Molecular Mechanisms of White Spot Syndrome Virus Infection and Perspectives on Treatments.

Published onlineJournal ArticleReviewSince its emergence in the 1990s, White Spot Disease (WSD) has had major economic and societal impact in the crustacean aquaculture sector. Over the years shrimp farming alone has experienced billion dollar losses through WSD. The disease is caused by the White Spot Syndrome Virus (WSSV), a large dsDNA virus and the only member of the Nimaviridae family. Susceptibility to WSSV in a wide range of crustacean hosts makes it a major risk factor in the translocation of live animals and in commodity products. Currently there are no effective treatments for this disease. Understanding the molecular basis of disease processes has contributed significantly to the treatment of many human and animal pathogens, and with a similar aim considerable efforts have been directed towards understanding host-pathogen molecular interactions for WSD. Work on the molecular mechanisms of pathogenesis in aquatic crustaceans has been restricted by a lack of sequenced and annotated genomes for host species. Nevertheless, some of the key host-pathogen interactions have been established: between viral envelope proteins and host cell receptors at initiation of infection, involvement of various immune system pathways in response to WSSV, and the roles of various host and virus miRNAs in mitigation or progression of disease. Despite these advances, many fundamental knowledge gaps remain; for example, the roles of the majority of WSSV proteins are still unknown. In this review we assess current knowledge of how WSSV infects and replicates in its host, and critique strategies for WSD treatment.This work was funded by the Open Innovation Platform at the University of Exeter (Open Innovation Fund Initiative PHSW029) and by the Centre for Environment, Fisheries and Aquaculture Science (Cefas) (under seedcorn project DP318 to GDS) under the Strategic Alliance partnership between the University of Exeter and Cefas

De novo assembly of the Carcinus maenas transcriptome and characterization of innate immune system pathways.

Journal ArticleCopyright © 2015 Verbruggen et al.BACKGROUND: The European shore crab, Carcinus maenas, is used widely in biomonitoring, ecotoxicology and for studies into host-pathogen interactions. It is also an important invasive species in numerous global locations. However, the genomic resources for this organism are still sparse, limiting research progress in these fields. To address this resource shortfall we produced a C. maenas transcriptome, enabled by the progress in next-generation sequencing technologies, and applied this to assemble information on the innate immune system in this species. RESULTS: We isolated and pooled RNA for twelve different tissues and organs from C. maenas individuals and sequenced the RNA using next generation sequencing on an Illumina HiSeq 2500 platform. After de novo assembly a transcriptome was generated encompassing 212,427 transcripts (153,699 loci). The transcripts were filtered, annotated and characterised using a variety of tools (including BLAST, MEGAN and RSEM) and databases (including NCBI, Gene Ontology and KEGG). There were differential patterns of expression for between 1,223 and 2,741 transcripts across tissues and organs with over-represented Gene Ontology terms relating to their specific function. Based on sequence homology to immune system components in other organisms, we show both the presence of transcripts for a series of known pathogen recognition receptors and response proteins that form part of the innate immune system, and transcripts representing the RNAi, Toll-like receptor signalling, IMD and JAK/STAT pathways. CONCLUSIONS: We have produced an assembled transcriptome for C. maenas that provides a significant molecular resource for wide ranging studies in this species. Analysis of the transcriptome has revealed the presence of a series of known targets and functional pathways that form part of their innate immune system and illustrate tissue specific differences in their expression patterns.Cefas Seedcorn Contract #DP318University of Exeter’s Open Innovation PlatformWellcome Trust Institutional Strategic Support Awar

Decoding the Cortical Dynamics of Sound-Meaning Mapping.

Comprehending speech involves the rapid and optimally efficient mapping from sound to meaning. Influential cognitive models of spoken word recognition (Marslen-Wilson and Welsh, 1978) propose that the onset of a spoken word initiates a continuous process of activation of the lexical and semantic properties of the word candidates matching the speech input and competition between them, which continues until the point at which the word is differentiated from all other cohort candidates (the uniqueness point, UP). At this point, the word is recognized uniquely and only the target word's semantics are active. Although it is well established that spoken word recognition engages the superior (Rauschecker and Scott, 2009), middle, and inferior (Hickok and Poeppel, 2007) temporal cortices, little is known about the real-time brain activity that underpins the computations and representations that evolve over time during the transformation from speech to meaning. Here, we test for the first time the spatiotemporal dynamics of these processes by collecting MEG data while human participants listened to spoken words. By constructing quantitative models of competition and access to meaning in combination with spatiotemporal searchlight representational similarity analysis (Kriegeskorte et al., 2006) in source space, we were able to test where and when these models produced significant effects. We found early transient effects ∼400 ms before the UP of lexical competition in left supramarginal gyrus, left superior temporal gyrus, left middle temporal gyrus (MTG), and left inferior frontal gyrus (IFG) and of semantic competition in MTG, left angular gyrus, and IFG. After the UP, there were no competitive effects, only target-specific semantic effects in angular gyrus and MTG. SIGNIFICANCE STATEMENT: Understanding spoken words involves complex processes that transform the auditory input into a meaningful interpretation. This effortless transition occurs on millisecond timescales, with remarkable speed and accuracy and without any awareness of the complex computations involved. Here, we reveal the real-time neural dynamics of these processes by collecting data about listeners' brain activity as they hear spoken words. Using novel statistical models of different aspects of the recognition process, we can locate directly which parts of the brain are accessing the stored form and meaning of words and how the competition between different word candidates is resolved neurally in real time. This gives us a uniquely differentiated picture of the neural substrate for the first 500 ms of word recognition.This work was supported by a European Research Council Advanced Investigator grant under the European Community’s Horizon 2020 Research and Innovation Programme (2014 –2020 ERC Grant 669820 to L.K.T.)

Interactive effects of inbreeding and endocrine disruption on reproduction in a model laboratory fish

This is the final version of the article. Available from Wiley via the DOI in this record.Inbreeding depression is expected to be more severe in stressful environments. However, the extent to which inbreeding affects the vulnerability of populations to environmental stressors, such as chemical exposure, remains unresolved. Here we report on the combined impacts of inbreeding and exposure to an endocrine disrupting chemical (the fungicide clotrimazole) on zebrafish (Danio rerio). We show that whilst inbreeding can negatively affect reproductive traits, not all traits are affected equally. Inbreeding depression frequently only became apparent when fish were additionally stressed by chemical exposure. Embryo viability was significantly reduced in inbred exposed fish and there was a tendency for inbred males to sire fewer offspring when in direct competition with outbred individuals. Levels of plasma 11-ketotestosterone, a key male sex hormone, showed substantial inbreeding depression that was unaffected by addition of the fungicide. In contrast, there was no effect of inbreeding or clotrimazole exposure on egg production. Overall, our data provide evidence that stress may amplify the effects of inbreeding on key reproductive traits, particularly those associated with male fitness. This may have important implications when considering the consequences of exposure to chemical pollutants on the fitness of wild populations.Thanks to NERC's Post Genomics & Proteomics Programme NE/F0077871/1 and AstraZeneca's Safety, Health and Environment Research Programme for funding this work. We thank Alexander Scott (11-ketotestosterone radioimmunoassay) at the Centre for Environment, Fisheries and Aquaculture Science, Jan Shears and Luanne Wilkes at University of Exeter, Gareth Readman, Vicki Cammack, Kate Hurd and Yohanna Glennon at Brixham Environmental Laboratory for their assistance

Bioavailability and kidney responses to Diclofenac in the fathead minnow (Pimephales promelas)

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this record.Diclofenac is one of the most widely prescribed nonsteroidal anti-inflammatory drugs worldwide. It is frequently detected in surface waters; however, whether this pharmaceutical poses a risk to aquatic organisms is debated. Here we quantified the uptake of diclofenac by the fathead minnow (Pimephales promelas) following aqueous exposure (0.2-25.0 μg L(-1)) for 21 days, and evaluated the tissue and biomolecular responses in the kidney. Diclofenac accumulated in a concentration- and time-dependent manner in the plasma of exposed fish. The highest plasma concentration observed (for fish exposed to 25 μg L(-1) diclofenac) was within the therapeutic range for humans. There was a strong positive correlation between exposure concentration and the number of developing nephrons observed in the posterior kidney. Diclofenac was not found to modulate the expression of genes in the kidney associated with its primary mode of action in mammals (prostaglandin-endoperoxide synthases) but modulated genes associated with kidney repair and regeneration. There were no significant adverse effects following 21 days exposure to concentrations typical of surface waters. The combination of diclofenac's uptake potential, effects on kidney nephrons and relatively small safety margin for some surface waters may warrant a longer term chronic health effects analysis for diclofenac in fish.This work was funded by Knowledge Transfer Partnership (KTP): Use of ‘omic’ technologies in the environmental risk assessment of pharmaceuticals (KTP007650) and AstraZeneca’s Safety, Health and Environment (SHE) Research Programme. We thank Lina Gunnarsson, Matt Winter and James Cresswell (Exeter University), and former members of the Brixham Environmental Laboratory for their advice and assistance. Authors declare no competing financial interest

How do abiotic environmental conditions influence shrimp susceptibility to disease? A critical analysis focussed on White Spot Disease

This is the author accepted manuscript (article in press version). The final version is available from the publisher via the DOI in this recordWhite Spot Syndrome Virus (WSSV) causes White Spot Disease (WSD) and is historically the most devastating disease in the shrimp industry. Global losses from this disease have previously exceeded $3 bn annually, having a major impact on a global industry worth US$19 bn per annum. Shrimp are cultured predominantly in enclosed ponds that are subject to considerable fluctuations in abiotic conditions and WSD outbreaks are increasingly linked to periods of extreme weather, which may cause major fluctuations in pond culture conditions. Combined with the intensity of production in these systems, the resulting suboptimal physicochemical conditions have a major bearing on the susceptibility of shrimp to infection and disease. Current knowledge indicates that pond temperature and salinity are major factors determining outbreak severity. WSSV appears to be most virulent in water temperatures between 25 and 28 °C and salinities far removed from the isoosmotic point of shrimp. Elevated temperatures (>30 °C) may protect against WSD, depending on the stage of infection, however the mechanisms mediating this effect have not been well established. Other factors relating to water quality that may play key roles in determining outbreak severity include dissolved oxygen concentration, nitrogenous compound concentration, partial pressure of carbon dioxide and pH, but data on their impacts on WSSV susceptibility in cultured shrimps is scarce. This illustrates a major research gap in our understanding of the influence of environmental conditions on disease. For example, it is not clear whether temperature manipulations can be used effectively to prevent or mitigate WSD in cultured shrimp. Therefore, developing our understanding of the impact of environmental conditions on shrimp susceptibility to WSSV may provide insight for WSD mitigation when, even after decades of research, there is no effective practical prophylaxis or treatment.Centre for Environment, Fisheries and Aquaculture Scienc

Resistance to white spot syndrome virus in the European shore crab is associated with suppressed virion trafficking and heightened immune responses

This is the final version. Available on open access from Frontiers Media via the DOI in this recordData availability statement: The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: https://www.ncbi.nlm.nih.gov/genbank/, SRR14278211 - SRR14278323 and https://doi.org/10.6084/m9.figshare.21225128, as well as https://doi.org/10.6084/m9.figshare.21435831.INTRODUCTION: All decapod crustaceans are considered potentially susceptible to White Spot Syndrome Virus (WSSV) infection, but the degree of White Spot Disease (WSD) susceptibility varies widely between species. The European shore crab Carcinus maenas can be infected with the virus for long periods of time without signs of disease. Given the high mortality rate of susceptible species, the differential susceptibility of these resistant hosts offers an opportunity to investigate mechanisms of disease resistance. METHODS: Here, the temporal transcriptional responses (mRNA and miRNA) of C. maenas following WSSV injection were analysed and compared to a previously published dataset for the highly WSSV susceptible Penaeus vannamei to identify key genes, processes and pathways contributing to increased WSD resistance. RESULTS: We show that, in contrast to P. vannamei, the transcriptional response during the first 2 days following WSSV injection in C. maenas is limited. During the later time points (7 days onwards), two groups of crabs were identified, a recalcitrant group where no replication of the virus occurred, and a group where significant viral replication occurred, with the transcriptional profiles of the latter group resembling those of WSSV-susceptible species. We identify key differences in the molecular responses of these groups to WSSV injection. DISCUSSION: We propose that increased WSD resistance in C. maenas may result from impaired WSSV endocytosis due to the inhibition of internal vesicle budding by dynamin-1, and a delay in movement to the nucleus caused by the downregulation of cytoskeletal transcripts required for WSSV cytoskeleton docking, during early stages of the infection. This response allows resistant hosts greater time to fine-tune immune responses associated with miRNA expression, apoptosis and the melanisation cascade to defend against, and clear, invading WSSV. These findings suggest that the initial stages of infection are key to resistance to WSSV in the crab and highlight possible pathways that could be targeted in farmed crustacean to enhance resistance to WSD.University of Exeter (UK) Open Innovation PlatformCentre for Environment, Fisheries and Aquaculture Science (Weymouth, UK)Wellcome TrustBiotechnology and Biological Sciences Research Council (BBSRC

Spatial Stereoresolution for Depth Corrugations May Be Set in Primary Visual Cortex

Stereo “3D” depth perception requires the visual system to extract binocular disparities between the two eyes' images. Several current models of this process, based on the known physiology of primary visual cortex (V1), do this by computing a piecewise-frontoparallel local cross-correlation between the left and right eye's images. The size of the “window” within which detectors examine the local cross-correlation corresponds to the receptive field size of V1 neurons. This basic model has successfully captured many aspects of human depth perception. In particular, it accounts for the low human stereoresolution for sinusoidal depth corrugations, suggesting that the limit on stereoresolution may be set in primary visual cortex. An important feature of the model, reflecting a key property of V1 neurons, is that the initial disparity encoding is performed by detectors tuned to locally uniform patches of disparity. Such detectors respond better to square-wave depth corrugations, since these are locally flat, than to sinusoidal corrugations which are slanted almost everywhere. Consequently, for any given window size, current models predict better performance for square-wave disparity corrugations than for sine-wave corrugations at high amplitudes. We have recently shown that this prediction is not borne out: humans perform no better with square-wave than with sine-wave corrugations, even at high amplitudes. The failure of this prediction raised the question of whether stereoresolution may actually be set at later stages of cortical processing, perhaps involving neurons tuned to disparity slant or curvature. Here we extend the local cross-correlation model to include existing physiological and psychophysical evidence indicating that larger disparities are detected by neurons with larger receptive fields (a size/disparity correlation). We show that this simple modification succeeds in reconciling the model with human results, confirming that stereoresolution for disparity gratings may indeed be limited by the size of receptive fields in primary visual cortex