Plasma proteomics of green turtles (\u3cem\u3eChelonia mydas\u3c/em\u3e) reveals pathway shifts and potential biomarker candidates associated with health and disease

Evaluating sea turtle health can be challenging due to an incomplete understanding of pathophysiologic responses in these species. Proteome characterization of clinical plasma samples can provide insights into disease progression and prospective biomarker targets. A TMT-10-plex-LC–MS/MS platform was used to characterize the plasma proteome of five, juvenile, green turtles (Chelonia mydas) and compare qualitative and quantitative protein changes during moribund and recovered states. The 10 plasma samples yielded a total of 670 unique proteins. Using ≥1.2-fold change in protein abundance as a benchmark for physiologic upregulation or downregulation, 233 (34.8%) were differentially regulated in at least one turtle between moribund and recovered states. Forty-six proteins (6.9%) were differentially regulated in all five turtles with two proteins (0.3%) demonstrating a statistically significant change. A principle component analysis showed protein abundance loosely clustered between moribund samples or recovered samples and for turtles that presented with trauma (n = 3) or as intestinal floaters (n = 2). Gene Ontology terms demonstrated that moribund samples were represented by a higher number of proteins associated with blood coagulation, adaptive immune responses and acute phase response, while recovered turtle samples included a relatively higher number of proteins associated with metabolic processes and response to nutrients. Abundance levels of 48 proteins (7.2%) in moribund samples significantly correlated with total protein, albumin and/or globulin levels quantified by biochemical analysis. Differentially regulated proteins identified with immunologic and physiologic functions are discussed for their possible role in the green turtle pathophysiologic response and for their potential use as diagnostic biomarkers. These findings enhance our ability to interpret sea turtle health and further progress conservation, research and rehabilitation programs for these ecologically important species

Support for the Slope Sea as a major spawning ground for Atlantic bluefin tuna: evidence from larval abundance, growth rates, and particle-tracking simulations

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Hernandez, C. M., Richardson, D. E., Rypina, I. I., Chen, K., Marancik, K. E., Shulzitski, K., & Llopiz, J. K. Support for the Slope Sea as a major spawning ground for Atlantic bluefin tuna: evidence from larval abundance, growth rates, and particle-tracking simulations. Canadian Journal of Fisheries and Aquatic Sciences, 79(5), (2021): 814-824, https://doi.org/10.1139/cjfas-2020-0444.Atlantic bluefin tuna (Thunnus thynnus) are commercially and ecologically valuable, but management is complicated by their highly migratory lifestyle. Recent collections of bluefin tuna larvae in the Slope Sea off northeastern United States have opened questions about how this region contributes to population dynamics. We analyzed larvae collected in the Slope Sea and the Gulf of Mexico in 2016 to estimate larval abundance and growth rates and used a high-resolution regional ocean circulation model to estimate spawning locations and larval transport. We did not detect a regional difference in growth rates, but found that Slope Sea larvae were larger than Gulf of Mexico larvae prior to exogenous feeding. Slope Sea larvae generally backtracked to locations north of Cape Hatteras and would have been retained within the Slope Sea until the early juvenile stage. Overall, our results provide supporting evidence that the Slope Sea is a major spawning ground that is likely to be important for population dynamics. Further study of larvae and spawning adults in the region should be prioritized to support management decisions.Ship time was supported by NOAA, the Bureau of Ocean Energy Management, and the US Navy through interagency agreements for Atlantic Marine Assessment Program for Protected Species (AMAPPS). CMH and JKL received funding from the Woods Hole Oceanographic Institution’s Ocean Life Institute (#13080700) and Academic Programs Office. CMH was additionally supported by the Adelaide and Charles Link Foundation and the J. Seward Johnson Endowment in support of the Woods Hole Oceanographic Institution’s Marine Policy Center. IIR, KC, and JKL were supported by a US National Science Foundation (NSF) grant (OCE-1558806). JKL was additionally supported by the Lenfest Fund for Early Career Scientists and the Early Career Scientist Fund at Woods Hole Oceanographic Institution

Whole-body transcriptome of selectively bred, resistant-, control-, and susceptible-line rainbow trout following experimental challenge with Flavobacterium psychrophilum

Genetic improvement for enhanced disease resistance in fish is an increasingly utilized approach to mitigate endemic infectious disease in aquaculture. In domesticated salmonid populations, large phenotypic variation in disease resistance has been identified but the genetic basis for altered responsiveness remains unclear. We previously reported three generations of selection and phenotypic validation of a bacterial cold water disease (BCWD) resistant line of rainbow trout, designated ARS-Fp-R. This line has higher survival after infection by either standardized laboratory challenge or natural challenge as compared to two reference lines, designated ARS-Fp-C (control) and ARS-Fp-S (susceptible). In this study, we utilized 1.1 g fry from the three genetic lines and performed RNA-seq to measure transcript abundance from the whole body of naive and Flavobacterium psychrophilum infected fish at day 1 (early time-point) and at day 5 post-challenge (onset of mortality). Sequences from 24 libraries were mapped onto the rainbow trout genome reference transcriptome of 46,585 predicted protein coding mRNAs that included 2633 putative immune-relevant gene transcripts. A total of 1884 genes (4.0% genome) exhibited differential transcript abundance between infected and mock-challenged fish (FDR \u3c 0.05) that included chemokines, complement components, tnf receptor superfamily members, interleukins, nod-like receptor family members, and genes involved in metabolism and wound healing. The largest number of differentially expressed genes occurred on day 5 post-infection between naive and challenged ARS-Fp-S line fish correlating with high bacterial load. After excluding the effect of infection, we identified 21 differentially expressed genes between the three genetic lines. In summary, these data indicate global transcriptome differences between genetic lines of naive animals as well as differentially regulated transcriptional responses to infection

Identification of larval sea basses (Centropristis spp.) using ribosomal DNA-specific molecular assays

This paper is not subject to U.S. copyright. The definitive version was published in Fishery Bulletin 106 (2008): 183-193.The identification of sea bass (Centropristis) larvae to species is difficult because of similar morphological characters, spawning times, and overlapping species ranges. Black sea bass (Centropristis striata) is an important fishery species and is currently considered to be overfished south of Cape Hatteras, North Carolina. We describe methods for identifying three species of sea bass larvae using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) assays based on species-specific amplification of rDNA internal transcribed spacer reg ions. The assays were tested against DNA of ten other cooccurring reef fish species to ensure the assay’s specificity. Centropristis larvae were collected on three cruises during cross-shelf transects and were used to validate the assays. Seventysix Centropristis larvae were assayed and 69 (91%) were identified successfully. DNA was not amplified from 5% of the larvae and identification was inconclusive for 3% of the larvae. These assays can be used to identify sea bass eggs and larvae and will help to assess spawning locations, spawning times, and larval dispersal.Collection of larvae at sea was supported by funding from the National Science Foundation through OCE 9876565 to C. Jones, S. Thorrold, A. Valle-Levinson, and J. Hare. Additional funding for this project was provided by Office of National Marine Sanctuaries and by Grays Reef National Marine Sanctuary

A Vulnerability Assessment of Fish and Invertebrates to Climate Change on the Northeast U.S. Continental Shelf

Climate change and decadal variability are impacting marine fish and invertebrate species worldwide and these impacts will continue for the foreseeable future. Quantitative approaches have been developed to examine climate impacts on productivity, abundance, and distribution of various marine fish and invertebrate species. However, it is difficult to apply these approaches to large numbers of species owing to the lack of mechanistic understanding sufficient for quantitative analyses, as well as the lack of scientific infrastructure to support these more detailed studies. Vulnerability assessments provide a framework for evaluating climate impacts over a broad range of species with existing information. These methods combine the exposure of a species to a stressor (climate change and decadal variability) and the sensitivity of species to the stressor. These two components are then combined to estimate an overall vulnerability. Quantitative data are used when available, but qualitative information and expert opinion are used when quantitative data is lacking. Here we conduct a climate vulnerability assessment on 82 fish and invertebrate species in the Northeast U.S. Shelf including exploited, forage, and protected species. We define climate vulnerability as the extent to which abundance or productivity of a species in the region could be impacted by climate change and decadal variability. We find that the overall climate vulnerability is high to very high for approximately half the species assessed; diadromous and benthic invertebrate species exhibit the greatest vulnerability. In addition, the majority of species included in the assessment have a high potential for a change in distribution in response to projected changes in climate. Negative effects of climate change are expected for approximately half of the species assessed, but some species are expected to be positively affected (e.g., increase in productivity or move into the region). These results will inform research and management activities related to understanding and adapting marine fisheries management and conservation to climate change and decadal variability

Proteomic characterization of the acute-phase response of yellow stingrays Urobatis jamaicensis after injection with a Vibrio anguillarum-ordalii bacterin

Systemic inflammatory responses of mammals and bony fish are primarily driven by coordinated up-regulation and down-regulation of plasma acute-phase proteins. Although this general principle is believed to be universal among vertebrates, it remains relatively unexplored in elasmobranchs. The objective of this study was to characterize acute changes in the plasma proteome of three yellow stingrays Urobatis jamaicensis following intraperitoneal injection with a commercial Vibrio bacterin. Changes in plasma protein levels were analyzed immediately prior to vaccination (time 0) and at 24 and 72 h post-injection by isobaric tags for relative and absolute quantitation (iTRAQ 4-plex) using shotgun-based nano liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis and de novo peptide sequencing. Pooled 2D-LC-MS/MS and de novo sequencing data revealed differential expression of 156 distinct plasma proteins between time 0 and at least one post-vaccination time point. Using 1.5-fold change in expression as physiologically significant, 14/156 (9.0%) proteins were upregulated in at least one stingray through at least one experimental timepoint. Upregulated proteins included complement factors, Mx-protein, hemopexin, factor X and prothrombin. Seventy-six of 156 (48.7%) proteins were downregulated in the acute-phase response, including transferrin, apolipoprotein B, heparin cofactor 2, alpha2-macroglobulin, and various growth factors. Other differentially upregulated or downregulated proteins included intracellular, cell binding and structural proteins, proteins involved in physiologic processes, and unknown/hypothetical proteins. Selected bioactive factors are discussed for their putative roles in the elasmobranchs acute-phase response. These findings contribute to our understanding of disease processes in elasmobranchs, immunologic phylogeny in vertebrates, and begin the search for potential biomarkers of disease in these ecologically important fish

Effects of ozone on post-smolt Atlantic salmon (Salmo salar) performance, health, and maturation in freshwater recirculation aquaculture systems

Steroid hormones accumulate in recirculation aquaculture systems (RAS) and may influence the reproductive physiology of farmed fish. Ozone reduces hormone concentrations in freshwater RAS used to rear Atlantic salmon, but its effect on reproductive development is unknown. Accordingly, an 8-month trial was carried out to evaluate the growth, health, and maturation of post-smolt Atlantic salmon (296 ± 4 g initial weight) reared in six replicated freshwater RAS (9.5 m3 total volume) operated with or without ozone (N = 3/treatment). Residual ozone was controlled with an oxidation reduction potential (ORP) of 300–320 mV, and mean water temperature was maintained at 14.7 °C. Atlantic salmon growth was generally faster in ozonated RAS. Salmon from RAS with and without ozone weighed 2156 ± 101 and 1810 ± 15 g, respectively, by the end of the study. Caudal, anal, and pelvic fin damage was greater (P < 0.05) for salmon in ozonated RAS early in the trial but improved thereafter. No statistical differences in gill, skin, and skeletal muscle histopathology were observed between treatments at the end of the study. Waterborne estradiol, testosterone, and 11-ketotestosterone levels were periodically lower (P < 0.05) in ozonated RAS, but maturing salmon were more prevalent in these systems. At the end of the trial, percent maturation of salmon populations reared in RAS with and without ozone was 63 ± 7 and 48 ± 1%, respectively; however, maturity appeared to be related to fish size. Improved water quality was observed in ozonated RAS including reduced dissolved copper, iron, and zinc levels, total heterotrophic bacteria counts, and true color, and increased ultraviolet transmittance, which may have supported improved Atlantic salmon growth. Overall, ozone did not inhibit the onset or prevalence of Atlantic salmon maturation, but significant improvements in water quality and salmon growth performance resulted from its use.publishedVersio

Long-Term Changes in the Distributions of Larval and Adult Fish in the Northeast U.S. Shelf Ecosystem

<div><p>Many studies have documented long-term changes in adult marine fish distributions and linked these changes to climate change and multi-decadal climate variability. Most marine fish, however, have complex life histories with morphologically distinct stages, which use different habitats. Shifts in distribution of one stage may affect the connectivity between life stages and thereby impact population processes including spawning and recruitment. Specifically, many marine fish species have a planktonic larval stage, which lasts from weeks to months. We compared the spatial distribution and seasonal occurrence of larval fish in the Northeast U.S. Shelf Ecosystem to test whether spatial and temporal distributions changed between two decades. Two large-scale ichthyoplankton programs sampled using similar methods and spatial domain each decade. Adult distributions from a long-term bottom trawl survey over the same time period and spatial area were also analyzed using the same analytical framework to compare changes in larval and adult distributions between the two decades. Changes in spatial distribution of larvae occurred for 43% of taxa, with shifts predominately northward (i.e., along-shelf). Timing of larval occurrence shifted for 49% of the larval taxa, with shifts evenly split between occurring earlier and later in the season. Where both larvae and adults of the same species were analyzed, 48% exhibited different shifts between larval and adult stages. Overall, these results demonstrate that larval fish distributions are changing in the ecosystem. The spatial changes are largely consistent with expectations from a changing climate. The temporal changes are more complex, indicating we need a better understanding of reproductive timing of fishes in the ecosystem. These changes may impact population productivity through changes in life history connectivity and recruitment, and add to the accumulating evidence for changes in the Northeast U.S. Shelf Ecosystem with potential to impact fisheries and other ecosystem services.</p></div

<i>A priori</i> expectations and results for change in spatial distribution and seasonal occurrence of larval fish in the Northeast U.S. Shelf Ecosystem.

<p>Based on recent meta-analyses [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137382#pone.0137382.ref034" target="_blank">34</a>], studies from other regions [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137382#pone.0137382.ref021" target="_blank">21</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137382#pone.0137382.ref022" target="_blank">22</a>] and studies in the Northeast U.S. Shelf Ecosystem [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137382#pone.0137382.ref004" target="_blank">4</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137382#pone.0137382.ref032" target="_blank">32</a>], we developed several <i>a priori</i> expectations.</p

Percent frequency of occurrence of directional shifts in distribution among taxa by life stage and regions of the Northeast U.S. Shelf Ecosystem.

<p>Occurrence of each type of distributional shift among larvae (A) and spring- (B) and fall-collected adults (C) were grouped by taxa that occurred primarily in the Mid-Atlantic Bight to Georges Bank (MAB to GB), Southern New England to Gulf of Maine (SNE to GOM), and Georges Bank to Gulf of Maine (GB to GOM) to examine patterns in relation to regional occurrence.</p