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At least 55 records · Page 3Linked to original sources

Progression of infection and detection of Pseudoloma neurophilia in zebrafish Danio rerio Hamilton by PCR and histology

Pseudoloma neurophilia is a critical threat to the zebrafish ( Danio rerio ) model, as it is the most common infectious agent found in research facilities. In this study, our objectives were two-fold: (1) compare the application of diagnostic tools for P. neurophilia and (2) track the progression of infection using PCR and histology. The first experiment showed that whole-body analysis by qPCR (WB-qPCR) can be a standardized process, providing a streamlined diagnostic protocol, without the need for extraction of specific tissues. Evaluating the course of infection in experimentally infected fish, we showed key dynamics in infection. Starting with a low dose exposure of 8000 spores/fish, the prevalence remained low until 92 days post-exposure (dpe), followed by a 30%–40% prevalence by histology or 40%–90% by PCR until the end of the experiment at 334 dpe. WB-qPCR positively detected infection in more fish than histology throughout the study, as WB-qPCR detected the parasite as early as 4 dpe, whereas it was undetected by histology until 92 dpe. We also added a second slide for histologic analyses, showing an increase in detection rate from 24% to 26% when we combined all data from our experiments, but this increase was not statistically significant.

Journal of Fish Diseases

Pseudoloma neurophilia infections in zebrafish Danio rerio: effects of stress on survival, growth, and reproduction.

Pseudoloma neurophilia (Microsporidia) is a common disease of zebrafish Danio rerio, including those used as research models. We conducted a study comprised of 4 separate experiments to determine the effects of husbandry stress on preexisting and experimental P. neurophilia infections and the subsequent effects on survival, infection onset and intensity, fish growth, and reproduction. In fish (AB strain) with preexisting infections, stress or feeding cortisol significantly increased mortality over 7 wk compared to no stress or cortisol treatment. In contrast, no mortality was observed in fish (TL strain) experimentally exposed to P. neurophilia over 10 wk. A third experiment involved experimental exposure of AB fish to P. neurophilia and exposure to crowding and handling stressors. No mortality was associated with P. neurophilia regardless of stress treatment over a period of 20 wk. However, the onset of infection occurred sooner in stress-treated fish. Stress significantly increased the mean intensity of infection (described as xenoma area/spinal cord area in histological sections) at Week 20 post-exposure (PE). In fish with preexisting infections, myositis was significantly greater in stressed and cortisol-treated fish than those not stressed. With experimental exposure of AB fish, stressed and infected groups weighed significantly less than the control group at Week 20 PE. Regarding fecundity, the number of larvae hatched at 5 d post fertilization was negatively associated with mean infection intensity among P. neurophilia-infected and stressed AB fish. These experiments are the first to show empirically that P. neurophilia can be associated with reduced weight and fecundity, and that stress can exacerbate the severity of the infection.

Diseases of Aquatic Organisms

High-throughput screening identifies bisphenol P as a potent cardiotoxin, inducing cardiotoxicity through apoptosis and NF-κB Pathway

The increasing use of plastic additives, particularly bisphenols (BPs), has raised significant concerns about their potential risks to human health, especially during critical developmental stages. In this study, we developed a novel high-throughput toxicity screening platform using zebrafish ( Danio rerio ) to identify and prioritize chemicals with cardiotoxic potential, which is based on multidimensional exposure pathways ranging from environmental to human levels. The platform quantitatively assesses heart rate changes based on multilevel exposure pathways (environmental, ecological, and human), effectively prioritizing the most relevant cardiotoxic compounds with potential health risks. Using this platform, we identified bisphenol P (BPP), a widely used substitute for bisphenol A, as a potent cardiotoxic compound. BPP exposure significantly inhibited heart development and function in zebrafish, inducing abnormal heart morphology, reduced heart rate, cardiac output, and hemodynamic disturbances. Additionally, NF-κB signaling pathway analysis, including morpholino knock-down and inhibitor experiments, confirmed that BPP mediates cardiac toxicity by inducing cardiomyocyte apoptosis. Our findings underscore the toxicity that BP substitutes can have, while highlighting the potential of this multidimensional screening platform in evaluating cardiovascular toxicity and guiding future toxicological assessments during critical developmental windows.

Environmental Science & Technology

Deconstructing cardiovascular and coagulation-related traits links dietary ecology to multi-functional snake venom specificity

Animal venoms vary greatly in compositional complexity, where complex venoms are hypothesized to be maintained by greater dietary breadth. Beyond explaining venom composition, the dietary breadth hypothesis predicts that these more complex venoms should show greater functional breadth in terms of overall toxicity and by disrupting multiple prey physiological processes. We evaluate these predictions with six distinct physiological assays of cardiac and blood clotting functions and compared the effects of venoms from snake species with a range of taxonomic dietary diversity levels. We compared the taxonomic dietary generalists Agkistrodon piscivorus and Sistrurus miliarius to taxa with varying taxonomic specialization, namely Ag. contortrix and Crotalus adamanteus , and Azemiops feae . Comparing fish thrombocyte and mammal platelet aggregation and fibrin clot formation, only species with broader diets disrupted both fish and mammal hemostatic function. Venom of Ag. piscivorus , a uniquely fish-eating species, was the most disruptive of zebrafish heart rate, thrombocyte activation, vascular permeability, and clotting after injury. Sistrurus miliarius was also highly toxic, whereas mammal-specialists’ venoms scarcely altered zebrafish physiology. Our results support the hypothesis that dietary breadth selects functionally complex venoms. Understanding venom gene evolution, snakebite symptoms, and searching for therapeutics in venom should be guided by evolutionary ecology.

Evolution

A novel antibody-based biomarker for chronic algal toxin exposure and sub-acute neurotoxicity

The neurotoxic amino acid, domoic acid (DA), is naturally produced by marine phytoplankton and presents a significant threat to the health of marine mammals, seabirds and humans via transfer of the toxin through the foodweb. In humans, acute exposure causes a neurotoxic illness known as amnesic shellfish poisoning characterized by seizures, memory loss, coma and death. Regular monitoring for high DA levels in edible shellfish tissues has been effective in protecting human consumers from acute DA exposure. However, chronic low-level DA exposure remains a concern, particularly in coastal and tribal communities that subsistence harvest shellfish known to contain low levels of the toxin. Domoic acid exposure via consumption of planktivorous fish also has a profound health impact on California sea lions ( Zalophus californianus ) affecting hundreds of animals yearly. Due to increasing algal toxin exposure threats globally, there is a critical need for reliable diagnostic tests for assessing chronic DA exposure in humans and wildlife. Here we report the discovery of a novel DA-specific antibody response that is a signature of chronic low-level exposure identified initially in a zebrafish exposure model and confirmed in naturally exposed wild sea lions. Additionally, we found that chronic exposure in zebrafish caused increased neurologic sensitivity to DA, revealing that repetitive exposure to DA well below the threshold for acute behavioral toxicity has underlying neurotoxic consequences. The discovery that chronic exposure to low levels of a small, water-soluble single amino acid triggers a detectable antibody response is surprising and has profound implications for the development of diagnostic tests for exposure to other pervasive environmental toxins.

PLoS ONE

Pilot Study of Sublethal Effects on Fish of Pesticides Currently Used and Proposed for Use on Maine Blueberries

Blueberry pesticides have been detected consistently in some Down East Maine rivers, yet little is known about the sublethal effects of these pesticides on fish early life stages. The Maine blueberry industry is proposing to replace the insecticide ImidanTM (active ingredient phosmet) and the herbicide VelparTM (active ingredient hexazinone), two of the pesticides found in these rivers, with candidate alternatives SpinTor TM (active ingredient spinosad) and Callistso TM (active ingredient mesotrione). Our objective is to evaluate potential sublethal effects of these four formulations before the industry adopts the two candidate alternatives. We exposed zebrafish (Danio rerio) early life stages, from fertilization through larval swim-up, to a range of pesticide concentrations and evaluated their response relative to untreated controls. In this report we provide preliminary data on immune function as well as on parameters in addition to those originally proposed: development and performance fitness. We also provide information on our progress towards optimizing chemical protocols for analyzing the concentration of active ingredient in each of our formulation dosing solutions, another new parameter we added to those originally proposed. Preliminary results indicate that at environmentally realistic concentrations, these pesticides may have no significant effect on innate immunity, development rate or behavior (spontaneous swimming), however further replication is needed to confirm these initial findings. We have also observed some degree of developmental abnormalities in both pesticide-treated and control zebrafish embryos; however, additional replication is underway to determine if these groups differ significantly.

Open-File Report

Dissolved saxitoxin causes transient inhibition of sensorimotor function in larval Pacific herring (Clupea harengus pallasi) Kathi A. Lefebvre , N

Herring ( Clupea harengus pallasi ) spawning sites in Puget Sound, Washington overlap spatially and temporally with blooms of Alexandrium catenella , a toxic dinoflagellate species responsible for paralytic shellfish poisoning. Consequently, newly hatched herring larvae may be regularly exposed to the suite of dissolved paralytic shellfish toxins that are released into the water column from toxic cells during blooms. To date, virtually nothing is known about the impacts of these neurotoxins on early developmental stages of marine fish. In the present study, herring larvae at three ages, 0 days post hatch (dph), 4 dph, and 11 dph, were exposed to dissolved saxitoxin (STX) in 24-h and multi-day exposures. All larvae were examined for sensorimotor function (i.e. spontaneous swimming behavior and touch response). Significant reductions in spontaneous and touch-activated swimming behavior occurred within 1 h of exposure. EC 50 s at 1 h of exposure were 1,500, 840, and 700 μg STX equiv. l −1 for larvae introduced to STX at 0, 4, and 11 dph, respectively. This progressive age-specific increase in STX-induced paralysis suggests that older larvae were more sensitive to the toxin than younger larvae. Interestingly, herring larvae at all ages exhibited a significant degree of neurobehavioral recovery within 4–24 h of continuous exposure relative to the 1-h time point. This recovery of normal motor behaviors was not observed in previous studies with freshwater zebrafish ( Danio rerio ) larvae under the same continuous exposure conditions, suggesting that an adaptive detoxification or toxin sequestration mechanism may have evolved in some species of marine fish larvae. Our data reveal that (1) dissolved STX is bioavailable to marine finfish larvae, (2) the toxin is a paralytic agent with potencies that differ between developmental stages, and (3) STX-induced sensorimotor inhibition occurs rapidly but is transient in marine larvae. Collectively, these results suggest that dissolved algal toxins may have important sublethal effects on marine fish populations.

Marine Biology

Evolutionary history of the ABCB2 genomic region in teleosts

Gene duplication, silencing and translocation have all been implicated in shaping the unique genomic architecture of the teleost MH regions. Previously, we demonstrated that trout possess five unlinked regions encoding MH genes. One of these regions harbors ABCB2 which in all other vertebrate classes is found in the MHC class II region. In this study, we sequenced a BAC contig for the trout ABCB2 region. Analysis of this region revealed the presence of genes homologous to those located in the human class II (ABCB2, BRD2, ??DAA), extended class II (RGL2, PHF1, SYGP1) and class III (PBX2, Notch-L) regions. The organization and syntenic relationships of this region were then compared to similar regions in humans, Tetraodon and zebrafish to learn more about the evolutionary history of this region. Our analysis indicates that this region was generated during the teleost-specific duplication event while also providing insight about potential MH paralogous regions in teleosts. ?? 2006 Elsevier Ltd. All rights reserved.

Developmental and Comparative Immunology

Effects of carbamazepine to visual function in early life stage fish

The frequent detection of pharmaceuticals and personal care products (PPCPs) in the environment raises concern for aquatic systems. Carbamazepine (CBZ), an antiepileptic drug, is among the most detected PPCP globally, with concentrations in surface water exceeding those that induce toxicity to aquatic organisms. Non-targeted transcriptomic profiling was conducted in zebrafish ( Danio rerio ) larvae exposed to 0, 1, 5, 10, or 50 μg/L CBZ from 2 h post fertilization (hpf) through hatching, and then sampled at 48, 72, or 144 hpf. Transcriptomic profiles were annotated and characterized with in silico bioinformatic software to assess top enriched pathways and identify targets of environmentally relevant concentrations of CBZ and anchor molecular effects to higher levels of biological organization. Based on this analysis, CBZ was predicted to impair visual perception and sensory system development. The number of eye saccades, determined with a visually mediated behavioral assay, optokinetic response, was significantly reduced in 144 hpf larvae exposed to concentrations as low as 1 μg/L CBZ. These results indicate that environmentally relevant concentrations of CBZ may target and impact processes involved in visual function in fish.

Environmental Research

Spatial and temporal patterns of surface water quality and ichthyotoxicity in urban and rural river basins in Texas

The Double Mountain Fork Brazos River (Texas, USA) consists of North (NF) and South Forks (SF). The NF receives urban runoff and twice-reclaimed wastewater effluent, whereas the SF flows through primarily rural areas. The objective of this study was to determine and compare associations between standard water quality variables and ichthyotoxicity at a landscape scale that included urban (NF) and rural (SF) sites. Five NF and three SF sites were sampled quarterly from March 2008 to March 2009 for specific conductance, salinity, hardness, pH, temperature, and turbidity; and a zebrafish ( Danio rerio ) embryo bioassay was used to determine ichthyotoxicity. Metal and nutrient concentrations at all sites were also measured in addition to standard water quality variables in spring 2009. Principal component analyses identified hardness, specific conductance, and salinity as the water variables that best differentiate the urban NF (higher levels) from rural SF habitat. Nutrient levels were also higher in the NF, but no landscape scale patterns in metal concentrations were observed. Ichthyotoxicity was generally higher in NF water especially in winter, and multiple regression analyses suggested a positive association between water hardness and ichthyotoxicity. To test for the potential influence of the toxic golden alga ( Prymnesium parvum ) on overall ichthyotoxicity, a cofactor known to enhance golden alga toxin activity was used in the bioassays. Golden alga ichthyotoxicity was detected in the NF but not the SF, suggesting golden alga may have contributed to overall ichthyotoxicity in the urban but not in the rural system. In conclusion, the physicochemistry of the urban-influenced NF water was conducive to the expression of ichthyotoxicity and also point to water hardness as a novel factor influencing golden alga ichthyotoxicity in surface waters.

Texas

Prolactin and teleost ionocytes: new insights into cellular and molecular targets of prolactin in vertebrate epithelia

The peptide hormone prolactin is a functionally versatile hormone produced by the vertebrate pituitary . Comparative studies over the last six decades have revealed that a conserved function for prolactin across vertebrates is the regulation of ion and water transport in a variety of tissues including those responsible for whole-organism ion homeostasis. In teleost fishes, prolactin was identified as the “freshwater-adapting hormone”, promoting ion-conserving and water-secreting processes by acting on the gill, kidney, gut and urinary bladder. In mammals, prolactin is known to regulate renal, intestinal, mammary and amniotic epithelia, with dysfunction linked to hypogonadism, infertility, and metabolic disorders. Until recently, our understanding of the cellular mechanisms of prolactin action in fishes has been hampered by a paucity of molecular tools to define and study ionocytes, specialized cells that control active ion transport across branchial and epidermal epithelia. Here we review work in teleost models indicating that prolactin regulates ion balance through action on ion transporters, tight-junction proteins, and water channels in ionocytes, and discuss recent advances in our understanding of ionocyte function in the genetically and embryonically accessible zebrafish ( Danio rerio ). Given the high degree of evolutionary conservation in endocrine and osmoregulatory systems, these studies in teleost models are contributing novel mechanistic insight into how prolactin participates in the development, function, and dysfunction of osmoregulatory systems across the vertebrate lineage.

General and Comparative Endocrinology

Estradiol-induced gene expression in largemouth bass (Micropterus salmoides)

Vitellogenin (Vtg) and estrogen receptor (ER) gene expression levels were measured in largemouth bass to evaluate the activation of the ER-mediated pathway by estradiol (E2). Single injections of E2 ranging from 0.0005 to 5 mg/kg up-regulated plasma Vtg in a dose-dependent manner. Vtg and ER mRNAs were measured using partial cDNA sequences corresponding to the C-terminal domain for Vtg and the ligand-binding domain of ER?? sequences. After acute E2-exposures (2 mg/kg), Vtg and ER mRNAs and plasma Vtg levels peaked after 2 days. The rate of ER mRNA accumulation peaked 36-42 h earlier than Vtg mRNA. The expression window for ER defines the primary response to E2 in largemouth bass and that for Vtg a delayed primary response. The specific effect of E2 on other estrogen-regulated genes was tested during these same time windows using differential display RT-PCR. Specific up-regulated genes that are expressed in the same time window as Vtg were ERp72 (a membrane-bound disulfide isomerase) and a gene with homology to an expressed gene identified in zebrafish. Genes that were expressed in a pattern that mimics the ER include the gene for zona radiata protein ZP2, and a gene with homology to an expressed gene found in winter flounder. One gene for fibrinogen ?? was down-regulated and an unidentified gene was transiently up-regulated after 12 h of exposure and returned to basal levels by 48 h. Taken together these studies indicate that the acute molecular response to E2 involves a complex network of responses over time. ?? 2002 Elsevier Science Ireland Ltd. All rights reserved.

Molecular and Cellular Endocrinology

Integrating contaminant source indicators, water quality measures, and ecotoxicity to characterize contaminant mixtures and per- and polyfluoroalkyl substances (PFAS) variability in an urban watershed

Thousands of chemical contaminants threaten watersheds but are time and cost prohibitive to monitor. Identifying their sources, transport, and ecological risk is limited in heterogeneous urban watersheds. We present an integrative watershed approach using source-specific indicator compounds, common water quality measures, and ecotoxicity assays to examine the distribution of contaminant mixtures in an urbanized watershed. Indicator compound concentrations were temporally and spatially distributed for treated/untreated sewage (sucralose, artificial sweetener), road runoff (diphenyl-guanidine [DPG] and 6PPD-quinone [6PPD-Q], automobile tire additives), and lawncare runoff (aminomethanephosphonic acid (AMPA), major degradant of the herbicide glyphosate). Sucralose was predominately sourced from treated wastewater; measurable concentrations in tributaries indicated raw sewage inputs. DPG and 6PPD-Q concentrations correlated to road density during base flow and were elevated during stormflow. AMPA was measurable spring through fall, especially where lawns were dense. When specific sources dominated flow, water quality measures correlated with wastewater (sulfate, potassium, chloride, and sodium) and road runoff (chromium and lead) indicators. The limited behavioral toxicity observed in exposed zebrafish ( Danio rerio ) (18%) was not well explained by source-indicators. PFAS concentrations were highly variable spatially but not well explained by our source-specific indicator compounds. More costly compound-specific monitoring may be necessary when multiple sources exist or when unexpected toxicity trends occur.

North Carolina

Cell-Based metabolomics for untargeted screening and prioritization of vertebrate-active stressors in streams across the United States

The U.S. Geological Survey and the U.S. Environmental Protection Agency have assessed contaminants in 38 streams across the U.S., using an extensive suite of target-chemical analysis methods along with a variety of biological effects tools. Here we report zebrafish liver (ZFL) cell-culture based NMR metabolomic analysis of these split stream samples. We used this untargeted approach to evaluate the sites according to overall impact on the ZFL metabolome and found that neither the total number of organics detected at the sites, nor their cumulative concentrations, were good predictors of these impacts. Further, we used partial-least squares regression to compare ZFL endogenous metabolite profiles to values for 455 potential stressors (organics, inorganics, and physical properties) measured in these waters and found that the profiles covaried with at most 280 of the stressors, which were subsequently ranked into quartiles based on the strength of their covariance. While contaminants of emerging concern (CECs) were well represented in the top, most strongly, covarying quartile – suggesting considerable potential for eliciting biological responses at these sites – there was even higher representation of various well-characterized legacy contaminants (e.g., PCBs). These results emphasize the importance of complementing chemical analysis with untargeted bioassays to help focus regulatory efforts on the most significant ecosystem threats.

Environmental Science & Technology

Protection from UV light is an evolutionarily conserved feature of the haematopoietic niche

Haematopoietic stem and progenitor cells (HSPCs) require a specific microenvironment, the haematopoietic niche, which regulates HSPC behaviour . The location of this niche varies across species, but the evolutionary pressures that drive HSPCs to different microenvironments remain unknown. The niche is located in the bone marrow in adult mammals, whereas it is found in other locations in non-mammalian vertebrates, for example, in the kidney marrow in teleost fish. Here we show that a melanocyte umbrella above the kidney marrow protects HSPCs against ultraviolet light in zebrafish. Because mutants that lack melanocytes have normal steady-state haematopoiesis under standard laboratory conditions, we hypothesized that melanocytes above the stem cell niche protect HSPCs against ultraviolet-light-induced DNA damage. Indeed, after ultraviolet-light irradiation, unpigmented larvae show higher levels of DNA damage in HSPCs, as indicated by staining of cyclobutane pyrimidine dimers and have reduced numbers of HSPCs, as shown by cmyb (also known as myb ) expression. The umbrella of melanocytes associated with the haematopoietic niche is highly evolutionarily conserved in aquatic animals, including the sea lamprey, a basal vertebrate. During the transition from an aquatic to a terrestrial environment, HSPCs relocated into the bone marrow, which is protected from ultraviolet light by the cortical bone around the marrow. Our studies reveal that melanocytes above the haematopoietic niche protect HSPCs from ultraviolet-light-induced DNA damage in aquatic vertebrates and suggest that during the transition to terrestrial life, ultraviolet light was an evolutionary pressure affecting the location of the haematopoietic niche.

Nature

Ecotoxicological effects of crude oil to early life stage Danio rerio: A molecular, morphological and behavioral approach focused on swim bladder development

Proper development and inflation of the swim bladder is essential for swimming and foraging behavior in fish. To characterize the effects of the Brazilian oil spill that occurred between 2019 and 2020 to early life stage fish, the expression of genes involved in swim bladder development were targeted, with biochemical assays, morphology, and behavior assessed. The swim bladder was focused on due to recent findings of being a target of polycyclic aromatic hydrocarbons (PAHs) present in oil. Zebrafish ( Danio rerio ) were exposed to oil water accommodated fractions at concentrations measured following the spill, 1.12–71.8 total PAHs (μg-ΣPAHt L −1 ). Larvae exposed to 4.49 and 17.9 μg-ΣPAHt L −1 had a downregulation in swim bladder development genes hb9 , sox2 , has2 , and elovl1a at 48 hr postfertilization (hpf). Downregulation in these genes was associated with a high frequency of uninflated swim bladders at 96 and 168 hpf, with uninflated swim bladders detected in 100% of 96 to 168 hpf larvae exposed to 35.9 and 71.8 μg-ΣPAHt L −1 . Superoxide dismutase and catalase levels were inhibited in larvae exposed to 1.12 and 2.24 μg-ΣPAHt L −1 , respectively. There was an increase in glutathione-S-transferase and glutathione levels in exposed larvae. Average swimming speed and distance were reduced in larvae exposed to 1.12 μg-ΣPAHt L −1 . This suggests that complex mixtures of PAHs from crude oil can inhibit the transcription of genes involved in swim bladder tissue development and proper swim-up behavior, which may have implications for the viability and success of developing larvae, affecting recruitment.

Environmental Toxicology and Chemistry

Exploring genetic variation and population structure in a threatened species, Noturus placidus, with whole-genome sequence data

The Neosho madtom ( Noturus placidus ) is a small catfish, generally less than 3 inches in length, unique to the Neosho-Spring River system within the Arkansas River Basin. It was federally listed as threatened in 1990, largely due to habitat loss. For conservation efforts, we generated whole-genome sequence data from 10 Neosho madtom individuals originating from 3 geographically separated populations to evaluate genetic diversity and population structure. A Neosho madtom genome was de novo assembled, and genome size and content were assessed. Single nucleotide polymorphisms were assessed from de Bruijn graphs, and via reference alignment with both the channel catfish ( Ictalurus punctatus) reference genome and Neosho madtom reference genome. Principal component analysis and structure analysis indicated weak population structure, suggesting fish from the 3 locations represent a single population. Using a novel method, genome-wide conservation and divergence between the Neosho madtom, channel catfish, and zebrafish ( Danio rerio ) was assessed by pairwise contig alignment, which demonstrated that genes important to embryonic development frequently had conserved sequences. This research in a threatened species with no previously published genomic resources provides novel genetic information to guide current and future conservation efforts and demonstrates that using whole-genome sequencing provides detailed information of population structure and demography using only a limited number of rare and valuable samples.

Arkansas, Kansas, Missouri, Oklahoma