Critical questions must be addressed to evaluate the potential of metabolomics for studying free-living wildlife. First, can metabolomics identify stress-induced phenotypes in animals experiencing a highly variable environment or must animals be stabilized in a controlled laboratory prior to sampling? Second, is knowledge of species and phenotype (gender and age) required to interpret metabolomics data? To address these questions, we characterized the metabolic variability of the mussel and determined if inherent variability masked the metabolic response to an environmental stressor, hypoxia. Specifically, we compared metabolic fingerprints of adductor muscle and mantle from four groups of Mytilus galloprovincialis: animals sampled directly from the field with and without hypoxia and those stabilized in a laboratory for 60 h, also with and without hypoxia. Contrary to expectation, laboratory stabilization increased metabolic variability in adductor muscle, thereby completely masking the response to hypoxia. The principal source of metabolic variability in mantle was shown to be gender-based, highlighting the importance of phenotypic anchoring of samples to known life history traits. We conclude that direct field sampling is recommended for environmental metabolomics since it minimizes metabolic variability and enables stress-induced phenotypic changes to be observed. Furthermore, we recommend that species and phenotype of the study organism must be known for meaningful interpretation of metabolomics data.
Tilapia lake virus (TiLV), a negative sense RNA virus with a 10 segment genome, is an emerging threat to tilapia aquaculture worldwide, with outbreaks causing over 90% mortality reported on several continents since 2014. Following a severe tilapia mortality event in July 2017, we confirmed the presence of TiLV in Bangladesh and obtained the near-complete genome of this isolate, BD-2017. Phylogenetic analysis of the concatenated 10 segment coding regions placed BD-2017 in a clade with the two isolates from Thailand, separate from the Israeli and South American isolates. However, phylogenetic analysis of individual segments gave conflicting results, sometimes clustering BD-2017 with one of the Israeli isolates, and splitting pairs of isolates from the same region. By comparing patterns of topological difference among segments of quartets of isolates, we showed that TiLV likely has a history of reassortment. Segments 5 and 6, in particular, appear to have undergone a relatively recent reassortment event involving Ecuador isolate EC-2012 and Israel isolate Til-4-2011. The phylogeny of TiLV isolates therefore depends on the segment sequenced. Our findings illustrate the need to exercise caution when using phylogenetic analysis to infer geographic origin and track the movement of TiLV, and we recommend using whole genomes wherever possible.
ABSTRACT:We assessed seasonal histological changes as markers of health status in mussels Mytilus spp. sampled from Southampton Water, Hampshire, UK and the River Exe, Devon, UK between November 2004 and October 2005. A total of 29 health parameters related to pathogens, inflammatory and non-specific pathologies, and reproductive and physiological condition were recorded monthly from individual mussels collected from these 2 sites. We then assessed the diffential prevalence of these health parameters according to species. M. edulis, M. galloprovincialis and their hybrids were identified using the Glu-5' gene and the ME15 and ME16 primer sets that distinguish alleles specific to M. edulis (180 bp), M. galloprovincialis (126 bp) and hybrids (180 bp/126 bp). Although no overall annual differences were observed between species with respect to median levels of adipogranular (ADG) tissue and reproductive status, specific differences in reproductive status were observed within individual months. During these months (August to October), M. edulis exhibited a relatively lower reproductive status compared to M. galloprovincialis and hybrids. With respect to all remaining health parameters (pathogens, inflammatory and non-specific pathology), principal components analysis revealed no overall differences between species throughout the year. However, greater differences were observed between species during the autumn and winter than during the spring and summer, thus indicating that species differences may be exacerbated by season. This study highlights how species can affect the accurate interpretation of histopathology data collected during biological effects monitoring programmes. Whether species can also affect the biomarker response of Mytilus mussels to contaminated environments remains to be shown. The results are discussed in the context of biological effects monitoring utilising mussels.
The link between environment, alteration in DNA methylation and cancer has been well established in humans; yet, it is under-studied in unsequenced non-model organisms. The occurrence of liver tumors in the flatfish dab collected at certain UK sampling sites exceeds 20%, yet the causative agents and the molecular mechanisms of tumor formation are not known, especially regarding the balance between epigenetic and genetic factors. Methylated DNA Immunoprecipitation (MeDIP) combined with de novo high-throughput DNA sequencing were used to investigate DNA methylation changes in dab hepatocellular adenoma tumors for the first time in an unsequenced species. Novel custom-made dab gene expression arrays were designed and used to determine the relationship between DNA methylation and gene expression. In addition, the confirmatory techniques of bisulfite sequencing PCR (BSP) and RT-PCR were applied. Genes involved in pathways related to cancer, including apoptosis, wnt/β-catenin signaling and genomic and non-genomic estrogen responses, were altered both in methylation and transcription. Global methylation was statistically significantly 1.8-fold reduced in hepatocellular adenoma and non-cancerous surrounding tissues compared with liver from non-cancer bearing dab. Based on the identified changes and chemical exposure data, our study supports the epigenetic model of cancer. We hypothesize that chronic exposure to a mixture of environmental contaminants contributes to a global hypomethylation followed by further epigenetic and genomic changes. The findings suggest a link between environment, epigenetics and cancer in fish tumors in the wild and show the utility of this methodology for studies in non-model organisms.
Marine environmental monitoring is undertaken to provide evidence that environmental management targets are being met. Moreover, monitoring also provides context to marine science and over the last century has allowed development of a critical scientific understanding of the marine environment and the impacts that humans are having on it. The seas around the UK are currently monitored by targeted, impact-driven, programmes (e.g., fishery or pollution based monitoring) often using traditional techniques, many of which have not changed significantly since the early 1900s. The advent of a new wave of automated technology, in combination with changing political and economic circumstances, means that there is currently a strong drive to move toward a more refined, efficient, and effective way of monitoring. We describe the policy and scientific rationale for monitoring our seas, alongside a comprehensive description of the types of equipment and methodology currently used and the technologies that are likely to be used in the future. We contextualize the way new technologies and methodologies may impact monitoring and discuss how whole ecosystems models can give an integrated, comprehensive approach to impact assessment. Furthermore, we discuss how an understanding of the value of each data point is crucial to assess the true costs and benefits to society of a marine monitoring programme.
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