothesis, DHA proportions increased in membranes from pectoralis muscle, muscle mitochondria and liver during post-hatch development. By contrast, elevated dietary DHA had no effect on resting metabolic rate, despite causing significant changes to membrane lipid composition. During cold challenges, higher metabolic rates were achieved by birds that had lower DHA and higher linoleic acid in membrane phospholipids. Given the mixed support for this hypothesis, we conclude that correlations between membrane DHA and metabolic rate are likely spurious, and should be attributed to a still-unidentified confounding variable.
Subject(s)
Basal Metabolism , Docosahexaenoic Acids/metabolism , Membrane Lipids/metabolism , Phospholipids/metabolism , Songbirds/physiology , Animal Feed/analysis , Animals , Animals, Newborn/growth & development , Animals, Newborn/physiology , Cold Temperature , Diet/veterinary , Dietary Supplements/analysis , Female , Liver/metabolism , Mitochondria, Muscle/metabolism , Pectoralis Muscles/metabolism , Songbirds/growth & developmentABSTRACT
Few studies have investigated the thyroid-disrupting effects of polybrominated diphenyl ethers (PBDEs) across multiple levels of biological organization in anurans, despite their suitability for the screening of thyroid disruptors. Therefore, the present study evaluated the effects of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) on development, thyroid histology and thyroid hormone-related gene expression in Xenopus laevis exposed to 0 (control), 50 (low), 500 (medium) or 5000µg BDE-47/g food (high) for 21days. Only the high dose of BDE-47 hindered growth and development; however, thyroid hormone-associated gene expression was downregulated in the brains of tadpoles regardless of dose. These results show that BDE-47 disrupts thyroid hormone signaling at the molecular and whole-organism levels and suggest that gene expression in the brain is a more sensitive endpoint than metamorphosis. Furthermore, the altered gene expression patterns among BDE-47-exposed tadpoles provide insight into the mechanisms of PBDE-induced thyroid disruption and highlight the potential for PBDEs to act as neurodevelopmental toxicants.
Subject(s)
Endocrine Disruptors/toxicity , Gene Expression/drug effects , Halogenated Diphenyl Ethers/toxicity , Larva/drug effects , Thyroid Gland/drug effects , Thyroid Hormones/genetics , Administration, Oral , Animals , Body Weight/drug effects , Dose-Response Relationship, Drug , Larva/genetics , Larva/growth & development , Larva/metabolism , Receptors, Thyroid Hormone/genetics , Thyroid Gland/metabolism , Thyroid Gland/pathology , Xenopus laevisABSTRACT
L-Phenylalanine ammonia-lyase (PAL) is the first enzyme in the biosynthesis of phenylpropanoid-derived plant compounds such as flavonoids, coumarins and the cell wall polymer lignin. The cell walls of grasses possess higher proportions of syringyl (S)-rich lignins and high levels of esterified coumaric acid compared with those of dicotyledonous plants, and PAL from grasses can also possess tyrosine ammonia-lyase (TAL) activity, the reason for which has remained unclear. Using phylogenetic, transcriptomic and in vitro biochemical analyses, we identified a single homotetrameric bifunctional ammonia-lyase (PTAL) among eight BdPAL enzymes in the model grass species Brachypodium distachyon. (13)C isotope labelling experiments along with BdPTAL1-downregulation in transgenic plants showed that the TAL activity of BdPTAL1 can provide nearly half of the total lignin deposited in Brachypodium, with a preference for S-lignin and wall-bound coumarate biosynthesis, indicating that PTAL function is linked to the characteristic features of grass cell walls. Furthermore, isotope dilution experiments suggest that the pathways to lignin from L-phenylalanine and L-tyrosine are distinct beyond the formation of 4-coumarate, supporting the organization of lignin synthesis enzymes in one or more metabolons.
Subject(s)
Brachypodium/genetics , Lignin/biosynthesis , Phenylalanine Ammonia-Lyase/genetics , Plant Proteins/genetics , Brachypodium/metabolism , Cell Wall/metabolism , Phenylalanine Ammonia-Lyase/metabolism , Phylogeny , Plant Proteins/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Sequence Analysis, DNAABSTRACT
2,2',4,4'-Tetrabromodiphenyl ether (BDE-47), a compound manufactured for use as a flame retardant, is a ubiquitous environmental contaminant and suspected endocrine disruptor. Though several studies have explored the reproductive effects of BDE-47 in adult fish, there is a paucity of data regarding the reproductive effects of early life stage exposure. The goal of this study was to assess the reproductive effects of early life stage BDE-47 exposure in fathead minnows (Pimephales promelas). To achieve this, minnows were exposed to either a low (57.68 µg BDE-47/g Artemia) or high (392.59 µg BDE-47/g Artemia) dose of BDE-47 from fertilization to 34 days postfertilization (dpf) via a combination of maternal transfer and dietary exposure. Larvae were then raised on a clean diet until sexual maturity (â¼184 dpf) when reproductive function was evaluated using a 21 day breeding study. Fish exposed to BDE-47 had significantly reduced clutch size and fecundity relative to controls. BDE-47 exposed groups also had female-biased sex ratios and exposed males had fewer tubercles. Overall, this study demonstrates that exposure to BDE-47 during early life stages can alter both sexual differentiation and reproductive function.
Subject(s)
Cyprinidae , Sex Differentiation , Animals , Endocrine Disruptors/pharmacology , Flame Retardants , Reproduction/drug effects , Water Pollutants, Chemical/pharmacologyABSTRACT
The present study examined the bioconcentration of 2 basic pharmaceuticals: verapamil (a calcium channel blocker) and clozapine (an antipsychotic compound) in 2 fresh water fishes, fathead minnow and channel catfish. In 4 separate bioconcentration factor (BCF) experiments (2 chemicals × 1 exposure concentration × 2 fishes), fathead minnow and channel catfish were exposed to 190 µg/L and 419 µg/L of verapamil (500 µg/L nominal) or 28.5 µg/L and 40 µg/L of clozapine (50 µg/L nominal), respectively. Bioconcentration factor experiments with fathead consisted of 28 d uptake and 14 d depuration, whereas tests conducted on catfish involved a minimized test design, with 7 d each of uptake and depuration. Fish (n = 4-5) were sampled during exposure and depuration to collect different tissues: muscle, liver, gills, kidneys, heart (verapamil tests only), brain (clozapine tests only), and blood plasma (catfish tests only). Verapamil and clozapine concentrations in various tissues of fathead and catfish were analyzed using liquid chromatography-mass spectrometry. In general, higher accumulation rates of the test compounds were observed in tissues with higher perfusion rates. Accumulation was also high in tissues relevant to pharmacological targets in mammals (i.e. heart in verapamil test and brain in the clozapine test). Tissue-specific BCFs (wet wt basis) for verapamil and clozapine ranged from 0.7 to 75 and from 31 to 1226, respectively. Tissue-specific concentration data were used to examine tissue-blood partition coefficients.
Subject(s)
Clozapine/analysis , Cyprinidae/metabolism , Ictaluridae/metabolism , Verapamil/analysis , Water Pollutants, Chemical/analysis , Animals , Chromatography, High Pressure Liquid , Clozapine/isolation & purification , Female , Gills/chemistry , Gills/metabolism , Kidney/chemistry , Kidney/metabolism , Liquid-Liquid Extraction , Liver/chemistry , Liver/metabolism , Male , Mass Spectrometry , Muscles/chemistry , Muscles/metabolism , Myocardium/chemistry , Myocardium/metabolism , Verapamil/isolation & purification , Water Pollutants, Chemical/isolation & purificationABSTRACT
The goal of the present study was to evaluate the reproductive function of fathead minnows (Pimephales promelas) exposed to brominated diphenyl ether-47 (BDE-47) at doses lower than those used in previous studies. This was accomplished by evaluating the impacts of BDE-47 exposures across multiple levels of biological organization. Breeding pairs were exposed to BDE-47 via diet for 21 d, during which reproductive success was monitored. At the conclusion of the exposure, fish were euthanized to assess the effects of BDE-47 on sex steroid-related and thyroid-related transcripts, plasma androgen levels, gonadosomatic index (GSI), and secondary sexual characteristics. Several alterations in gene expression were noted including a >2.1-fold decrease in hepatic estrogen receptor α (erα) and a 2.9-fold decrease in ovarian aromatase (arom). In addition, BDE-47-exposed males experienced increases in deiodinase 2 (dio2) expression in brain tissue (â¼1.5-fold) and decreases in hepatic transthyretin (ttr) expression (â¼1.4-fold). Together, these gene expression alterations suggest the potential for BDE-47 to disrupt endocrine signaling. There were no significant differences in plasma hormone levels, GSI, secondary sexual characteristics, or reproductive success. Overall, the present study demonstrates that exposure to BDE-47 is capable of altering both sex steroid-related and thyroid-related transcripts but that these observed alterations do not necessarily manifest themselves at higher levels of biological organization for the endpoints selected. Environ Toxicol Chem 2016;35:2048-2057. © 2016 SETAC.
Subject(s)
Cyprinidae/metabolism , Endocrine Disruptors/toxicity , Environmental Monitoring/methods , Halogenated Diphenyl Ethers/toxicity , Reproduction/drug effects , Animal Feed , Animals , Brain/drug effects , Brain/metabolism , Breeding , Cyprinidae/blood , Cyprinidae/genetics , Dose-Response Relationship, Drug , Female , Gene Expression/drug effects , Gonadal Steroid Hormones/metabolism , Liver/drug effects , Liver/metabolism , Male , Thyroid Hormones/genetics , Thyroid Hormones/metabolismABSTRACT
The steroid hormone medroxyprogesterone acetate (MPA), commonly used in oral and injectable contraceptives, has been detected in surface and wastewaters near urban and agricultural areas in several rivers of the world. The objectives of this study were to examine the accumulative potential and tissue distribution of MPA in fish. A freshwater species, the common carp (Cyprinus carpio), was exposed to 100 µg/L of MPA for a 7-day period followed by a depuration phase in which fish were maintained in dechlorinated tap water for an additional 7 days. Tissues (muscle, brain, plasma, and liver) were sampled during the uptake (days 1, 3, and 7) and depuration (day 14) phases of the experiment. Tissue-specific bioconcentration factors (BCF) ranged from 4.3 to 37.8 and uptake was greatest in the liver>brain>plasma and lowest in the muscle. From a regulatory standpoint, MPA shows little tendency to bioaccumulate in fish.