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A reo-like virus, CsRV1, is found in blue crabs, Callinectes sapidus, from every North American location assessed, including Chesapeake Bay and the Atlantic and Gulf coasts, USA and associated with blue crabs in softshell production. CsRV1-associated crab mortality is prevalent in captive crabs, but it is still unknown how CsRV1 is transmitted. The purpose of this study was to examine the role that conspecific predation or scavenging may play in per os transmission in single exposure and repeated exposure experiments. For viruses without cell culture propagation, repeated exposure experiments have the challenge of presenting the virus consistently during the experiment and across time replicates. In a single-exposure experiment, none of the crabs fed muscle tissue of crabs carrying intense infections of CsRV1 developed CsRV1 infections. In a repeated-exposure trial, using infected muscle tissue prepared in alginate blocks, CsRV1 was detected in 11% of the crabs fed infected tissue but was not significantly different from the control group fed alginate lacking CsRV1. For repeated per os exposure experiments, the study demonstrated the utility of using alginate to present the same homogenous sample of virus, both injected and per os, over time for oral challenge experiments. Conspecific predation and scavenging could be a transmission route, but future work into this and other possible routes of transmission for CsRV1 is important to better understand the role this virus plays in wild crab populations and the soft-shell crab industry.more » « less
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The movement of viruses in aquatic systems is rarely studied over large geographic scales. Oceanic currents, host migration, latitude-based variation in climate, and resulting changes in host life history are all potential drivers of virus connectivity, adaptation, and genetic structure. To expand our understanding of the genetic diversity of Callinectes sapidus reovirus 1 (CsRV1) across a broad spatial and host life history range of its blue crab host (Callinectes sapidus), we obtained 22 complete and 96 partial genomic sequences for CsRV1 strains from the US Atlantic coast, Gulf of Mexico, Caribbean Sea, and the Atlantic coast of South America. Phylogenetic analyses of CsRV1 genomes revealed that virus genotypes were divided into four major genogroups consistent with their host geographic origins. However, some CsRV1 sequences from the US mid-Atlantic shared high genetic similarity with the Gulf of Mexico genotypes, suggesting potential human-mediated movement of CsRV1 between the US mid-Atlantic and Gulf coasts. This study advances our understanding of how climate, coastal geography, host life history, and human activity drive patterns of genetic structure and diversity of viruses in marine animals and contributes to the capacity to infer broadscale host population connectivity in marine ecosystems from virus population genetic data.more » « less
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The National Oceanic and Atmospheric Administration (NOAA) Living Marine Resources Cooperative Science Center (LMRCSC) was established in 2001 as a multi-institutional collaborative program with the goal of preparing a diverse future STEM workforce in marine and fisheries sciences. Using best practices for recruiting, mentoring, and training students, the Center has had a significant impact on the number of individuals from underrepresented groups earning degrees in NOAA-related sciences. From 2001 to 2022, the LMRCSC recruited and trained 1,092 students (516 BS, 241 MS, 89 PhD, and 246 non-degree students), and graduated 630 (401 BS/BA, 173 MS/MA, 56 PhD) students. About 77% of the graduates belong to underrepresented groups. Of the LMRCSC graduates, 41 work for NOAA or NOAA contractors. Since 2016, 67 graduate students have taken part in internships at NOAA Laboratories across the United States. Institutional capacities built using NOAA funds have enabled Historically Black Colleges and Universities collaborating with the Center to leverage funds to develop several programs, including a National Science Foundation Research Experiences for Undergraduates site in marine and estuarine science. The NOAA LMRCSC has become a model collaborative educational partnership that should be replicated elsewhere to enhance diversity in STEM disciplines, particularly the geosciences.more » « less
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The advancement of high throughput sequencing has greatly facilitated the exploration of viruses that infect marine hosts. For example, a number of putative virus genomes belonging to the Totiviridae family have been described in crustacean hosts. However, there has been no characterization of the most newly discovered putative viruses beyond description of their genomes. In this study, two novel double-stranded RNA (dsRNA) virus genomes were discovered in the Atlantic blue crab ( Callinectes sapidus ) and further investigated. Sequencing of both virus genomes revealed that they each encode RNA dependent RNA polymerase proteins (RdRps) with similarities to toti-like viruses. The viruses were tentatively named Callinectes sapidus toti-like virus 1 (CsTLV1) and Callinectes sapidus toti-like virus 2 (CsTLV2). Both genomes have typical elements required for −1 ribosomal frameshifting, which may induce the expression of an encoded ORF1–ORF2 (gag-pol) fusion protein. Phylogenetic analyses of CsTLV1 and CsTLV2 RdRp amino acid sequences suggested that they are members of two new genera in the family Totiviridae . The CsTLV1 and CsTLV2 genomes were detected in muscle, gill, and hepatopancreas of blue crabs by real-time reverse transcription quantitative PCR (RT-qPCR). The presence of ~40 nm totivirus-like viral particles in all three tissues was verified by transmission electron microscopy, and pathology associated with CsTLV1 and CsTLV2 infections were observed by histology. PCR assays showed the prevalence and geographic range of these viruses, to be restricted to the northeast United States sites sampled. The two virus genomes co-occurred in almost all cases, with the CsTLV2 genome being found on its own in 8.5% cases, and the CsTLV1 genome not yet found on its own. To our knowledge, this is the first report of toti-like viruses in C. sapidus . The information reported here provides the knowledge and tools to investigate transmission and potential pathogenicity of these viruses.more » « less
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Matthijnssens, Jelle (Ed.)ABSTRACT This report describes the sequence of a reovirus genome, discovered in Callinectes sapidus in Brazil. The genome sequence of Callinectes sapidus reovirus 2 (CsRV2) consists of 12 segments that encode 13 putative proteins. The predicted RNA-dependent RNA polymerase is highly similar to that of Eriocheir sinensis reovirus 905, suggesting that CsRV2 also belongs to the genus Cardoreovirus .more » « less
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