Adaptive capabilities and fitness consequences associated with pollution exposure in fish

Many fish populations are exposed to harmful levels of chemical pollution and selection pressures associated with these exposures have led to the evolution of tolerance. Our understanding of the physiological basis for these adaptations is limited, but they are likely to include processes involved with the absorption, distribution, metabolism and/or excretion of the target chemical. Other potential adaptive mechanisms include enhancements in antioxidant responses, an increased capacity for DNA and/or tissue repair and alterations to the life cycle of fish that enable earlier reproduction. Analysis of single-nucleotide polymorphism frequencies has shown that tolerance to hydrocarbon pollutants in both marine and estuarine fish species involves alteration in the expression of the xenobiotic metabolism enzyme CYP1A. In this review, we present novel data showing also that variants of the CYP1A gene have been under selection in guppies living in Trinidadian rivers heavily polluted with crude oil. Potential costs associated with these adaptations could reduce fitness in unpolluted water conditions. Integrating knowledge of local adaptation to pollution is an important future consideration in conservation practices such as for successful restocking, and improving connectivity within river systems. This article is part of the themed issue ‘Human influences on evolution, and the ecological and societal consequences’

A laboratory investigation into features of morphology and physiology for their potential to predict reproductive success in male frogs

Amphibian populations are declining globally, however, the contribution of reduced reproduction to declines is unknown. We investigated associations between morphological (weight/snout-vent length, nuptial pad colour/size, forelimb width/size) and physiological (nuptial pad/testis histomorphology, plasma hormones, gene expression) features with reproductive success in males as measured by amplexus success and fertility rate (% eggs fertilised) in laboratory maintained Silurana/Xenopus tropicalis. We explored the robustness of these features to predict amplexus success/fertility rate by investigating these associations within a sub-set of frogs exposed to anti-androgens (flutamide (50 μg/L)/linuron (9 or 45 μg/L)). In unexposed males, nuptial pad features (size/colour/number of hooks/androgen receptor mRNA) were positively associated with amplexus success, but not with fertility rate. In exposed males, many of the associations with amplexus success differed from untreated animals (they were either reversed or absent). In the exposed males forelimb width/nuptial pad morphology were also associated with fertility rate. However, a more darkly coloured nuptial pad was positively associated with amplexus success across all groups and was indicative of androgen status. Our findings demonstrate the central role for nuptial pad morphology in reproductive success in S. tropicalis, however, the lack of concordance between unexposed/exposed frogs complicates understanding of the utility of features of nuptial pad morphology as biomarkers in wild populations. In conclusion, our work has indicated that nuptial pad and forelimb morphology have potential for development as biomarkers of reproductive health in wild anurans, however, further research is needed to establish this

Evidence suggesting that di-n-butyl phthalate has anti-androgenic effects in fish

This article is the pre-print version of the full and final published article.Phthalate ester plasticizers are anti-androgenic in mammals. High doses of certain phthalates consistently interfere with the normal development of male offspring exposed in utero, causing disrupted sperm production, abnormal development of the genitalia, and in some cases infertility. In the environment, phthalates are considered ubiquitous and are commonly measured in aquatic ecosystems at low ng to mu g per litre concentrations. Given the similarity between mammalian and teleost endocrine systems, phthalate esters may be able to cause anti-androgenic endocrine disruption in fish in the wild. In the present study, adult male three-spined sticklebacks (Gasterosteus aculetaus) (n = 8) were exposed to di-n-butyl phthalate (DBP) (0, 15, and 35 mu g DBP/L) for 22 d and analyzed for changes in nesting behavior, plasma androgen concentrations, spiggin concentrations, and steroidogenic gene expression. Plasma testosterone concentrations were significantly higher in males from the 35 mu g DBP/L group compared with the solvent control, whereas plasma 11-ketotestosterone concentrations were not significantly affected. Expression of steroid acute regulatory protein and 3 beta-hydroxysteroid dehydrogenase remained unchanged. Spiggin concentrations were significantly lower in the males exposed to 35 mu g DBP/L. Nest building appeared to be slower in some males exposed to DBP, but this was not statistically significant. These results suggest that DBP has anti-androgenic effects in fish. However, further research is required to firmly establish the consequences of chronic DBP exposure in fish

Immune-Related Transcriptional Responses to Parasitic Infection in a Naturally Inbred Fish: Roles of Genotype and Individual Variation

Parasites are strong drivers of evolutionary change and the genetic variation of both host and parasite populations can co-evolve as a function of parasite virulence and host resistance. The role of transcriptome variation in specific interactions between host and parasite genotypes has been less studied and can be confounded by differences in genetic variation. We employed two naturally inbred lines of a self-fertilizing fish to estimate the role of host genotype in the transcriptome response to parasite infection using RNA-seq. In addition, we targeted several differentially expressed immune-related genes to further investigate the relative role of individual variation in the immune response using RT-qPCR, taking advantage of the genomic uniformity of the self-fertilizing lines. We found significant differences in gene expression between lines in response to infection both in the transcriptome and in individual gene RT-qPCR analyses. Individual RT-qPCR analyses of gene expression identified significant variance differences between lines for six genes but only for three genes between infected and control fish. Our results indicate that although the genetic background plays an important role in the transcriptome response to parasites, it cannot fully explain individual differences within genetically homogeneous lines, which can be important for determining the response to parasites

Hepatic transcriptional responses to copper in the three-spined stickleback are affected by their pollution exposure history

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this record.Some fish populations inhabiting contaminated environments show evidence of increased chemical tolerance, however the mechanisms contributing to this tolerance, and whether this is heritable, are poorly understood. We investigated the responses of two populations of wild three-spined stickleback (Gasterosteus aculeatus) with different histories of contaminant exposure to an oestrogen and copper, two widespread aquatic pollutants. Male stickleback originating from two sites, the River Aire, with a history of complex pollution discharges, and Siblyback Lake, with a history of metal contamination, were depurated and then exposed to copper (46μg/L) and the synthetic oestrogen ethinyloestradiol (22ng/L). The hepatic transcriptomic response was compared between the two populations and to a reference population with no known history of exposure (Houghton Springs, Dorset). Gene responses included those typical for both copper and oestrogen, with no discernable difference in response to oestrogen between populations. There was, however, some difference in the magnitude of response to copper between populations. Siblyback fish showed an elevated baseline transcription of genes encoding metallothioneins and a lower level of metallothionein induction following copper exposure, compared to those from the River Aire. Similarly, a further experiment with an F1 generation of Siblyback fish bred in the laboratory found evidence for elevated transcription of genes encoding metallothioneins in unexposed fish, together with an altered transcriptional response to 125μg/L copper, compared with F1 fish originating from the clean reference population exposed to the same copper concentration. These data suggest that the stickleback from Siblyback Lake have a differential response to copper, which is inherited by the F1 generation in laboratory conditions, and for which the underlying mechanism may include an elevation of baseline transcription of genes encoding metallothioneins. The genetic and/or epigenetic mechanisms contributing to this inherited alteration of metallothionein transcription have yet to be established.This work was funded by the UK NERC postgenomic and proteomic programme grant NE/C507661/1 and by a Fisheries Society of the British Isles research grant to EMS. Birmingham functional genomics facilities were funded by BBSRC grant 6/JIF13209. We thank R.E. Godfrey, S. Jondhale, A. Jones, and L. Klovrza for technical assistance, J.K. Chipman for help and support, and the Environment Agency for provision of water chemistry data

Investigating Mechanisms of Environmental Chemical Tolerance and Toxicity in Brown Trout using RNA-seq

Brown trout (Salmo trutta) are an ecologically and economically important native European species, known to be sensitive to environmental stressors. Compared to other model species, there is little information available on the toxicological responses of this species to environmental pollutants. High-throughput RNA-sequencing (RNA-seq) is emerging as a sensitive and accurate tool for conducting transcriptomics, but is yet to be widely used in ecotoxicology. A major advantage of RNA-seq is that it can be used to conduct non-biased, global gene expression analysis in species without existing genomic sequence information. Therefore, during this PhD I set out to investigate global mechanisms of toxicity for a selection of the most environmentally relevant chemicals likely to impact upon natural brown trout populations. By using RNA-seq, I also aimed to demonstrate the potential application of this technology as a valuable tool in ecotoxicology. To address these objectives, I conducted transcriptomic profiling, both on wild brown trout and on those exposed to agricultural pollutants in a laboratory setting. Using RNA-seq in combination with analysis of tissue metal concentration I found evidence of a high degree of metal tolerance in a chronically exposed wild population of brown trout from the river Hayle. The main molecular mechanisms responsible for this metal-tolerance included regulation of metal- and ion-homeostasis pathways. In the laboratory exposures, I found evidence of considerable transcriptomic changes in male brown trout exposed to 34.38 ng/L E2, including up-regulation of typical oestrogen-responsive transcripts (vitellogenins, zona pellucida proteins and estrogen receptor 1), as well as hepatic processes that can be associated with vitellogenesis such as lipid metabolism, cell proliferation and ribosome biogenesis. This concentration is within a range measured in sewage effluent and, more occasionally, in surface waters. I also exposed male brown trout to linuron, a widely used pesticide, and observed a striking down-regulation of enzymes involved in the cholesterol biosynthesis pathway and up-regulation of transcripts involved in cellular stress response following exposure to 250 µg linuron/L. There was also some evidence of similar responses occurring at the lower, environmentally relevant concentration (2.5 µg/L). I then compared the mechanisms of toxicity of glyphosate, the most widely used herbicide in the world, and its commercial formulation Roundup in juvenile brown trout. I found evidence of a cellular stress response consistent with generation of oxidative stress at concentrations of 10 µg/L and above which, importantly, is within the range of concentrations measured in the environment. To investigate the potential reproductive toxicity of these compounds I also conducted an exposure of breeding zebrafish to glyphosate and Roundup, and found evidence of reproductive toxicity, but only at a very high concentration (10 mg/L). This work therefore provides valuable information on the toxicological effects of these environmentally relevant chemicals in brown trout, which can potentially be used to assess the risk they pose to natural populations and therefore contribute to the sustainable management of this species. I have successfully employed RNA-seq to achieve the main objective of this PhD and, in so doing, have demonstrated the value of this technology in ecotoxicology. Specifically, we have demonstrated the ability of RNA-seq to identify conserved responses typically associated with oestrogen exposure. We also highlight the importance of optimising the experimental design and strategy for RNA-seq data analysis to improve the quality of transcript expression analysis. Throughout the course of this work we have benefited from improvements in sequencing technology and the tools available for data analysis. This technology is continuing to develop rapidly, and it is likely that RNA-seq will become the dominant tool for conducting transcriptomics in ecotoxicology in the future.NERCThe Salmon and Trout Associatio

Impact of ocean acidification on the intestinal microbiota of the marine sea bream (Sparus aurata L.)

Within a scenario of increasing atmospheric CO2 and ocean acidification (OA), it is highly relevant to investigate its impacts not only on fish performance but also on fish intestinal microbiome and how that reflects on host performance and health. The main objective of this study was to establish if the intestinal microbiota of the sea bream (Sparus aurata) was affected by high level of CO2 in line with the predictions for this century. The bacterial communities of the intestinal fluid were characterized in animals kept at the present-day level of CO2 (400 μatm) and in animals switched to high CO2 (1200 μatm) for 1 month. Bacterial taxa identification was based on molecular methods, using the DNA coding for the 16S ribosomal RNA and primers targeting the regions V1-V3. Amplicons obtained from DNA samples of animals in the same tank were combined, cloned to obtain a bacterial DNA library, and the clones were sequenced. No significant differences were found between the two treatments for alpha diversity. However, beta diversity analysis revealed distinct dysbiosis in response to hypercapnia, with phylum Firmicutes absent from the bacterial communities of fish exposed to 1200 μatm CO2, whereas Proteobacteria relative abundance was increased at elevated CO2, due to the presence of Gammaproteobacteria (Vibrionaceae and Alteromonadaceae), a class not present in the control samples. This study provides a first glimpse at the impact of OA in fish intestinal microbiota and highlights potential downstream effects to the general condition of fishes under hypercapnia.Funding Agency Portuguese Foundation for Science and Technology PTDC/MAR-BIO/3034/2014 Portuguese Foundation for Science and Technology UID/Multi/04326/2019 Ministry of Science and Higher Education, Polandinfo:eu-repo/semantics/publishedVersio

Bisphenol A causes reproductive toxicity, decreasesdnmt1transcription, and reduces global DNA methylation in breeding zebrafish(Danio rerio)

Bisphenol A (BPA) is a commercially important high production chemical widely used in epoxy resins and polycarbonate plastics, and is ubiquitous in the environment. Previous studies demonstrated that BPA activates estrogenic signaling pathways associated with adverse effects on reproduction in vertebrates and that exposure can induce epigenetic changes. We aimed to investigate the reproductive effects of BPA in a fish model and to document its mechanisms of toxicity. We exposed breeding groups of zebrafish (Danio rerio) to 0.01, 0.1, and 1 mg/L BPA for 15 days. We observed a significant increase in egg production, together with a reduced rate of fertilization in fish exposed to 1 mg/L BPA, associated with significant alterations in the transcription of genes involved in reproductive function and epigenetic processes in both liver and gonad tissue at concentrations representing hotspots of environmental contamination (0.1 mg/L) and above. Of note, we observed reduced expression of DNA methyltransferase 1 (dnmt1) at environmentally relevant concentrations of BPA, along with a significant reduction in global DNA methylation, in testes and ovaries following exposure to 1 mg/L BPA. Our findings demonstrate that BPA disrupts reproductive processes in zebrafish, likely via estrogenic mechanisms, and that environmentally relevant concentrations of BPA are associated with altered transcription of key enzymes involved in DNA methylation maintenance. These findings provide evidence of the mechanisms of action of BPA in a model vertebrate and advocate for its reduction in the environment.We thank the Aquatic Resources Centre technical team for support with zebrafish husbandry. This work was funded by a PhD studentship from the Fisheries Society of the British Isles (http://www.fsbi.org.uk/) and the University of Exeter (http://www.exeter.ac.uk/) to LVL and EMS. TMUW was funded by a Natural Environment Research Council CASE PhD studentship (grant no. NE/I528326/1) and the Salmon & Trout Association (http://www.salmon-trout.org/)

Shaken not stirred: Mixing semantics into XPDL

Ubiquitous computing requires lightweight approaches to coordinating tasks distributed across smart devices. We are currently developing a semantic workflow modelling approach that blends the proven robustness of XPDL with semantics to support proactive behaviour. We illustrate the potential of the model through an example based on mixing a dry martini

Benefits of the microalgae Spirulina and Schizochytrium in fish nutrition: a meta-analysis

Use of microalgae in fish nutrition can relieve pressure on wild fish stocks, but there is no systematic quantitative evaluation of microalgae benefits. We conducted a metanalysis on the nutritional benefits of Spirulina and Schizochytrium as replacements of fishmeal and fish or plant oil, respectively. We reviewed 50 peer-reviewed studies involving 26 finfish species and 144 control vs microalgae replacement comparisons. Inclusion of Spirulina in the fish diet significantly improved growth compared to controls (SMD = 1.21; 95% CI 0.71–1.70), while inclusion of Schizochytrium maintained the content of omega-3 PUFA of the fish fillet compared to fish fed on fish or plant oils (SMD = 0.62; 95% CI − 0.51–1.76). Benefits were apparent at replacement levels as low as 0.025% in the case of Spirulina and 10% in the case of Schizochytrium oil. Dose-dependent effects were found for Spirulina replacement on growth, but not for Schizochytrium on omega-3 fillet content. Subgroup analysis and meta-regression revealed that ~ 24–27% of variation in effect sizes can be accounted by variation between fish families, the rest likely reflecting variation in experimental conditions. Overall, the evidence indicates that Spirulina and Schizochytrium replacement in aquafeeds can be used to improve fish growth and maintain fillet quality, respectively, but considerable uncertainty exists on the predicted responses. To reduce uncertainty and facilitate the transition towards more sustainable aquafeeds, we recommend that feeding trials using microalgae are conducted under commercially relevant conditions and that greater care is taken to report full results to account for sources of heterogeneity