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Aquac. J., Volume 4, Issue 2 (June 2024) – 4 articles

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16 pages, 2820 KiB  
Article
A Thorough Insight into the Biofilm Properties of Potential Fish Pathogen Tenacibaculum discolor Strain FMCC B487
by Eirini Schoina, Laetitia Marchand, Agata Zykwinska, Corinne Sinquin, Françoise Leroi, Christine Delbarre-Ladrat and George-John E. Nychas
Aquac. J. 2024, 4(2), 76-91; https://doi.org/10.3390/aquacj4020006 - 11 Jun 2024
Viewed by 805
Abstract
Tenacibaculum discolor develops biofilm in marine aquaculture production tanks and is identified as one of the causative agents of tenacibaculosis, a bacterial disease that causes significant losses in marine aquaculture production. In this study, the biofilm characteristics of T. discolor strain FMCC B487 [...] Read more.
Tenacibaculum discolor develops biofilm in marine aquaculture production tanks and is identified as one of the causative agents of tenacibaculosis, a bacterial disease that causes significant losses in marine aquaculture production. In this study, the biofilm characteristics of T. discolor strain FMCC B487 were evaluated. Cell growth and biofilm formation and development were studied in miniaturized assays to assess the effect of different levels of environmental factors temperature and salinity, as well as the presence of monosaccharides potentially found in aquaculture hatcheries. The ability of the strain to grow and develop strong biofilms in ambient to high temperatures and at salinities above 20 g/L was shown. Mannose was the monosaccharide with the most prominent impact on the T. discolor strain FMCC B487 biofilm. The composition of planktonic cell extract, biofilm extracts, and extracellular polymeric substances (EPS) produced by T. discolor strain FMCC B487 were investigated by means of colorimetric and fluorometric assays as well as analyses by electrophoresis, gas chromatography, and high-performance size-exclusion chromatography coupled with a multiangle light scattering detector, revealing the dominance of proteins and lipids and the absence of high-molecular-weight polysaccharides. This information may serve as a basis for considering anti-biofilm strategies against the pathogen T. discolor. Full article
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<p>Effect of environmental conditions on the growth (<b>a</b>,<b>c</b>) and biofilm formation (<b>b</b>,<b>d</b>) of <span class="html-italic">T. discolor</span> strain FMCC B487 under different salinities (<b>a</b>,<b>b</b>) and temperatures (<b>c</b>,<b>d</b>). OD 590 nm was normalized by dividing by OD 600. Biofilm formation is ranged as absent (-), moderate (++), and high (+++).</p>
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<p>Effect of monosaccharides glucose (Glc), mannose (Man), galactose (Gal), glucosamine (GlcN), and fructose (Fru) at 10, 20, and 30 g/L on the biofilm formation of <span class="html-italic">T. discolor</span> strain FMCC B487. Y axis represents OD 590 nm values from the crystal violet assay. Bars show mean ± SD values. Asterisk (*) denotes significant difference from the control (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Mass yield of prepared extracts. The extract names (ToT, C, Sn, A) are followed by B for biofilm-adhered cells, P for planktonic cells, and G for glucose or M for mannose added to the medium. Blue bars represent planktonic and adhered cell extracts from the biofilm experiments. Green bars stand for cultures in Erlenmeyer flasks. Ethyl acetate extracts were ToT for total extract from the whole broth, and C refers to cell pellets. Sn refers to the broth supernatant; A contains the substances bound to the cell surface that were extracted using NaOH.</p>
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<p>Compositional analysis of planktonic, biofilm, and EPS extracts (<span class="html-italic">w</span>/<span class="html-italic">w</span> %).</p>
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<p>Electrophoretic analyses of <span class="html-italic">T. discolor</span> strain FMCC B487 extracts: agarose gel (<b>a</b>) and PAGE (<b>b</b>) show the presence of medium- and low-molecular-weight polysaccharides with stains all staining, and SDS-PAGE stained with Coomassie Blue (<b>c</b>) shows a variety of proteins (two separate gels). Extracts: supernatants of planktonic (SnP) and adhered cell (SnB), supernatants of cultures in Erlenmeyer in presence of glucose (SnG) or mannose (SnM), and at the cell surface of planktonic (AP) and adhered cell extracts (AB), as well as glucose-supplemented (AG) and mannose-supplemented cultures (AM). PM: protein MW marker (peq GOLD Marker I 116–14.4 kDa); GY785: high-molecular-weight polysaccharide.</p>
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<p>Osidic composition (<span class="html-italic">w</span>/<span class="html-italic">w</span> %) of <span class="html-italic">T. discolor</span> strain FMCC B487 extracts. GalNAc: N-Acetylgalactosamine; GlcNAc: N-Acetylglucosamine; GalA: galacturonic acid; GlcA: Glc: glucose; Gal: galactose; Man: mannose.</p>
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<p>Combined chromatograms of RI (refractive index) and UV profiles from the HPSEC-MALS analysis of SnB and SnM extracts.</p>
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21 pages, 291 KiB  
Article
Issues and Needs for the Sustainable Development of Shellfish Farming in Italy
by Lucia Tudini and Andrea Forgione
Aquac. J. 2024, 4(2), 55-75; https://doi.org/10.3390/aquacj4020005 - 7 Jun 2024
Viewed by 1090
Abstract
The Italian shellfish industry mainly comprises clams, mussels, and oysters. While clam production thrives and Italy leads Europe, mussel farming faces economic challenges. Oyster production is relatively new and holds potential. Sustainable development is crucial for meeting growing seafood demand while ensuring resource [...] Read more.
The Italian shellfish industry mainly comprises clams, mussels, and oysters. While clam production thrives and Italy leads Europe, mussel farming faces economic challenges. Oyster production is relatively new and holds potential. Sustainable development is crucial for meeting growing seafood demand while ensuring resource conservation and food safety. This paper, part of the VALUE-SHELL project funded by the Italian Ministry of Agriculture, Food Sovereignty and Forests (MASAF) as part of support activities for the National Strategic Plan of Aquaculture, combines desk research on industry structure and public interventions using a collaborative approach involving stakeholder interviews and focus groups with producers and local entities to assess the sector’s most compelling needs. The collaborative process highlighted key challenges across the following four sustainability pillars: environmental (global warming, predator control, and pollution mitigation are critical concerns for ecosystem balance); economic (increased production costs and limited diversification opportunities affect profitability); social (fear of generational decline due to limited training and education opportunities poses a threat to the industry’s future); and institutional (lack of a dedicated aquaculture law and fragmented governance hinder long-term planning). The project identifies policy bottlenecks hindering the sector’s potential, caused by not fully addressing producer needs. Sustainable development strategies are essential for addressing these challenges and ensuring the Italian shellfish industry’s future. Full article
11 pages, 1840 KiB  
Article
Sex Determination, Evolution of Gonadal Stage in Females, and Seasonal Evolution of Sperm Production in Chelon labrosus (Risso, 1827)
by Ignacio Eduardo Martín-Montero, Juan Manuel Martínez-Vázquez, Inmaculada Rasines Perez, Miguel Torres, Juan Antonio Martos-Sitcha and Felipe Aguado-Giménez
Aquac. J. 2024, 4(2), 44-54; https://doi.org/10.3390/aquacj4020004 - 4 May 2024
Viewed by 1162
Abstract
Chelon labrosus represents a promising species for current aquaculture to lean towards a more sustainable model. The control of reproduction in captivity is essential to develop the industrial production of new species. This work adds to the knowledge of the reproductive aspects of [...] Read more.
Chelon labrosus represents a promising species for current aquaculture to lean towards a more sustainable model. The control of reproduction in captivity is essential to develop the industrial production of new species. This work adds to the knowledge of the reproductive aspects of males and females of this species. We aim to (1) develop a methodology for sex identification of reproductive and prereproductive specimens of the studied species based on the plasma levels of steroid hormones—estradiol (E2) and 11-ketotestosterone (11-KT); (2) determine the maturity evolution of the females throughout the spawning season by gonadal biopsy; and (3) describe sperm quality throughout the breeding season (volume, motility, density). The results show that an 11-KT/E2 ratio > 0.4 is 100% effective in distinguishing males from females, provided that individuals present a size greater than that of first sexual maturity. A double trend was observed in the development of oocytes, with an increase in the initial diameter until the middle of the spawning season, followed by a divergence of the group, with some females continuing the maturation process and others going into regression. Sperm data show stable quality at the beginning and in the middle of the spawning season and a significant loss of quality at the end of the reproductive period. The present study also shows that, between the beginning and the middle of the reproductive season, males of this species can recover sperm volume removed by stripping. Full article
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<p>Biplot showing the levels of estradiol and 11-ketotestosterone measured in each specimen. Orange circles indicate those identified as females with the proposed ratio 11-KT/E2 &lt; 0.4. Blue diamonds indicate those identified as males. Green triangles correspond to the specimens misidentified as males with the ratio 11-KT/E2 &lt; 0.4.</p>
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<p>The development of oocyte diameter (mean ± SE) in <span class="html-italic">Chelon labrosus</span> females during the 2023 breeding season. n indicates the number of females from which a sample was obtained by gonadal biopsy. The red section of the line indicates the period with all samples increasing in diameter. The blue section indicates the samples that continued to increase the diameter. The yellow section indicates the samples that began to decrease in diameter.</p>
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<p>Mean ± SE at the beginning, middle, and end of the 2023 breeding season for volume, motility, and density of the analyzed samples. The different letters and numbers in superscript over the error bars indicate significant differences for the volume and density, respectively (<span class="html-italic">p</span> &lt; 0.05; paired <span class="html-italic">t</span>-test for related samples). Volume is expressed in 10× mL, motility in %, and density in 10<sup>9</sup> cells per mL.</p>
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<p>Volume, motility, and density values studied at the beginning, in the middle, and at the end of the spawning season for each male. Volume is expressed in 10× mL, motility in %, and density in 10<sup>9</sup> cells per mL.</p>
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<p>Mean ± SE motility loss of <span class="html-italic">C. labrosus</span> sperm over time after activation with seawater 35‰. The dotted line corresponds to the projection of the logarithmic function model associated with motility loss.</p>
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<p>Sperm quality parameters after activation of samples with seawater diluted with ultrapure water in different ratios (35, 26, and 18‰). Percentages are represented in percentage per one and average velocities in µm/sg. The different letters and symbols in superscript over the error bars indicate significant differences in % medium speed cells and the mean velocity, respectively.</p>
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16 pages, 2834 KiB  
Article
Single-Cell Transcriptome Profiling of Scale Drop Disease Virus-Infected Asian Seabass (Lates calcarifer)
by Zhixuan Loh, Ting Wei Lim, Shanshan Wu Howland, Sunita Awate, Laurent Renia, Jinmiao Chen and Ee Chee Ren
Aquac. J. 2024, 4(2), 28-43; https://doi.org/10.3390/aquacj4020003 - 7 Apr 2024
Viewed by 1337
Abstract
The study aims to characterize the immune cell landscape in convalescent Asian seabass (Lates calcarifer) blood samples after exposure to scale-drop disease virus (SDDV). Traditional immunophenotyping approaches used in human and mouse studies are impractical for non-model organisms like the Asian [...] Read more.
The study aims to characterize the immune cell landscape in convalescent Asian seabass (Lates calcarifer) blood samples after exposure to scale-drop disease virus (SDDV). Traditional immunophenotyping approaches used in human and mouse studies are impractical for non-model organisms like the Asian seabass due to the lack of specific antibody-based reagents. To overcome this challenge, 10x Genomics single-cell RNA sequencing was employed. The analysis of blood samples revealed 24 distinct leukocyte clusters, with elevated proportions of B cells, granulocytes, and T cells in the convalescent group compared to the uninfected group. While distinguishing granulocyte and macrophage subsets was challenging, the analysis of differential gene expression in the macrophage population indicated that the upregulated genes were linked to inflammatory processes. Specific T cell clusters showed notable expressions of cd4-1, cd8a, perforin-1 and il-2rβ, suggesting the presence of CD4+ T helper (Th), CD8+ cytotoxic T (Tc) cells, immature T cells, and naive T cells. Attempts to categorize CD4+ T cells into Th subtypes lacked clear distinctions, while CD8+ T cells exhibited three clusters, predominantly Tc1 cells. Furthermore, comparisons between convalescent and uninfected groups revealed increased percentages of activated and antibody-secreting B cells in the convalescent group. This single-cell analysis provides vital insights into the immune cell dynamics in convalescent and uninfected Asian seabass, providing valuable information on potential immune responses to SDDV infection. Full article
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<p>Single-cell RNA sequencing (scRNA-seq) analysis of Asian seabass whole-blood leukocytes. (<b>A</b>) Experimental procedures for the scRNA-seq of leukocytes extracted from the whole blood of Asian seabass, involving cell isolation, 10× Genomics Chromium 3′ scRNA-seq, and subsequent data visualization. (<b>B</b>) The presence of the SDDV MCP gene using the PCR assay. DNA was extracted from plasma. C, convalescent; U, uninfected; N; no template control; P, positive control using plasmid. GeneRuler 50 bp DNA ladder was used to determine the band sizes given in base pairs. A product of approximately 200 bp was amplified from all samples. (<b>C</b>) A collective of 24 cell clusters were identified and represented in a uniform manifold approximation and projection (UMAP) space. (<b>D</b>) Bubble chart depicting the marker genes associated with B cells, T cells, macrophages, granulocytes, dendritic cells (DCs), and erythrocytes across the cell clusters. Dot size corresponds to the percentage of gene expression, while the color intensity reflects the average expression levels of the cells within each cluster. (<b>E</b>) Identification of potential cell types through the expression of marker genes observed in both mammals and fish. (<b>F</b>) UMAP clustering of convalescent and uninfected samples. (<b>G</b>) The numbers of cells (<b>left</b>) and percentage (<b>right</b>) of each cell type in convalescent and uninfected samples.</p>
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<p>Granulocyte diversity in response to SDDV infection. (<b>A</b>) UMAP plot displaying 10 distinct clusters. Clusters labelled from 0 to 9 represent granulocytes with distinct gene signatures identified through the application of the Seurat algorithm. (<b>B</b>) Violin plots showing the expression of specific genes in granulocytes. The y axis represents the normalized and log-transformed average gene expression, while the x axis indicates the granulocytes sub-cluster. (<b>C</b>) Volcano plot illustrating the differentially expressed genes in granulocytes, with each dot representing an individual gene based on both the <span class="html-italic">p</span>-value and fold differences.</p>
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<p>Immune response of monocytes/macrophages against SDDV infection. (<b>A</b>) UMAP plot displaying 11 distinct clusters. Clusters labeled from 0 to 10 represent monocytes/macrophages with distinct gene signatures identified through the application of the Seurat algorithm. (<b>B</b>) Violin plots showing the expression of specific genes in monocytes/macrophages. The y axis represents the normalized and log-transformed average gene expression, while the x axis indicates the granulocytes sub-cluster. (<b>C</b>) Volcano plot illustrating the differentially expressed genes in monocytes/macrophages, with each dot representing an individual gene based on both <span class="html-italic">p</span>-value and fold differences.</p>
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<p>Characterization of T cell subpopulations in SDDV-infected convalescent Asian seabass. (<b>A</b>) The identified T cells were categorized into 7 distinct clusters and visualized with UMAP. Clusters 0, 5, and 6 represent naïve T cells, while clusters 2, 4, and 7 represent immature T cells. Cluster 1 represents CD4+ T helper cells, and cluster 3 represents CD8+ cytotoxic T cells. (<b>B</b>) Violin plots showing the expression of specific genes in T cells. The y axis represents the normalized and log-transformed average gene expression, while the x axis indicates the granulocytes sub-cluster. (<b>C</b>, <b>top</b>) Cluster 2 were sub-categorized into 4 distinct clusters and visualized with UMAP in convalescent and uninfected samples. (<b>C</b>, <b>bottom</b>) Violin plots showing the expression of specific genes in CD4<sup>+</sup> T cells. (<b>D</b>, <b>top</b>) Cluster 3 and cluster 4 were sub-categorized into 3 distinct clusters and visualized with UMAP in convalescent and uninfected samples. (<b>D</b>, <b>bottom</b>) Violin plots showing the expression of specific genes in CD8<sup>+</sup> T cells. (<b>E</b>) Pseudotime trajectory of T cell subsets estimated using Monocle. (<b>F</b>) Proportion of T cell subsets in convalescent and uninfected samples. (<b>G</b>) Volcano plot illustrating the differentially expressed genes in T cells, with each dot representing an individual gene based on both <span class="html-italic">p</span>-value and fold differences.</p>
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<p>Classification of B cells after SDDV infection in Asian seabass. (<b>A</b>) Violin plots showing the expression of specific genes in B cells. The y axis represents the normalized and log-transformed average gene expression, while the x axis indicates the granulocytes sub-cluster. (<b>B</b>) Pseudotime trajectory of B cell subsets estimated using Monocle. (<b>C</b>) Proportion of B cell subsets in convalescent and uninfected samples. (<b>D</b>) Volcano plot illustrating the differentially expressed genes in B cells, with each dot representing an individual gene based on both <span class="html-italic">p</span>-value and fold differences.</p>
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