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BIOLOGY (1529 journals)                  1 2 3 4 5 6 7 8 | Last

Showing 1 - 200 of 1720 Journals sorted alphabetically
AAPS Journal     Hybrid Journal   (Followers: 23)
Achievements in the Life Sciences     Open Access   (Followers: 5)
ACS Synthetic Biology     Hybrid Journal   (Followers: 25)
Acta Biologica Colombiana     Open Access   (Followers: 7)
Acta Biologica Hungarica     Full-text available via subscription   (Followers: 4)
Acta Biologica Sibirica     Open Access   (Followers: 2)
Acta Biologica Turcica     Open Access  
Acta Biomaterialia     Hybrid Journal   (Followers: 27)
Acta Biotheoretica     Hybrid Journal   (Followers: 4)
Acta Chiropterologica     Full-text available via subscription   (Followers: 6)
acta ethologica     Hybrid Journal   (Followers: 4)
Acta Fytotechnica et Zootechnica     Open Access   (Followers: 1)
Acta Limnologica Brasiliensia     Open Access   (Followers: 3)
Acta Médica Costarricense     Open Access   (Followers: 2)
Acta Musei Silesiae, Scientiae Naturales     Open Access  
Acta Neurobiologiae Experimentalis     Open Access  
Acta Parasitologica     Hybrid Journal   (Followers: 11)
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Acta Scientifica Naturalis     Open Access   (Followers: 3)
Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis     Open Access   (Followers: 1)
Actualidades Biológicas     Open Access   (Followers: 1)
Advanced Health Care Technologies     Open Access   (Followers: 4)
Advanced Journal of Graduate Research     Open Access  
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Advanced Studies in Biology     Open Access  
Advances in Antiviral Drug Design     Full-text available via subscription   (Followers: 2)
Advances in Bioinformatics     Open Access   (Followers: 17)
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African Journal of Range & Forage Science     Hybrid Journal   (Followers: 8)
AFRREV STECH : An International Journal of Science and Technology     Open Access   (Followers: 1)
Ageing Research Reviews     Hybrid Journal   (Followers: 11)
Aging Cell     Open Access   (Followers: 19)
Agrokémia és Talajtan     Full-text available via subscription   (Followers: 2)
Agrokreatif Jurnal Ilmiah Pengabdian kepada Masyarakat     Open Access  
AJP Cell Physiology     Hybrid Journal   (Followers: 17)
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AJP Lung Cellular and Molecular Physiology     Hybrid Journal   (Followers: 3)
Al-Kauniyah : Jurnal Biologi     Open Access  
Alasbimn Journal     Open Access   (Followers: 1)
Alces : A Journal Devoted to the Biology and Management of Moose     Open Access  
AMB Express     Open Access   (Followers: 1)
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American Biology Teacher     Full-text available via subscription   (Followers: 14)
American Fern Journal     Full-text available via subscription   (Followers: 1)
American Journal of Agricultural and Biological Sciences     Open Access   (Followers: 8)
American Journal of Bioethics     Hybrid Journal   (Followers: 13)
American Journal of Human Biology     Hybrid Journal   (Followers: 14)
American Journal of Medical and Biological Research     Open Access   (Followers: 8)
American Journal of Plant Sciences     Open Access   (Followers: 19)
American Journal of Primatology     Hybrid Journal   (Followers: 16)
American Malacological Bulletin     Full-text available via subscription   (Followers: 3)
American Naturalist     Full-text available via subscription   (Followers: 76)
Amphibia-Reptilia     Hybrid Journal   (Followers: 6)
Anadol University Journal of Science and Technology B : Theoritical Sciences     Open Access  
Anadolu University Journal of Science and Technology : C Life Sciences and Biotechnology     Open Access  
Anaerobe     Hybrid Journal   (Followers: 4)
Analytical Methods     Full-text available via subscription   (Followers: 11)
Anatomical Science International     Hybrid Journal   (Followers: 3)
Animal Cells and Systems     Hybrid Journal   (Followers: 4)
Animal Models and Experimental Medicine     Open Access  
Annales de Limnologie - International Journal of Limnology     Hybrid Journal   (Followers: 1)
Annales françaises d'Oto-rhino-laryngologie et de Pathologie Cervico-faciale     Full-text available via subscription   (Followers: 3)
Annales Henri Poincaré     Hybrid Journal   (Followers: 3)
Annals of Applied Biology     Hybrid Journal   (Followers: 7)
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Annals of Science and Technology     Open Access  
Annual Review of Biomedical Engineering     Full-text available via subscription   (Followers: 14)
Annual Review of Biophysics     Full-text available via subscription   (Followers: 23)
Annual Review of Cancer Biology     Full-text available via subscription   (Followers: 2)
Annual Review of Cell and Developmental Biology     Full-text available via subscription   (Followers: 37)
Annual Review of Food Science and Technology     Full-text available via subscription   (Followers: 13)
Annual Review of Genomics and Human Genetics     Full-text available via subscription   (Followers: 24)
Annual Review of Phytopathology     Full-text available via subscription   (Followers: 12)
Anthropological Review     Open Access   (Followers: 23)
Anti-Infective Agents     Hybrid Journal   (Followers: 3)
Antibiotics     Open Access   (Followers: 9)
Antioxidants     Open Access   (Followers: 4)
Antioxidants & Redox Signaling     Hybrid Journal   (Followers: 8)
Antonie van Leeuwenhoek     Hybrid Journal   (Followers: 5)
Anzeiger für Schädlingskunde     Hybrid Journal   (Followers: 1)
Apidologie     Hybrid Journal   (Followers: 4)
Apmis     Hybrid Journal   (Followers: 1)
APOPTOSIS     Hybrid Journal   (Followers: 9)
Applied Biology     Open Access  
Applied Bionics and Biomechanics     Open Access   (Followers: 8)
Applied Vegetation Science     Full-text available via subscription   (Followers: 10)
Aquaculture Environment Interactions     Open Access   (Followers: 4)
Aquaculture International     Hybrid Journal   (Followers: 26)
Aquaculture Reports     Open Access   (Followers: 3)
Aquaculture, Aquarium, Conservation & Legislation - International Journal of the Bioflux Society     Open Access   (Followers: 7)
Aquatic Biology     Open Access   (Followers: 6)
Aquatic Ecology     Hybrid Journal   (Followers: 36)
Aquatic Ecosystem Health & Management     Hybrid Journal   (Followers: 15)
Aquatic Science and Technology     Open Access   (Followers: 3)
Aquatic Toxicology     Hybrid Journal   (Followers: 23)
Archaea     Open Access   (Followers: 3)
Archiv für Molluskenkunde: International Journal of Malacology     Full-text available via subscription   (Followers: 3)
Archives of Biological Sciences     Open Access  
Archives of Microbiology     Hybrid Journal   (Followers: 9)
Archives of Natural History     Hybrid Journal   (Followers: 7)
Archives of Oral Biology     Hybrid Journal   (Followers: 3)
Archives of Virology     Hybrid Journal   (Followers: 5)
Archivum Immunologiae et Therapiae Experimentalis     Hybrid Journal   (Followers: 2)
Arid Ecosystems     Hybrid Journal   (Followers: 2)
Arquivos do Instituto Biológico     Open Access   (Followers: 1)
Arquivos do Museu Dinâmico Interdisciplinar     Open Access  
Arthropod Structure & Development     Hybrid Journal   (Followers: 2)
Arthropods     Open Access   (Followers: 1)
Artificial DNA: PNA & XNA     Hybrid Journal   (Followers: 3)
Asian Bioethics Review     Full-text available via subscription   (Followers: 3)
Asian Journal of Biodiversity     Open Access   (Followers: 4)
Asian Journal of Biological Sciences     Open Access   (Followers: 3)
Asian Journal of Cell Biology     Open Access   (Followers: 5)
Asian Journal of Developmental Biology     Open Access   (Followers: 2)
Asian Journal of Medical and Biological Research     Open Access   (Followers: 4)
Asian Journal of Nematology     Open Access   (Followers: 4)
Asian Journal of Poultry Science     Open Access   (Followers: 4)
Atti della Accademia Peloritana dei Pericolanti - Classe di Scienze Medico-Biologiche     Open Access  
Australian Life Scientist     Full-text available via subscription   (Followers: 2)
Australian Mammalogy     Hybrid Journal   (Followers: 6)
Autophagy     Hybrid Journal   (Followers: 3)
Avian Biology Research     Full-text available via subscription   (Followers: 4)
Avian Conservation and Ecology     Open Access   (Followers: 11)
Bacteriology Journal     Open Access   (Followers: 1)
Bacteriophage     Full-text available via subscription   (Followers: 3)
Bangladesh Journal of Bioethics     Open Access  
Bangladesh Journal of Plant Taxonomy     Open Access  
Bangladesh Journal of Scientific Research     Open Access   (Followers: 1)
Batman Üniversitesi Yaşam Bilimleri Dergisi     Open Access  
Berita Biologi     Open Access   (Followers: 1)
Between the Species     Open Access   (Followers: 1)
Bio Tribune Magazine     Hybrid Journal  
BIO Web of Conferences     Open Access  
BIO-Complexity     Open Access  
Bio-Grafía. Escritos sobre la Biología y su enseñanza     Open Access  
Bioanalytical Reviews     Hybrid Journal   (Followers: 2)
Biocatalysis and Biotransformation     Hybrid Journal   (Followers: 6)
BioCentury Innovations     Full-text available via subscription   (Followers: 1)
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Biochimie     Hybrid Journal   (Followers: 7)
BioControl     Hybrid Journal   (Followers: 5)
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BioDiscovery     Open Access   (Followers: 2)
Biodiversidade e Conservação Marinha : Revista CEPSUL     Open Access  
Biodiversitas : Journal of Biological Diversity     Open Access  
Biodiversity Data Journal     Open Access   (Followers: 4)
Biodiversity Informatics     Open Access   (Followers: 1)
Biodiversity Information Science and Standards     Open Access  
Biodiversity: Research and Conservation     Open Access   (Followers: 27)
Bioedukasi : Jurnal Pendidikan Biologi FKIP UM Metro     Open Access  
Bioeksperimen : Jurnal Penelitian Biologi     Open Access  
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BioEssays     Hybrid Journal   (Followers: 10)
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BioéthiqueOnline     Open Access  
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Biogeosciences (BG)     Open Access   (Followers: 10)
Biogeosciences Discussions (BGD)     Open Access   (Followers: 2)
Bioinformatics     Hybrid Journal   (Followers: 317)
Bioinformatics and Biology Insights     Open Access   (Followers: 11)
Bioinspiration & Biomimetics     Hybrid Journal   (Followers: 7)
Biointerphases     Open Access   (Followers: 1)
Biojournal of Science and Technology     Open Access  
BioLink : Jurnal Biologi Lingkungan, Industri, Kesehatan     Open Access   (Followers: 1)
Biologia     Hybrid Journal  
Biologia on-line : Revista de divulgació de la Facultat de Biologia     Open Access  
Biological Bulletin     Partially Free   (Followers: 6)
Biological Control     Hybrid Journal   (Followers: 4)
Biological Invasions     Hybrid Journal   (Followers: 21)
Biological Journal of the Linnean Society     Hybrid Journal   (Followers: 18)

        1 2 3 4 5 6 7 8 | Last

Journal Cover
Aquatic Toxicology
Journal Prestige (SJR): 1.456
Citation Impact (citeScore): 4
Number of Followers: 23  
  Hybrid Journal Hybrid journal (It can contain Open Access articles)
ISSN (Print) 0166-445X - ISSN (Online) 1879-1514
Published by Elsevier Homepage  [3157 journals]
  • Impact of erythromycin on a non-target organism: cellular effects on the
           freshwater microalga Pseudokirchneriella subcapitata
    • Abstract: Publication date: Available online 16 January 2019Source: Aquatic ToxicologyAuthor(s): Manuela D. Machado, Eduardo V. Soares The increasing and indiscriminate use of antibiotics is the origin of their introduction in aquatic systems through domestic and livestock effluents. The occurrence of erythromycin (ERY), a macrolide antibiotic, in water bodies raises serious concerns about its potential toxic effect in aquatic biota (non-target organisms), particularly in microalgae, the first organisms in contact with aquatic contaminants. This study aimed to evaluate the possible toxic effects of ERY on relevant cell targets of the freshwater microalga Pseudokirchneriella subcapitata. Algal cells incubated with significant environmental ERY concentrations presented disturbance of the photosynthetic apparatus (increased algal autofluorescence and reduction of chlorophyll a content) and mitochondrial function (hyperpolarization of mitochondrial membrane). These perturbations can apparently be attributed to the similarity of the translational machinery of these organelles (chloroplasts and mitochondria) with the prokaryotic cells. P. subcapitata cells treated with ERY showed a modification of metabolic activity (increased esterase activity) and redox state (alteration of intracellular levels of reactive oxygen species and reduced glutathione content) and an increased biovolume. ERY induced an algistatic effect: reduction of growth rate without loss of cell viability (plasma membrane integrity). The present study shows that chronic exposure (72 h), at low (µg L-1) ERY concentrations (within the range of concentrations detected in surface and ground waters), induce disturbances in the physiological state of the alga P. subcapitata. Additionally, this work alerts to the possible negative impact of the uncontrolled use of ERY on the aquatic systems.
  • Molecular signaling pathways elicited by 17α-ethinylestradiol in Japanese
           medaka male larvae undergoing gonadal differentiation
    • Abstract: Publication date: Available online 16 January 2019Source: Aquatic ToxicologyAuthor(s): Ahmed Abdelmoneim, Amira Abdu, Shuai Chen, Maria S. Sepúlveda Estrogenic contaminants released into water bodies are potentially affecting the reproduction of aquatic organisms. Exposure to 17α-ethinylestradiol (EE2), a synthetic estrogen agonist commonly found in sewage effluents, has been shown to cause gonadal changes in male gonochoristic fish ranging from gonadal intersex to complete sex reversal. Although these gonadal changes have been well studied in Japanese medaka Oryzias latipes, the molecular mechanisms behind them are poorly understood. Our objective was to study the signaling pathways elicited by exposure to different concentrations of EE2 in this species. Embryos and larvae were sexed by the presence of leucophores and dmy expression (only in males). Male medaka were exposed to two EE2 concentrations (30 and 300 ng/L) during their gonadal differentiation period (7 – 22 dpf). The transcriptome of larvae was analyzed using RNA sequencing followed by pathway analysis. Genes involved in sex differentiation and gonadal development (e.g., cldn19, ctbp1, hsd17b4) showed a female-like expression pattern in EE2-exposed males with some genes changing in expression in an EE2 concentration-dependent manner. However, not all genes known to be involved in sex differentiation and gonadal development (e.g., wnt4b) were altered by EE2. Several of the prominently affected signaling pathways involved genes associated with steroidogenesis, steroid receptor signaling and steroid metabolism, such as cyp2b3, cyp3b40, cyp1a, hsd17b4. We also report on novel genes and pathways affected that might play a role in gonadal changes, including several genes associated with FXR/RXR and LXR/RXR activation networks. This study is the first to examine the transcriptomic changes in male fish resulting from exposure to EE2 during the gonadal differentiation period, providing new insights on the signaling pathways involved in the development of gonadal changes in gonochoristic fish.
  • Estrogen sensitive liver transgenic zebrafish (Danio rerio) line
           (Tg(vtg1:mCherry)) suitable for the direct detection of estrogenicity in
           environmental samples
    • Abstract: Publication date: Available online 15 January 2019Source: Aquatic ToxicologyAuthor(s): Katalin Bakos, Robert Kovacs, Erna Balogh, Dora Kanaine Sipos, Marta Reining, Orsolya Gyomorei-Neuberger, Adrienn Balazs, Balazs Kriszt, Dora Bencsik, Andrea Csepeli, Gyongyi Gazsi, Yavor Hadzhiev, Bela Urbanyi, Ferenc Mueller, Balazs Kovacs, Zsolt Csenki Environmental estrogens are a serious concern worldwide due to their ubiquity and adverse ecotoxicological and health effects. Chemical structure of these substances is highly diverse, therefore estrogenicity cannot be predicted on the basis of molecular structure. Furthermore, estimation of estrogenicity of environmental samples based on chemical analytics of suspects is difficult given the complex interaction of chemicals and the impact on estrogenicity. The full estrogenic impact of an environmental sample can thus only be revealed by a series of sensitive in vitro and in vivo ecotoxicological tests. Herein we describe a vitellogenin reporter transgenic zebrafish line (Tg(vtg1:mCherry)) that enables the detection of estrogenicity in the environmentally relevant, low concentration ranges in embryonic tests that are in accordance with 3Rs and relevant animal welfare regulations.The transgene construct used for the development of Tg(vtg1:mCherry) carried a long (3.4 kbp) natural vitellogenin-1 promoter sequence with a high number of ERE sites.A test protocol was developed based on our finding that the endogenous vitellogenin and the reporter show similar spatial expression pattern and both endogenous and vitellogenin reporter is only produced in the left hepatic lobe of 5 dpf zebrafish embryos.Seven generations of Tg(vtg1:mCherry) have been established, and the estrogen responsiveness was tested with different estrogenic substances and wastewater samples. Embryos were exposed from 3 to 5 days post fertilization (dpf). Fluorescence in embryos could be detected upon treatment with 17-ß-estradiol from a concentration of 100 ng/L, 17-α-ethynilestradiol from 1 ng/L, zearalenone from 100 ng/L and bisphenol-A from 1 mg/L. In the adult stage transgene activity appeared to be more sensitive to estrogen treatment, with detectable transgene activity from 5 ng/L 17-ß-estradiol concentration. The transgenic line Tg(vtg1:mCherry) was also suitable for the direct measurement of estrogenicity in wastewater samples without sample extraction. The detection of estrogenic activity using the reporter line was confirmed by the bioluminescent yeast estrogen screen.
  • ROS changes are responsible for tributyl phosphate (TBP)-induced toxicity
           in the alga Phaeodactylum tricornutum
    • Abstract: Publication date: Available online 15 January 2019Source: Aquatic ToxicologyAuthor(s): Qian Liu, Xuexi Tang, You Wang, Yingying Yang, Wei Zhang, Yunchen Zhao, Xinxin Zhang As a newly emerging environmental contaminant, tributyl phosphate (TBP) is an additive flame retardant of high production volume that is frequently detected in biota and the environment. Despite evidence that TBP is a potential threat to marine organisms, ecotoxicology data for TBP in marine organisms at low trophic levels are scarce. In this study, the acute toxicological effect of TBP on the marine phytoplankton Phaeodactylum tricornutum was thoroughly investigated, and the possible mechanism was explored. The results showed that TBP at concentrations ≥ 0.2 mg L-1 significantly inhibited P. tricornutum growth in a clear dose-response manner, with 72-h EC10, EC20, EC50 and EC90 values of 0.067, 0.101, 0.219 and 0.716 mg L-1, respectively. Algal cells treated with TBP exhibited distorted shapes, ruptured cell membranes and damaged organelles, especially mitochondria. Additionally, apoptosis was triggered, followed by a decrease in mitochondrial membrane potential, indicating that cellular damage occurred during exposure. Although the activities of two antioxidant enzymes, superoxide peroxidase and catalase, were upregulated by TBP at 1.2 mg L-1, excess reactive oxygen species (ROS) and malondialdehyde still accumulated in algal cells after exposure, suggesting that the cells experienced oxidative stress. Moreover, both growth inhibition and apoptosis were positively correlated with ROS levels and were ameliorated by pretreatment with the ROS scavenger N-acetyl-L-cysteine. Taken together, the results indicate that TBP exposure leads to growth inhibition and cellular damage in P. tricornutum, and a ROS-mediated pathway might contribute to these observed toxicological effects.
  • Subtle Morphometric, Behavioral and Gene Expression Effects in Larval
           Zebrafish Exposed to PFHxA, PFHxS and 6:2 FTOH
    • Abstract: Publication date: Available online 14 January 2019Source: Aquatic ToxicologyAuthor(s): Kate M. Annunziato, Carrie E. Jantzen, Melissa C. Gronske, Keith R. Cooper Recent studies of perfluoroalkylated substances (PFASs) have focused on the toxicity of long chain PFASs, such as PFOS or PFOA, which have been demonstrated to cause an array of developmental and behavioral effects. However, less is known about low molecular weight PFASs and alternatives. This study examined the morphometric and behavioral effects in zebrafish following developmental exposures of C6 PFASs: perfluorohexanoic acid, PFHxA, perfluorohexane sulfonate, PFHxS, and 6:2 fluorotelomer alcohol, 6:2 FTOH. Embryos were exposed to 0.02 to 20 µM concentrations of these compounds from the high stage (˜3 hours post fertilization, hpf) until 120 hpf. Morphometric and gene expression endpoints were examined at 120 hpf. Genes selected for analysis were previously shown to be altered in zebrafish developmentally exposed to PFOS and PFOA. Additionally, exposed larvae were transferred to clean water and reared until 14 days post fertilization, dpf, when behavioral assays were completed and morphometric endpoints examined. While PFHxA was found to be the most acutely toxic at 120 hpf, few morphometric effects were observed. Gene expression was the most sensitive endpoint with significant increased tgfb1a, bdnf, and ap1s1 expression observed with PFHxA exposure. PFHxS exposure produced morphometric effects in the larvae, specifically increased length and yolk sac area at 2 and 20 µM. This phenotype persisted to the 14 dpf time point, where these larvae additionally displayed decreased distance traveled and crosses through the center of the arena of the behavioral assay. Exposure to 6:2 FTOH caused no morphometric effects at 120 hpf, and this compound was the least acutely toxic. However, expression of both tgfb1a and bdnf were increased by greater than 2 fold change at this time point. Effects also persisted to 14 dpf where a significant increase in distance traveled and velocity were observed in the behavioral assay. This study demonstrates effects on behavioral, morphometric and gene expression endpoints with developmental PFHxA, PFHxS, and 6:2 FTOH exposures in zebrafish.
  • Carbamazepine disrupts molting hormone signaling and inhibits molting and
           growth of Eriocheir sinensis at environmentally relevant concentrations
    • Abstract: Publication date: Available online 14 January 2019Source: Aquatic ToxicologyAuthor(s): Huihui Chen, Xiaohong Gu, Qingfei Zeng, Zhigang Mao, Xuefang Liang, Christopher J. Martyniuk Carbamazepine (CBZ), one of the most frequently detected pharmaceutical compounds in aquatic environments, has recently been shown to cause chronic toxicity and endocrine disruption in a variety of non-target aquatic organisms. However, neither the effects nor the specific mechanism of CBZ action on the molting of crustaceans is well understood. The aim of this study was to investigate the effects of CBZ on the molting and growth of the Chinese mitten crab Eriocheir sinensis, a native and economically important species in China, and to elucidate the specific mechanisms through which molt inhibition occurs. Juvenile E. sinensis were treated with four nominal environmentally relevant concentrations (0.01, 0.1, 1, or 10 µg/L) of CBZ for acute (4 days) and chronic (40 days) exposures. After acute exposure, chitinase activity in the epidermis and 20-hydroxyecdysone (20-HE) concentration in the hemolymph were significantly decreased (p 
  • Expression of immune, antioxidant and stress related genes in different
           organs of common carp exposed to indoxacarb
    • Abstract: Publication date: Available online 14 January 2019Source: Aquatic ToxicologyAuthor(s): Melika Ghelichpour, Ali Taheri Mirghaed, Seyed Hossein Hoseinifar, Mohsen Khalili, Morteza Yousefi, Hien Van Doan, Amalia Perez Jimenez The aim of the present study was to investigate the effects of chronic exposure of common carp (Cyprinus carpio) to indoxacarb on immune, antioxidant and stress gene expression. After 21 days exposure to 0, 0.75, 1.5 and 3 ppm indoxacarb, expression of IL-1β, IL-8, IL-10, TNF-α, IFN-γ, SOD, CAT, HSP70, IGF-I and IGF-II were assessed in liver, kidney and gills. In general, exposure to low concentration of indoxacarb increased inflammatory cytokine gene expression (IL-1β, IL-8, IL-10, TNF-α and IFN-γ) and inhibits inflammatory cytokines’ expression at higher concentrations. The assessment of antioxidant gene expression (SOD and CAT) in different organs indicate that they were increased by low concentrations of indoxacarb to deal with primary oxidative situation. However, higher concentrations of indoxacarb caused reduction in oxidative gene expression. IGF genes expression in liver significantly increased at a concentration of 0.75 ppm treatment, then it decreased at 1.5 ppm indoxacarb and increased again by increasing in the indoxacarb concentration to 3 ppm. The expression of HSP70 in kidney showed a significant elevation in 0.75 and 1.5 ppm treatments compared with 3 ppm treatment and the control group. The expression of this gene in liver was significantly increased in 1.5 and 3 ppm treatments. The same pattern of expression was also observed in gill. Overall, indoxacarb exposure affects common carp health at transcription levels. Changes in the genes expression generally suggest that indoxacarb exposure led to interference in inflammation, oxidative stress and tissue damage.
  • The endocrine disruptor, 17α-ethinyl estradiol, alters male mate
           choice in a freshwater fish
    • Abstract: Publication date: Available online 9 January 2019Source: Aquatic ToxicologyAuthor(s): Minna Saaristo, Christopher P. Johnstone, Kun Xu, Mayumi Allinson, Bob B.M. Wong Among the handful of studies on the behavioural effects of endocrine disrupting chemicals (EDCs), only a few have set out to disentangle the mechanisms underpinning behavioural changes. In fish, previous studies have shown that both visual and chemical cues play an important role in mate choice. As such, contaminant-induced changes in either transmission or perception of mate choice cues could have direct implications for individual’s fitness. One widespread contaminant of environmental concern is 17α-ethinyl estradiol (EE2), a synthetic estrogen used in the contraceptive pill. Here, we investigated the impacts of EE2 exposure (28 days; measured concentration 14 ng/L) on visual and chemical communication in wild guppies (Poecilia reticulata). Using a standard dichotomous mate choice assay, we first gave individual males (either control or EE2-exposed) the opportunity to court two size-matched females (one control and one EE2-exposed) using only visual cues. We then introduced chemical cues of females (control and EE2-exposed) to the trial tank. We found that there was no significant effect of EE2-treatment on total time males spent associating with the females, when given only visual cues. There was, however, a significant effect on male courtship behaviour, with both control and EE2-exposed males spending more time performing ‘sigmoid’ displays towards the visual cues of control females compared to EE2-exposed females. When males were presented with both visual and chemical female cues simultaneously, we found that males spent more time courting control females that were paired with EE2-chemical cues. Not only does our study uncover a previously unknown behavioural impact of EE2-exposure on chemical cues, but demonstrates that EE2-exposure can exert complex effects on visual and chemical communication in a mate choice context. Finally, we contribute to the discussion of intraspecific variability by providing data on the potential trade-offs underpinning contaminant-induced behavioural changes.
  • Recovery of Alexandrium tamarense under chronic exposure of TiO2
           nanoparticles and possible mechanisms
    • Abstract: Publication date: Available online 9 January 2019Source: Aquatic ToxicologyAuthor(s): Manlu Li, Yuelu Jiang, Chia-Ying Chuang, Jin Zhou, Xiaoshan Zhu, Daoyi Chen Harmful algal blooms (HAB), heavily influenced by human activities, pose serious hazard to aquatic ecology and human health. In this study, we monitored the physiological responses and paralytic shellfish poisoning toxins (PSTs) of the toxin-producing HAB species Alexandrium tamarense under titanium dioxide nanoparticles (nTiO2) exposure in the concentration range of 2-320 mg L-1 over a period of 13 days. The results showed the acute inhibition of nTiO2 on the algal growth, photosynthetic efficiency and esterase activity at all concentrations except 2 mg L-1. Nonetheless, they recovered after 13 days nTiO2 exposure from 20 to 80 mg L-1. The EC50 value increased from 85.1 mg L-1 in Day 4 to 140.9 mg L-1 in Day 13. The physiological recovery after prolonged exposure may result from the elimination of excess reactive oxygen species (ROS), a combined outcome of increased nTiO2 aggregation and algal antioxidant defense mechanisms. This observation is supported by the immediately increased antioxidant enzyme activities, including the superoxide dismutase (SOD) and catalase (CAT) activities upon nTiO2 exposure. Moreover, the production of PSTs in A. tamarense significantly increased by 1.41-1.76 folds after chronic nTiO2 exposure at all tested concentrations (p 
  • Assessing the effect of human pharmaceuticals (carbamazepine, diclofenac
           and ibuprofen) on the marine clam Ruditapes philippinarum: an integrative
           and multibiomarker approach
    • Abstract: Publication date: Available online 9 January 2019Source: Aquatic ToxicologyAuthor(s): Chiara Trombini, Miriam Hampel, Julián Blasco The presence of pharmaceuticals in the aquatic ecosystem has become a topic of growing interest in recent years. In this study, the marine clam Ruditapes philippinarum was exposed during 14 days to concentrations close to those found in the environment: (15 µg L-1) of carbamazepine (CBZ), diclofenac (DCF) and ibuprofen (IBU), three pharmaceuticals widely used worldwide and commonly found within the aquatic environment. Additionally, exposure was followed by a depuration phase (7 days). A battery of biomarkers (superoxide dismutase SOD, catalase CAT, glutathione reductase GR, total glutathione peroxidase T-GPx, glutathione transferase GST, lipid peroxidation LPO, acetylcholinesterase AChE and metallothionein MT) was evaluated throughout the exposure and depuration. The Integrated Biomarker Response index was calculated with all selected biomarkers and used as a complementary tool in the evaluation of the organisms’ health status. Exposure induced changes in the clams’ biochemical responses that led to the hypothesis of the harmful role of the pharmaceuticals resulting in negative effects (changes in enzyme activities, LPO and MT levels, related to ROS production) particularly after short-term exposure. However, the clams showed the ability to cope with these imbalances by recovering their general oxidative status by the end of exposure.
  • Molecular characterization of zebrafish Gstr1, the only member of
           teleost-specific glutathione S- transferase class
    • Abstract: Publication date: Available online 9 January 2019Source: Aquatic ToxicologyAuthor(s): Branka Bašica, Ivan Mihaljević, Nikola Maraković, Radmila Kovačević, Tvrtko Smital Glutathione S-transferases (GSTs) are multifunctional phase II detoxification enzymes with primary function of glutathione conjugation of various endogenous and exogenous compounds. Teleost-specific Gstr1 in zebrafish (Danio rerio) was previously shown to have high expression in toxicologically relevant tissues and high activity towards model substrates. The aim of this study was a detailed functional characterization of zebrafish Gstr1. Molecular docking analyses were used to get novel insight into structural characteristics of Gstr1 and elucidation of the mechanistic interactions with both GSH and various Gstr1 substrates or inhibitors. An initial screening inhibition assay performed using model fluorescence substrate monochlorobimane (MCB) revealed interactions of different endogenous compounds and environmentally relevant xenobiotics with zebrafish Gstr1. All interacting compounds were further analyzed to determine their inhibition type and Ki values. Our data revealed that pregnenolone, progesterone, testosterone, DHEAS and corticosterone competitively inhibited transformation of MCB by Gstr1 with the calculated Ki values in the range 14-26 μM, implying that these hormones are physiological substrates of zebrafish Gstr1. Estrogens had no effect on Gstr1 activity. Taurochenodeoxycholate (TCDC) expressed lower inhibition potency toward Gstr1 with the Ki value of 33 μM. Among tested xenobiotics tributyltin chloride and rifampicin non-enzymatically bound Gstr1 enzyme (the calculated Ki values are 0.26 μM and 65 μM, respectively) and inhibited its activity, showing that these compounds are reversible noncompetitive inhibitors of zebrafish Gstr1. Insecticide diazinon competitively inhibited Gstr1 activity with calculated Ki value of 27 μM, while other Gstr1-interacting insecticides, chlorpyrifos-methyl (CPF-methyl) and malathion, showed allosteric activation-like effect. Among tested pharmaceuticals, tetracycline, erythromycin and methotrexate demonstrated competitive type of inhibition with the calculated Ki values of 17.5, 36.5 and 29 μM, respectively. In summary, we suggest that zebrafish Gstr1 has an important role in steroidogenesis, metabolism and/or physiological actions of androgens, but not estrogens in fish. Finally, our results imply the role of Gstr1 in metabolism of xenobiotics and protection of fish against deleterious environmental contaminants such as organophosphate insecticides and pharmaceuticals.
  • Metal accumulation, biochemical and behavioral responses on the
           Mediterranean clams Ruditapes decussatus exposed to two photocatalyst
           nanocomposites (TiO2 NPs and AuTiO2NPs)
    • Abstract: Publication date: Available online 7 January 2019Source: Aquatic ToxicologyAuthor(s): Wiem Saidani, Sellami Badreddine, Abdelhafidh Khazri, Amine Mezni, Mouhamed Dellali, Olivier Joubert, David Sheehan, Hamouda Beyrem Nanoparticle decoration with noble metal represents a promising alternative to improve their photocatalytic and photovoltaic properties. However, toxicity can be influenced by such modification, as the bioavailability of these substances may be influenced. To understand how decoration influences the NP impacts in marine ecosystems, we exposed suspension-feeding clams, Ruditapes decussatus, to two photocatalyst nanocomposites, TiO2 NPs and AuTiO2 NPs, over 2 concentrations, 50 µg L−1and 100 µg L-1, in a laboratory experiment.Accumulation of Au and Ti in gills and digestive gland was noted in clams after exposure to TiO2 NPs and AuTiO2 NPs using inductively coupled plasma optic emission spectroscopy (ICP-OES).TiO2 and AuTiO2 NPs alter the behavior of the clams Ruditapes decussatus by reducing filtration and respiration rates. Furthermore, the highest concentration of TiO2NPs induces an overproduction of H2O2 in gills and digestive gland and NO production only in gills. Superoxide dismutase (SOD), Catalase (CAT), Glutathione-S-transferase (GST) and acetylcholinesterase (AChE) activities were induced in gills and digestives gland in concentration and nanocomposite type dependent manner.Decorated form presented higher Malondialdehyde (MDA) levels in gills and digestive gland than the undecorated form, suggesting different mechanisms of action that may be mediated through oxidative stress.In conclusion, the considered parameters could represent reliable biomarkers for the assessment of NP toxicity on R. decussatus as biological biomonitoring model. In addition, based on the obtained results, nanoparticle decoration influences the toxicity of metal nanoparticles in marine organism.
  • The Retinoic Acid Receptor (RAR) in molluscs: function, evolution and
           endocrine disruption insights
    • Abstract: Publication date: Available online 7 January 2019Source: Aquatic ToxicologyAuthor(s): Ana André, Raquel Ruivo, Elza Fonseca, Elsa Froufe, L. Filipe C. Castro, Miguel M. Santos Retinoid acid receptor (RAR)-dependent signalling pathways are essential for the regulation and maintenance of essential biological functions and are recognized targets of disruptive anthropogenic compounds. Recent studies put forward the inability of mollusc RARs to bind and respond to the canonical vertebrate ligand, retinoic acid: a feature that seems to have been lost during evolution. Yet, these studies were carried out in a limited number of molluscs. Therefore, using an in vitro transactivation assay, the present work aimed to characterize phylogenetically relevant mollusc RARs, as monomers or as functional units with RXR, not only in the presence of vertebrate bone fine ligands but also known endocrine disruptors, described to modulate retinoid-dependent pathways. In general, none of the tested mollusc RARs were able to activate reporter gene transcription when exposed to retinoic acid isomers, suggesting that the ability to respond to retinoic acid was lost across molluscs. Similarly, the analysed mollusc RAR were unresponsive towards organochloride pesticides. In contrast, transcriptional repressions were observed with the RAR/RXR unit upon exposure to retinoids or RXR-specific ligands. Loss-of-function and gain-of-function mutations further corroborate the obtained results and suggest that the repressive behaviour, observed with mollusc and human RAR/RXR heterodimers, is possibly mediated by ligand biding to RXR.
  • Molecular characterization of thioredoxin reductase in waterflea Daphnia
           magna and its expression regulation by polystyrene microplastics
    • Abstract: Publication date: Available online 4 January 2019Source: Aquatic ToxicologyAuthor(s): Jinghong Tang, Xuan Wang, Jun Yin, Yiran Han, Jian Yang, Xiaoyu Lu, Tianchen Xie, Siddiq Akbar, Kai Lyu, Zhou Yang Global scale concerns regarding rise in microplastics pollution in the environment have recently aroused. Ingestion of microplastics by biota, including freshwater zooplankton has been well studied, however, despite keystone species in freshwater food webs, the molecular response (e.g. oxidative defense) of zooplankton in response to microplastics is still in its infancy. The thioredoxin (TRx) system has a vital function in cellular antioxidative defense via eliminating the excessive generation of reactive oxygen species (ROS). Therefore, it is necessary to investigate the effects of thioredoxin reductase (TRxR), due to its triggering the TRx catalysis cascade. The present study identified TRxR in Daphnia magna (Dm-TRxR) for the first time, and found that the full-length cDNA was 1862 bp long, containing an 1821-bp open reading frame. Homologous alignments showed the presence of conserved catalytic domain CVNVGC and the seleocysteine (SeCys) residue (U) located in the N- and C- terminal portions. Subsequently, the expression of Dm-TRxR, together with permease, arginine kinase (AK), was investigated by approach of quantitative real-time PCR after exposure to four (1.25-μm) polystyrene (PS) microbeads concentrations: 0 (control), 2, 4 and 8 mg L-1 for 10 days. Dm-TRxR, permease and AK mRNA were significantly upregulated after exposure to 2, 4 mg L-1 of PS, but then declined in the presence of 8 mg L-1 PS. The gene expression results suggested that oxidative defense, energy production and substance extra cellular transportation were significantly regulated by microplastic exposure. Collectively, the present study will advance our knowledge regarding the biological effects of microplastic pollution on zooplankton, and builds a foundation for freshwater environmental studies on mechanistic and biochemical responses to microplastics.
  • Amphibian (Rana nigromaculata)exposed to cyproconazole: changes in growth
           index, behavioral endpoints, antioxidant biomarkers, thyroid and gonad
    • Abstract: Publication date: Available online 3 January 2019Source: Aquatic ToxicologyAuthor(s): Wenjun Zhang, Li Chen, Yuanyuan Xu, Yue Deng, Luyao Zhang, Yinan Qin, Zikang Wang, Rui Liu, Zhiqiang Zhou, Jinling Diao Pesticides are a major cause of reduction in the global population of amphibians. This study investigates the effect of varying concentrations of cyproconazole (1 and 10 mg/L) on Rana nigromaculata during a chronic 90days exposure period. High levels of cyproconazole (10 mg/L) induced declined body weight, short snout–vent length, slow metamorphic development and abnormal behavioral endpoints in R. nigromaculata tadpoles. Tadpoles exposed to 10 mg/L did not survive beyond 42days. Abnormal behaviors were observed more frequently with exposure to the highest concentration of cyproconazole.Compared with controls, the concentrations of dismutase (SOD), catalase (CAT) and glutathione (GSH) were significantly increased in tadpoles exposed to 1 mg/L cyproconazole. However, when the concentration of cyproconazole increased to 10 mg/L, concentrations of SOD, GSH and CAT activity began to decline. In addition, thyroid and gonad development were also affected at the gene and hormone level, with varied effects observed with different exposure levels and days. Exposure to cyproconazole at the lower level of 1 mg/L induced damage to histological structures of the thyroid gland. Stereoselective tissue distribution and bioaccumulation of cyproconazole was observed in tadpoles. The ranked order of bioaccumulation was: enantiomer -4 > 3> 2 > 1, with the level of cyproconazole highest in the gut. These findings reflect the toxicity of cyproconazole to R. nigromaculata and further our understanding of the effects of pesticide exposure on global amphibian population declines.Graphical abstractGraphical abstract for this article
  • Fenobucarb induces heart failure and cerebral hemorrhage in zebrafish
    • Abstract: Publication date: Available online 3 January 2019Source: Aquatic ToxicologyAuthor(s): Xiao-Yu Zhu, Bo Xia, Yu-Ying Wu, Hua Yang, Chun-Qi Li, Ping LiABSTRACTThe potential risk and toxic mechanisms of fenobucarb (2-sec-butylphenyl methylcarbamate, BPMC) to animals and humans have not been fully elucidated. In this study, zebrafish embryos were exposed to various concentrations of BPMC from 48 hpf (hour post fertilization, hpf) to 72 hpf. We found that BPMC induced severe heart failure with bradycardia, reduced heart contractions, cardiac output and blood flow dynamics;and myocardial apoptosis. BPMC also induced cerebral hemorrhages and blood erythrocyte reduction in a dose-dependent manner. Also observed were increased ROS production and capase 9 and 3/7 activation. The mRNA levels of the ATPase-related gene (atp2a1l), calcium channel-related gene (cacna1ab), sodium channel-related gene (scn5Lab), potassium channel-related gene (kcnq1), the regulatory gene (tnnc1a) for cardiac troponin C, and several apoptosis-related genes were significantly downregulated in zebrafish following BPMC exposure. These results suggest that exposure to BPMC is a possible risk factor to cardiovascular and cerebrovascular systems in animals.
  • Genotoxic and Cytotoxic Effects of 50 Hz 1 mT Electromagnetic Field on
           Larval Rainbow Trout (Oncorhynchus mykiss), Baltic Clam (Limecola
           balthica) and Common Ragworm (Hediste diversicolor)
    • Abstract: Publication date: Available online 2 January 2019Source: Aquatic ToxicologyAuthor(s): Milda Stankevičiūtė, Magdalena Jakubowska, Janina Pažusienė, Tomas Makaras, Zbigniew Otremba, Barbara Urban-Malinga, Dariusz P. Fey, Martyna Greszkiewicz, Gintarė Sauliutė, Janina Baršienė, Eugeniusz Andrulewicz The aim of this research was to assess genotoxicity and cytotoxicity responses in aquatic animals exposed to 50 Hz 1 m T electromagnetic field (EMF). Rainbow trout (Oncorhynchus mykiss) at early stages of development were exposed to EMF for 40 days, whereas marine benthic invertebrates – the common ragworm Hediste diversicolor and the Baltic clam Limecola balthica – for 12 days. To define genotoxicity and cytotoxicity responses in selected animals, assays of nuclear abnormalities in peripheral blood erythrocytes of O. mykiss, coelomocytes of H. diversicolor and gill cells of L. balthica were performed. Induction of formation of micronuclei (MN), nuclear buds (NB), nuclear buds on filament cells (NBf) and cells with blebbed nuclei (BL) were assessed as genotoxicity endpoints, and 8-shaped nuclei, fragmented (Fr), apoptotic (Ap) and binucleated (BN) cells as cytotoxicity endpoints. Exposure to EMF affected all studied species but with varying degrees. The strongest responses to EMF treatment were elicited in L. balthica, in which six out of the total eight analyzed geno- and cytotoxicity endpoints were significantly elevated. Significantly induced frequencies of MN were detected in O. mykiss and H. diversicolor cells, NBf and BL only in gill cells of L. balthica, and NB in analyzed tissues of all the test species. As cytotoxicity endpoints, a significant elevation in frequencies of cells with 8-shaped nuclei was found in O. mykiss and L. balthica, while Ap and BN was observed only in L. balthica. EMF exposure did not induce any significant cytotoxic activity in H. diversicolor coelomocytes. The present study is the first to reveal the genotoxic and cytotoxic activity of 1 m T EMF in aquatic animals, and, consequently, the first one to report the adverse effect of this factor on common marine invertebrates and early life stages of fish.
  • Examining multi- and transgenerational behavioral and molecular
           alterations resulting from parental exposure to an environmental PCB and
           PBDE mixture
    • Abstract: Publication date: Available online 28 December 2018Source: Aquatic ToxicologyAuthor(s): Sébastien Alfonso, Mélanie Blanc, Lucette Joassard, Steffen H. Keiter, Catherine Munschy, Véronique Loizeau, Marie-Laure Bégout, Xavier Cousin Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) are persistent organic pollutants extensively used during the 20th century and still present in aquatic environments despite their ban. Effects of exposure to these compounds over generations are poorly documented. Therefore, our aims were to characterize behavioral responses and underlying molecular mechanisms in zebrafish exposed to an environmentally relevant mixture of PCBs and PBDEs as well as in four unexposed offspring generations. Zebrafish (F0) were chronically exposed from the first meal onward to a diet spiked with a mixture containing 22 PCB and 7 PBDE congeners in proportions and concentrations reflecting environmental situations (ΣPCBs = 1991 and ΣPBDEs = 411 ng/g). Four offspring generations (F1 to F4) were obtained from this F0 and were not further exposed. Behavior was assessed at both larval and adult stages. Mechanisms related to behavioral defects (habenula maturation and c-fos transcription) and methylation (dnmts transcription) were monitored in larvae. Exposed adult F0 as well as F1 and F3 adults displayed no behavioral change while F2 expressed anxiety-like behavior. Larval behavior was also disrupted, i.e. hyperactive after light to dark transition in F1 or hypoactive in F2, F3 and F4. Behavioral disruptions may be related to defect in habenula maturation (observed in F1) and change in c-fos transcription (observed in F1 and F2). Transcription of the gene encoding DNA methyltransferase (dnmt3ba) was also modified in all generations. Our results lead us to hypothesize that chronic dietary exposure to an environmentally relevant mixture of PCB and PBDE triggers multigenerational and transgenerational molecular and behavioral disruptions in a vertebrate model.
  • Effects of a common pharmaceutical, atorvastatin, on energy metabolism and
           detoxification mechanisms of a marine bivalve Mytilus edulis
    • Abstract: Publication date: Available online 28 December 2018Source: Aquatic ToxicologyAuthor(s): Halina Falfushynska, Eugene P. Sokolov, Fouzia Haider, Christina Oppermann, Udo Kragl, Wolfgang Ruth, Marius Stock, Sabrina Glufke, Eileen J. Winkel, Inna M. Sokolova
  • Effects of polluted seawater on oxidative stress, mortality, and
           reproductive parameters in the marine rotifer Brachionus koreanus and the
           marine copepod Tigriopus japonicus
    • Abstract: Publication date: Available online 24 December 2018Source: Aquatic ToxicologyAuthor(s): Chang-Bum Jeong, Hye-Min Kang, Min-Chul Lee, Eunjin Byeon, Heum Gi Park, Jae-Seong Lee Although many efforts have been made to understand the toxic effects of metals in aquatic invertebrates, there are limited data regarding metal toxicity in natural ecosystems, as most previous studies were conducted under controlled laboratory conditions. To address this data gap, we analyzed toxic effects and molecular responses in the marine rotifer Brachionus koreanus and the marine copepod Tigriopus japonicus following in vivo exposure to a seawater sample collected from a polluted region in South Korea. Inductively coupled plasma-mass spectrometry (ICP-MS) analysis of the field seawater sample found a variety of metals. Exposure to several dilutions of the field seawater sample impacted several endpoints in both species, including mortality and reproduction. Interestingly, the rotifer and copepod test species exhibited different patterns of effects on reactive oxygen species (ROS) and antioxidant enzymatic activities, suggesting that different regulatory mechanisms may be activated in the two species in response to exposure to toxic chemicals. Our study helps to better understand the defense mechanisms activated in aquatic invertebrates in response to metal-induced oxidative stress induced by contaminated seawater.
  • New insight into the toxic effects of chloramphenicol and roxithromycin to
           algae using FTIR spectroscopy
    • Abstract: Publication date: February 2019Source: Aquatic Toxicology, Volume 207Author(s): Qian Xiong, Li-Xin Hu, You-Sheng Liu, Tuan-Tuan Wang, Guang-Guo Ying Antibiotics have been frequently detected in the aquatic environment, and they may affect aquatic organisms such as algae. Here we investigated toxicity of chloramphenicol (CAP) and roxithromycin (ROX) on four species of green algae (Pseudokirchneriella subcapitata, Scenedesmus quadricauda, Scenedesmus obliquus, and Scenedesmus acuminatus) at biochemical level by Fourier transform infrared spectroscopy (FTIR). The results revealed that both CAP and ROX had negative effects on algal growth and caused alterations of biochemical components. The toxic effects varied among the four algal species and S. acuminatus was found to be less sensitive than the other three species to the antibiotics. Even with similar mechanism of action, ROX displayed more adverse effects to algae than CAP. Both antibiotics could affect algae by inhibiting fatty acid synthesis and promoting protein and DNA aggregation, thus leading to accumulation of lipid peroxidation products, increment of the loose β-sheet structure protein and transformation of B-DNA to Z-DNA. The findings from this study revealed the toxic mechanism of antibiotics to algae at the biochemical level.Graphical abstractGraphical abstract for this article
  • Physiological responses of eelgrass (Zostera marina) to ambient stresses
           such as herbicide, insufficient light, and high water temperature
    • Abstract: Publication date: Available online 23 December 2018Source: Aquatic ToxicologyAuthor(s): Kazuhiko Mochida, Takeshi Hano, Toshimitsu Onduka, Katsutoshi Ito, Goro YoshidaABSTRACTThis study aimed to elucidate the biological responses of eelgrass (Zostera marina) to artificially induced stresses such as herbicide (Irgarol 1051, Irg) exposure, insufficient light, and high water temperature (27 ± 1.0 °C) by evaluating growth inhibition, photosynthetic activity, and metabolomic profiles. After 14 days, all treatments inhibited growth, but photosynthetic activity was only reduced in the Irg-exposed group. In the Irg-exposed and insufficient light groups, the metabolomic profiles were characterized by decreased levels of sugar (sucrose) and increased levels of amino acids such as glutamine, glycine, and leucine. Biochemical and ultrastructural analyses revealed that the loss of sugar-derived metabolic energy was compensated for by energy generated during autophagic protein degradation. Furthermore, the level of myo-inositol, which has various biological roles and participates in several cellular processes such as cell wall synthesis, stress response, and mineral nutrient storage, was markedly increased in the Irg-exposed and insufficient light groups. A combination of metabolomic analysis with other analyses such as measurement of photosynthetic activity might further elucidate the response of eelgrass to ambient stresses in the natural environment.
  • Control of Invasive Sea Lampreys Using the Piscicides TFM and Niclosamide:
           Toxicology, Successes & Future Prospects
    • Abstract: Publication date: Available online 21 December 2018Source: Aquatic ToxicologyAuthor(s): Michael P. Wilkie, Terrance D. Hubert, Michael A. Boogaard, Oana Birceanu The invasion of the Laurentian Great Lakes of North America by sea lampreys (Petromyzon marinus) in the early 20th century contributed to the depletion of commercial, recreational and culturally important fish populations, devastating the economies of communities that relied on the fishery. Sea lamprey populations were subsequently controlled using an aggressive integrated pest-management program which employed barriers and traps to prevent sea lamprey from migrating to their spawning grounds and the use of the piscicides (lampricides) 3-trifluoromethyl-4-nitrophenol (TFM) and niclosamide to eliminate larval sea lampreys from their nursery streams. Although sea lampreys have not been eradicated from the Great Lakes, populations have been suppressed to less than 10 % of their peak numbers in the mid-1900s. The ongoing use of lampricides provides the foundation for sea lamprey control in the Great Lakes, one of the most successful invasive species control programs in the world. Yet, significant gaps remain in our understanding of how lampricides are taken-up and handled by sea lampreys, how lampricides exert their toxic effects, and how they adversely affect non-target invertebrate and vertebrates species. In this review we examine what has been learned about the uptake, handling and elimination, and the mode of TFM and niclosamide toxicity in lampreys, and in non-target invertebrates and vertebrates, particularly in the last 10 years. It is now clear that the mode of TFM toxicity is the same in non-target fishes and lampreys, in which TFM interferes with oxidative phosphorylation by the mitochondria leading to decreased ATP production. Vulnerability to TFM is related to abiotic factors such as water pH and alkalinity, which we propose changes the relative amounts of the bioavailable un-ionized form of TFM in the gill microenvironment. Niclosamide, which is also a molluscicide used to control snails in areas prone to schistosomiasis infections of humans, likely works in a similar manner, but less is known about other aspects of its toxicology. The effects of TFM include reductions in energy stores, particularly glycogen and high energy phosphagens. However, non-target fishes readily recover from sub-lethal TFM exposure as demonstrated by the rapid restoration of energy stores and clearance of TFM. Although both TFM and niclosamide are non-persistent in the environment and critical for sea lamprey control, increasing public and institutional concerns about pesticides in the environment makes it imperative to explore other means of sea lamprey control. Accordingly, we also address possible “next-generation” strategies of sea lamprey control including genetic tools such as RNA interference and CRISPR-Cas9 to impair critical physiological processes (e.g. reproduction, digestion, metamorphosis) in lamprey, and the use of green chemistry to develop more environmentally benign chemical methods of sea lamprey control.
  • The role of delta-1-pyrroline-5-carboxylate dehydrogenase (P5CDh) in the
           Pacific white shrimp (Litopenaeus vannamei) during biotic and abiotic
    • Abstract: Publication date: Available online 19 December 2018Source: Aquatic ToxicologyAuthor(s): QingJian Liang, XuJian Wu, Pan Yang, JinRong Kong, Wei Wei, Xueli Qiao, Yuan Liu, Weina Wang Proline (Pro) metabolism is intimately associated with stress adaptation. The catabolism of Pro includes two dehydrogenation reactions catalyzed by proline dehydrogenase (ProDH) and Δ1-pyrroline-5-carboxylate dehydrogenase (P5CDh). P5CDh is a mitochondrial matrix NAD+-dependent dehydrogenase that is critical in preventing P5C-Pro intensive cycling and avoiding ROS production from electron run-off. Little is known about the roles of P5CDh in invertebrates, however. We cloned the P5CDh sequence in the Pacific white shrimp, Litopenaeus vannamei, and found that LvP5CDh is expressed predominantly in pleopod, hepatopancreas and gill. Subcellular localization analysis revealed that LvP5CDh protein was mainly found in the cytoplasm. In addition, overexpressing LvP5CDh in cells reduced ROS formation and inhibited apoptosis induced by LC50 Cd2+. Shrimp were exposed to various stress factors including infection with Vibrio alginolyticus, (½ LC50 and LC50) Cd2+, acid (pH 5.6) and alkali stress (pH 9.3). Both biotic and abiotic stress resulted in increased LvP5CDh expression and Pro accumulation; V. alginolyticus infection, pH 9.3 and LC50 Cd2+ stress apparently stimulated the Glu pathway of Pro synthesis, while pH 5.6 and ½ LC50 Cd2+ stress promoted the Orn pathway of Pro synthesis. Silencing of Lvp53 in shrimp attenuated LvP5CDh expression during Cd2+ stress, but had no effect on LvP5CDh mRNA levels if no Cd2+ stress was imposed. Our study contributes to the functional characterization of LvP5CDh in biotic and abiotic stress and reveals it to protect against ROS generation, damage to the cell, including the mitochondria, and apoptosis. Thus, LvP5CDh plays a critical role in immune defense and antioxidant responses.
  • Influence of differently functionalized polystyrene microplastics on the
           toxic effects of P25 TiO2 NPs towards marine algae Chlorella sp
    • Abstract: Publication date: Available online 18 December 2018Source: Aquatic ToxicologyAuthor(s): Vignesh Thiagarajan, V. Iswarya, Abraham Julian P., R. Seenivasan, N. Chandrasekaran, Amitava Mukherjee Increased utilization of titanium dioxide nanoparticles (TiO2 NPs) for commercial as well as industrial purposes resulted in the accumulation of nanoparticles in the marine system. Microplastics being an emerging secondary pollutant in the marine ecosystem have an impact on the toxic effects of TiO2 NPs which has not been evaluated up to date. So it is important to assess the toxic effects of both these pollutants on the marine environment. The present study examines the impact of differently functionalized microplastics on the toxic effects of P25 TiO2 NPs to marine algae Chlorella sp. The tendency of nanoparticles to undergo aggregation in artificial seawater was observed with increase in time. The median effective concentration for TiO2 NPs was found to be 81 µM which indicates higher toxic effects of NPs toward algae. In contrast, microplastics irrespective of their difference in functionalization had minimal toxic effect of about 15% at their higher concentration tested, 1000 mg L-1. Plain and aminated polystyrene microplastics enhanced the TiO2 NPs toxicity which was further validated with oxidative stress determination studies like reactive oxygen species and lipid peroxidation assays. Negatively charged carboxylated polystyrene microplastics decreased the TiO2 NPs toxicity with possible hetero-aggregation between TiO2 NPs and microplastics in the system. The toxicity data obtained for the mixture was further corroborated with Abbott’s mathematical model.
  • CuZnSOD and MnSOD from freshwater planarian Dugesia japonica: cDNA
           cloning, mRNA expression and enzyme activity in response to environmental
    • Abstract: Publication date: Available online 18 December 2018Source: Aquatic ToxicologyAuthor(s): He-Cai Zhang, Ke-Xue Ma, Yu-Juan Yang, Chang-Ying Shi, Guang-Wen Chen, De-Zeng Liu As an important antioxidant enzyme, the superoxide dismutase (SOD) can protect aerobic organisms from oxidative damage through catalyzing the dismutation of superoxide into hydrogen peroxide and oxygen. The SODs have been cloned in some species and their dynamic expression or enzymatic activity in response to environmental stressors were investigated. In the current study, the full-length cDNA of two SODs from freshwater planarian Dugesia japonica were firstly cloned (named as DjCuZnSOD and DjMnSOD, respectively). The complete cDNA of DjCuZnSOD consists of 661 nucleotides encoding 186 amino acids while the 765 bp DjMnSOD encodes a polypeptide of 226 residues. Sequence analysis and multiple alignment showed that DjCuZnSOD possesses two CuZnSOD family signature motifs and an N-terminal signal peptide suggesting it is an extracellular secretory protein. DjMnSOD possesses the MnSOD family signature sequence and is predicted to be located in mitochondrion with a mitochondrial targeting sequence. Phylogenetic analysis based on CuZnSOD and MnSOD orthologs from representative species further verified that DjCuZnSOD is an extracellular CuZnSOD while DjMnSOD is a mitochondrial MnSOD. For the purpose of studying their potential role against environmental pollutants, D. japonica were exposed to glyphosate or 1-decyl-3-methylimidazolium bromide ([C10mim]Br), and the mRNA expression levels of DjCuZnSOD and DjMnSOD along with total SOD activity were measured. The results showed that DjCuZnSOD exhibited more sensitive expression profiles in response to environmental pollutants in contrast with DjMnSOD, and the total SOD activity in response to both pollutants was more related to the expression level of DjCuZnSOD than to DjMnSOD, indicating that the mRNA expression of CuZnSOD would be a more sensitive biomarker than MnSOD in monitoring the pollution of aquatic environment and CuZnSOD might play more important role than MnSOD in eliminating superoxide anions caused by pollutants in D. japonica.
  • Environmentally relevant concentrations of bifenthrin affects the
           expression of estrogen and glucocorticoid receptors in brains of female
           western mosquitofish
    • Abstract: Publication date: Available online 17 December 2018Source: Aquatic ToxicologyAuthor(s): Isaac Ligocki In recent decades, pyrethroid pesticides have been deemed a safer alternative to previously used pesticides. While some evidence supports this assumption in mammals and birds, exposure to certain pyrethroids can affect concentrations of hormones vital to reproduction in fish. Thus, we hypothesized that pyrethroid exposure impacts fish reproductive behavior and the expression of genes associated with reproduction. We tested our hypothesis by examining effects of the widely used pyrethroid pesticide, bifenthrin, on the reproductive behaviors of the broadly distributed livebearing western mosquitofish, Gambusia affinis. We exposed sexually mature female fish to one of five environmentally relevant concentrations of bifenthrin and conducted behavioral assays to assess reproductive, social, and space use behaviors before and after exposure. We did not detect changes in behaviors measured in response to bifenthrin. However, exposure was associated with increased expression of an estrogen receptor gene (ER-α) and glucocorticoid receptor (GR) in brain tissue at bifenthrin concentrations at concentrations of 5.90 and 24.82 ng/L, and 5.90 and 12.21 ng/L, respectively. Our study supports the perspective that the use of multiple endpoints through integrative approaches is essential for understanding the cumulative impact of pollutants. Integrating physiological, morphological, and behavioral investigations of nonlethal concentrations of pollutants like bifenthrin may heighten our potential to predict their impact on individuals, populations, and communities.
  • Bisphenol A disturbed the lipid metabolism mediated by sterol regulatory
           element binding protein 1 in rare minnow Gobiocypris rarus
    • Abstract: Publication date: Available online 14 December 2018Source: Aquatic ToxicologyAuthor(s): Yongjing Guan, Ting Zhang, Jiafa He, Jia Jia, Long Zhu, Zaizhao Wang Bisphenol A (BPA), a representative endocrine disrupting compound, exists ubiquitously in the aquatic environment. Several studies on fish have validated the role of BPA in the lipid metabolism. However, the action mechanisms of BPA on lipid metabolism have been little studied. To clarify how BPA regulates lipid metabolism, Gobiocypris rarus were exposed to 15 μg/L BPA for 3 and 6 weeks. Results showed that BPA altered lipid content by regulating some metabolism-related genes. The BPA’s inhibiting effect on fatty acid β-oxidation might be stronger than on lipid synthesis. BPA disturbed the expression of acaca (acetyl-CoA carboxylase), fasn (fatty acid synthase) and cpt1α (carnitine palmitoyltransferase 1α) by altering the sterol regulatory element binding protein 1 (SREBP-1) binding to their sterol regulatory elements (SREs). Our result also revealed that DNA methylation in the 5’ flanking regions of cpt1α could perturb the SREBP-1 binding adjacent to its SRE in females under BPA exposure. Besides, BPA exposure led to gender-specific effect on fatty acid β-oxidation in G. rarus. This will contribute to our understanding of the regulation mechanisms of BPA on lipid metabolism in fish.
  • The synergistic potential of azole fungicides does not directly correlate
           to the inhibition of cytochrome P450 activity in aquatic invertebrates
    • Abstract: Publication date: Available online 14 December 2018Source: Aquatic ToxicologyAuthor(s): Michele Gottardi, Nina Cedergreen The ability of azole fungicides to inhibit cytochrome P450 dependent metabolism is proposed to be the main mechanism for their synergizing effect on pyrethroid insecticide toxicity in aquatic invertebrates. This study investigates the correlation between inhibition strength and synergistic potential of azole fungicides in the crustacean Daphnia magna and the insect larvae Chironomus riparius. Inhibition strength was measured in vivo toward the cytochrome P450 catalysed conversion of 7-ethoxycoumarin to 7-hydroxycoumarin (ECOD). Synergistic potentials were determined as the ratio between predicted and observed toxicity of mixtures based on the model of concentration addition (CA) and independent action (IA). Azoles (n = 9 - 11) enhanced the toxicity of α-cypermethrin in D. magna (Synergy ratios CA: 0.8 – 16; IA: 1.1 – 22) and inhibited cytochrome P450 activity by different degrees (IC50: 0.0023 – 36 μM for D. magna and 0.08 – 24 μM for C. riparius). Inhibition strengths were strongly correlated in the two organisms (r: 0.937 p: 0.019 for triazoles and r: 0.903 p: 0.097 for imidazoles). Lipophilicity governed the inhibition strength of triazoles in both species (r> 0.9, p 
  • Variation in Metal Tolerance Associated with Population Exposure History
           in Southern Toads (Anaxyrus terrestris)
    • Abstract: Publication date: Available online 13 December 2018Source: Aquatic ToxicologyAuthor(s): R. Wesley Flynn, Cara N. Love, Austin Coleman, Stacey L. Lance Human activities have radically shaped the global landscape, affecting the structure and function of ecosystems. Habitat loss is one of the most visible changes to the landscape and a primary driver of species declines; however, anthropogenic environmental contamination also threatens population persistence, but is not as readily observed. Aquatic organisms are especially susceptible to chemical perturbations, which can negatively impact survival and fitness related traits. Some populations have evolved tolerance to chemical stressors, which could mitigate the consequences associated with contamination. Amphibians are experiencing global declines due to multiple stressors and are particularly at risk to aquatic chemical stressors due to their permeable skin and reliance on wetlands for reproduction and larval development. However, amphibians also have substantial plasticity in response to environmental variation. We designed our study to examine whether tolerance to heavy metals is greater in Southern toad (Anaxyrus terrestris) larvae from wetlands with a history of contamination. Considering many of the most common trace elements elicit acute toxicity by disrupting osmotic- and ionic-regulation, we hypothesized that alterations to these aspects of physiology resulting from multigenerational exposure to trace element mixtures would be the most likely routes by which tolerance would evolve. We used copper (Cu) as a proxy for heavy metal exposure because it is a widely distributed aquatic stressor known to cause osmotic stress that can also cause mortality at levels commonly encountered in the environment. We found considerable within and among population variation in Cu tolerance, as measured by time to death. Larvae from populations living in sites contaminated with mixtures of heavy metals associated with coal fly ash were no more tolerant to Cu than those from reference sites. However, larvae from a population inhabiting a constructed wetland complex with high Cu levels were significantly more tolerant; having half the risk of mortality as reference animals. This wetland complex was created < 20 years ago, thus if elevated Cu tolerance in this population is due to selection in the aquatic habitat, such adaptation may occur rapidly (i.e. ~10 generation). Our results provide evidence that amphibians may be able to evolve tolerance in response to trace element contamination, though such tolerance may be specific to the combination of contaminants present.
  • Predicting cadmium toxicity with the kinetics of phytochelatin induction
           in a marine diatom
    • Abstract: Publication date: Available online 11 December 2018Source: Aquatic ToxicologyAuthor(s): Yun Wu, Yue Yuan, Hezhong Yuan, Wei Zhang, Li Zhang Phytochelatin (PC) synthesis is thought to be a rapid and specific response to metal exposure in marine phytoplankton, but its potential as a predictor of metal toxicity is far from conclusive. Thus this research examines the bioaccumulation, PC induction, and toxicity of Cadmium (Cd) in Thalassiosira weissflogii, a coastal diatom under varying nutrient conditions. Nitrogen limitation strongly inhibited Cd uptake and PC induction at the same [Cd2+] level, and increased metal sensitivity. Conversely, phosphorus limitation had little influence on Cd accumulation and PC induction, yet also enhanced metal effect on growth. Differential growth inhibitions were correlated with [Cd2+], intracellular Cd concentration, PC concentration, the kinetics of Cd uptake and PC induction, respectively. It was found that stronger interrelations existed between kinetic rates (both Cd uptake and PC synthesis) and Cd sensitivity than between the static concentrations (Cd and PC) and growth inhibition. Moreover, according to the calculated median inhibition concentration (IC50), median effective uptake rate of Cd, as well as median effective induction rate of PCs, the latter two showed the smallest variation when nutrients were varied (1.4–1.9 fold). Our study set out the first step toward considering the use of PC synthesis kinetics to predict metal toxicity for phytoplankton.
  • Transcriptional and physiological responses of Dunaliella salina to
           cadmium reveals time-dependent turnover of ribosome, photosystem, and
           ROS-scavenging pathways
    • Abstract: Publication date: Available online 7 December 2018Source: Aquatic ToxicologyAuthor(s): Qing-Ling Zhu, Sai-Nan Guo, Fang Wen, Xiao-Lin Zhang, Cheng-Cheng Wang, Lan-Fang Si, Jia-Lang Zheng, Jianhua Liu Effects on short-term (6 h) and long-term (96 h) exposure to cadmium (Cd) at 0.1, 0.5 and 2.5 mg/L in microalga Dunaliella salina were assessed using both physiological end points and gene expression analysis. Different physiological responses between the short-term and long-term exposures were observed. Upon 6 h after Cd exposure, lipid peroxidation and cell ultrastructure remained unchanged, while contents of chlorophyll a, chlorophyll b, carotenoids were increased at 0.5 and 2.5 mg/L Cd. Contrarily, 96 h after Cd exposure, lipid peroxidation levels were increased, while pigments content was decreased, and damaged cell ultrastructure was apparent at 2.5 mg/L Cd. Activities of antioxidant enzymes (APX, SOD, GST, GPX, and GR) changed differently both at 6 h and 96 h after Cd exposure. Upon 6 h after Cd exposure, SOD and GST activity increased at all three doses, GR and GPX activity increased at 0.5 mg/L Cd while APX activity increased at 0.1 mg/L Cd. Contrarily, 96 h after Cd exposure, activities of all the antioxidant enzymes increased both at 0.1 and 0.5 mg/L Cd; but there was a decrease in SOD and GR activity in D. salina exposed to 2.5 mg/L Cd. RNA-seq and qRT-PCR analyses indicated that genes involved in ROS-scavenge, photosystem, and ribosome functions were differentially expressed. The most significantly enriched function was the ribosome, in which more than 30 ribosome genes were up-regulated at 6 h but down-regulated at 96 h after Cd exposure at 2.5 mg/L. Our study indicated for the first time that genes encoding ribosomal proteins are the primary target for Cd in microalgae, which allowed gaining new insights into temporal dynamics of toxicity and adaptive response pathways in microalgae exposed to metals.
  • Oxidative damage induced by copper in testis of the red swamp crayfish
           Procambarus clarkii and its underlying mechanisms
    • Abstract: Publication date: Available online 6 December 2018Source: Aquatic ToxicologyAuthor(s): Dan Zhao, Xiaona Zhang, Xuefu Li, Shaoguo Ru, Yanwen Wang, Jinbin Yin, Dasheng Liu Copper (Cu) is one of the most widespread environmental pollutants and is known to exert multiple toxic effects including reproductive toxicity. In this study, we investigated the toxic effect of Cu on reproduction of the red swamp crayfish (Procambarus clarkii), an economic crustacean species, by exposing adult male crayfish to 0.03 and 3.00 mg/L Cu2+ for 7 days. The results showed that Cu2+ exposure induced oxidative stress accompanied by elevated reactive oxygen species (ROS) and malondialdehyde (MDA) levels in testes, and resulted in decreased sperm quality and abnormal testicular structures with apoptotic germ cells and vacuolisation in Sertoli cells. To reveal the molecular mechanism of Cu2+-induced oxidative damage in crayfish testes, we sequenced, assembled and annotated the transcriptome for crayfish testes, using the Illumina sequencing approach. After the 3.00 mg/L Cu2+ treatment, 6,745 genes with differentially expressed profile were identified, of which many genes were involved in cellular response to ROS based on Gene Ontology enrichment analysis. Further, KEGG analysis demonstrated that genes with up-regulated expression levels significantly enriched in mitochondria oxidative phosphorylation pathway, suggesting disturbed mitochondrial electron transport chain was probably a main source of Cu2+-induced ROS production in testes. This study represented the first use of transcriptome to investigate the toxic effect of Cu2+ on male crayfish reproduction, and the pathways identified underlying Cu2+ toxicity at molecular level provide a novel insight into the reproductive toxicity of Cu in crustaceans.
  • QSAR for baseline toxicity and classification of specific modes of action
           of ionizable organic chemicals in the zebrafish embryo toxicity test
    • Abstract: Publication date: Available online 5 December 2018Source: Aquatic ToxicologyAuthor(s): Nils Klüver, Kai Bittermann, Beate I. Escher The fish embryo toxicity (FET) test with the zebrafish Danio rerio is widely used to assess the acute toxicity of chemicals thereby serving as animal alternative to the acute fish toxicity test. The minimal toxicity of neutral chemicals in the FET can be predicted with a previously published Quantitative Structure-Activity Relationship (QSAR) based on the liposome-water partition coefficient Klipw. Such a QSAR may serve to plan toxicity testing and to evaluate whether an observed effect is caused by a specific mode of action (MoA). The applicability domain of this QSAR was extended to ionizable organic chemicals (IOC) without any modification of slope and intercept simply by replacing the Klipw with the speciation-corrected liposome-water distribution ratio (Dlipw(pH)) as descriptor for the uptake into the embryo. FET LC50 values of IOCs were extracted from an existing FET database and published literature. IOCs were selected that are present concomitantly as neutral and charged, species, i.e., acids with an acidity constant pKa 5. IOCs were grouped according to their putative MoA of acute aquatic toxicity. The toxic ratios (TR) in the FET were derived by of the experimental FET-LC50 in comparison with the baseline toxicity QSAR. Baseline toxicants were confirmed to align well with the FET baseline toxicity QSAR (TR  10) were generally consistent with MoA classification for acute fish toxicity with a few exceptions that were suspected to have had issues with the stability of the pH during testing. One critical aspect for the effect analysis of ionizable chemicals is the pH, since the difference between pH and pKa determines the speciation and thereby the Dlipw(pH).
  • In vivo effects of serotonin and fluoxetine on cardio-ventilatory
           functions in the shore crab Carcinus maenas (L. 1758)
    • Abstract: Publication date: Available online 5 December 2018Source: Aquatic ToxicologyAuthor(s): Alexandrine Robert, Tiphaine Monsinjon, Romain Péden, Virginie Rasoamampianina, Jean-Claude Le Mével, Thomas Knigge Serotonin (5-HT) takes a key position in regulating vital functions, such as cardio-ventilatory activity, locomotion and behaviour. Selective serotonin reuptake inhibitors (SSRIs) modulate the serotonergic system and thus affect these functions. Rhythmic behaviours, such as cardio-ventilatory activity, are controlled by central pattern generators, which in turn are regulated by 5-HT. In crustaceans, 5-HT also regulates the synthesis and secretion of crustacean hyperglycaemic hormone, a pleiotropic hormone involved in the mobilisation and release of glucose into the haemolymph, thus stimulating the animal’s activity. As a matter of consequence, SSRIs may affect cardio-ventilatory activity. In order to examine how the SSRIs affect fundamental physiological parameters based on rhythmic behaviour in decapods, cardio-respiratory activity in the shore crab Carcinus maenas was assessed after pericardial injection of a single dose of either 0.5 μM, 0.75 μM or 1 µM fluoxetine, respectively. Simultaneous recordings of heart and scaphognathite movements in both brachial chambers were conducted by measuring impedance changes in the respective body compartments. Injection of 5-HT had an immediate effect on cardio-ventilatory activities and strongly upregulated both cardiac and ventilatory activities. Fluoxetine showed similar effects, entailing moderate tachycardia and increased ventilation rates. Compared to 5-HT, these effects were delayed in time and much less pronounced. Metabolism of fluoxetine into the active compound nor-fluoxetine might account for the delayed action, whereas compensatory regulation of cardio-ventilatory frequencies and amplitudes are likely to explain the attenuation of the responses compared to the strong and immediate increase by 5-HT. Overall, the results suggest increased 5-HT levels in invertebrates following fluoxetine exposure, which are able to disturb physiological functions regulated by 5-HT, such as cardiac and respiratory activity.
  • Excess copper promotes photoinhibition and modulates the expression of
           antioxidant-related genes in Zostera muelleri
    • Abstract: Publication date: Available online 5 December 2018Source: Aquatic ToxicologyAuthor(s): Pimchanok Buapet, Nasim Shah Mohammadi, Mathieu Pernice, Manoj Kumar, Unnikrishnan Kuzhiumparambil, Peter J. Ralph Copper (Cu) is an essential micronutrient for plants and as such is vital to many metabolic processes. Nevertheless, when present at elevated concentrations, Cu can exert toxic effects on plants by disrupting protein functions and promoting oxidative stress. Due to their proximity to the urbanised estuaries, seagrasses are vulnerable to chemical contamination via industrial runoff, waste discharges and leachates. Zostera muelleri is a common seagrass species that forms habitats in the intertidal areas along the temperate coast of Australia. Previous studies have shown the detrimental effects of Cu exposure on photosynthetic efficiency of Z. muelleri. The present study focuses on the impacts of sublethal Cu exposure on the physiological and molecular responses. By means of a single addition, plants were exposed to 250 and 500 µg Cu L-1 (corresponding to 3.9 and 7.8 µM, respectively) as well as uncontaminated artificial seawater (control) for 7 days. Chlorophyll fluorescence parameters, measured as the effective quantum yield ( φ PSII), the maximum quantum yield (Fv/Fm) and non-photochemical quenching (NPQ) were assessed daily, while Cu accumulation in leaf tissue, total reactive oxygen species (ROS) and the expression of genes involved in antioxidant activities and trace metal binding were determined after 1, 3 and 7 days of exposure. Z. muelleri accumulated Cu in the leaf tissue in a concentration-dependent manner and the bioaccumulation was saturated by day 3. Cu exposure resulted in an acute suppression of φ PSII and Fv/Fm. These two parameters also showed a concentration- and time-dependent decline. NPQ increased sharply during the first few days before subsequently decreasing towards the end of the experiment. Cu accumulation induced oxidative stress in Z. muelleri as an elevated level of ROS was detected on day 7. Lower Cu concentration promoted an up-regulation of genes encoding Cu/Zn-superoxide dismutase (Cu/Zn-sod), ascorbate peroxidase (apx), catalase (cat) and glutathione peroxidase (gpx), whereas no significant change was detected with higher Cu concentration. Exposure to Cu at any concentration failed to induce regulation in the expression level of genes encoding metallothionein type 2 (mt2), metallothionein type 3 (mt3) and cytochrome c oxidase copper chaperone (cox17). It is concluded that chlorophyll fluorescence parameters provide timely probe of the status of photosynthetic machinery under Cu stress. In addition, when exposed to a moderate level of Cu, Z. muelleri mitigates any induced oxidative stress by up-regulating transcripts coding for antioxidant enzymes.
  • Immunotoxic effects of 4-nonylphenol on Clarias gariepinus:
           cytopathological changes in hepatic melanomacrophages
    • Abstract: Publication date: Available online 3 December 2018Source: Aquatic ToxicologyAuthor(s): Alaa El-Din H. Sayed, Mahmoud Abd-Elkareem, Nasser S. Abou Khalil Melanomacrophage centres (MMCs) play a key role in the immune response in fish. They are considered sensitive bio-monitoring structures with roles in the assessment of toxicant impacts. The aim of this study was to examine the potential histopathological effect of 4-nonylphenol (4-NP) on hepatic MMCs in Clarias gariepinus. To achieve this objective, adult male fish were divided randomly and equally into two groups: a control group and a group that was exposed to 4-NP (dissolved in water at a dose of 0.1 mg/L) for 21 days. The 4-NP-intoxicated hepatic MMCs contained numerous necrotic macrophages. Superoxide dismutase 2 was immuno-expressed in the hepatic MMCs in both groups, with no significant difference. Histomorphometric examination revealed that the sizes and numbers of MMCs were dramatically higher in the livers of 4-NP-exposed C. gariepinus than in control fish. Following 4-NP challenge, in the liver, the abundance of lipofuscin and haemosiderin pigments increased, and single-pigmented macrophages, aggregated groups of deformed red blood cells (RBCs) and macrophages were present near blood vessels and hepatic sinusoids. These results reveal that 4-NP exerts immunological effects on hepatic MMCs in C. gariepinus and support the utility of MMCs as a cytological biomarker for aquatic exposure to 4-NP.Graphical abstractGraphical abstract for this article
  • Investigating tissue bioconcentration and the behavioural effects of two
           pharmaceutical pollutants on sea trout (Salmo trutta) in the laboratory
           and field
    • Abstract: Publication date: Available online 1 December 2018Source: Aquatic ToxicologyAuthor(s): Erin S. McCallum, Anna Sundelin, Jerker Fick, Anders Alanärä, Jonatan Klaminder, Gustav Hellström, Tomas Brodin Pharmaceuticals entering aquatic ecosystems via wastewater effluents are of increasing concern for wild animals. Because some pharmaceuticals are designed to modulate human behaviour, measuring the impacts of exposure to pharmaceuticals on fish behaviour has become a valuable endpoint. While laboratory studies have shown that pharmaceuticals can affect fish behaviour, there is a lack of understanding if behaviour is similarly affected in natural environments. Here, we exposed sea trout (Salmo trutta) smolts to two concentrations of two pharmaceutical pollutants often detected in surface waters: temazepam (a benzodiazepine, anxiolytic) or irbesartan (an angiotensin II receptor blocker, anti-hypertensive). We tested the hypothesis that changes to behavioural traits (anxiety and activity) measured in laboratory trials following exposure are predictive of behaviour in the natural environment (downstream migration). Measures of anxiety and activity in the laboratory assay did not vary with temazepam treatment, but temazepam-exposed fish began migrating faster in the field. Activity in the laboratory assay did predict overall migration speed in the field. In contrast to temazepam, we found that irbesartan exposure did not affect behaviour in the laboratory, field, or the relationship between the two endpoints. However, irbesartan was also not readily taken up into fish tissues (i.e. below detection levels in the muscle tissue), while temazepam bioconcentrated (bioconcentration factor 7.68) rapidly (t1/2 < 24 h). Our findings add to a growing literature showing that benzodiazepine pollutants can modulate fish behaviour and that laboratory assays may be less sensitive at detecting the effects of pollutants compared to measuring effects in natural settings. Therefore, we underscore the importance of measuring behavioural effects in the natural environment.
  • Physiological insights into largemouth bass (Micropterus salmoides)
           survival during long-term exposure to high environmental ammonia
    • Abstract: Publication date: Available online 30 November 2018Source: Aquatic ToxicologyAuthor(s): Nathan Egnew, Nilima Renukdas, Yathish Ramena, Amit K. Yadav, Anita M. Kelly, Rebecca T. Lochmann, Amit Kumar Sinha Waterborne ammonia is an environmental pollutant that is toxic to all aquatic animals. However, ammonia induced toxicity as well as compensatory mechanisms to defend against high environmental ammonia (HEA) are not well documented at present for largemouth bass (Micropterus salmoides), a high value fish for culture and sport fisheries in the United States. To provide primary information on the sensitivity of this species to ammonia toxicity, a 96 h-LC50 test was conducted. Thereafter, responses at physiological, ion-regulatory and transcript levels were determined to get insights into the underlying adaptive strategies to ammonia toxicity. For this purpose, fish were progressively exposed to HEA (8.31 mg/L representing 25% of 96 h-LC50) for 3, 7, 14, 21 and 28 days. Temporal effects of HEA on oxygen consumption rate (MO2), ammonia and urea dynamics, plasma ions (Na+, Cl− and K+), branchial Na+/K+-ATPase (NKA) and H+-ATPase activity, muscle water content (MWC), energy store (glycogen, lipid and protein) as well as branchial mRNA expression of Rhesus (Rh) glycoproteins were assessed. Probit analysis showed that 96 h-LC50 of (total) ammonia (as NH4HCO3) at 25 °C and pH 7.8 was 33.24 mg/L. Results from sub-lethal end-points shows that ammonia excretion rate (Jamm) was strongly inhibited after 7 days of HEA, but was unaffected at 3, 14 and 21 days. At 28 days fish were able to increase Jamm efficiently and concurrently, plasma ammonia re-established to the basal level. Urea production was increased as evidenced by a considerable elevation of plasma urea, but urea excretion rate remained unaltered. Expression of Rhcg isoform (Rhcg2) mRNA was up-regulated in parallel with restored or increased Jamm, suggesting its ammonia excreting role in largemouth bass. Exposure to HEA also displayed pronounced augmentations in NKA activity, exemplified by a rise in plasma [Na+]. Furthermore, [K+], [Cl−] and MWC homeostasis were disrupted followed by recovery to the control levels. H+-ATPase activity was elevated but NKA did not appear to function preferentially as a Na+/NH4+-ATPase. From 14 days onwards MO2 was depressed, potentially an attempt towards minimizing catabolism. Glycogen content in liver and muscle were temporarily depleted, whereas a remarkable increment in protein was evident at the last exposure period. Overall, these data suggest that ammonia induced toxicity can disturb several biological processes in largemouth bass, however, it can adapt to the long-term sub-lethal ammonia concentrations by activating various components of ammonia excretory, ion-regulatory and metabolic pathways.
  • The responses of Oncorhynchus mykiss coping with BDE-47 stress via
           PXR-mediated detoxification and Nrf2-mediated antioxidation system
    • Abstract: Publication date: Available online 30 November 2018Source: Aquatic ToxicologyAuthor(s): Chunchen Liu, Boyuan Wang, Bin Zhou, Xiaoyang Jian, Xinxin Zhang, You Wang The low brominated polybrominated diphenyl ether (PBDE) 2,2’,4,4’-tetrabromodiphenyl ether (BDE-47) is ubiquitous in the marine environment. To elucidate the stress response and possible mechanisms underlying BDE-47, the rainbow trout fish Oncorhynchus mykiss were selected and orally fed bait with BDE-47 concentrations of 50 ng/g and 500 ng/g. BDE-47 was found to be mainly accumulated in head kidney and caused lipid peroxidation after prolonged exposure. We studied the detoxification system genes pregnane X receptor (PXR) and downstream genes (cytochrome 3 A, CYP3 A; glutathione S-transferase, GST) and their corresponding enzyme activity and found that the above indicators in the treatment groups increased first and then decreased with time, while the 500 ng/g group showed more significant changes. Further, the antioxidant system gene expression levels of the NF-E2-related factor 2 (Nrf2) and downstream genes (superoxide dismutase, SOD; catalase, CAT) were found significantly up-regulated with concentration and time. The change in the enzyme activity of SOD and CAT showed the same tendency as that of indicators of detoxifying system. The results showed that BDE-47 can accumulated in head kidney and caused activate and fast increase of genes and enzymes of detoxification and antioxidant system in the short-term and then damage the response systems in longer times. After Pearson correlation analysis, the Integrated Biomarker Response (IBR) Index was established with malondialdehyde (MDA) content; PXR, Nrf2, SOD, and CAT gene expression; and CYP3 A, GST, and CAT enzymatic activity, which were significantly related to BDE-47 bioaccumulation (P 
  • Primary green turtle (Chelonia mydas) skin fibroblasts as an in vitro
           model for assessing genotoxicity and oxidative stress
    • Abstract: Publication date: February 2019Source: Aquatic Toxicology, Volume 207Author(s): Kimberly A. Finlayson, Frederic D.L. Leusch, Jason P. van de Merwe Little is known about the effects of contaminants that accumulate in sea turtles. When in vivo exposure studies have ethical and logistical barriers, as is the case with sea turtles, in vitro tools can provide important information on the effects of contaminants. Several in vitro studies have assessed cytotoxicity of contaminants to sea turtles cells, however to gain a more refined mechanistic understanding of the effects of contaminants, sub-lethal effects also require investigation. Considering the complex mixture of contaminants that sea turtles are potentially exposed to, high throughput testing methods are necessary so that a large number of contaminants (and mixtures) can be rapidly tested. This study examined oxidative stress (reactive oxygen species production) and genotoxicity (micronucleus formation) in primary green turtle skin fibroblasts in response to 16 organic and inorganic contaminants found in coastal environments. Significant induction of oxidative stress was found with Cu, Co, Cr, and Hg. Significant effects on genotoxicity were found with Cu, Co, Cr, Hg, Pb and metolachlor. Effect concentrations from the bioassays were used in a simple risk assessment of turtles worldwide using accumulation values from the literature to identify populations at risk. Cu, Co, Cr and Hg were identified as posing the biggest threat to sea turtles. This study demonstrated the validity of using primary turtle cell cultures in the assessment of risk associated with a large number of contaminants using a high-throughput toxicity testing format.
  • Behavioral Consequences of Dietary Exposure to Crude Oil Extracts in the
           Siamese Fighting Fish (Betta splendens)
    • Abstract: Publication date: Available online 29 November 2018Source: Aquatic ToxicologyAuthor(s): Naim M. Bautista, Tanushri Pothini, Kelly Meng, Warren W. Burggren Uptake by fishes of crude oil and its polycyclic aromatic hydrocarbons (PAHs) components occurs via gills, dietary intake, or diffusion through the skin. Dietary exposure to crude oil and its components is environmentally relevant, and induces physiological and morphological disruptions in fish. However, the impacts of crude oil on fish social and reproductive behaviors and thus the possible influences on reproductive success are poorly understood. As a part of their intraspecific interactions, male Siamese fighting fish (Betta splendens) exhibit highly stereotypic behavioral and territorial displays. This makes this species a tractable model for testing crude oil effects on behavior. After 2 weeks of acclimation at 29 °C, male adult betta fish were divided into three groups and fed for 4 weeks with food spiked with water (control), low oil concentrations or high oil concentrations (∑Total PAH concentrations 340, 3960 or 8820 ng/g dw, respectively) to determine subsequent alterations in behavioral displays. Compared with control fish, the aggressive display of “opercular flaring” was significantly increased (P 
  • Altered non-reproductive behavior and feminization caused by developmental
           exposure to 17α-ethinylestradiol persist to adulthood in three-spined
           stickleback (Gasterosteus aculeatus)
    • Abstract: Publication date: Available online 28 November 2018Source: Aquatic ToxicologyAuthor(s): Tove Porseryd, Josefine Larsson, Martin Kellner, Tomas Bollner, Patrik Dinnétz, Inger Porsch Hällström The synthetic estrogen 17α-ethinylestradiol (EE2), ubiquitous in the aquatic environment and commonly detected in sewage effluents, interferes with the endocrine system in multiple ways. Exposure during sensitive windows of development causes persistent effects on fertility, reproductive and non-reproductive behavior in mammals and fish. In the present study, three-spined stickleback (Gasterosteus aculeatus) were exposed to nominal 0 and 20 ng/L EE2 from fertilization to 7 weeks post-hatch. After 8 months of remediation in clean water three non-reproductive behaviors, not previously analyzed in developmentally EE2-exposed progeny of wild-caught fish, were evaluated. Chemical analysis revealed that the nominal 0 and 20 ng/L exposure contained 5 and 30 ng/L EE2, respectively. Therefore, the use of control fish from previous experiments was necessary for comparisons. Fish exposed during development showed significant concentration-dependent reduction in anxiety-like behavior in the scototaxis (light/dark preference) test by means of shorter latency to first entrance to the white compartment, more visits in white, and longer total time in white compared to unexposed fish. In the novel tank test, developmental exposure significantly increased the number of transitions to the upper half of the aquaria. Exposure to EE2 during development did not alter shoal cohesion in the shoaling test compared with unexposed fish but fish exposed to 30 ng/L EE2 had significantly longer latency to leave the shoal and fewer transitions away from the shoal compared to fish exposed to 5 ng/L EE2. Skewed sex ratio with more females, sex reversal in genetic males as well as intersex in males was observed after exposure to 30, but not 5 ng/L EE2. In conclusion, EE2 exposure during development in three-spined stickleback resulted in persistent effects on anxiety-like behaviors. These long-term effects from developmental exposure are likely to be of higher relevance for natural populations than are short-term effects from adult exposure.
  • Investigating a Transcriptomic Approach on Marine Mussel Hemocytes Exposed
           to Carbon Nanofibers: an in vitro/in vivo Comparison
    • Abstract: Publication date: Available online 27 November 2018Source: Aquatic ToxicologyAuthor(s): Andrew Barrick, Nicolas Manier, Pierre Lonchambon, Emmanuel Flahaut, Nisrine Jrad, Catherine Mouneyrac, Amélie Châtel Manufactured nanomaterials are an ideal test case of the precautionary principle due to their novelty and potential environmental release. In the context of regulation, it is difficult to implement for manufactured nanomaterials as current testing paradigms identify risk late into the production process, slowing down innovation and increasing costs. One proposed concept, namely safe(r)-by-design, is to incorporate risk and hazard assessment into the design process of novel manufactured nanomaterials by identifying risks early. When investigating the manufacturing process for nanomaterials, differences between products will be very similar along key physicochemical properties and biological endpoints at the individual level may not be sensitive enough to detect differences whereas lower levels of biological organization may be able to detect these variations. In this sense, the present study used a transcriptomic approach on Mytilus edulis hemocytes following an in vitro and in vivo exposure to three carbon nanofibers created using different production methods. Integrative modeling was used to identify if gene expression could be in linked to physicochemical features. The results suggested that gene expression was more strongly associated with the carbon structure of the nanofibers than chemical purity. With respect to the in vitro/in vivo relationship, results suggested an inverse relationship in how the physicochemical impact gene expression.
  • Physiological and biochemical effect of silver on the aquatic plant Lemna
           gibba L.: evaluation of commercially available product containing
           colloidal silver
    • Abstract: Publication date: Available online 27 November 2018Source: Aquatic ToxicologyAuthor(s): Martina Varga, Janja Horvatić, Lara Barišić, Zdenko Lončarić, Maja Dutour Sikirić, Ina Erceg, Aleksandra Kočić, Ivna Štolfa Čamagajevac This paper aims to evaluate the effects of a product containing colloidal silver in the aquatic environment, using duckweed Lemna gibba as a model plant. Therefore, growth parameters, photosynthetic pigments content and protein content as physiological indices were evaluated. Changes in the content of non-enzymatic antioxidants and activity of several antioxidant enzymes, alongside with the accumulation of hydrogen peroxide and lipid peroxidation end-products were assessed to explore the potential of colloidal silver to induce oxidative stress. The commercially available colloidal silver product contained a primary soluble form of silver. The treatment with colloidal silver resulted in significant physiological and biochemical changes in L. gibba plants and a consequent reduction of growth. Accumulation of silver caused altered nutrient balance in the plants as well as a significant decrease in photosynthetic pigments content and protein concentration. The antioxidative response of L. gibba plants to treatment with colloidal silver was inadequate to protect the plants from oxidative stress caused by metal accumulation. Silver caused concentration-dependent and time-dependent hydrogen peroxide accumulation as well as the elevation of lipid peroxidation levels in L. gibba plants. The use of commercially available products containing colloidal silver, and consequent accumulation of silver, both ionic and nanoparticle form in the environment, represents a potential source of toxicity to primary producers in the aquatic ecosystem.
  • Individual variation in aquatic toxicology: not only unwanted noise
    • Abstract: Publication date: Available online 26 November 2018Source: Aquatic ToxicologyAuthor(s): Mikko Nikinmaa, Katja Anttila The mean value of any parameter and its changes are usually discussed, when ecotoxicological studies are carried out. However, also the variation of any parameter and its changes can be important components of the responses to environmental contamination. Although the homogeneity of variances is commonly tested, testing is done for the use of correct statistical methods, not because of exploring the possibility that variability and its changes could be important components of environmental responses. We evaluated recent aquatic toxicological literature and found that in the majority of articles indicating that homogeneity of variances was tested and giving the result of testing, the assumption of homogeneity was not fulfilled. Further, it was observed that in some studies experimental treatment clearly affected the variability. In this commentary we discuss the reasons for variability: measurement errors, experimental design, genetic heterogeneity and phenotypic plasticity, and conclude that even after accounting for experimental design and genetic makeup significant variability remains. This plasticity may change in environmental responses as suggested by a hypothetical example, and as confirmed by experimental data. As a consequence, the changes of variability can be significant, even when the means do not differ. Because of this, variability and its changes should always be analysed and reported. This will be easy, since the datasets are exactly the same for comparing the variances and means, and as normally variances are tested for homogeneity. It is likely that much new information about the responses of organisms to environmental contamination will be obtained. However, the present journal practices tend to discourage one from concentrating on anything but the mean. In contrast, we think it is imperative that variability is always included as an endpoint in data analysis in the future.
  • Estrogenic activity of multicyclic aromatic hydrocarbons in rainbow trout
           (Oncorhynchus mykiss) in vitro assays
    • Abstract: Publication date: Available online 26 November 2018Source: Aquatic ToxicologyAuthor(s): Richard C. Kolanczyk, Jeffrey S. Denny, Barbara R. Sheedy, Patricia K. Schmieder, Mark A. Tapper A representative group of multicyclic aromatic hydrocarbons (MAHC) which can be further classified as bridged-ring (bridged-MAHC) or fused-ring (fused-MAHC) were examined for their ability to interact with the estrogen receptor of rainbow trout (rtER) in a hepatic cytosolic estrogen receptor competitive binding assay (cyto rtERαβ) and the vitellogenin (Vtg) mRNA gene activation liver slice assay. All five fused-MAHCs; naphthalene (NAFT), fluorene (FE), Fluoranthene (FAT), pyrene (PY), and 9,10-dihydroanthracene (DAC) had no estrogenic activity in the in vitro assays used. Five of the eight bridged-MAHCs; triphenylethylene (3PE), o-terphenyl (OTP), triphenylmethane (TPM), 1,1-diphenylethylene (DPE), and cis-stilbene (CSB) were positive in the rtER-binding assay. The additional three bridged-MAHC’s; trans-stilbene (TSB), tetraphenylethylene (4PE), and 4,4-di-tertbutylphenyl (DtBB) were determined to be non-binders due to isomeric configuration, solubility limitation, and possible steric hinderance. It is possible that the bridged-MAHCs bind to the rtER through a proposed aromatic-aromatic stacking (π-π interaction) facilitated by perpendicular ring orientation achieved through free rotation of the bridged rings. The fused-ring structures are locked in a planar configuration which doesn’t allow for rotation of rings perpendicular to one another. This first report of the rtER-binding of bridged-MAHCs in fish demonstrates binding for a class of chemicals normally not thought of as having an affinity for the estrogen receptor and further supports the versatility or promiscuity of ER ligand selectivity.
  • Toxicokinetics and bioaccumulation of polycyclic aromatic compounds in
           wood frog tadpoles (Lithobates sylvaticus) exposed to Athabasca oil sands
    • Abstract: Publication date: Available online 17 November 2018Source: Aquatic ToxicologyAuthor(s): J.C. Bilodeau, J.M. Gutierrez Villagomez, L.E. Kimpe, P.J. Thomas, B.D. Pauli, V.L. Trudeau, J.M. Blais We performed accumulation-elimination experiments of polycyclic aromatic compounds (PACs) in wood frog tadpoles (Lithobates sylvaticus) using river sediment from Canada’s Athabasca oil sands region. The PACs in wood frog tadpoles were ~2x higher on average when the animals were in direct contact with PAC-contaminated sediment than when they were separated from the sediment with a screen and exposed only to aqueous PACs. These results suggest that sediment exposure/ingestion contributes as much to PAC accumulation in tadpoles as exposure via aqueous pathways. Alkyl-substituted PAC concentrations in exposed tadpoles exceeded those of the unsubstituted (parent) PACs by about 10 × . Bioaccumulation factors ranged between 0.01 and 4.93, with parent PACs having higher bioaccumulation factors than alkylated PACs. Wood frog tadpoles efficiently eliminated and metabolized most parent and alkyl-substituted PACs, though some compounds (e.g., C4-naphthalenes) had higher bioaccumulation potential and may serve as effective markers of exposure. Here we present a comprehensive analysis of the toxicokinetics and bioaccumulation of PACs (52 analytes) in amphibian larvae, and highlight the importance of sediment exposure when considering the bioaccumulation and potential biological impact of PACs in benthic and epibenthic organisms.
  • Subcellular accumulation and source of O2 •– and H2O2 in submerged
           plant Hydrilla verticillata (L.f.) Royle under NH4 +-N stress condition
    • Abstract: Publication date: Available online 13 November 2018Source: Aquatic ToxicologyAuthor(s): Kai Zhuang, Danlu Shi, Zhubing Hu, Fuliu Xu, Yahua Chen, Shen Zhenguo In this study, the effects of excess NH4+-N on the subcellular accumulation of O2•– and H2O2 in submerged plant Hydrilla verticillata (L.f.) Royle were investigated using both histochemical and cytochemical methods. Treatments with ≥ 2.00 and ≥ 5.00 mg L-1 NH4+-N for 5 d significantly increased production of O2•– and H2O2, respectively. The activities of plasma membrane-bound NADPH (nicotinamide adenine dinucleotide phosphate) oxidases and antioxidant enzymes (superoxide dismutase, peroxidase, ascorbate peroxidase, catalase, dehydroascorbate reductase and glutathione reductase) were also increased correspondingly. This study also provides the first cytochemical evidence of subcellular accumulation of O2•– and H2O2 in the submerged plants. In the leaves of H. verticillata treated with 20.0 mg L-1 NH4+-N, O2•– dependent DAB precipitates were found primarily on the inner side of the plasma membrane, extracellular space and chloroplasts. H2O2-CeCl3 precipitates were mainly localized on the inner side of the plasma membrane and extracellular space of the mesophyll cells. Treatments with the inhibitors of NADPH oxidase (diphenylene iodonium and imidazole) indicate that NH4+-N-induced production of O2•– and H2O2 in H. verticillata leaves may involve plasma membrane-bound NADPH oxidase. Moreover, low-light treatment decreased NH4+-induced O2•– production, suggesting that alterations in the photosynthetic electron transfer chain due to NH4+ toxicity could lead to O2•– production.
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