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Donald E. Tillitt

Publications and source records attributed to Donald E. Tillitt.

At least 19 recordsLinked to original sources

Immunomodulation in adult largemouth bass (Micropterus salmoides) exposed to a model estrogen or mixture of endocrine disrupting contaminants during early gonadal recrudescence

Disease outbreaks, skin lesions, fish kill events, and reproductive abnormalities have been observed in wild populations of Centrarchids in watersheds throughout the United States. Occurrence of synthetic and natural hormones from wastewater treatment plants and livestock operations, pesticides from agricultural land use, and phytoestrogens have been implicated as potential causes of these adverse effects. Our objective was to investigate possible immunomodulation in adult largemouth bass ( Micropterus salmoides ) in response to a seasonal exposure to environmentally relevant contaminants in outdoor experimental ponds. Exposures included 17α-ethinylestradiol (EE2; 3.6 ng/L) or a binary mixture of endocrine-active substances commonly detected in surface waters, estrone (E1; 85.6 ng/L) and atrazine (ATR; 5.4 µg/L). The 4-month exposure was conducted from July to November. Functional immune responses of anterior kidney-derived leukocytes were evaluated in December in the week following the end of the dosing period, and in the following April, four months after dosing ended and just prior to spawning. Concentrations of EE2 and E1 in the ponds fell below detectable levels in December, but detectable concentrations of ATR (2.9 µg/L) persisted at least through May. For each sampling time, anterior kidney leukocytes were isolated and grown in primary culture for the assessment of zymosan-stimulated respiratory burst and lectin-stimulated mitogenic responses. We observed seasonal differences in respiratory burst stimulation over time and treatment with a significantly greater response in April relative to December. Respiratory burst activity was also significantly greater in April for fish exposed to the E1+ATR relative to control. In April, prior to spawning, we observed a significantly dampened mitogenic response to PHAP (a T cell mitogen) and LPS (a B cell mitogen) in the EE2 treatment relative to control fish. There were no significant differences in mitogenic responses or respiratory burst between sexes. However, there was significantly higher alternative complement pathway hemolytic activity in males compared to females in both the control and E1+ATR treatment groups. Our results demonstrate that environmentally relevant concentrations of contaminants can alter immune function in a socioeconomically important fish species.

Comparative Immunology Reports

Reevaluation of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalency factors for dioxin-like polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and polychlorinated biphenyls for fishes

An expert meeting was organized by the World Health Organization (WHO) in 1997 to streamline assessments of risk posed by mixtures of dioxin-like chemicals (DLCs) through development of 2,3,7,8-tetrachlorodibenzo- p -dioxin (2,3,7,8-TCDD) equivalency factors (TEFs) for mammals, birds, and fishes. No reevaluation has been performed for fish TEFs. Therefore, the objective of the present study was to reevaluate the TEFs for fishes based on an updated database of relative potencies (RePs) for DLCs. Selection criteria consistent with the WHO meeting resulted in 53 RePs across 14 species of fish ultimately being considered. Of these RePs, 70% were not available at the time of the WHO meeting. These RePs were used to develop updated TEFs for fishes based on a similar decision process as used at the WHO meeting. The updated TEF for 16 DLCs was greater than the WHO TEF, but only four differed by more than an order of magnitude. Measured concentrations of DLCs in four environmental samples were used to compare 2,3,7,8-TCDD equivalents (TEQs) calculated using the WHO TEFs relative to the updated TEFs. The TEQs for none of these environmental samples differed by more than an order of magnitude. Therefore, present knowledge supports that the WHO TEFs are suitable potency estimates for fishes. However, the updated TEFs pull from a larger database with a greater breadth of data and as a result offer greater confidence relative to the WHO TEFs. Risk assessors will have different criteria in the selection of TEFs, and the updated TEFs are not meant to immediately replace the formal WHO TEFs; but those who value a larger database and increased confidence in TEQs could consider using the updated TEFs.

Environmental Toxicology and Chemistry

Thiamine supplementation improves survival and body condition of hatchery-reared steelhead (Oncorhynchus mykiss) in Oregon

Early rearing of steelhead ( Oncorhynchus mykiss) in Oregon hatcheries is often problematic; fry can become emaciated and die during the period between hatch and first feed. Thiamine (vitamin B1) deficiency has caused early mortality in salmonids; however, the thiamine status of Oregon’s steelhead populations is unknown, to date. Of the 26 egg samples from three Oregon hatcheries in 2019, 20 (77%) had thiamine levels < 10 nmol/g, and 13 of those samples (50%) had levels <6.5 nmol/g, suggesting the thiamine deficiency of adult, female steelhead. To investigate if thiamine deficiency was causally related to fry survival, females were injected with buffered thiamine HCl 50 mg/kg prior to spawning; additionally, a subset of eggs were supplemented via bath treatment with thiamine mononitrate (1000 ppm) at spawning. Cumulative fry mortality at 8 weeks post-hatch from thiamine-injected females was 2.9% compared to 13.8% mortality of fry without thiamine supplementation. Fry treated only with the thiamine via bath as eggs had a mortality rate of 6.9%. There were no additional improvements for the survival of fry from injected females that also received a thiamine bath. Furthermore, condition factors were greater in thiamine-supplemented fry than in those that received no thiamine. These data identify thiamine deficiency in Oregon steelhead and suggest supplementation with thiamine can mitigate early rearing mortality.

Oregon

Genetic basis of thiaminase I activity in a vertebrate, zebrafish Danio rerio

Thiamine (vitamin B 1 ) metabolism is an important driver of human and animal health and ecological functioning. Some organisms, including species of ferns, mollusks, and fish, contain thiamine-degrading enzymes known as thiaminases, and consumption of these organisms can lead to thiamine deficiency in the consumer. Consumption of fish containing thiaminase has led to elevated mortality and recruitment failure in farmed animals and wild salmonine populations around the world. In the North American Great Lakes, consumption of the non-native prey fish alewife ( Alosa pseudoharengus ) by native lake trout ( Salvelinus namaycush ) led to thiamine deficiency in the trout, contributed to elevated fry mortality, and impeded natural population recruitment. Several thiaminases have been genetically characterized in bacteria and unicellular eukaryotes, and the source of thiaminase in multicellular organisms has been hypothesized to be gut microflora. In an unexpected discovery, we identified thiaminase I genes in zebrafish ( Danio rerio ) with homology to bacterial tenA thiaminase II. The biochemical activity of zebrafish thiaminase I (GenBank NP_001314821.1) was confirmed in a recombinant system. Genes homologous to the zebrafish tenA-like thiaminase I were identified in many animals, including common carp ( Cyprinus carpio ), zebra mussel ( Dreissena polymorpha ) and alewife. Thus, the source of thiaminase I in alewife impacting lake trout populations is likely to be de novo synthesis.

Scientific Reports

Understanding and mitigating thiaminase activity in silver carp

A deficiency of thiamine (vitamin B1), an essential cofactor for enzymes involved in metabolic processes, can be caused by the enzyme thiaminase. Thiaminase in food stocks has been linked to morbidity and mortality due to thiamine depletion in many ecologically and economically important species. Thiaminase activity has been detected in certain bacteria, plants, and fish species, including carp. The invasive silver carp ( Hypophthalmichthys molitrix) presents an enormous burden to ecosystems throughout the Mississippi River watershed. Its large biomass and nutritional content offer an attractive possibility as a food source for humans, wild animals, or pets. Additionally, harvesting this fish could alleviate some of the effects of this species on waterways. However, the presence of thiaminase would detract from its value for dietary consumption. Here we confirm the presence of thiaminase in several tissues from silver carp, most notably the viscera, and systematically examine the effects of microwaving, baking, dehydrating, and freeze-drying on thiaminase activity. Certain temperatures and durations of baking and microwaving reduced thiaminase activity to undetectable levels. However, caution should be taken when carp tissue is concentrated by processes without sufficient heat treatment, such as freeze-drying or dehydration, which results in concentration, but not inactivation of the enzyme. The effects of such treatments on the ease of extracting proteins, including thiaminase, and the impact on data interpretation using the 4-nitrothiophenol (4-NTP) thiaminase assay were considered.

Food Science

Factors influencing egg thiamine concentrations of Lake Ontario lake trout: 2019–2020

In the Great Lakes region, thiamine deficiency is considered a recruitment bottleneck for lake trout Salvelinus namaycush and has been correlated with the consumption of non-native alewife Alosa pseudoharengus . While alewife, the most abundant forage fish in Lake Ontario, are the predominant prey for lake trout, they also consume benthic prey such as round goby Neogobius melanostomus . Because variation in the proportion of alewife in lake trout diets is linked to variation in egg thiamine concentrations, understanding how factors such as region of capture and hatchery-strain of lake trout influence diet, are key to understanding the patterns of variation in egg thiamine concentrations observed in this species. With recent increases in natural recruitment of lake trout being observed in the western region of the lake, understanding if egg thiamine is a potential driver is crucial to the rehabilitation of lake trout. In this study, we evaluated egg thiamine concentrations in lake trout during 2019–2020. We found no significant difference in egg thiamine concentrations among regions. However, a stocked Lake Superior deepwater morphotype (Superior Klondike Wild – SKW) showed significantly higher egg thiamine concentrations compared to the lean morphotype including Seneca (SEN) and Lake Champlain Domestic (LCD) strains. An analysis of fatty acid signatures of each hatchery-strain suggested that the SKW strain consumed a higher proportion of round goby than lean strains. Overall, these results suggest that morphotypic differences in the feeding ecology of lake trout can result in biochemical changes which may influence the effectiveness of restoration efforts.

Lake Ontario

Declines in reproductive condition of male largemouth bass (Micropterus salmoides) following seasonal exposure to estrogenic endocrine-disrupting compounds

Reproductive abnormalities, that could lead to possible effects at the population level, have been observed in wild fish throughout the United States, with high prevalence in largemouth bass (LMB; Micropterus salmoides ) and smallmouth bass ( Micropterus dolomieu ). Estrone (E1) and atrazine (ATR) are common environmental contaminants often associated with agricultural land use. 17alpha-ethinylestradiol (EE2) is a contaminant associated with wastewater treatment effluent, and a representative, well-studied estrogen commonly used for fish toxicity testing. Our objective was to assess whether early gonad recrudescence in adult fish was a period of sensitivity for alterations in reproductive condition and function. Adult male LMB were exposed from post-spawning to early gonad recrudescence to either a mixture of E1 (47.9 ng/L) + ATR (5.4 µg/L), or EE2 (2.4 ng/L) in outdoor experimental ponds. Gonad samples were collected from fish just prior to the start of exposure (July), at the end of the exposure period (December), the following spring just prior to spawning (April), and post spawning (May). Gonadosomatic index (GSI) was significantly reduced in E1 + ATR-exposed and EE2-exposed males compared to control at every post-exposure time point. Reduced sperm count and sperm motility were observed in the mixture treatment (E1 + ATR) compared to the control. Sperm motility was also reduced in the EE2 treatment. These data together indicate that estrogenic endocrine-disrupting compounds can lessen the reproductive condition of adult male LMB, and that effects of exposure during early gonad recrudescence can persist at least through the subsequent spawning cycle.

International Journal of Molecular Sciences

Exposure to 17α-ethinylestradiol results in differential susceptibility of largemouth bass (Micropterus salmoides) to bacterial infection

Disease outbreaks, skin lesions, mortality events, and reproductive abnormalities have been observed in wild populations of centrarchids. The presence of estrogenic endocrine disrupting compounds (EEDCs) has been implicated as a potential causal factor for these effects. The effects of prior EEDC exposure on immune response were examined in juvenile largemouth bass ( Micropterus salmoides ) exposed to a potent synthetic estrogen (17α-ethinylestradiol, EE2) at a low (EE2 Low , 0.87 ng/L) or high (EE2 High , 9.08 ng/L) dose for 4 weeks, followed by transfer to clean water and injection with an LD 40 dose of the Gram-negative bacteria Edwardsiella piscicida . Unexpectedly, this prior exposure to EE2 High significantly increased survivorship at 10 d post-infection compared to solvent control or EE2 Low -exposed, infected fish. Both prior exposure and infection with E. piscicida led to significantly reduced hepatic glycogen levels, indicating a stress response resulting in depletion of energy stores. Additionally, pathway analysis for liver and spleen indicated differentially expressed genes associated with immunometabolic processes in the mock-injected EE2 High treatment that could underlie the observed protective effect and metabolic shift in EE2 High -infected fish. Our results demonstrate that exposure to a model EEDC alters metabolism and immune function in a fish species that is ecologically and economically important in North America.

Environmental Science and Technology

Egg hatchability and egg thiamine concentrations in Lake Trout from the Northern Refuge of Lake Michigan

The Northern Refuge of Lake Michigan was established in 1985 under the auspices of the Great Lakes Fishery Commission as part of an intensely managed effort to restore a self‐sustaining Lake Trout Salvelinus namaycush population to the lake. Despite the stocking of millions of Lake Trout into the refuge and survival of these stocked fish to adulthood, signs of natural recruitment by Lake Trout in the refuge have been very sparse. To ascertain whether low egg viability or poor fry survival could be limiting natural recruitment by Lake Trout in the refuge, hatching rates of eggs taken from Lake Trout caught in the Northern Refuge during October 2019 were determined in the laboratory. Moreover, survival rates of the hatched‐out fry were determined along with the thiamine concentrations in the fry. In addition, thiamine concentrations in eggs of ovulating Lake Trout caught in the Northern Refuge during 2019–2021 were determined. Egg hatching rate in the laboratory averaged 93%. Survival rate of Lake Trout fry up to 5 weeks after hatching averaged 97%. Mean (SE) thiamine concentrations in eggs of Lake Trout caught in 2019, 2020, and 2021 were 19.1 (3.6), 5.9 (1.3), and 6.2 (0.4) nmol/g, respectively; these levels are well above the threshold concentration of 2.3 nmol/g, above which the likelihood of experiencing mortality due to thiamine deficiency complex (TDC) is low. Thiamine concentrations in the Lake Trout fry at the end of the laboratory experiment averaged 11.2 nmol/g, and abnormal behavior in the fry was not observed. We conclude that TDC does not appear to represent a major impediment to Lake Trout rehabilitation in the Northern Refuge at present, although TDC may have contributed to the lack of detectable natural recruitment in the past.

Lake Michigan, Northern Refuge

Response to Gard et al.'s (2021) Comments on the Critical Review “Polychlorinated Biphenyl Tissue-Concentration Thresholds for Survival, Growth, and Reproduction in Fish”

This response is offered to the critique by Gard et al. ( 2021 ) of our meta-analysis of polychlorinated biphenyl (PCB)-induced toxicity data in fish (Berninger and Tillitt 2019 ). Gard et al. ( 2021 ) offered numerous comments, the most substantive suggesting that 1) we should have added no-observable–adverse effect residue (NOAER) data from additional studies and all data points from selected studies, and 2) the uncertainty of aggregating data from different PCB mixtures, different species, and different life stages is too great based on a limited data set. The additional studies Gard et al. suggested either were not designed to produce toxicological data, had experimental design issues, were confounded by co-contaminants, or did not contain paired exposure–effects data and as such were not appropriate to add to the data set. Lowest-observable–adverse effect residue (LOAER) values were selected for our analysis because they represent population sensitivities from the central portions of a frequency distribution (the linear portion of dose–response curves). As a consequence, there is less uncertainty in these input data (LOAER values) and greater confidence that they accurately represent the response of fish populations tested. Modeling NOAER values is in the extrapolation portion of a dose–response relationship and subject to enhanced uncertainty. The Gard et al. ( 2021 ) critique ignores this fundamental principle of toxicology and adds/deletes data points from our data set without clear selection criteria, which artificially enhances the uncertainty of their models that ultimately are not useful. We reject the premise that it is better to use individual study data as opposed to aggregation of PCB-induced toxicity thresholds in fish.

Environmental Toxicology and Chemistry

Reproduction

Lake charr Salvelinus namaycush are typically fall spawners although one ecotype has populations that spawn during spring and fall (siscowets in Lake Superior). Lake charr are iteroparous (reproduce more than once in a lifetime) with group-synchronous ovarian development and typically spawn once per year. However, lake charr may not reproduce every year, a phenomenon known as skipped spawning. Free embryos are active on spawning reefs, make diurnal vertical movements from spawning substrate, and feed exogenously much earlier than previously assumed. The abundance of food and predators strongly affects the rate of development, yolk sac absorption, and duration of residence on spawning sites. The necessity for, and timing of, gas bladder inflation, and mechanisms for inflation without access to the surface, need further study. The low survival of free embryos due to thiamine deficiency has likely contributed to the lack of recruitment of lake charr in the Laurentian Great Lakes for decades. Thiaminase, a thiamine-degrading enzyme, appears to be the causal agent for thiamine deficiency in Great Lakes lake charr.

Great Lakes

Effects of early life stage exposure of largemouth bass to atrazine or a model estrogen (17α-ethinylestradiol)

Endocrine disrupting contaminants are of continuing concern for potentially contributing to reproductive dysfunction in largemouth and smallmouth bass in the Chesapeake Bay watershed (CBW) and elsewhere. Exposures to atrazine (ATR) have been hypothesized to have estrogenic effects on vertebrate endocrine systems. The incidence of intersex in male smallmouth bass from some regions of CBW has been correlated with ATR concentrations in water. Fish early life stages may be particularly vulnerable to ATR exposure in agricultural areas, as a spring influx of pesticides coincides with spawning and early development. Our objectives were to investigate the effects of early life stage exposure to ATR or the model estrogen 17α-ethinylestradiol (EE2) on sexual differentiation and gene expression in gonad tissue. We exposed newly hatched largemouth bass (LMB, Micropterus salmoides ) from 7 to 80 days post-spawn to nominal concentrations of 1, 10, or 100 µg ATR/L or 1 or 10 ng EE2/L and monitored histological development and transcriptomic changes in gonad tissue. We observed a nearly 100% female sex ratio in LMB exposed to EE2 at 10 ng/L, presumably due to sex reversal of males. Many gonad genes were differentially expressed between sexes. Multidimensional scaling revealed clustering by gene expression of the 1 ng EE2/L and 100 µg ATR/L-treated male fish. Some pathways responsive to EE2 exposure were not sex-specific. We observed differential expression in male gonad in LMB exposed to EE2 at 1 ng/L of several genes involved in reproductive development and function, including star , cyp11a2 , ddx4 (previously vasa ), wnt5b , cyp1a and samhd1 . Expression of star , cyp11a2 and cyp1a in males was also responsive to ATR exposure. Overall, our results confirm that early development is a sensitive window for estrogenic endocrine disruption in LMB and are consistent with the hypothesis that ATR exposure induces some estrogenic responses in the developing gonad. However, ATR-specific and EE2-specific responses were also observed.

New York, Pennsylvania, Maryland, West Virginia, V

Bisphenol A and 17α-ethinylestradiol-induced transgenerational gene expression differences in the brain–pituitary–testis axis of medaka, Oryzias latipes

Endocrine disrupting chemicals (EDCs), such as bisphenol A (BPA) and 17α-ethinylestradiol (EE2), can have far reaching health effects, including transgenerational abnormalities in offspring that never directly contacted either chemical. We previously reported reduced fertilization rates and embryo survival at F2 and F3 generations caused by 7-day embryonic exposure (F0) to 100 μg/L BPA or 0.05 μg/L EE2 in medaka. Crossbreeding of fish in F2 generation indicated subfertility in males. To further understand the mechanisms underlying BPA or EE2-induced adult onset and transgenerational reproductive defects in males, the present study examined the expression of genes regulating the brain–pituitary–testis (BPT) axis in the same F0 and F2 generation male medaka. Embryonic exposure to BPA or EE2 led to hyperactivation of brain and pituitary genes, which are actively involved in reproduction in adulthood of the F0 generation male fish, and some of these F0 effects continued to the F2 generation (transgenerational effects). Particularly, the F2 generation inherited the hyperactivated state of expression for kisspeptin ( kiss1 and kiss2 ) and their receptors ( kiss1r and kiss2r), and gnrh and gnrh receptors. At F2 generation, expression of DNA methyltransferase 1 ( dnmt1 ) decreased in brain of the BPA treatment lineage, while EE2 treatment lineage showed increased dnmt3bb expression. Global hypomethylation pattern was observed in the testis of both F0 and F2 generation fish. Taken together, these results demonstrated that BPA or EE2-induced transgenerational reproductive impairment in the F2 generation was associated with alterations of reproductive gene expression in brain and testis and global DNA methylation in testis.

Biology of Reproduction

Biological effects of hydrocarbon degradation intermediates: Is the total petroleum hydrocarbon analytical method adequate for risk assessment?

In crude oil contaminant plumes, the dissolved organic carbon (DOC) is mainly hydrocarbon degradation intermediates only partly quantified by the diesel range total petroleum hydrocarbon (TPHd) method. To understand potential biological effects of degradation intermediates, we tested three fractions of DOC: (1) solid-phase extract (HLB); (2) dichloromethane (DCM-total) extract used in TPHd; and (3) DCM extract with hydrocarbons isolated by silica gel cleanup (DCM-SGC). Bioactivity of extracts from five wells spanning a range of DOC was tested using an in vitro multiplex reporter system that evaluates modulation of the activity of 46 transcription factors; extracts were evaluated at concentrations equivalent to the well water samples. The aryl hydrocarbon receptor (AhR) and pregnane X receptor (PXR) transcription factors showed the greatest upregulation, with HLB exceeding DCM-total, and no upregulation in the hydrocarbon fraction (DCM-SGC). The HLB extracts were further studied with HepG2 chemically activated luciferase expression (CALUX) in vitro assays at nine concentrations ranging from 40 to 0.01 times the well water concentrations. Responses decreased with distance from the source but were still present at two wells without detectable hydrocarbons. Thus, our in vitro assay results indicate that risks associated with degradation intermediates of hydrocarbons in groundwater will be underestimated when protocols that remove these chemicals are employed.

Minnesota

Factors affecting sampling strategies for design of an effects‐directed analysis for endocrine‐active chemicals

Effects‐directed analysis (EDA) is an important tool for identifying unknown bioactive components in a complex mixture. Such an analysis of endocrine‐active chemicals (EACs) from water sources has promising regulatory implications but also unique logistical challenges. We propose a conceptual EDA (framework) based on a critical review of EDA literature and concentrations of common EACs in waste and surface waters. Required water volumes for identification of EACs under this EDA framework were estimated based on bioassay performance (in vitro and in vivo bioassays), limits of quantification by mass spectrometry (MS), and EAC water concentrations. Sample volumes for EDA across the EACs showed high variation in the bioassay detectors, with genistein, bisphenol A, and androstenedione requiring very high sample volumes and ethinylestradiol and 17β‐trenbolone requiring low sample volumes. Sample volume based on the MS detector was far less variable across the EACs. The EDA framework equation was rearranged to calculate detector “thresholds,” and these thresholds were compared with the literature EAC water concentrations to evaluate the feasibility of the EDA framework. In the majority of instances, feasibility of the EDA was limited by the bioassay, not MS detection. Mixed model analysis showed that the volumes required for a successful EDA were affected by the potentially responsible EAC, detection methods, and the water source type, with detection method having the greatest effect on the EDA of estrogens and androgens. The EDA framework, equation, and model we present provide a valuable tool for designing a successful EDA.

Environmental Toxicology and Chemistry

Neonicotinoid insecticide concentrations in agricultural wetlands and associations with aquatic invertebrate communities

Neonicotinoids are considered a superior insecticide for agricultural pest management, although their impacts on non-target insects is a rising concern. Aside from laboratory and mesocosm studies, limited research has been directed towards the role neonicotinoids may have in structuring aquatic invertebrate communities in field settings. Therefore, we simultaneously collected aquatic invertebrate and surface water samples from 26 wetlands within a highly modified agricultural landscape of Nebraska’s Rainwater Basin during spring 2015. Water samples were tested for six different neonicotinoids, nutrients, and physical properties. Trace levels of clothianidin and imidacloprid were the only neonicotinoids detected, occurring in 85% and 15%, respectively, of wetlands sampled. All measurements for clothianidin and imidacloprid were below chronic toxicity benchmarks set by the United States Environmental Protection Agency. Neonicotinoid concentrations were significantly lower (W 26, 0.05 = 42.5) at wetlands with vegetative buffer strips > 50 m wide compared to wetlands with vegetative buffers strips <50 m. Although neonicotinoids were below benchmark concentrations proposed by government regulations, a significant negative association between neonicotinoid concentrations and aquatic invertebrate biomass was observed across all wetlands studied (Parameter Estimate = -0.031; SE = 0.014).

Nebraska

Acute toxicity and clotting times of anticoagulant rodenticides to red-toothed (Odonus niger) and black (Melichthys niger) triggerfish, fathead minnow (Pimephales promelas), and largemouth bass (Micropterus salmoides)

Anticoagulant rodenticides (ARs) areused in rateradication efforts on island wildlife refuges. ARbait pellets can get into coralreefareasduring broadcasting and leadto exposure ofnon-target organisms, such as marine fishes. The objective of this study was to determine the sensitivity of representative saltwater fishes, Red-toothed triggerfish (Odonus niger) and Black triggerfish (Melichthys niger), and common freshwater fishes, fathead minnow (Pimephales promelas), and largemouth bass (Micropterus salmoides) to first generation ARs, diphacinone (DPN) and chlorophacinone (CPN), as well as a second-generation AR, brodifacoum (BROD). Acute toxicity of ARs was evaluated by single dose, intraperitoneal injections. The median lethal dose (LD50) ranges were 137−175μg DPN/g, 155−182μg CPN/g, and 36−48μg BROD/g for Red-toothed triggerfish and 90−122μg DPN/g, 125−164μg CPN/g, and 50−75μg BROD/g for black triggerfish. Laboratory surrogate test fish species fathead minnow and largemouth bass were of similar sensitivity toward AR-induced toxicity compared to triggerfish based on LD50 values. Sublethal effects on elevated clotting time occurred in dose-dependent fashion in all fish tested. Fish appear to have low sensitivity to AR chemicals as compared to other taxa, in particular mammals and birds, based on across-taxa comparisons of species sensitivity distributions of whole body, single dose acute lethality (LD50 values). The sensitivity of fish to waterborne exposures of ARs has yet to be fully evaluated and indeed may prove more hazardous to fish.

Aquatic Toxicology

Transcriptome analysis of testis reveals the effects of developmental exposure to bisphenol a or 17α-ethinylestradiol in medaka (Oryzias latipes)

Endocrine disrupting chemicals (EDCs) can induce abnormalities in organisms via alteration of molecular pathways and subsequent disruption of endocrine functions. Bisphenol A (BPA) and 17α-ethinylestradiol (EE2) are ubiquitous EDCs in the environment. Many aquatic organisms, including fish, are often exposed to varying concentrations of BPA and EE2 throughout their lifespan. Both BPA and EE2 can activate estrogenic signaling pathways and cause adverse effects on reproduction via alteration of pathways associated with steroidogenesis. However, transcriptional pathways that are affected by chronic exposure to these two ubiquitous environmental estrogens during embryonic, larval, and juvenile stages are not clearly understood. In the present study, we examined transcriptional alterations in the testis of medaka fish ( Oryzias latipes ) chronically exposed to a low concentration of BPA or EE2. Medaka were exposed to BPA (10 μg/L) or EE2 (0.01 μg/L) from 8 h post-fertilization (as embryos) to adulthood 50 days post fertilization (dpf), and transcriptional alterations in the testis were examined by RNA sequencing (RNA-seq). Transcriptomic profiling revealed 651 differentially expressed genes (DEGs) between BPA-exposed and control testes, while 1475 DEGs were found between EE2-exposed and control testes. Gene ontology (GO) analysis showed a significant enrichment of “intracellular receptor signaling pathway”, “response to steroid hormone” and “hormone-mediated signaling pathway” in the BPA-induced DEGs, and of “cilium organization”, “microtubule-based process” and “organelle assembly” in the EE2-induced DEGs. Pathway analysis showed significant enrichment of “integrin signaling pathway” in both treatment groups, and of “cadherin signaling pathway”, “Alzheimer disease-presenilin pathway” in EE2-induced DEGs. Single nucleotide polymorphism (SNP) and insertion-deletion (Indel) analysis found no significant differences in mutation rates with either BPA or EE2 treatments. Taken together, global gene expression differences in testes of medaka during early stages of gametogenesis were responsive to chronic BPA and EE2 exposure.

Aquatic Toxicology