Life history traits and genotoxic effects on Daphnia magna exposed to waterborne uranium and to a uranium mine effluent - a transgenerational study Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-19 Paulo Reis, Ruth Pereira, F.P. Carvalho, J. Oliveira, M. Malta, Sónia Mendo, Joana Lourenço
Influence of pH on the uptake and toxicity of β-blockers in embryos of zebrafish, Danio rerio Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-28 Lisa Bittner, Elisabet Teixido, Bettina Seiwert, Beate I. Escher, Nils Klüver
ß-Blockers are weak bases with acidity constants related to their secondary amine group. At environmental pH they are protonated with the tendency to shift to their neutral species at more alkaline pH. Here we studied the influence of pH from 5.5 to 8.6 on the toxicity of the four ß-blockers atenolol, metoprolol, labetalol and propranolol in zebrafish embryos, relating toxicity not only in a conventional way to external aqueous concentrations but also to measured internal concentrations. Besides lethality, we evaluated changes in swimming activity and heartbeat, using the Locomotor Response (LMR) method and the Vertebrate Automated Screening Technology (VAST) for high throughput imaging. Effects of metoprolol, labetalol and propranolol were detected on phenotype, heart rate and swimming activity. External effect concentrations decreased with increasing neutral fraction for all three pharmaceuticals, attributed by an enhanced uptake of the neutral species in comparison to the corresponding charged form. The LC50 of metoprolol decreased by a factor of 35 from 1.91 mM with almost complete cationic state at pH 7.0 to 0.054 mM with 8% neutral fraction at pH 8.6. For propranolol the LC50 of 2.42 mM at pH 5.5 was even 100 fold higher than the LC50 at pH 8 with 0.023 mM where 3% were neutral fraction. No effects were detected in the zebrafish embryo exposed to atenolol. The internal concentrations for metoprolol and propranolol were quantified at non-toxic concentrations and at the LC10. Apparent bioconcentration factors (BCF) ranged from 1.96 at pH 7.0 to 32.0 at pH 8.6 for metoprolol and from 1.86 at pH 5.5 to 169 at pH 8.0 for propranolol. The BCFs served to predict the internal effect concentrations from the measured external effect concentrations. Internal effect concentrations of metoprolol and propranolol were in a similar range for all pH-values and for all endpoints. Interestingly, the internal effect concentrations were in the internal concentration range of baseline toxicity, which suggests that the effects of the ß-blockers are rather unspecific, even for sublethal effects on heart rate. In summary, our data confirm that the pH-dependent toxicity related to external concentrations can be explained by toxicokinetic effects and that the internal effect concentrations are pH-independent.
Copper oxide nanoparticles induce the transcriptional modulation of oxidative stress-related genes in Arbacia lixula embryos Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-14 Alessia Giannetto, Tiziana Cappello, Sabrina Oliva, Vincenzo Parrino, Giuseppe De Marco, Salvatore Fasulo, Angela Mauceri, Maria Maisano
Copper oxide nanoparticles (CuO NPs) are widely used in various industrial applications, i.e. semiconductor devices, batteries, solar energy converter, gas sensor, microelectronics, heat transfer fluids, and have been recently recognized as emerging pollutants of increasing concern for human and marine environmental health. Therefore, the toxicity of CuO NPs needs to be thoroughly understood. In this study, we evaluated the potential role of oxidative stress in CuO NP toxicity by exploring the molecular response of Arbacia lixula embryos to three CuO NP concentrations (0.7, 10, 20 ppb) by investigating the transcriptional patterns of oxidative stress-related genes (catalase and superoxide dismutase) and metallothionein, here cloned and characterized for the first time. Time- and concentration-dependent changes in gene expression were detected in A. lixula embryos exposed to CuO NPs, up to pluteus stage (72 hours post-fertilization, hpf), indicating that oxidative stress is one of the toxicity mechanisms for CuO NPs. These findings provide new insights into the comprehension of the molecular mechanisms underlying copper nanoparticle toxicity in A. lixula sea urchin and give new tools for monitoring of aquatic areas, thus corroborating the suitability of this embryotoxicity assay for future evaluation of impacted sites.
Transcriptomics provides mechanistic indicators of fluoride toxicology on endochondral ossification in the hind limb of Bufo gargarizans Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-10 Wu Chao, Yuhui Zhang, Lihong Chai, Hongyuan Wang
Endochondral ossification, the process by which most of the bone is formed, is regulated by many specific groups of molecules and extracellular matrix components. Hind limb of Bufo gargarizans is a model to study endochondral ossification during metamorphosis. Chinese toad (Bufo gargarizans) were exposed to different fluoride concentrations (0, 1, 5, 10 and 20 mg L-1) from G3 to G42. The development of hind limb of B. gargarizans was observed using the double staining methodology. The transcriptome of hind limb of B. gargarizans was conducted using RNA-seq approach, and differentially expressed gene was also validated. In addition, the location of Sox9 and Ihh in the growth cartilage was determined using in situ hybridization. Our results showed that 5 mg L-1 stimulated bone mineralization, while 10 and 20 mg L-1 exposure could inhibit the tibio-fibula, tarsus and metacarpals ossification. Besides, 10 mg F/L treatment could down-regulate Ihh, Sox9, D2, D3, TRα, TRβ, Wnt10, FGF3 and BMP6 expression, while up-regulate ObRb and HHAT mRNA expression in the hind limb of B. gargarizans. Transcript level changes of Ihh, Sox9, D2, D3, TRα, TRβ, Wnt10, FGF3 and BMP6 were consistent with the results of RT-qPCR. In situ hybridization revealed that Ihh was expressed in prehypertrophic chondrocytes, while Sox9 was abundantly expressed in proliferous, prehypertrophic and hypertrophic chondrocytes. However, 10 mg F-/L did not cause any affect in the location of the Ihh and Sox9 mRNA. Therefore, high concentration of fluoride could affect the ossification-related genes mRNA expression and then inhibit the endochondral ossification. The present study thus will greatly contribute to our understanding of the effect of environmental contaminant on ossification in amphibian.
Phosphate alleviation of glyphosate-induced toxicity in Hydrocharis dubia (Bl.) Backer Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-30 Guidi Zhong, Zhonghua Wu, Nian Liu, Jun Yin
Glyphosate, as a broad-spectrum herbicide, is frequently detected in water, and phosphorus widely enters the water due to the extensive use of phosphorus-containing substances in agriculture, industries and daily life. Thus, aquatic ecosystems are exposed to both glyphosate and phosphorus, which may affect aquatic organisms. In the present research, we studied the physiological responses of the floating aquatic plant species H. dubia to different concentrations of glyphosate (0, 1, 5, 15 mg/L) with different levels of phosphate (0, 50, 100 mg/L) after 14 days (d) of treatment. We explored glyphosate toxicity in H. dubia and investigated whether phosphate addition mitigates glyphosate toxicity in this species, which will provide a theoretical basis for the ecotoxicological study of aquatic plants. The results show that glyphosate significantly reduced the chlorophyll content, leaf number and root length of H. dubia, while it significantly increased the malondialdehyde (MDA), hydrogen peroxide (H2O2), shikimate, proline, and soluble protein content and enzyme activities (superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX) and polyphenol oxidase (PPO)) in H. dubia. After phosphate supplement, the MDA, H2O2, proline, and soluble protein contents and enzyme activities in the plants treated with glyphosate decreased. These results indicate that the concentration of glyphosate investigated in our study can cause oxidative stress and affect the growth of H. dubia. Phosphate can alleviate glyphosate-induced oxidative stress in H. dubia.
Temporal variations in kidney metal concentrations and their implications for retinoid metabolism and oxidative stress response in wild yellow perch (Perca flavescens) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-09 Michel A. Defo, Louis Bernatchez, Peter G.C. Campbell, Patrice Couture
The objective of this study was to determine if temporal variations in tissue metal concentrations are related to biomarkers of retinoid metabolism and oxidative stress responses in juvenile yellow perch (Perca flavescens). To this end, kidney metal (Cd, Cu and Zn) concentrations were measured in fish sampled in spring and fall 2012 in four lakes representing a wide range of water and sediment metal contamination in the Rouyn-Noranda (Quebec) region. Lakes Opasatica and Hélène were considered as reference lakes while lakes Dufault and Marlon were metal-contaminated. Kidney concentrations of Cd, Cu and Zn varied widely between spring and fall in fish from both clean and metal-contaminated lakes. An inter-lake difference in renal metal concentrations was only observed for Cd, with fish from Lake Marlon consistently displaying higher concentrations. In the spring, the concentrations of liver dehydroretinol, dehydroretinyl palmitate and total vitamin A esters were higher in fish sampled in the most contaminated lake.Strong temporal variations in the concentrations of these metabolites, as well as in the percentage of liver free dehydroretinol and the epidermal retinol dehydrogenase 2 transcription levels, were observed in fish living in the most metal-impacted lake, with generally higher values in the spring. In contrast to liver, in muscle, no clear seasonal variations in the concentrations of dehydroretinol, dehydroretinyl stearate or in the percentage of free dehydroretinol were observed in fish captured in the most contaminated lake. Temporal variations of traditional biomarkers of oxidative stress response were also observed in the most metal-impacted lake. For example, the transcription level of the gene encoding Cu/Zn superoxide dismutase-1 in liver and muscle catalase activity of perch sampled in the most contaminated lake were higher in spring than in fall. Positive relationships were found between kidney Cd concentrations and the transcription level of the gene encoding glucose 6-phosphate dehydrogenase, and all forms of retinoid concentrations in liver in spring, except with the percentage of free dehydroretinol where the correlation was negative. Our results translate to a state of stress caused by Cd and illustrate that temporal variations in tissue metal concentrations affect retinoid metabolism and antioxidant capacities in juvenile wild yellow perch. Overall this study contributes to evidence the importance of considering temporal variations when investigating the consequences of metal contamination on the physiology of wild fish.
New toxicogenetic insights and ranking of the selected pharmaceuticals belong to the three different classes: a toxicity estimation to confirmation approach Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-09 Yi Liu, Muhammad Junaid, Yan Wang, Yu-Mei Tang, Wan-Ping Bian, Wen-Xu Xiong, Hai-Yang Huang, Chun-Di Chen, De-Sheng Pei
Comparative effects of cadmium, zinc, arsenic and chromium on olfactory-mediated neurobehavior and gene expression in larval zebrafish (Danio rerio) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-28 Kevin Heffern, Keith Tierney, Evan P. Gallagher
Studies have shown that olfactory-mediated behaviors that are critical to survival can be disrupted by exposure to certain metals. Polluted waterways often contain elevated levels of metals, yet only a subset have been characterized for their potential to cause olfactory toxicity. A larval zebrafish behavioral assay was developed to characterize concentration-response curves for zinc (Zn), hexavalent chromium (Cr), and arsenate (As) olfaction inhibition. Cadmium (Cd), an established olfactory toxicant, was used as a positive control. As expected, following a 24-hour exposure to Cd, we observed a reduced response to taurocholic acid (TCA), a substrate for ciliated olfactory sensory neurons (OSNs), thus validating the behavioral assay. Zn exposure similarly decreased the olfactory response toward TCA, (IC50: 36 μg/L and 76 μg/L, for Cd and Zn, respectively). The response towards a secondary odorant L-cysteine (Cys), a substrate for ciliated and microvillous OSNs, was significantly altered by both Cd and Zn exposure, although the response to Cys was not completely removed in Zn treated larvae, suggesting preferential toxicity towards ciliated OSNs. No significant changes in olfactory responses were observed following Cr and As exposures. Exposures to binary mixtures of Cd and Zn indicated that Zn had a protective effect against Cd toxicity at low Zn concentrations. QuantiGene (QDP) RNA analysis revealed Cd to be a potent inducer of metallothionein 2 (mt2) mRNA in zebrafish larvae, and Zn to be a weak mt2 inducer, suggesting a protective role of mt2 in Cd and Zn olfactory injury. By contrast, QDP analysis of eight other genes important in mitigating the effects of oxidative stress suggested an antioxidant response to Cd, but not Zn, As, and Cr suggesting that oxidative stress was not a primary mechanism of Zn-induced olfactory dysfunction. In summary, our study indicates that Zn inhibits zebrafish olfaction at environmental concentrations and may potentially mitigate Cd induced olfactory dysfunction when present in mixtures. The zebrafish behavioral trough assay incorporating the odorants L-cysteine and TCA is an effective assay to assess the effects of metals on olfactory function.
Transcriptome Analysis Reveals Novel Insights Into the Response of Low-dose Benzo(a)pyrene Exposure in Male Tilapia Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-08 Reyna Cristina Colli-Dula, Xiefan Fang, David Moraga-Amador, Nacira Albornoz-Abud, Roberto Zamora-Bustillos, Ana Conesa, Omar Zapata-Perez, Diego Moreno, Emanuel Hernandez-Nuñez
Despite a wide number of toxicological studies that describe benzo[a]pyrene (BaP) effects, the metabolic mechanisms that underlie these effects in fish are largely unknown. Of great concern is the presence of BaP in aquatic systems, especially those in close proximity to human activity leading to consumption of potentially contaminated foods. BaP is a known carcinogen and it has been reported to have adverse effects on the survival, development and reproduction of fish. The purpose of this study was to investigate if a low dose of BaP can alter genes and key metabolic pathways in the liver and testis in male adult tilapia, and whether these could be associated with biological endpoints disruption. We used both high-throughput RNA-Sequencing to assess whole genome gene expression following repeated intraperitoneal injections of 3 mg/Kg of BaP (every 6 days for 26 days) and morphometric endpoints as indicators of general health. Condition factor (K) along with hepatosomatic and gonadosomatic indices (morphometric parameters) were significantly lower in BaP-treated fish than in controls. BaP exposure induced important changes in the gene expression pattern in liver and testis as revealed by both Pathway and Gene Ontology (GO) analyses. Alterations that were shared by both tissues included arachidonic acid metabolism, androgen receptor to prostate-specific antigen signaling, and insulin-associated effects on lipogenesis. The most salient liver-specific effects included: biological processes involved in detoxification, IL6-associated insulin resistance, mTOR hyperactivation, mitotic cytokinesis, spindle pole and microtubule binding. BaP effects that were confined to the testis included: immune system functions, inflammatory response, estrogen and androgen metabolic pathways. Taken together, gene expression and morphometric end point data indicate that the reproductive success of adult male tilapia could be compromised as a result of BaP exposure. These results constitute new insights on the mechanism of action of low dose BaP in a non-model organism (tilapia).
RNA-Seq analysis of transcriptome responses in Atlantic cod (Gadus morhua) precision-cut liver slices exposed to benzo[a]pyrene and 17α-ethynylestradiol Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-07 Fekadu Yadetie, Xiaokang Zhang, Eileen Marie Hanna, Libe Aranguren-Abadía, Marta Eide, Nello Blaser, Morten Brun, Inge Jonassen, Anders Goksøyr, Odd André Karlsen
Polycyclic aromatic hydrocarbons such as benzo[a]pyrene (BaP) that activate the aryl hydrocarbon receptor (Ahr) pathway, and endocrine disruptors acting through the estrogen receptor pathway are among environmental pollutants of major concern. In this work, we exposed Atlantic cod (Gadus morhua) precision-cut liver slices (PCLS) to BaP (10 nM and 1000 nM), ethynylestradiol (EE2) (10 nM and 1000 nM), and equimolar mixtures of BaP and EE2 (10 nM and 1000 nM) for 48 hours, and performed RNA-Seq based transcriptome mapping followed by systematic bioinformatics analyses. Our gene expression analysis showed that several genes were differentially expressed in response to BaP and EE2 treatments in PCLS. Strong up-regulation of genes coding for the cytochrome P450 1a (Cyp1a) enzyme and the Ahr repressor (Ahrrb) was observed in BaP treated PCLS. EE2 treatment of liver slices strongly up-regulated genes coding for precursors of vitellogenin (Vtg) and eggshell zona pellucida (Zp) proteins. As expected, pathway enrichment and network analysis showed that the Ahr and estrogen receptor pathways are among the top affected by BaP and EE2 treatments, respectively. Interestingly, two genes coding for fibroblast growth factor 3 (Fgf3) and fibroblast growth factor 4 (Fgf4) were up-regulated by EE2 in this study. To our knowledge, the fgf3 and fgf4 genes have not previously been described in relation to estrogen signaling in fish liver, and these results suggest the modulation of the FGF signaling pathway by estrogens in fish. The signature expression profiles of top differentially expressed genes in response to the single compound (BaP or EE2) treatment were generally maintained in the expression responses to the equimolar binary mixtures. However, in the mixture-treated groups, BaP appeared to have anti-estrogenic effects as observed by lower number of differentially expressed putative EE2 responsive genes. Our in-depth quantitative analysis of changes in liver transcriptome in response to BaP and EE2, using PCLS tissue culture provides further mechanistic insights into effects of the compounds. Moreover, the analyses demonstrate the usefulness of PCLS in cod for omics experiments.
Photo-induced antibacterial activity of a porphyrin derivative isolated from the harmful dinoflagellate Heterocapsa circularisquama Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-07 Li Wencheng, Kichul Cho, Yasuhiro Yamasaki, Satoshi Takeshita, Kiju Hwang, Daekyung Kim, Tatsuya Oda
The dinoflagellate Heterocapsa circularisquama is highly toxic to bivalves. However, significant toxicity to finfish species has not been reported. We previously found that H. circularisquama has light-dependent haemolytic agents. Purification and chemical structural analyses revealed that the haemolytic agent H2-a is a porphyrin derivative, which exhibits light-dependent cytotoxicity toward tumour cells. To clarify the biological activity of H2-a further, its antibacterial activities against Gram-positive and Gram-negative bacteria were investigated in this study. A fraction (F5) equivalent to H2-a purified from the methanol extract of H. circularisquama showed potent light-dependent bactericidal activity toward Staphylococcus aureus, and the activity was concentration- and light illumination time-dependent; however, Escherichia coli was highly resistant to F5. Electron microscopic observation suggested that F5 induces morphological changes in S. aureus in a light-dependent manner. Further analysis using other bacterial species showed that the Gram-positive bacterium Bacillus subtilis was more sensitive than the Gram-negative bacteria Pseudomonas aeruginosa and Vibrio alginolyticus. These results indicate that F5 is a photo-induced antibacterial agent with relatively higher specificity to Gram-positive bacteria. Iodometric assay suggested that singlet oxygen was generated from light-illuminated F5. Histidine, a specific singlet oxygen scavenger, markedly inhibited the photosensitising antibacterial activity of F5 against S. aureus, suggesting the involvement of singlet oxygen in antibacterial activity. The antibacterial spectrum of F5 was evidently different from that of 5,10,15,20-tetra (N,N,N-trimethylanilinium) porphyrin tetratosylate, a commercially available porphyrin compound with antibacterial activity. Our results demonstrate that H. circularisquama has a novel antibacterial photosensitiser, a porphyrin derivative, with relatively higher specificity to Gram-positive bacteria. To the best of our knowledge, this is the first study to discover a porphyrin derivative with antibacterial activity in marine microalga.
Genome-wide identification of 99 autophagy-related (Atg) genes in the monogonont rotifer Brachionus spp. and transcriptional modulation in response to cadmium Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-30 Hye-Min Kang, Jin-Sol Lee, Min-Sub Kim, Young Hwan Lee, Jee-Hyun Jung, Atsushi Hagiwara, Bingsheng Zhou, Jae-Seong Lee, Chang-Bum Jeong
Autophagy originated from the common ancestor of all life forms, and its function is highly conserved from yeast to humans. Autophagy plays a key role in various fundamental biological processes including defense, and has developed through serial interactions of multiple gene sets referred to as autophagy-related (Atg) genes. Despite their significance in metazoan life and evolution, few studies have been conducted to identify these genes in aquatic invertebrates. In this study, we identified whole Atg genes in four Brachionus rotifer spp., namely B. calyciflorus, B. koreanus, B. plicatilis, and B. rotundiformis, through searches of their entire genomes; and we annotated them according to the yeast nomenclature. Twenty-four genes orthologous to yeast genes were present in all of the Brachionus spp. while three additional gene duplicates were identified in the genome of B. koreanus, indicating that these genes had diversified during the speciation. Also, their transcriptional responses to cadmium exposure indicated regulation by cadmium-induced oxidative-stress–related signaling pathways. This study provides valuable information on 99 conserved Atg genes involved in autophagosome formation in Brachionus spp., with transcriptional modulation in response to cadmium, in the context of the role of autophagy in the damage response.
Immune response induced by major environmental pollutants through altering neutrophils in zebrafish larvae Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-06 Hongyan Xu, Xiaoyan Zhang, Hankun Li, Caixia Li, Xiao-Jing Huo, Li-Ping Hou, Zhiyuan Gong
Environmental pollutants may cause adverse effects on the immune system of aquatic organisms. However, the cellular effects of pollutants on fish immune system are largely unknown. Here, we exploited the transgenic zebrafish Tg(lysC:DsRed2) larva as a preliminary screening system to evaluate the potential inflammatory effects of environmental pollutants. Tg(lysC:DsRED2) larvae aged 7-day-postfertilization (7 dpf) were treated with selected environmental chemicals for 24 hours (24 h) and the number of neutrophils were quantified using both image analysis and fluorescence activated cell sorting (FACS). We found that the numbers of neutrophils in the Tg(lysC:DsRED2) larvae were significantly increased by most of the organic chemicals tested, including E2 (17β-estradiol), BPA (Bisphenol-A), NDEA (N-nitrosodiethylamine), 4-NP (4-Nitrophenol) and Lindane (γ-hexachlorocyclohexane). Neutrophil numbers were also increased by all the metals tested (Na2HAsO4· 7H2O, Pb(NO3)2, HgCl2, CdCl2, CuSO4·5H2O, ZnSO4, and K2Cr2O7). The only exception was TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin), which significantly reduced the number of neutrophils after exposure. Additionally, the transcription of genes (lyz, mpo, tnfα and il8) related to fish immune system were significantly modulated upon exposure to some of the selected chemicals such as E2, TCDD, Cu and Cd. This study revealed that representatives of major categories of environmental pollutants could cause an acute inflammatory response in zebrafish larvae as shown by alterations in the neutrophils, which may imply a common immunotoxicity mechanism for most environmental pollutants. This study has also demonstrated that Tg(lyz:DsRed2) transgenic zebrafish is an excellent tool for screening environmental chemicals with potential inflammatory effects through FACS-facilitated neutrophil counting.
PROTEOMIC RESPONSE OF GILL MICROSOMES OF CRASSOSTREA BRASILIANA EXPOSED TO DIESEL FUEL WATER-ACCOMMODATED FRACTION Aquat. Toxicol. (IF 4.129) Pub Date : 2018-06-06 Gabrielle do Amaral e Silva Müller, Karim Hahn Lüchmann, Guilherme Razzera, Guilherme Toledo-Silva, Maria João Bebianno, Maria Risoleta Freire Marques, Afonso Celso Dias Bainy
Diesel fuel water-accommodated fraction (diesel-WAF) is a complex mixture of organic compounds that may cause harmful effects to marine invertebrates. Expression of microsomal proteins can be changed by oil exposure, causing functional alterations in endoplasmic reticulum (ER). The aim of this study was to investigate changes in protein expression signatures in microsomes of oysterl Crassostrea brasiliana (=C.gasar) gill after exposure to 10% diesel-WAF for 24 and 72 h. Protein expression signatures of gills of oysters exposed to diesel-WAF were compared to those of unexposed oysters using two–dimensional electrophoresis (2-DE) to identify differentially expressed proteins. A total of 458 protein spots with molecular weights between 30–75 kDa were detected by 2-DE in six replicates of exposed oyster proteomes compared to unexposed ones. Fourteen differentially expressed proteins (six up-regulated and eight down-regulated) were identified. They are: proteins related to xenobiotic biotransformation (cytochrome P450 6 A, NADPH-cytochrome P450 reductase); cytoskeleton (α-tubulin, β-tubulin, gelsolin); processing and degradation of proteins pathways (thioredoxin domain-containing protein E3 ubiquitin-protein ligase MIB2); involved in the biosynthesis of glycolipids and glycoproteins (beta-1,3-galactosyltransferase 1); associated with stress responses (glutamate receptor 4 and 14-3-3 protein zeta, corticotropin-releasing factor-binding protein); plasmalogen biosynthesis (fatty acyl-CoA reductase 1), and sodium-and chloride-dependent glycine transporter 2 and glyoxylate reductase/hydroxypyruvate reductase. Different patterns of protein responses were observed between 24 and 72 h-exposed groups. Expression pattern of microsomal proteins provided a first insight on the potential diesel-WAF effects at protein level in microsomal fraction of oyster gills and indicated new potential biomarkers of exposure and effect. The present work can be a basis for future ecotoxicological studies in oysters aiming to elucidate the molecular mechanisms behind diesel-WAF toxicity and for environmental monitoring programs.
Chronic environmentally relevant levels of simvastatin disrupt embryonic development, biochemical and molecular responses in zebrafish (Danio rerio) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-25 Susana Barros, Rosa Montes, José Benito Quintana, Rosario Rodil, Ana André, Ana Capitão, Joana Soares, Miguel M. Santos, Teresa Neuparth
Comparative toxicity of three phenolic compounds on the embryo of fathead minnow, Pimephales promelas Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-29 Erchao Li, Derek G. Bolser, Kevin J. Kroll, Erica K. Brockmeier, Francesco Falciani, Nancy D. Denslow
Phenols are classified as polar narcotics, which are thought to cause toxicity by non-specific mechanisms, possibly by disrupting membrane structure and function. Here we test three phenolic chemicals, phenol, 2,4-dichlorphenol and pentachlorophenol on embryo development, heartbeat rate and mitochondrial respiration in fathead minnow (Pimephales promelas). While these chemicals have been used on isolated mitochondria, they have not yet been used to verify respiration in intact embryos. Mitochondrial respiration in intact embryos was measured after optimizing the Seahorse XFe24 Extracellular Flux Analyzer. Heartbeat rate and mitochondrial respiration patterns of fathead minnow embryos at different developmental stages were also characterized. Exposures of embryos at developmental stage 20 occurred for 24 h with five concentrations of each phenolic compound ranging from 0.85 to 255 μM for phenol, 0.49 to 147 μM for 2,4-dichlorophenol and 0.3 to 90 μM for pentachlorophenol. Exposure to phenol at the concentrations tested had no effects on development, heartbeat or mitochondrial respiration. However, both 2,4-dichlorophenol and pentachlorophenol showed dose-dependent effects on development, heartbeat rate, and mitochondrial respiration, with the effects occurring at lower concentrations of pentachlorophenol, compared to 2,4-dichlorophenol, highlighting the higher toxicity of the more chlorinated phenols. Both 2,4-dichlorophenol and pentachlorophenol decreased basal mitochondrial respiration of embryos and ATP production. These results indicate that higher chlorinated phenolic chemicals cause developmental toxicity in fathead minnow embryos by decreasing mitochondrial respiration and heartbeat rate.
Styrene impairs normal embryo development in the Mediterranean mussel (Mytilus galloprovincialis) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-30 Rajapaksha Haddokara Gedara Rasika Wathsala, Silvia Franzellitti, Morena Scaglione, Elena Fabbri
This study analysed the effects of styrene, a main monomer in plastic manufacturing and acknowledged to be amongst the most common plastic leachates, on early embryo development of the Mediterranean mussel. Embryotoxicity tests showed that styrene impaired normal embryo development at concentrations (0.01 μg/L–1 mg/L) encompassing the environmental range. Occurrence of normal D-veligers was significantly reduced up to 40% of the total, and larval size was reduced of about 20%. D-veligers grown in the presence of styrene (0.1 and 10 μg/L) showed significant reduction of total Multixenobiotic resistance (MXR) efflux activity that was not apparently related to transcriptional expression of genes encoding P-glycoprotein (ABCB) and Mrp (ABCC), the two main ABC transporters of embryonal MXR system. Indeed, ABCB transcription was not affected by styrene, while ABCC was up-regulated. At these same concentrations, transcriptional profiles of 15 genes underlying key biological functions in embryo development and potential targets of adverse effects of styrene were analysed. Main transcriptional effects were observed for genes involved in shell biogenesis and lysosomal responses (down-regulation), and in neuroendocrine signaling and immune responses (up-regulation). On the whole, results indicate that styrene may affect mussel early development through dysregulation of gene transcription and suggest the possible conservation of styrene mode of action across bivalve life cycle and between bivalves and humans, as well as through unpredicted impacts on protective systems and on shell biogenesis.
High-throughput assessment of oxidative respiration in fish embryos: Advancing adverse outcome pathways for mitochondrial dysfunction Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-27 Christopher L. Souders, Xuefang Liang, Xiaohong Wang, Naomi Ector, Yuan H. Zhao, Christopher J. Martyniuk
Mitochondrial dysfunction is a prevalent molecular event that can result in multiple adverse outcomes. Recently, a novel high throughput method to assess metabolic capacity in fish embryos following exposure to chemicals has been adapted for environmental toxicology. Assessments of oxygen consumption rates using the Seahorse XF(e) 24/96 Extracellular Flux Analyzer (Agilent Technologies) can be used to garner insight into toxicant effects at early stages of development. Here we synthesize the current state of the science using high throughput metabolic profiling in zebrafish embryos, and present considerations for those wishing to adopt high throughput methods for mitochondrial bioenergetics into their research. Chemicals that have been investigated in zebrafish using this metabolic platform include herbicides (e.g. paraquat, diquat), industrial compounds (e.g. benzo-[a]-pyrene, tributyltin), natural products (e.g. quercetin), and anti-bacterial chemicals (i.e. triclosan). Some of these chemicals inhibit mitochondrial endpoints in the μM-mM range, and reduce basal respiration, maximum respiration, and spare capacity. We present a theoretical framework for how one can use mitochondrial performance data in zebrafish to categorize chemicals of concern and prioritize mitochondrial toxicants. Noteworthy is that our studies demonstrate that there can be considerable variation in basal respiration of untreated zebrafish embryos due to clutch-specific effects as well as individual variability, and basal oxygen consumption rates (OCR) can vary on average between 100 and 300 pmol/min/embryo. We also compare OCR between chorionated and dechorionated embryos, as both models are employed to test chemicals. After 24 h, dechorionated embryos remain responsive to mitochondrial toxicants, although they show a blunted response to the uncoupling agent carbonylcyanide-4-trifluoromethoxyphenylhydrazone (FCCP); dechorionated embryos are therefore a viable option for investigations into mitochondrial bioenergetics. We present an adverse outcome pathway framework that incorporates endpoints related to mitochondrial bioenergetics. High throughput bioenergetics assays conducted using whole embryos are expected to support adverse outcome pathways for mitochondrial dysfunction.
Effects of environmentally relevant sub-chronic atrazine concentrations on African clawed frog (Xenopus laevis) survival, growth and male gonad development Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-23 Cornelius Rimayi, David Odusanya, Jana M. Weiss, Jacob de Boer, Luke Chimuka, Felix Mbajiorgu
Sub-chronic toxicity of environmentally relevant atrazine concentrations on exposed tadpoles and adult male African clawed frogs (Xenopus laevis) was evaluated in a quality controlled laboratory for 90 days. The aim of this study was to determine the effects of atrazine on the survival, growth and gonad development of African clawed frogs. After exposure of tadpoles to atrazine concentrations of 0 (control), 0.01, 200 and 500 μg L−1 in water, mortality rates of 0, 0, 3.3 and 70% respectively were recorded for the 90 day exposure period. Morphometry showed significantly reduced tadpole mass in the 500 μg L−1 atrazine exposed tadpoles (p < 0.05). Light microscopy on testes of adult frogs exposed to the same atrazine concentrations using hematoxylin and eosin (H&E) and Van Gieson staining techniques revealed gonadal atrophy, disruption of germ cell lines, seminiferous tubule structure damage and formation of extensive connective tissue around seminiferous tubules of frogs exposed to 200 μg L−1 and 500 μg L−1 atrazine concentrations. Ultrastructural analysis of the cellular organelles using transmission electron microscopy (TEM) revealed significant amounts of damaged mitochondria in testosterone producing Leydig cells as well as Sertoli cells. Biochemical analysis revealed reduced serum testosterone levels in adult frogs at all exposure levels as well as presence of six atrazine metabolites in frog serum and liver. The results indicate that atrazine concentrations greater than the calculated LC50 of 343.7 μg L−1 cause significant mortality in tadpoles, while concentrations ≥200 μg L−1 adversely affect reproductive health of adult frogs and development of tadpoles sub-chronically exposed to atrazine.
Oxidative stress and mitochondrial dysfunction mediated Cd-induced hepatic lipid accumulation in zebrafish Danio rerio Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-17 Ya-Xiong Pan, Zhi Luo, Mei-Qing Zhuo, Chuan-Chuan Wei, Guang-Hui Chen, Yu-Feng Song
The present study was performed to determine the effect of waterborne CdCl2 exposure influencing lipid deposition and metabolism, oxidative stress and mitochondrial dysfunction, and explore the underlying molecular mechanism of cadmium (Cd)-induced disorder of hepatic lipid metabolism in fish. To this end, adult zebrafish were exposed to three waterborne CdCl2 concentrations (0(control), 5 and 25 μg Cd/l, respectively) for 30 days. Lipid accumulation, the activities of enzymes related to lipid metabolism and oxidative stress, as well as the expression level of genes involved in lipid metabolism and mitophagy were determined in the liver of zebrafish. Waterborne CdCl2 exposure increased hepatic triglyceride (TG) and Cd accumulation, the activities of fatty acid synthase (FAS), 6-phosphogluconate dehydrogenase (6PGD), glucose 6-phosphate dehydrogenase (G6PD) and malic enzyme (ME), and the mRNA level of fatty acid synthase (fas), acetyl-CoA carboxylase alpha (acaca), glucose 6-phosphate dehydrogenase (g6pd) and malic enzyme (me), but reduced the mRNA level of carnitine palmitoyl transferase 1 (cpt1), hormone-sensitive lipase alpha (hsla), and adipose triacylglyceride lipase (atgl). The activities of superoxide dismutase (SOD), glutathoinine peroxidase (GPx) and cytochrome c oxidase (COX) and the ATP level were significantly reduced after CdCl2 exposure. CdCl2 exposure significantly increased the mRNA level of genes (microtubule-associated protein light chain 3 alpha (lc3a), PTEN-induced putative kinase 1 (pink1), NIP3-like protein X (nix) and PARKIN (parkin)) related to mitophagy. To elucidate the mechanism, reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC) and the mitochondrial permeability transition (MPT) inhibitor cyclosporine A (CsA) were used to verify the role of ROS and mitochondrial dysfunction in Cd-induced disorder of lipid metabolism. NAC pretreatment reversed the Cd-induced up-regulation of TG accumulation and activities of lipogenic enzymes, and the Cd-induced down-regulation of mRNA levels of lipolytic genes. Meanwhile, NAC pretreatment also blocked the mitochondrial membrane potential (MMP) collapse and decreased the ATP level, suggesting that ROS played a crucial role in regulating the Cd-induced mitochondrial dysfunction. Taken together, our findings, for the first time, highlight the importance of the oxidative stress and mitochondrial dysfunction in Cd-induced disorder of hepatic lipid metabolism, which proposed a novel mechanism for elucidating metal element exposure inducing the disorder of lipid metabolism in vertebrates.
Effects of bisphenol A and its analogs bisphenol F and S on life parameters, antioxidant system, and response of defensome in the marine rotifer Brachionus koreanus Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-22 Jun Chul Park, Min-Chul Lee, Deok-Seo Yoon, Jeonghoon Han, Moonkoo Kim, Un-Ki Hwang, Jee-Hyun Jung, Jae-Seong Lee
To understand the adverse outcome in response to bisphenol A and its analogs bisphenol F and S (BPA, BPF, and BPS), we examined acute toxicity, life parameter, and defensome in the marine rotifer Brachionus koreanus. Among the bisphenol analogs, BPA showed the highest acute toxicity and then BPF and BPS, accordingly in the view of descending magnitude of toxicity. In life parameters including life span and reproduction, BPA, BPF, and BPS were found to cause adverse effect. Both intracellular ROS level and GST activity were significantly increased (P < 0.05) in response to each dosage of bisphenol analogs exposures. In response to bisphenol analogs, defensomes of phase I, II, and III detoxification mechanism demonstrated inverse relationship between the lipophilicity of bisphenol analogs and the expression patterns of defensomes. BPA and BPF were found to have significant modulation (P < 0.05) in the expression of cytochrome P450 (CYP) and GST genes. In phase III, BPS with comparatively lower lipophilicity demonstrated highly diversified expressional pattern, suggesting that BPS is likely caused less toxicity compared to BPA and BPF. In this study, via phase I, II, and III detoxification mechanism, bisphenol A and its analogs F and S demonstrated specific detoxification mechanism in rotifer.
Physiological impacts and bioaccumulation of dietary Cu and Cd in a model teleost: The Amazonian tambaqui (Colossoma macropomum) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-19 Marina Giacomin, Gisele C. Vilarinho, Katia F. Castro, Márcio Ferreira, Rafael M. Duarte, Chris M. Wood, Adalberto L. Val
Increasing anthropogenic activities in the Amazon have led to elevated metals in the aquatic environment. Since fish are the main source of animal protein for the Amazonian population, understanding metal bioaccumulation patterns and physiological impacts is of critical importance. Juvenile tambaqui, a local model species, were exposed to chronic dietary Cu (essential, 500 μg Cu/g food) and Cd (non-essential, 500 μg Cd/g food). Fish were sampled at 10–14, 18–20 and 33–36 days of exposure and the following parameters were analyzed: growth, voluntary food consumption, conversion efficiency, tissue-specific metal bioaccumulation, ammonia and urea-N excretion, O2 consumption, Pcrit, hypoxia tolerance, nitrogen quotient, major blood plasma ions and metabolites, gill and gut enzyme activities, and in vitro gut fluid transport. The results indicate no ionoregulatory impacts of either of the metal-contaminated diets at gill, gut, or plasma levels, and no differences in plasma cortisol or lactate. The Cd diet appeared to have suppressed feeding, though overall tank growth was not affected. Bioaccumulation of both metals was observed. Distinct tissue-specific and time-specific patterns were seen. Metal burdens in the edible white muscle remained low. Overall, physiological impacts of the Cu diet were minimal. However dietary Cd increased hypoxia tolerance, as evidenced by decreased Pcrit, increased time to loss of equilibrium, a lack of plasma glucose elevation, decreased plasma ethanol, and decreased NQ during hypoxia. Blood O2 transport characteristics (P50, Bohr coefficient, hemoglobin, hematocrit) were unaffected, suggesting that tissue level changes in metabolism accounted for the greater hypoxia tolerance in tambaqui fed with a Cd-contaminated diet.
A protective role of autophagy in TDCIPP-induced developmental neurotoxicity in zebrafish larvae Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-15 Ruiwen Li, Ling Zhang, Qipeng Shi, Yongyong Guo, Wei Zhang, Bingsheng Zhou
Tris (1, 3-dichloro-2-propyl) phosphate (TDCIPP), an extensively used organophosphorus flame retardant, is frequently detected in various environmental media and biota, and has been demonstrated as neurotoxic. Autophagy has been proposed as a protective mechanism against toxicant-induced neurotoxicity. The purpose of the present study was to investigate the effect of TDCIPP exposure on autophagy, and its role in TDCIPP-induced developmental neurotoxicity. Zebrafish embryos (2–120 h post-fertilization [hpf]) were exposed to TDCIPP (0, 5, 50 and 500 μg/l) and a model neurotoxic chemical, chlorpyrifos (CPF, 100 μg/l). The developmental endpoints, locomotive behavior, cholinesterase activities, gene and protein expression related to neurodevelopment and autophagy were measured in the larvae. Our results demonstrate that exposure to TDCIPP (500 μg/l) and CPF causes developmental toxicity, including reduced hatching and survival rates and increased malformation rate (e.g., spinal curvature), as well as altered locomotor behavior. The expression of selected neurodevelopmental gene and protein markers (e.g., mbp, syn2a, and α1-tubulin) was significantly down-regulated in CPF and TDCIPP exposed zebrafish larvae. Treatment with CPF significantly inhibits AChE and BChE, while TDCIPP (0–500 μg/l) exerts no effects on these enzymes. Furthermore, the conversion of microtubule-associated protein I (LC3 I) to LC3 II was significantly increased in TDCIPP exposed zebrafish larvae. In addition, exposure to TDCIPP also activates transcription of several critical genes in autophagy (e.g. Becn1, atg3, atg5, map1lc3b and sqstm1). To further investigate the role of autophagy in TDCIPP induced developmental neurotoxicity, an autophagy inducer (rapamycin, Rapa, 1 nM) and inhibitor (chloroquine, CQ, 1 μM) were used. The results demonstrate that the hatching rate, survival rate, and the expression of mbp and а1-tubulin proteins were all significantly increased in larvae treated with TDCIPP (500 μg/l) and Rapa compared to TDCIPP alone. In contrast, co-treatment with the autophagy inhibitor CQ results in exacerbated neurodevelopmental toxicity. Taken together, our results confirm that exposure to TDCIPP induces autophagy, which plays a protective role in TDCIPP-induced developmental neurotoxicity in zebrafish embryos and larvae.
Does fluoxetine exposure affect hypoxia tolerance in the Gulf toadfish, Opsanus beta? Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-23 Molly H.B. Amador, Kevin L. Schauer, M. Danielle McDonald
Due to ineffective wastewater treatment technologies, pharmaceuticals such as the selective serotonin reuptake inhibitors (SSRIs)—a common class of antidepressants which inhibit the serotonin transporter (SERT)—can be found in surface waters and marine receiving waters near wastewater effluents. Understanding how exposure to these chemicals might impact non-target organisms, especially combined with other environmental stressors like hypoxia, is essential in order to thoroughly evaluate environmental risk. It was hypothesized that both acute and chronic exposure to the SSRI fluoxetine (FLX) would interfere with the metabolic hypoxia response of the Gulf toadfish, Opsanus beta. Here we demonstrate that acute intraperitoneal treatment with 50 μg g−1 FLX significantly reduces the regulation index, or degree of metabolic regulation, in toadfish. Acute FLX exposure significantly reduced SERT mRNA expression in the first and third gill arches, but mRNA expression was not affected in heart tissues or in the second gill arch. In contrast, the regulation index was unaffected by 14–17 day waterborne FLX exposure to environmentally relevant (0.01 μg L−1) and approximately 1000-fold higher (8.5 μg L−1) concentrations. However, the higher concentration was sufficient to induce a systemic elevation in plasma serotonin concentrations. Chronic FLX exposure did not alter SERT mRNA expression in heart or gill tissues. The results of this study implicate the involvement of 5-HT pathways in hypoxia tolerance but demonstrate that current environmental levels of FLX are insufficient to impair the metabolic hypoxia response in marine fish.
Effects of oil spill response technologies on the physiological performance of the Arctic copepod Calanus glacialis Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-27 Kirstine Toxværd, Marina Pančić, Helene O. Eide, Janne E. Søreide, Camille Lacroix, Stéphane Le Floch, Morten Hjorth, Torkel Gissel Nielsen
A mesocosm study with oil in ice was performed in Van Mijenfjorden in Svalbard to compare effects of the oil spill responses (OSR) in situ burning, chemical dispersion and natural attenuation on the physiological performance of the Arctic copepod Calanus glacialis. Seawater collected from the mesocosms in winter and spring was used in laboratory incubation experiments, where effects on fecal pellet production, egg production and hatching success were investigated over a period of 14 days. Polycyclic aromatic hydrocarbon (PAH) seawater concentrations were lowest in winter. Brine channel formation in spring resulted in an 18 times increase in PAH concentration in the chemical dispersion treatment (1.67 μg L−1), and a 3 fold increase in the natural attenuation (0.36 μg L−1) and in situ burning (0.04 μg L−1) treatments. The physiological performance of female C. glacialis was unaffected by the PAH seawater concentrations. However, a higher mortality and deformity of nauplii was observed in the chemical dispersion treatment, highlighting the importance of considering secondary effects on next generation in future environmental risk assessment of OSR. This study shows that during the ice-covered period, chemical dispersion of oil spills leads to higher PAH exposure than natural attenuation and in situ burning, with potential consequences for recruitment of Arctic copepods.
Part B: Morphometric and transcriptomic responses to sub-chronic exposure to the polycyclic aromatic hydrocarbon phenanthrene in the fathead minnow (Pimephales promelas) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-23 Jennifer R. Loughery, Karen A. Kidd, Angella Mercer, Chris J. Martyniuk
Phenanthrene is a tricyclic polycyclic aromatic hydrocarbon and environmental contaminant found in high concentrations around urban catchments and in the vicinity of oil extraction activities. Fish exposed to phenanthrene can exhibit altered reproductive hormone profiles and/or differences within gonadosomatic index and altered gamete proportions, but the mechanisms underlying these changes are not fully understood. In this study, we conducted a sub-chronic bioassay and measured transcriptional responses in the liver, the major tissue involved in generating lipids for oocyte growth. Adult male and female fathead minnow (Pimephales promelas) were exposed to an average measured concentration of 202 μg phenanthrene/L for a 7 week period. Condition factor was reduced in both males and females, while female fish also showed decreased gonadosomatic index relative to control females. In females exposed to phenanthrene, perinucleolar proportions were increased ∼1.9-fold relative to the control group whereas the proportions of vitellogenic oocytes decreased ∼8.8 fold. In males exposed to phenanthrene, spermatogonia proportions were increased ∼2.3 fold in testicular tissues compared to control fish. Thus, gametes were at an earlier stage of maturation in phenanthrene-treated fish compared to controls. However, no differences were detected in the production of 17β-estradiol or testosterone from the gonad in either sex. Catalase activity was also assessed in the liver as a measure of oxidative stress and this biomarker did not change in activity in either sex. In addition to endpoints in the ovary, the female hepatic transcriptome was measured, as this tissue produces lipids for oocyte maturation. Transcriptomic responses to phenanthrene exposure suggested a reduction in vitellogenin mRNA, and lipid metabolism and immune system pathways. Comparisons of hepatic transcriptome responses with Part A (72 h phenanthrene exposure) showed that energy homeostasis pathways were consistently altered following phenanthrene exposure over multiple durations and concentrations. We suggest that altered energy homeostasis may be adversely affecting reproductive efforts, as impaired reproduction has been observed in other studies investigating polycyclic aromatic hydrocarbons.
Part A: Temporal and dose-dependent transcriptional responses in the liver of fathead minnows following short term exposure to the polycyclic aromatic hydrocarbon phenanthrene Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-22 Jennifer R. Loughery, Karen A. Kidd, Angella Mercer, Christopher J. Martyniuk
Phenanthrene is a low molecular weight polycyclic aromatic hydrocarbon (PAH) that is composed of three fused benzene rings. PAHs are formed naturally through incomplete combustion of organic materials, and are environmental contaminants due to anthropogenic activities (e.g. oil extraction and refining, industrial and municipal effluents, fossil fuel burning). Fish exposed to PAHs such as phenanthrene have been reported to exhibit altered reproductive axis endpoints, however the mechanisms that underlie these responses are not fully characterized. To better understand effects at the mechanistic level, we applied transcriptomics to identify molecular pathways altered after acute exposure to phenanthrene on both a dose and temporal scale. Female fathead minnow (Pimephales promelas) were exposed to an average measured concentration of either 0, 29.8, 389 or 943 μg phenanthrene/L for 24, 48, and 72 h in a static-renewal bioassay. Ovaries were assessed for oocyte distribution as well as in vitro 17β-estradiol production and gene expression for transcripts related to steroidogenesis and estrogen signalling. In addition, the liver transcriptome was measured as this tissue is the primary source of the egg yolk precursor protein vitellogenin. Exposure to 29.8 μg phenanthrene/L increased proportions of the cortical alveolar stage in the ovaries after 48 h while the proportion of cortical alveolar oocyte were decreased in fish exposed to 943 μg phenanthrene/L for 48 h. Phenanthrene did not affect 17β-estradiol production at any time or dose, and did not affect transcripts associated with hormone synthesis nor signalling pathways. In the liver, the transcriptome showed fewer genes in common across time when compared to those transcripts affected by concentration at a single time point. Cholesterol metabolism was the only pathway perturbed in the liver following all comparisons in both the dose and time course experiments. Our data suggest that transcriptome networks associated with hepatic lipid metabolism are rapidly affected by phenanthrene, and this may indirectly reduce resources available for reproductive efforts.
Characterization and comparison of transcriptional activities of the retinoid X receptors by various organotin compounds in three prosobranch gastropods; Thais clavigera, Nucella lapillus and Babylonia japonica Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-26 Hiroshi Urushitani, Yoshinao Katsu, Hiroyuki Kagechika, Ana C.A. Sousa, Carlos M. Barroso, Yasuhiko Ohta, Hiroaki Shiraishi, Taisen Iguchi, Toshihiro Horiguchi
Two cDNAs of RXR were isolated, for the first time, from the ivory shell, Babylonia japonica, and the transcriptional activities were tested in vitro to compare with other gastropod (Thais clavigera and Nucella lapillus) RXR isoforms. The transcriptional activities of all of these RXR isoforms were significantly induced by mammalian RXR agonist, 9-cis retinoic acid (9cRA). The transcriptional activity of T. clavigera RXR-1 was also examined by using 9cRA and 16 organotin compounds, and significant ligand-dependent transactivations were observed by 9cRA and 5 organotins (tributyltin (TBT), tetrabutyltin (TeBT), tripropyltin (TPrT), tricyclohexyltin (TcHT) and triphenyltin (TPhT)). These 5 organotins also induced significant transcriptional activities in N. lapillus and B. japonica RXR isoforms. These 4 organotins, except for TeBT, have been reported to promote the development of imposex after a month of a single injection each, using female T. clavigera. To investigate the function of gastropod RXR isoforms, the effects of mammalian specific pan-agonist, PA024, and pan-antagonist, HX531, were examined, and significant induction of transcriptional activity by PA024 was demonstrated in these gastropod RXR isoforms. The additions of HX531 significantly suppressed the transcriptional activities of these gastropod RXR isoforms by 9cRA and 5 organotins. Using the mammalian two retinoic acid response elements, the transcriptional activities by 2 agonists, 9cRA and PA024, were different among the RXR isoforms of each gastropod species. With retinoid X response element (RXRE), transcriptional activities of TcRXR-1, BjRXR-1, and NlRXRa were significantly higher than those of TcRXR-2, BjRXR-2, and NlRXRb. Transcriptional activities of TcRXR-2, BjRXR-2, and NlRXRb, however, were significantly higher than those of TcRXR-1, BjRXR-1, and NlRXRa with thyroid hormone response element, TREpal. Thus, induction of imposex in prosobranch gastropods is strongly suggested to be triggered by 9cRA and certain organotins, such as TBT and TPhT through the activation of RXRs. These gastropod RXRs might control the different gene transcription by forming homo- or heterodimer complex with their own isoforms. These findings will contribute to our understanding of the fundamentals of the endocrine system in molluscs, particularly on RXR signaling pathway.
Microcystin-leucine arginine mediates apoptosis and engulfment of Leydig cell by testicular macrophages resulting in reduced serum testosterone levels Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-17 Yabing Chen, Jing Wang, Xiang Chen, Dongmei Li, Xiaodong Han
Microcystin-leucine arginine (MC-LR) causes decline of serum testosterone levels resulting in impaired spermatogenesis; however, the underlying molecular mechanisms are not fully understood. In this study, we aimed to investigate the effects of MC-LR exposure on the number of Leydig cells (LCs) in testis. Following chronic low dose exposure to MC-LR, the number of LCs was markedly decreased while macrophages were significantly increased. Then, we established a co-culture system to study the interaction between macrophages and LCs in the presence of MC-LR. No significant apoptosis of LCs cultured alone was observed after MC-LR (< 5 000 nM) treatment; however, apoptosis was robustly increased when LCs were co-cultured with macrophages in the presence of MC-LR. Further studies identified that MC-LR could stimulate macrophage to produce TNF-α, and secreted TNF-α induced LC apoptosis by binding to the tumor necrosis factor receptor 1 (TNFR1) on the LCs and thus activating reactive oxygen species (ROS)-p38MAPK signaling pathway. Furthermore, we also examined increased expression of Axl receptor and growth arrest-specific 6 (Gas6) in macrophages after MC-LR treatment. GAS6 mediates phagocytosis of apoptotic LCs by binding to the Axl receptor on macrophages and phosphatidylserine (PtdSer) on apoptotic LCs. Together, these results suggested that reduced serum testosterone levels may be associated with decrease of LCs as a result of LC apoptosis and phagocytosis by immune cells in MC-LR-treated mice.
Oyster transcriptome response to Alexandrium exposure is related to saxitoxin load and characterized by disrupted digestion, energy balance, and calcium and sodium signaling Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-27 Audrey M. Mat, Christophe Klopp, Laura Payton, Céline Jeziorski, Morgane Chalopin, Zouher Amzil, Damien Tran, Gary H. Wikfors, Hélène Hégaret, Philippe Soudant, Arnaud Huvet, Caroline Fabioux
Harmful Algal Blooms are worldwide occurrences that can cause poisoning in human seafood consumers as well as mortality and sublethal effets in wildlife, propagating economic losses. One of the most widespread toxigenic microalgal taxa is the dinoflagellate Genus Alexandrium, that includes species producing neurotoxins referred to as PST (Paralytic Shellfish Toxins). Blooms cause shellfish harvest restrictions to protect human consumers from accumulated toxins. Large inter-individual variability in toxin load within an exposed bivalve population complicates monitoring of shellfish toxicity for ecology and human health regulation. To decipher the physiological pathways involved in the bivalve response to PST, we explored the whole transcriptome of the digestive gland of the Pacific oyster Crassostrea gigas fed experimentally with a toxic Alexandrium minutum culture. The largest differences in transcript abundance were between oysters with contrasting toxin loads (1098 transcripts), rather than between exposed and non-exposed oysters (16 transcripts), emphasizing the importance of toxin load in oyster response to toxic dinoflagellates. Additionally, penalized regressions, innovative in this field, modeled accurately toxin load based upon only 70 transcripts. Transcriptomic differences between oysters with contrasting PST burdens revealed a limited suite of metabolic pathways affected, including ion channels, neuromuscular communication, and digestion, all of which are interconnected and linked to sodium and calcium exchanges. Carbohydrate metabolism, unconsidered previously in studies of harmful algal effects on shellfish, was also highlighted, suggesting energy challenge in oysters with high toxin loads. Associations between toxin load, genotype, and mRNA levels were revealed that open new doors for genetic studies identifying genetically-based low toxin accumulation.
Transcriptomics provides mechanistic indicators of mixture toxicology for IMX-101 and IMX-104 formulations in fathead minnows (Pimephales promelas) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-23 Kurt A. Gust, Guilherme R. Lotufo, Jacob K. Stanley, Mitchell S. Wilbanks, Pornsawan Chappell, Natalie D. Barker
Within the US military, new insensitive munitions (IMs) are rapidly replacing conventional munitions improving safety from unintended detonation. Toxicity data for IM chemicals are expanding rapidly, however IM constituents are typically deployed in mixture formulations, and very little is known about their mixture toxicology. In the present study we sought to characterize the mixture effects and toxicology of the two predominant IM formulations IMX-101 and IMX-104 in acute (48 h) larval fathead minnow (Pimephales promelas) exposures. IMX-101 consists of a mixture of 2,4-dinitroanisole (DNAN), 3-nitro-1,2,4-triazol-5-one (NTO), and nitroguanidine (NQ) while IMX-104 is composed of DNAN, NTO, and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). DNAN was the most potent constituent in IMX-101 eliciting an LC50 of 36.1 mg/L, whereas NTO and NQ did not elicit significant mortality in exposures up to 1040 and 2640 mg/L, respectively. Toxic unit calculations indicated that IMX-101 elicited toxicity representative of the component concentration of DNAN within the mixture. Toxicogenomic responses for the individual constituents of IMX-101 indicated unique transcriptional expression and functional responses characteristic of: oxidative stress, impaired energy metabolism, tissue damage and inflammatory responses in DNAN exposures; impaired steroid biosynthesis and developmental cell-signaling in NQ exposures; and altered mitogen-activated protein kinase signaling in NTO exposures. Transcriptional responses to the IMX-101 mixture were driven by the effects of DNAN where expression and functional responses were nearly identical comparing DNAN alone versus the fractional equivalent of DNAN within IMX-101. Given that each individual constituent of the IMX-101 mixture elicited unique functional responses, and NTO and NQ did not interact with DNAN within the IMX-101 mixture exposure, the overall toxicity and toxicogenomic responses within acute exposures to the IMX-101 formulation are indicative of “independent” mixture toxicology. Alternatively, in the IMX-104 exposure both DNAN and RDX were each present at concentrations sufficient to elicit lethality (RDX LC50 = 28.9 mg/L). Toxic-unit calculations for IMX-104 mixture formulation exposures indicated slight synergistic toxicity (ΣTU LC50 = 0.82, 95% confidence interval = 0.73–0.90). Unique functional responses relative to DNAN were observed in the IMX-104 exposure including responses characteristic of RDX exposure. Based on previous transcriptomics responses to acute RDX exposures in fathead minnow larvae, we hypothesize that the potentially synergistic responses within the IMX-104 mixture are related to interactive effects of each DNAN and RDX on oxidative stress mitigation pathways.
Transcriptional effects of phospholipid fatty acid profile on rainbow trout liver cells exposed to methylmercury Aquat. Toxicol. (IF 4.129) Pub Date : 2018-03-22 Aline Ferain, Chloé Bonnineau, Ineke Neefs, Krishna Das, Yvan Larondelle, Jean-François Rees, Cathy Debier, Benjamin Lemaire
Lipids, and their constitutive fatty acids, are key nutrients for fish health as they provide energy, maintain cell structure, are precursors of signalling molecules and act as nuclear receptor ligands. These specific roles may be of crucial importance in a context of exposure to pollutants. We recently showed that the fatty acid profile of rainbow trout liver cell phospholipids modulates sensitivity to an acute methylmercury challenge. In order to investigate mechanisms of effects, we herein tested whether specific polyunsaturated fatty acids (PUFAs) may protect cells from methylmercury through decreasing intracellular mercury accumulation and/or enhancing cellular defences (e.g. via modulation of gene expression patterns). We also investigated the inverse relationship and assessed the impact of methylmercury on cellular fatty acid metabolism. To do so, the fatty acid composition of rainbow trout liver cell phospholipids was first modified by incubating them in a medium enriched in a specific PUFA from either the n-3 family (alpha-linolenic acid, ALA; eicosapentaenoic acid, EPA) or the n-6 family (linoleic acid, LA; arachidonic acid, AA). Cells were then exposed to methylmercury (0.15 or 0.50 μM) for 24 h and sampled thereafter for assessing phospholipid fatty acid profile, intracellular total mercury burden, and expression pattern of genes involved in fatty acid metabolism, synthesis of PUFA-derived signalling molecules and stress response. We observed that cells incorporated the given PUFA and some biotransformation products in their phospholipids. Methylmercury had few impacts on this cellular phospholipid composition. None of the PUFA enrichments affected the cellular mercury burden, suggesting that the previously observed cytoprotection conferred by ALA and EPA was not linked to a global decrease in cellular accumulation of mercury. Fatty acid enrichments and methylmercury exposure both modulated gene expression patterns. Genes involved in the synthesis of PUFA-derived signalling molecules, in stress response and the orphan cytochrome P450 20A1 were identified as possible sites of interaction between fatty acids and methylmercury in rainbow trout liver cells.
Retrospective analyses of archive phytotoxicity test data can help in assessing internal dynamics and stability of growth in laboratory duckweed cultures Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-28 Viktor Oláh, Anna Hepp, Norma Yolanda Gaibor Vaca, Marianna Tamás, Ilona Mészáros
High growth potential of duckweed species (Lemnaceae family) has been utilized in wide range of research and practical applications. Based on literature data, however, it can be assumed that duckweed populations maintain constant growth rates only when short periods are considered but can vary over longer time scales. This intrinsic instability in growth can affect the interpretation of growth data. Duckweed phytotoxicity tests are usually performed according to highly standardized protocols. Therefore the archive data provide an opportunity for retrospective comparisons. In the present study we collected growth (frond number- and frond area-based relative growth rates) and morphology (average frond and colony sizes) data from control treatments of phytotoxicity tests. All the analyzed tests were carried out with the same Spirodela polyrhiza (L.) Schleid. (giant duckweed) clone (RDSC ID No. 5501) under the same experimental conditions over more than four years. We aimed to assess the overall variability of the above parameters and to test if intrinsic growth patterns affect growth data in short-term. In general, the results reflected high stability of the measured parameters in long term but also indicated that some temporal variability is inevitable which can bias the comparability of growth tests. The frond area-based relative growth rate resulted in smaller coefficient of variation than the usually preferred frond number-based one. The results also revealed a negative correlation between mean growth rates and their coefficients of variation. Therefore, it would be advisable to introduce higher minimal growth rates and/or maximized tolerable coefficients of variation for control cultures into the standard duckweed growth inhibition tests. Analyses of growth data aggregated on seasonal basis indicated faster growth and larger mean frond size in laboratory duckweed cultures from mid-autumn till mid-spring than during summer and early autumn. But, in shorter term (∼50 days) we did not observe distinct trends in growth suggesting that the successive frond generations have no effect on growth traits within this time-scale. Our results point to the importance of assessing intrinsic growth dynamics in duckweed cultures and also to the re-usability of the already collected phytotoxicity data in addressing new research questions.
Can the inhibition of cytochrome P450 in aquatic invertebrates due to azole fungicides be estimated with in silico and in vitro models and extrapolated between species? Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-26 Michele Gottardi, Jonathan D. Tyzack, Andreas Bender, Nina Cedergreen
Azole fungicides, designed to halt fungal growth by specific inhibition of fungal cytochrome P450 (CYP51), inhibit cytochrome P450s involved in the metabolism of xenobiotics in several non-target organisms thus raising environmental concern. The present study investigates the degree by which inhibition strengths of azoles toward cytochrome P450 in rat liver, the insect Chironomus riparius larvae and the snail Lymnaea stagnalis can be extrapolated from estimated in silico affinities. Azoles’ affinities toward human cytochrome P450 isoforms involved in xenobiotic metabolism (CYP3A4, CYP2C9 and CYP2D6) as well as fungal CYP51 were estimated with a ligand-protein docking model based on the ChemScore scoring function. Estimated affinities toward the selected enzymatic structures correlated strongly with measured inhibition strengths in rat liver (ChemScore vs. logIC50 among cytochrome P450 isoforms: −0.662 < r < −0.891, n = 17 azoles), while weaker correlations were found for C. riparius larvae (−0.167 < r < −0.733, n = 9) and L. stagnalis (−0.084 < r < −0.648, n = 8). Inhibition strengths toward C. riparius and rat liver activities were found to be highly correlated to each other (r: 0.857) while no significant relationship was found between either of the species and L. stagnalis. The inhibition of cytochrome P450 due to azole fungicides could be estimated in vitro and to a lesser extent in silico for C. riparius but not for L. stagnalis, possibly due to different enzymatic susceptibility toward azole inhibition among the species.
Dose- and age-specific antioxidant responses of the mysid crustacean Neomysis awatschensis to metal exposure Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-28 Md. Niamul Haque, Do-Hee Lee, Bo-Mi Kim, Sang-Eun Nam, Jae-Sung Rhee
Waterborne metals can adversely affect an organism’s innate defenses through oxidative stress. In the present study, the marine mysid Neomysis awatchensis was exposed to sublethal concentrations (1/10 and 1/5 of the median LC50s) of As, Cd, Cu, Pb, and Zn for 48 or 96 h at the juvenile and adult developmental stages, and the dose- and age-specific antioxidant defense system responses were characterized. Metal accumulation and modulation of four key antioxidant biomarkers, malondialdehyde (MDA), glutathione (GSH), superoxide dismutase, and catalase, were measured in juvenile and adult mysids. Based on LC50, level of metal toxicity was dependent on metal concentration, developmental stage, and the exposure duration. Intracellular MDA content was increased in the As-, Cu-, Pb-, and Zn-exposed juvenile mysids after exposure for 48 and/or 96 h, while increases in MDA content were observed in adult mysids following Cu and Zn exposure. Interestingly, GSH content was differentially modulated, where intracellular GSH levels decreased in juvenile mysids following Cd, Cu, Pb, and Zn exposure, but significantly increased in metal-exposed adult mysids, except for Pb exposure. The catalase and superoxide dismutase activities displayed similar stage-specific increases or decreases as also observed for the different GSH levels, suggesting that the susceptibility to and defense against metal-induced oxidative stress differed based on stage. Modulations in MDA and GSH content and enzymatic activity of the antioxidant defense system indicate that mysid antioxidant defense system factors are intimately connected during control of oxidative imbalances with different capacities at different developmental stages.
The progestin norethindrone affects sex differentiation and alters transcriptional profiles of genes along the hypothalamic–pituitary–gonadal and hypothalamic–pituitary–adrenal axes in juvenile zebrafish Dario renio Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-28 Li-ping Hou, Hongxing Chen, Chang-en Tian, Wen-Jun Shi, Ye Liang, Rong-rong Wu, Xu-wen Fang, Cui-ping Zhang, Yan-qiu Liang, Lingtian Xie
Natural and synthetic progestins may pose a threat to wild fish populations living in receiving waters. In this study, the effects of norethindrone (NET) on the sex differentiation of zebrafish (Dario renio) and the mechanisms underlying these effects were investigated. Juvenile zebrafish (20 days post fertilization, pdf) were exposed to environmentally relevant concentrations (5, 50, 500, and 1000 ng L−1) for 45 d. Sex ratio of the NET-exposed populations, the histology of the gonads and the transcriptional profile of the regulatory genes involved in sex differentiation and steroidogenesis were examined. The results showed that a significantly higher ratio of male/female was induced in the zebrafish populations exposed to NET at concentrations higher than 32.3 ng L−1. Exposure to NET caused acceleration of sexual mature in males and a delay in ovary maturation in female zebrafish. Among the genes regulating sexual differentiation, transcripts of Dmrt1 showed a dose-dependent increase while transcripts of Figa and Fox12 showed a dose-dependent decrease in response to exposure to NET. For genes regulating the steroidogenesis, the expressions of Cyp11a1, Cyp17, Cyp19a1a, and Cyp11b were significantly down-regulated by exposure to NET, while Hsd17b3 expression was significantly up-regulated by exposure to NET at 421.3 and 892.9 ng L−1. For the receptor genes in the gonads, the transcriptional expression of Pgr, Ar, and Mr was significantly up-regulated at 421.3 and 892.9 ng L−1 of NET. For genes involved in the hypothalamic–pituitary axis, the transcriptional expression of Gnrh3 and Pomc was significantly up-regulated by exposure to NET with the exception for Gnrh3 at 4.2 ng L−1. The results demonstrated that exposure to NET at the juvenile stage could affect gonad differentiation and sex ratio, which might be accounted for by the alterations of the transcriptional expressions of genes along the hypothalamic–pituitary–gonadal (HPG) and hypothalamic–pituitary–adrenal (HPA) axes.
Accumulation of perfluorobutane sulfonate (PFBS) and impairment of visual function in the eyes of marine medaka after a life-cycle exposure Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-25 Lianguo Chen, Mirabelle M.P. Tsui, Qipeng Shi, Chenyan Hu, Qi Wang, Bingsheng Zhou, Paul K.S. Lam, James C.W. Lam
As an alternative to perfluorooctane sulfonate (PFOS), increasing usage of perfluorobutane sulfonate (PFBS) has led to ubiquitous presence in the environment. PFBS is also shown to potently disrupt the thyroid endocrine system. Considering the regulation of thyroid hormones in visual development, PFBS is likely to adversely affect the development and function of visual systems, which is a sensitive target of environmental pollutants. Therefore, the present study exposed marine medaka embryos to environmentally realistic concentrations of PFBS (0, 1.0, 2.9 and 9.5 μg/L) for an entire life-cycle. After exposure until sexual maturity, eyes of adult medaka were dissected to directly investigate the ocular accumulation and toxicity of PFBS. For the first time, substantial accumulation of an environmental pollutant (i.e., PFBS) was observed in the eye tissue. PFBS exposure was also found to impair the visual development and function in a sex-dependent manner. In female medaka, weight of eyes was significantly decreased, while content of water was increased, probably resulting in higher intraocular pressure. Multiple neural signaling processes were also disturbed by PFBS life-cycle exposure, including cholinergic, glutamatergic, GABAergic and monoaminergic systems. Increased levels of norepinephrine and epinephrine neurotransmitters may adaptively decrease the intraocular hypertension in female eyes. In addition, proteomic profiling identified the visual proteins of differential expressions (e.g., beta and gamma crystallins, arrestin and lumican), which were significantly associated with visual perception and motor activity of eyes. Overall, this study found that PFBS was able to accumulate in the eyes and induce ocular toxicities. The susceptibility and sex-specific responses of visual systems to environmental pollutants warrants more works for a comprehensive risk assessment.
Role of endocytotic uptake routes in impacting the ROS-related toxicity of silver nanoparticles to Mytilus galloprovincialis: A redox proteomic investigation Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-23 Younes Bouallegui, Ridha Ben Younes, Ridha Oueslati, David Sheehan
Oxidative stress is often implicated in nanoparticle toxicity. Several studies have highlighted the role of internalization routes in determining nanotoxicity. Here, we investigate how two endocytotic mechanisms (clathrin- and caveolae-mediated) impact on redox balance in gill and digestive gland of the mussel, Mytilus galloprovincialis. Animals were exposed (for 3, 6 and 12 h) to two sizes of silver nanoparticles (AgNP: <50 nm and <100 nm) prior to and after blockade of two endocytic pathways (amantadine blocks clathrin-mediated endocytosis while nystatin blocks caveolae-mediated endocytosis). Redox-proteomic tools were used to determine effects. Our results demonstrate the ability of both sizes of AgNP (<50 and <100 nm) to cause protein thiol oxidation and/or protein carbonylation. However, blockade of endocytotic routes mitigated AgNP toxicity. Differential ROS-related toxicity of AgNP to mussel tissues seemed to be linked to tissue-specific mode of action requirements. Cell uptake mechanism strongly influences toxicity of AgNPs in this filter-feeder.
Stage-dependent effects of chlorpyrifos on medaka (Oryzias latipes) swimming behavior using a miniaturized swim flume Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-21 Salvador Sastre, Carlos Fernández Torija, Irene Atiénzar Pertusa, Eulalia María Beltrán, María Victoria Pablos, Miguel González-Doncel
By considering chlorpyrifos (CPF), an organophosphorus pesticide with known mechanisms of action that affect neurobehavioral development, we assessed the validity and sensitivity of a miniaturized swim flume by investigating the effects of the insecticide on swimming behavior in medaka (Oryzias latipes) fish growing stages. Medaka in three developmental periods, namely 0, 20 and 40 day-old post-hatch (i.e. time points 0, 20 and 40, respectively), were exposed to CPF (12.5, 25, 50 and 100 μg/L) for 48 h under semi-static conditions. The CPF half-lives during exposures were evaluated and the swimming patterns in a flume section (arena) were presented on two-dimensional gradient maps of forced movement of fish against water current. A comparative numerical analysis of fish residence times between each time point control and the corresponding CPF groups was performed by dividing arenas into 15 proportional areas. The time point 0 control group gradient map showed a noticeably different swim pattern from those of the ≥12.5 μg CPF/L groups, which was statistically supported by the differences for residence times seen in ≥12 corresponding areas. The control group gradient maps for time points 20 and 40 differed from those of the respective ≥12.5 μg CPF/L groups. The comparative analysis of the residence times in the corresponding 15 areas revealed differences in ≥5 areas for time point 20 and in ≥3 areas for time point 40. The integrative analysis of the gradient maps and the numerical statistics revealed stage-specific effects and a concentration-response relationship between CPF and alterations on forced medaka swimming despite the dissipation of CPF from the water column. These results indicate the validity of the miniaturized swim flume toward a more environmentally realistic scenario for the evaluation of neurodevelopmental and behavioral toxicity in small fish models.
Negative impacts of elevated nitrate on physiological performance are not exacerbated by low pH Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-10 Daniel F. Gomez Isaza, Rebecca L. Cramp, Craig E. Franklin
Multiple environmental stressors, including nutrient effluents (i.e. nitrates [ N O 3 − ]) and altered pH regimes, influence the persistence of freshwater species in anthropogenically disturbed habitats. Independently, nitrate and low pH affect energy allocation by increasing maintenance costs and disrupting oxygen uptake, which ultimately results in impacts upon whole animal performance. However, the interaction between these two stressors has not been characterised. To address this, the effects of nitrate and pH and their interaction on aerobic scope and physiological performance were investigated in the blueclaw crayfish, Cherax destructor. Crayfish were exposed to a 2 × 3 factorial combination, with two pH levels (pH 5.0 and 7.0) and three nitrate concentrations (0, 50 and 100 mg L−1 N O 3 − ). Crayfish were exposed to experimental conditions for 65 days and growth and survival were monitored. Aerobic scope (i.e. maximal – standard oxygen uptake) was measured at six time points (1, 3, 5, 7, 14, and 21 days) during exposure to experimental treatments. Crayfish performance was assessed after 28 days, by measuring chelae strength and whole animal activity capacity via the righting response. Survival was reduced in crayfish exposed to pH 5.0, but there was no exacerbation of this effect by exposure to high nitrate levels. Aerobic scope was compromised by the interaction between low pH and nitrate and resulted in prolonged elevations of standard oxygen uptake rates. Exposure to nitrate alone affected aerobic scope, causing a 59% reduction in maximum oxygen uptake. Reduced aerobic capacity translated to reduced chelae strength and righting capacity. Together, these data show that low pH and elevated nitrate levels reduce aerobic scope and translate to poorer performance in C. destructor, which may have the potential to affect organismal fitness in disturbed habitats.
Neurotoxic impact of acute TiO2 nanoparticle exposure on a benthic marine bivalve mollusk, Tegillarca granosa Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-26 Xiaofan Guan, Wei Shi, Shanjie Zha, Jiahuan Rong, Wenhao Su, Guangxu Liu
The release of nanoparticles (NPs) into the ocean inevitably poses a threat to marine organisms. However, to date, the neurotoxic effects of NPs remains poorly understood in marine bivalve species. Therefore, in order to gain a better understanding of the physiological effects of NPs, the impact of acute (96 h) TiO2 NP exposure on the in vivo concentrations of three major neurotransmitters, the activity of AChE, and the expression of neurotransmitter-related genes was investigated in the blood clam, Tegillarca granosa. The obtained results showed that the in vivo concentrations of the three tested neurotransmitters (DA, GABA, and ACh) were significantly increased when exposed to relatively high doses of TiO2 NPs (1 mg/L for DA and 10 mg/L for ACh and GABA). Additionally, clams exposed to seawater contaminated with TiO2 NP had significantly lower AChE activity. In addition, the expression of genes encoding modulatory enzymes (AChE, GABAT, and MAO) and receptors (mAChR3, GABAD, and DRD3) for the neurotransmitters tested were all significantly down-regulated after TiO2 NP exposure. Therefore, this study has demonstrated the evident neurotoxic impact of TiO2 NPs in T. granosa, which may have significant consequences for a number of the organism’s physiological processes.
Enhancement of coral calcification via the interplay of nickel and urease Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-19 T. Biscéré, C. Ferrier-Pagès, R. Grover, A. Gilbert, C. Rottier, A. Wright, C. Payri, F. Houlbrèque
Corals are the main reef builders through the formation of calcium carbonate skeletons. In recent decades, coral calcification has however been impacted by many global (climate change) and local stressors (such as destructive fishing practices and changes in water quality). In this particular context, it is crucial to identify and characterize the various factors that promote coral calcification. We thus performed the first investigation of the effect of nickel and urea enrichment on the calcification rates of three coral species. These two factors may indeed interact with calcification through the activity of urease, which catalyzes the hydrolysis of urea to produce inorganic carbon and ammonia that are involved in the calcification process. Experiments were performed with the asymbiotic coral Dendrophyllia arbuscula and, to further assess if urea and/or nickel has an indirect link with calcification through photosynthesis, results were compared with those obtained with two symbiotic corals, Acropora muricata and Pocillopora damicornis, for which we also measured photosynthetic rates. Ambient and enriched nickel (0.12 and 3.50 μg L−1) combined with ambient and enriched urea concentrations (0.26 and 5.52 μmol L−1) were tested during 4 weeks in aquaria. We demonstrate in the study that a nickel enrichment alone or combined with a urea enrichment strongly stimulated urea uptake rates of the three tested species. In addition, this enhancement of urea uptake and hydrolysis significantly increased the long-term calcification rates (i.e. growth) of the three coral species investigated, inducing a 1.49-fold to 1.64-fold increase, respectively for D. arbuscula and P. damicornis. Since calcification was greatly enhanced by nickel in the asymbiotic coral species – i.e. in absence of photosynthesis – we concluded that the effect of increased urease activity on calcification was mainly direct. According to our results, it can be assumed that corals in some fringing reefs, benefiting from seawater enriched in nickel may have advantages and might be able to use urea more effectively as a carbon and nitrogen source. It can also be suggested that urea, for which hotspots are regularly measured in reef waters may alleviate the negative consequences of thermal stress on corals.
Experimental evidence of dietary ciguatoxin accumulation in an herbivorous coral reef fish Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-26 Rachel J. Clausing, Barbara Losen, Francois R. Oberhaensli, H. Taiana Darius, Manoella Sibat, Philipp Hess, Peter W. Swarzenski, Mireille Chinain, Marie-Yasmine Dechraoui Bottein
A multibiomarker approach highlights effects induced by the human pharmaceutical gemfibrozil to gilthead seabream Sparus aurata Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-17 A. Barreto, L.G. Luis, P. Paíga, L.H.M.L.M. Santos, C. Delerue-Matos, A.M.V.M. Soares, K. Hylland, S. Loureiro, M. Oliveira
Exposure to tris(1,3-dichloro-2-propyl) phosphate for Two generations decreases fecundity of zebrafish at environmentally relevant concentrations Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-15 Yongkang Zhang, Meng Li, Shuying Li, Qiangwei Wang, Guonian Zhu, Guanyong Su, Robert J. Letcher, Chunsheng Liu
Previous studies reported that exposure to environmentally relevant concentrations of TDCIPP significantly decreased the number of cumulative eggs in zebrafish, but effects on the quantity of eggs and sperms remained unknown. Therefore, in this study, effects of TDCIPP on yolk diameter, surface morphology of eggs, sperm density and total motility were evaluated. First generation (F0) zebrafish larvae (Danio rerio) were exposed to 0, 50, 500 or 5000 ng/L tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) from 14 days post fertilization (dpf) to 120 dpf. The F0 generation of zebrafish were paired and F1 generation of embryos were collected and continuously exposed to the same concentrations of TDCIPP until 150 dpf. TDCIPP bioconcentration in the whole body as well as effects on survival and fecundity were evaluated in F1 generation. Exposure to TDCIPP resulted in an accumulation of the chemical and decreased survival of F1 generation of zebrafish. TDCIPP decreased cumulative production and changed surface morphology of eggs in females. In males, TDCIPP decreased total motility of sperm but did not affect sperm density. These effects on quality of egg and sperm might be responsible for the decreased hatching rates observed in cross mating experiments. Furthermore, TDCIPP exposure resulted in down-regulated gene expression related to gonadal development and maturation of germ cells in females or/and males, and the down-regulation was correlated to decreased fecundity. Taken together, the results suggested that exposure to TDCIPP could decrease the quantity of eggs and sperms by down-regulating the expression of genes related to gonadal development and maturation of germ cells in zebrafish.
Effects of microcystin-LR on the tissue growth and physiological responses of the aquatic plant Iris pseudacorus L. Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-27 Naiyu Wang, Can Wang
Environmentally relevant concentrations of tramadol and citalopram alter behaviour of an aquatic invertebrate Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-14 M. Buřič, K. Grabicová, J. Kubec, A. Kouba, I. Kuklina, P. Kozák, R. Grabic, T Randák
Environmental pollution by pharmaceutically active compounds, used in quantities similar to those of pesticides and other organic micropollutants, is increasingly recognized as a major threat to the aquatic environment. These compounds are only partly removed from wastewaters and, despite their low concentrations, directly and indirectly affect behaviour of freshwater organisms in natural habitats. The aim of this study was to behaviourally assess the effects of an opioid painkiller (tramadol) and antidepressant drug (citalopram) on behaviour patterns of a clonal model species, marbled crayfish. Animals exposed to environmentally relevant concentrations of both tested compounds (~1 µg l-1) exhibited significantly lower velocity and shorter distance moved than controls. Crayfish exposed to tramadol spent more time in shelters. Results were obtained by a simple and rapid method recommended as suitable for assessment of behaviour in aquatic organisms exposed to single pollutants and combinations.
Tributyltin impaired reproductive success in female zebrafish through disrupting oogenesis, reproductive behaviors and serotonin synthesis Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-12 Wei-Yang Xiao, Ying-Wen Li, Qi-Liang Chen, Zhi-Hao Liu
Tributyltin (TBT), an organotin acting as aromatase (Cyp19a1) inhibitor, has been found to disrupt gametogenesis and reproductive behaviors in several fish species. However, few studies addressing the mechanisms underlying the impaired gametogenesis and reproduction have been reported. In this study, female adults of zebrafish (Danio rerio) were continuously exposed to two nominal concentrations of TBT (100 and 500 ng/L, actual concentrations: 90.8 ± 1.3 ng/L and 470.3 ± 2.7 ng/L, respectively) for 28 days. After exposures, TBT decreased the total egg number, reduced the hatchability and elevated the mortality of the larvae. Decreased gonadosomatic index (GSI) and altered percentages of follicles in different developmental stages (increased early-stage follicles and reduced mid/late-stage follicles) were also observed in the ovary of TBT-treated fish. TBT also lowered the plasma level of 17β-estradiol and suppressed the expressions of cyp19a1a in the ovary. In treated fish, up-regulated expressions of aldhla2, sycp3 and dmc1 were present in the ovary, indicating an enhanced level of meiosis. The mRNA level of vtg1 was dramatically suppressed in the liver of TBT-treated fish, suggesting an insufficient synthesis of Vtg protein, consistent with the decreased percentage of mid/late-stage follicles in the ovaries. Moreover, TBT significantly suppressed the reproductive behaviors of the female fish (duration of both sexes simultaneously in spawning area, the frequency of meeting and the visit in spawning area) and down-regulated the mRNA levels of genes involved in the regulation of reproductive behaviors (cyp19a1b, gnrh-3 and kiss 2) in the brain. In addition, TBT significantly suppressed the expressions of serotonin-related genes, such as tph2 (encoding serotonin synthase), pet1 (marker of serotonin neuron) and kiss 1 (the modulator of serotonin synthesis), suggesting that TBT might disrupt the non-reproductive behaviors of zebrafish. The present study demonstrated that TBT may impair the reproductive success of zebrafish females probably through disrupting oogenesis, disturbing reproductive behaviors and altering serotonin synthesis. The present study greatly extends our understanding on the reproductive toxicity of TBT on fish.
Combined effects of metal mixtures and predator stress on the freshwater isopod Asellus aquaticus Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-30 M. Van Ginneken, R. Blust, L. Bervoets
Mitochondria-rich cells adjustments and ionic balance in the Neotropical fish Prochilodus lineatus exposed to titanium dioxide nanoparticles Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-09 Talita L.L. Carmo, Vinícius C. Azevedo, Priscila R. Siqueira, Tiago D. Galvão, Fabrício A. Santos, Cláudia B.R. Martinez, Carlos R. Appoloni, Marisa N. Fernandes
Manufactured titanium dioxide nanoparticles (TiO2-NP) have been intensely applied in numerous industrial products and may be a risk for aquatic systems as they are not completely removed from domestic and industrial wastes after water treatment. This study evaluated the osmo- and ionic balance, Na+/K+-ATPase, H+-ATPase and carbonic anhydrase activities and the mitochondria-rich cells (MRC) in the gills and kidney of the Neotropical fish Prochilodus lineatus after 2 (acute) and 14 (subchronic) days of exposure to nominal 0, 1, 5, 10 and 50 mg L-1 TiO2-NP. The nominal concentrations corresponded to 0.0, 0.6, 1.6, 2.7 and 18.1 mg L-1 suspended TiO2-NP, respectively, in the water column one hour after NP introduction and were maintained for at least 24 h. Acute exposure to TiO2-NP decreased plasma osmolality and Ca2+ levels. Na+/K+-ATPase, H+-ATPase and carbonic anhydrase activities were inhibited in the gills, but not in the kidney. Total MRC density did not change in gills and kidneys. At gill surface, total MRC density decreased in fish exposed to 50 mg L-1 TiO2-NP and the total MRC fractional surface area unchanged although, there were some changes in the fractional area of MRC with apical microvilli (MRCm) and MRC with apical sponge-like structure (MRCs). MRCm was more abundant than MRCs. After subchronic exposure, there was no change in plasma osmolality, ionic balance and enzyme activities. Total gill MRC density increased in the filament epithelium and renal tubules. In the gills, MRC contacting water exhibited some adjustments. Total MRC and fractional surface area unchanged, but there was an increase of MRCs contacting water at gill surface after exposure to10 and 50 mg L-1 TiO2-NP. MRC proliferation in filament epithelium and in renal tubules as well as the increasing MRCs at gill surface may have contributed to avoid change in plasma osmolality, ionic balance and enzyme activities and suggested a cellular physiological and morphological response to restore and maintain osmotic and ionic homeostasis after subchronic exposure.
Tritiated water exposure disrupts myofibril structure and induces mis-regulation of eye opacity and DNA repair genes in zebrafish early life stages Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-27 Caroline Arcanjo, Olivier Armant, Magali Floriani, Isabelle Cavalie, Virginie Camilleri, Olivier Simon, Daniel Orjollet, Christelle Adam-Guillermin, Béatrice Gagnaire
Tritium (3H) is a radioactive isotope of hydrogen. In the environment, the most common form of tritium is tritiated water (HTO). The present study aimed to identify early biomarkers of HTO contamination through the use of an aquatic model, the zebrafish (Danio rerio). We used the zebrafish embryo-larvae model to investigate the modes of action of HTO exposure at dose rates of 0.4 and 4 mGy/h, dose rates expected to induce deleterious effects on fish. Zebrafish were exposed to HTO from 3 hpf (hours post fertilization) to 96 hpf. The transcriptomic effects were investigated 24 h and 96 h after the beginning of the contamination, using mRNAseq. Results suggested an impact of HTO contamination, regardless of the dose rate, on genes involved in muscle contraction (tnnt2d, tnni2a.4, slc6a1a or atp2a1l) and eye opacity (crygm2d9, crygmxl1, mipb or lim2.3) after 24 h of contamination. Interestingly, an opposite differential expression was highlighted in genes playing a role in muscle contraction and eye opacity in 24 hpf embryos when comparing dose rates, suggesting an onset of DNA protective mechanisms. The expression of h2afx and ddb2 involved in DNA repair was enhanced in response to HTO exposure. The entrainment of circadian clock and the response to H2O2 signalling pathways were enriched at 96 hpf at 0.4 mGy/h and in both stages after 4 mGy/h. Genes involved in ROS scavenging were differentially expressed only after 24 h of exposure for the lowest dose rate, suggesting the onset of early protective mechanisms against oxidative stress. Effects highlighted on muscle at the molecular scale were confirmed at a higher biological scale, as electron microscopy observations revealed sarcomere impairments in 96 hpf larvae for both dose rates. Together with other studies, the present work provides useful data to better understand modes of action of tritium on zebrafish embryos-larvae.
Adverse effects and immune dysfunction in response to oral administration of weathered Iranian heavy crude oil in the rockfish Sebastes schlegeli Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-24 Eun-Hee Lee, Moonkoo Kim, Young-Sun Moon, Un Hyuk Yim, Sung Yong Ha, Chang-Bum Jeong, Jae-Seong Lee, Jee-Hyun Jung
Chlorophyll a ﬂuorescence and transcriptome reveal the toxicological effects of bisphenol A on an invasive cyanobacterium, Cylindrospermopsis raciborskii Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-09 Rong Xiang, Junqiong Shi, Hongbo Zhang, Congcong Dong, Li Liu, JunKe Fu, Xinyu He, Yanjun Yan, Zhongxing Wu
Bisphenol A has attracted worldwide attention due to its harmful effects on humans, animals and plants. In this study, the toxicological effects of BPA on Cylindrospermopsis raciborskii were assessed based on chlorophyll a ﬂuorescence and transcriptome analyses. The results showed that the growth of C. raciborskii was significantly inhibited when BPA exceeded 0.1 mg L-1. A marked rise of phase J was observed at a concentration greater than 0.1 mg L-1, while a K phase appeared at 20 mg L-1. The chlorophyll a ﬂuorescence parameters of RC/CS0, F0, φP0, φE0, and ψ0, underwent a significant decline under all treatments of BPA, whereas a significant increase in both VJ and M0 occurred under all concentrations of BPA. Additionally, ABS/RC and DIo/RC markedly increased at 10 mg L-1 and 20 mg L-1. The transcriptome analysis revealed that the genes of photosynthesis, including psbA, psbB, psbC, psbD, apcA, apcB, cpcA, and cpcB, as well as those of chlorophyll and carotenoid biosynthesis, namely hemN, acsF, chlL, chlN, chlP, crtB, pds, were all down-regulated. Moreover, BPA also inhibited the oxidative phosphorylation, glycolysis/gluconeogenesis, citrate cycle (TCA cycle), and fatty acid metabolism in C. raciborskii. Taken together, these results suggest BPA can negatively affect the expression of multiple genes and the vital energy metabolism process to arrest the growth and photosynthesis of C. raciborskii.
Interactions of oxidized multiwalled carbon nanotube with cadmium on zebrafish cell line: the influence of two co-exposure protocols on in vitro toxicity tests Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-05 Mariana Morozesk, Lidiane S. Franqui, Adrislaine S. Mansano, Diego Stéfani T. Martinez, Marisa N. Fernandes
An in vivo analysis of Cr6+ induced biochemical, genotoxicological and transcriptional profiling of genes related to oxidative stress, DNA damage and apoptosis in liver of fish, Channa punctatus (Bloch, 1793) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-05-05 Yashika Awasthi, Arun Ratn, Rajesh Prasad, Manoj Kumar, Sunil P. Trivedi
Present study was designed to assess the hexavalent chromium (Cr6+) mediated oxidative stress that induces DNA damage and apoptosis in adult fish, Channa punctatus (35 ± 3.0 g; 14.5 ± 1.0 cm; Actinopterygii). Fishes were maintained in three groups for 15, 30 and 45 d of exposure periods. They were treated with 5% (Group T1) and 10% (Group T2) of 96 h-LC50 of chromium trioxide (Cr6+). Controls were run for the similar duration. A significant (p < 0.05) increment in the activities of antioxidant enzymes, SOD and CAT in liver tissues of the exposed fish evinces the persistence of oxidative stress. A significant (p < 0.05) increase in induction of micronuclei (MN) coupled with transcriptional responses of target genes related to antioxidant enzymes, DNA damage and apoptosis (sod, cat, gsr, nox-1, p53, bax, bcl-2, apaf-1 and casp3a) establishes the impact of oxidative stress due to in vivo, Cr6+ accumulation in liver as compared to control (0 mg/L), in a dose and exposure-dependent manner. Initially, the increased level of reactive oxygen species (ROS) in liver coincided with that of enhanced mRNA expression of antioxidant enzymes, sod, cat, gsr and nox-1 but, later, the overproduction of ROS, after 45 d of exposure of Cr6+, resulted in a significant (p < 0.05) up-regulation of p53. Our findings also unveil that the up-regulation of bax, apaf-1 and casp3a and down-regulation of bcl-2 are associated with Cr6+-induced oxidative stress mediated-apoptosis in liver of test fish. Aforesaid molecular markers can, thus, be efficiently utilized for bio-monitoring of aquatic regimes and conservation of fish biodiversity.
Silver nanoparticles impair zebrafish skeletal and cardiac myofibrillogenesis and sarcomere formation Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-30 Qin-Han Xu, PengPeng Guan, Ting Zhang, Chang Lu, GuoLiang Li, Jing-Xia Liu
Metal nanoparticles from industries contaminate the environment and affect the normal development of fish even human health. However, little is known about their biological effects on fish embryogenesis and the potential mechanisms. In this study, zebrafish embryos exposed to/injected with silver nanopaticles (AgNPs) exhibited shorter body, reduced heartbeats, and dysfunctional movements. Less, loose, and unassembled myofibrils were observed in AgNPs-treated embryos, and genes in myofibrillogenesis and sarcomere formation were found to be down-regulated in treated embryos. Down-regulated calcium (Ca2+) signaling and loci-specific DNA methylation in specific muscle genes, such as bves, shroom1, and arpc1a, occurred in AgNPs-treated embryos, which might result in the down-regulated expression of myofibrillogenesis genes and muscle dysfunctions in the treated embryos. Our results for the first time reveal that through down-regulating Ca2+ signaling and myogenic loci-specific DNA methylation in zebrafish embryos, AgNPs might induce defects of myofibril assembly and sarcomere formation via their particles mostly, which may subsequently cause heartbeat reduction and behavior dysfunctions.
Basis of genetic adaptation to heavy metal stress in the acidophilic green alga Chlamydomonas acidophila Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-27 Fernando Puente-Sánchez, Silvia Díaz, Vanessa Penacho, Angeles Aguilera, Sanna Olsson
To better understand heavy metal tolerance in Chlamydomonas acidophila, an extremophilic green alga, we assembled its transcriptome and measured transcriptomic expression before and after Cd exposure in this and the neutrophilic model microalga Chlamydomonas reinhardtii. Genes possibly related to heavy metal tolerance and detoxification were identified and analyzed as potential key innovations that enable this species to live in an extremely acid habitat with high levels of heavy metals. In addition we provide a data set of single orthologous genes from eight green algal species as a valuable resource for comparative studies including eukaryotic extremophiles. Our results based on differential gene expression, detection of unique genes and analyses of codon usage all indicate that there are important genetic differences in C. acidophila compared to C. reinhardtii. Several efflux family proteins were identified as candidate key genes for adaptation to acid environments. This study suggests for the first time that exposure to cadmium strongly increases transposon expression in green algae, and that oil biosynthesis genes are induced in Chlamydomonas under heavy metal stress. Finally, the comparison of the transcriptomes of several acidophilic and non-acidophilic algae showed that the Chlamydomonas genus is polyphyletic and that acidophilic algae have distinctive aminoacid usage patterns.
Effects of environmental stressors on lipid metabolism in aquatic invertebrates Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-25 Min-Chul Lee, Jun Chul Park, Jae-Seong Lee
Lipid metabolism is crucial for the survival and propagation of the species, since lipids are an essential cellular component across animal taxa for maintaining homeostasis in the presence of environmental stressors. This review aims to summarize information on the lipid metabolism under environmental stressors in aquatic invertebrates. Fatty acid synthesis from glucose via de novo lipogenesis (DNL) pathway is mostly well-conserved across animal taxa. The structure of free fatty acid (FFA) from both dietary and DNL pathway could be transformed by elongase and desaturase. In addition, FFA can be stored in lipid droplet as triacylglycerol, upon attachment to glycerol. However, due to the limited information on both gene and lipid composition, in-depth studies on the structural modification of FFA and their storage conformation are required. Despite previously validated evidences on the disturbance of the normal life cycle and lipid homeostasis by the environmental stressors (e.g., obesogens, salinity, temperature, pCO2, and nutrients) in the aquatic invertebrates, the mechanism behind these effects are still poorly understood. To overcome this limitation, omics approaches such as transcriptomic and proteomic analyses have been used, but there are still gaps in our knowledge on aquatic invertebrates as well as the lipidome. This paper provides a deeper understanding of lipid metabolism in aquatic invertebrates.
Effects of ozonated sewage effluent on reproduction and behavioral endpoints in zebrafish (Danio rerio) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-25 Johannes Pohl, Berndt Björlenius, Tomas Brodin, Gunnar Carlsson, Jerker Fick, D.G. Joakim Larsson, Leif Norrgren, Stefan Örn
Pharmaceutical residues and other micro-contaminants may enter aquatic environments through effluent from sewage treatment plants (STPs) and could cause adverse effects in wild fish. One strategy to alleviate this situation is to improve wastewater treatment by ozonation. To test the effectiveness of full-scale wastewater effluent ozonation at a Swedish municipal STP, the added removal efficiency was measured for 105 pharmaceuticals. In addition, gene expression, reproductive and behavioral endpoints were analyzed in zebrafish (Danio rerio) exposed on-site over 21 days to ozonated or non-ozonated effluents as well as to tap water. Ozone treatment (7 g O3/m3) removed pharmaceuticals by an average efficiency of 77% in addition to the conventional treatment, leaving 11 screened pharmaceuticals above detection limits. Differences in biological responses of the exposure treatments were recorded in gene expression, reproduction and behavior. Hepatic vitellogenin gene expression was higher in male zebrafish exposed to the ozonated effluent compared to the non-ozonated effluent and tap water treatments. The reproductive success was higher in fish exposed to ozonated effluent compared to non-ozonated effluent and to tap water. The behavioral measurements showed that fish exposed to the ozonated STP effluent were less active in swimming the first minute after placed in a novel vessel. Ozonation is a capable method for removing pharmaceuticals in effluents. However, its implementation should be thoroughly evaluated for any potential biological impact. Future research is needed for uncovering the factors which produced the in vivo responses in fish.
Genome-wide scan reveals signatures of selection related to pollution adaptation in non-model estuarine Atlantic killifish (Fundulus heteroclitus) Aquat. Toxicol. (IF 4.129) Pub Date : 2018-04-25 J.S. Osterberg, K.M. Cammen, T.F. Schultz, B.W. Clark, R.T. Di Giulio
In many human-altered ecosystems, organisms are increasingly faced with more diverse and complex environmental stressors and pollutant mixtures, to which the adaptations necessary to survive exposure are likely to be numerous and varied. Improving our understanding of the molecular mechanisms that underlie complex polygenic adaptations in natural settings requires significant toxicological, biochemical, physiological, and genomic data rarely available for non-model organisms. Here, we build upon two decades of study of adaptation to anthropogenic pollutants in a population of Atlantic killifish (Fundulus heteroclitus) that inhabits the creosote-contaminated Atlantic Wood Industries Superfund (AW) site on the Elizabeth River, Virginia in the United States. To better understand the genotypes that underlie previously characterized resistance to PCBs and PAHs, we performed Restriction site-Associated DNA sequencing (RADseq) on killifish from AW and two relatively clean reference sites (King’s Creek–KC, and Mains Creek–MC). Across the genome, we analyzed over 83,000 loci and 12,000 single nucleotide polymorphisms (SNPs). Shared across both comparisons of killifish from polluted (AW) and relatively unpolluted (KC and MC) sites, we found eight genomic regions with smoothed FST values significantly (p < 0.001) elevated above background. Using the recently published F. heteroclitus reference genome, we identified candidate genes in these significant regions involved in the AHR pathway (e.g. AIP, ARNT1c), as well as genes relating to cardiac structure and function. These genes represent both previously characterized and potentially novel molecular adaptations involved with various aspects of resistance to these environmental toxins.
Some contents have been Reproduced by permission of The Royal Society of Chemistry.
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