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15 pages, 1727 KiB  
Article
Oxygen Consumption and Carbon Budget in Groundwater-Obligate and Surface-Dwelling Diacyclops Species (Crustacea Copepoda Cyclopoida) Under Temperature Variability
by Tiziana Di Lorenzo, Agostina Tabilio Di Camillo, Sanda Iepure, Diana M. P. Galassi, Nataša Mori and Tatjana Simčič
Environments 2025, 12(1), 32; https://doi.org/10.3390/environments12010032 - 20 Jan 2025
Viewed by 391
Abstract
This study explores the metabolic response and carbon budget of two cyclopoid copepod species, Diacyclops belgicus Kiefer, 1936 (a stygobitic, groundwater-adapted species) and Diacyclops crassicaudis crassicaudis (Sars G.O., 1863) (a stygophilic, predominantly surface-associated species). We measured oxygen consumption rates (OCRs), carbon requirements (CRs), [...] Read more.
This study explores the metabolic response and carbon budget of two cyclopoid copepod species, Diacyclops belgicus Kiefer, 1936 (a stygobitic, groundwater-adapted species) and Diacyclops crassicaudis crassicaudis (Sars G.O., 1863) (a stygophilic, predominantly surface-associated species). We measured oxygen consumption rates (OCRs), carbon requirements (CRs), ingestion (I) rates, and egestion (E) rates at 14 °C and 17 °C, representing current and predicted future conditions in the collection habitats of the two species. Diacyclops belgicus displayed OCRs (28.15 and 18.32 µL O2/mg DW × h at 14 and 17 °C, respectively) and carbon budget (CR: 0.14 and 0.10 µg C/mg × d at 14 and 17 °C) lower than those of D. crassicaudis crassicaudis (OCR: 55.67 and 47.93 µL O2/mg DW × h at 14 and 17 °C; CR: 0.3 and 0.27 µg C/mg × d at 14 and 17 °C). However, D. belgicus exhibited metabolic rates and carbon requirements comparable to those of other epigean species, challenging the assumption that low metabolic rates are universal among stygobitic species. Temperature variations did not significantly affect the metabolic responses and carbon requirements of the two species, suggesting that they may cope with moderate temperature increases. Full article
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<p>Phraetobiological net used to collect <span class="html-italic">Diacyclops belgicus</span> (<b>a</b>) and <span class="html-italic">Diacyclops crassicaudis crassicaudis</span> (<b>b</b>).</p>
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<p>The respirometric setup for oxygen consumption measurements of CV copepodids of <span class="html-italic">Diacyclops belgicus</span> and <span class="html-italic">Diacyclops crassicaudis crassicaudis</span> at 14 °C and 17 °C. One hour after collection, the specimens were kept in darkness for 21 days, transitioning through three media: 100% bore water (black beaker), a 50% bore and standard water mix (green beaker), and 100% standard water (blue beaker). Individual CV copepodids were placed in 80 μL glass wells with oxygen sensor spots, housed in a microplate, and monitored for oxygen levels over 18 h. At the end of the measurements, the specimens were measured, and their body volume was computed based on body dimensions.</p>
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<p>A heatmap of oxygen consumption rates (OCRs; μL O<sub>2</sub>/mg DW × h) of freshwater cyclopoid species across a temperature gradient: (<b>a</b>) all stages; (<b>b</b>) nauplii; (<b>c</b>) copepodids; (<b>d</b>) adults. Colour intensity indicates OCR magnitude. Mvi: <span class="html-italic">Megacyclops viridis</span> (Jurine, 1820); Mbr: <span class="html-italic">Mesocyclops brasilianus</span> Kiefer, 1933; Ese: <span class="html-italic">Eucyclops serrulatus serrulatus</span> (Fischer, 1851); Eag: <span class="html-italic">Eucyclops agilis agilis</span> (Koch, 1838); Dcr: <span class="html-italic">Diacyclops crassicaudis crassicaudis</span> (Sars G.O., 1863); Dbi: <span class="html-italic">Diacyclops bicuspidatus bicuspidatus</span> (Claus, 1857); Mva: <span class="html-italic">Microcyclops varicans varicans</span> (Sars G.O., 1863); Cvi: <span class="html-italic">Cyclops vicinus vicinus</span> Uljanin, 1875; Dbe: <span class="html-italic">Diacyclops belgicus</span> Kiefer, 1936.</p>
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18 pages, 4648 KiB  
Article
Potential Strategies Applied by Metschnikowia bicuspidata to Survive the Immunity of Its Crustacean Hosts
by Ji Zhang, Bingyu Li, Bingnan Zuo and Xiaodong Li
Pathogens 2025, 14(1), 95; https://doi.org/10.3390/pathogens14010095 - 18 Jan 2025
Viewed by 457
Abstract
Metschnikowia bicuspidata is the specific pathogen for “milky disease” in the Chinese mitten crab (Eriocheir sinensis), accounting for huge losses to the industry. And yet, there is no precise study describing the pathogenesis of M. bicuspidata, largely hindering the development [...] Read more.
Metschnikowia bicuspidata is the specific pathogen for “milky disease” in the Chinese mitten crab (Eriocheir sinensis), accounting for huge losses to the industry. And yet, there is no precise study describing the pathogenesis of M. bicuspidata, largely hindering the development of novel control methods against its causing diseases. Here, we compared the transcriptomes of M. bicuspidata cells collected from a control group (cultured without E. sinensis hemocytes) and a treatment group (cultured with E. sinensis hemocytes), using RNA sequencing. Through comprehensively analyzing the differentially expressed genes (DEGs), both the most regulated ones and the ones involved in crucial enriched KEGG pathways, we found that certain processes might be required for M. bicuspidata’s survival under hemocyte stress. Key genes involved in oxidative phosphorylation, fatty acid metabolism, upper glycolysis, and gluconeogenesis were upregulated, and those for β-glucan unmasking, autophagy, and cell polarity were downregulated, in the treatment group. Our results suggest that M. bicuspidata colonizes and therefore establishes an infection in E. sinensis via enhancing aerobic respiration, glucose-6-phosphate accumulation, and cell-wall masking. In addition, we applied multiple means to evaluate a series of candidate reference genes and found that PMA1 in combination with ACT1 is the most suitable choice for accurate normalization in quantitative real-time PCR (qRT-PCR) assays. Thus, we used this combination as the reference and performed qRT-PCR verification of several DEGs. It is shown that the expression trends of these tested DEGs in qRT-PCR assays are the same as those in RNA-Seq assays. This study not only provides insights into strategies facilitating M. bicuspidata’s survival within E. sinensis, initially elucidating the pathogenesis of this yeast, but also recommends a useful molecular tool regarding qRT-PCR assays in this pathogen. Full article
(This article belongs to the Section Fungal Pathogens)
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<p>Initial analyses of DEGs according to the RNA-Seq data. (<b>A</b>) A volcano plot showing the number, distribution, and difference significance of DEGs. The gene-expression difference and its statistical significance was measured as Log<sub>2</sub>FC and -Log<sub>10</sub>FDR, respectively. Red dots, upregulated DEGs; green dots, downregulated DEGs; black dots, unchanged genes. (<b>B</b>,<b>C</b>) are charts showing KEGG pathways enriched among the up- and downregulated DEGs, respectively. The sizes of the triangles represent the number of DEGs, and the <span class="html-italic">p</span> values were calculated for enrichment significance.</p>
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<p>Evaluation of the candidate reference genes. (<b>A</b>) The electrophoresis result for the respective PCR amplifications of <span class="html-italic">18S</span>, <span class="html-italic">ACT1</span>, <span class="html-italic">GAPDH</span>, <span class="html-italic">RIP</span>, <span class="html-italic">LSC2</span>, <span class="html-italic">PMA1</span>, and <span class="html-italic">TAF10</span> (shown in Lane 2–Lane 8). Lane 1, DNA ladder. (<b>B</b>) A boxplot showing the expression levels of the seven candidate reference genes in all samples (see Materials and Methods <a href="#sec2dot3-pathogens-14-00095" class="html-sec">Section 2.3</a>) by qRT-PCR. Gene expression levels were measured as Ct values. (<b>C</b>) A chart showing the M values calculated for the candidate genes via geNorm, and the stability of these genes was sorted as indicated. (<b>D</b>) A chart showing the pairwise variation (V) values calculated for different V<span class="html-italic"><sub>n</sub></span>/V<span class="html-italic"><sub>n</sub></span><sub>+1</sub> combinations as indicated.</p>
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<p>Verification of the expression patterns of certain DEGs via qRT-PCR. (<b>A</b>) A chart showing the relative expression levels of genes as indicated, in assays of qRT-PCR (gray bar) and RNA-Seq (black bar). The relative mRNA abundance (fold change, FC) for each gene in qRT-PCR was measured as described in <a href="#sec2dot6-pathogens-14-00095" class="html-sec">Section 2.6</a> (see Materials and Methods), and the Log<sub>2</sub>FC values were calculated as mean ± SEM. (<b>B</b>) The fold change in the expression of these DEGs in qRT-PCR was plotted against that in RNA-Seq, and shown as a scatter chart. A trendline for these scatters was drawn and its R<sup>2</sup> was calculated.</p>
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<p>A hypothesized regulatory network based on the transcriptomic analysis and literature review. I–V, the complex I–V in the electron transport chain; words in green, genes (gene products) or pathways that are upregulated in the presence of crab hemocytes according to the RNA-Seq results; words in red, genes (gene products) or pathways that are downregulated; words in purple, genes (gene products) that can either be upregulated or downregulated; red cross, process inhibition; S/F transporter, succinate/fumarate mitochondrial transporter; HXT, hexose transporter. This cartoon was created in BioRender (<a href="https://BioRender.com/p31n089" target="_blank">https://BioRender.com/p31n089</a>, accessed on 5 November 2024).</p>
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19 pages, 1653 KiB  
Article
Bioactive Molecules from the Invasive Blue Crab Callinectes sapidus Exoskeleton: Evaluation of Reducing, Radical Scavenging, and Antitumor Activities
by Francesco Longo, Alessandro Attanzio, Laura Marretta, Claudio Luparello, Serena Indelicato, David Bongiorno, Giampaolo Barone, Luisa Tesoriere, Ilenia Concetta Giardina, Giulia Abruscato, Manuela Perlotti, Lucie Branwen Hornsby, Vincenzo Arizza, Mirella Vazzana, Aiti Vizzini, Chiara Martino, Angelica Listro, Vinicius Queiroz, Antonio Fabbrizio, Paolo Salvatore Francesco Ciaccio, Stella Maria Cascioferro, Francesca Di Gaudio and Manuela Mauroadd Show full author list remove Hide full author list
Mar. Drugs 2025, 23(1), 45; https://doi.org/10.3390/md23010045 - 17 Jan 2025
Viewed by 444
Abstract
In recent years, the invasive Atlantic blue crab (Callinectes sapidus) has increased its spread throughout the Mediterranean Sea, threatening native biodiversity and local economies. This study aimed to valorize C. sapidus sampled in Sicily by utilizing its exoskeleton as a source [...] Read more.
In recent years, the invasive Atlantic blue crab (Callinectes sapidus) has increased its spread throughout the Mediterranean Sea, threatening native biodiversity and local economies. This study aimed to valorize C. sapidus sampled in Sicily by utilizing its exoskeleton as a source of chitosan, astaxanthin, and bio-phenolic compounds. These biomolecules were evaluated for their reducing, radical scavenging, and antitumor activity. The ferric ion reducing antioxidant power (FRAP) and the free radical scavenging activity against radical 2,2-Diphenyl-1-picrylhydrazyl (DPPH) were significantly higher for chitosan (3.16 ± 0.10 mg AAE/g and 8.1 ± 0.10 µmol TE/g). No significant differences were observed among the tested biomolecules in their activity in scavenging the radical 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). Both bio-phenolic compounds and astaxanthin exhibited dose-dependent cytotoxicity on CaCo-2 (IC50 = 12.47 and 18 µg/mL) and HepG2 (IC50 = 10.25 and 1.26 µg/mL) cell lines, while only bio-phenols showed no cytotoxic effect on differentiated CaCo-2 cells up to 20 µg/mL. These findings highlight the value of blue crab by-products in supporting a circular economy, offering a sustainable approach to managing this invasive species while providing bioactive compounds with promising medical and nutraceutical applications. Full article
(This article belongs to the Special Issue Marine Drug Research in Italy)
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<p>FT-IR spectrum of chitosan obtained from <span class="html-italic">C. sapidus</span> exoskeleton.</p>
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<p>Spectrum of astaxanthin of <span class="html-italic">C. sapidus</span> (<b>A</b>) and spectrum reported on the MZCloud database (<b>B</b>). The green line represents <span class="html-italic">m</span>/<span class="html-italic">z</span> ratio of [M+H]<sup>+</sup> astaxanthin precursor ion on database.</p>
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<p>Reducing (<b>A</b>) and radical scavenging activity (<b>B</b>,<b>C</b>) of chitosan, astaxanthin, and phenolic extract of <span class="html-italic">C. sapidus</span> exoskeleton. AAE: acid ascorbic equivalent; TE; trolox equivalent. Values are expressed as a mean ± SD. Bars within the same graph with different letters are significantly different for <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Dose-response effect of polyphenolic extract (<b>A</b>) and astaxanthin (<b>B</b>) from <span class="html-italic">C. sapidus</span> exoskeleton on the viability of HepG2 and CaCo-2 cells, both tumoral and differentiated, after 24 h of exposure. The error bars correspond to the standard error of the mean (s.e.m.) of three independent measurements. * <span class="html-italic">p</span> &lt; 0.05.</p>
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18 pages, 2085 KiB  
Article
Crustacean Zooplankton Ingestion of Potentially Toxic Microcystis: In Situ Estimation Using mcyE Gene Gut Content Detection in a Large Temperate Eutrophic Lake
by Helen Agasild, Margarita Esmeralda Gonzales Ferraz, Madli Saat, Priit Zingel, Kai Piirsoo, Kätlin Blank, Veljo Kisand, Tiina Nõges and Kristel Panksep
Toxins 2025, 17(1), 42; https://doi.org/10.3390/toxins17010042 - 16 Jan 2025
Viewed by 395
Abstract
Grazing by zooplankton can regulate bloom-forming cyanobacteria but can also transfer toxin-producing cells, as well as toxic metabolites, to the food web. While laboratory investigations have provided extensive knowledge on zooplankton and toxic cyanobacteria interactions, information on zooplankton feeding on toxin-producing cyanobacteria in [...] Read more.
Grazing by zooplankton can regulate bloom-forming cyanobacteria but can also transfer toxin-producing cells, as well as toxic metabolites, to the food web. While laboratory investigations have provided extensive knowledge on zooplankton and toxic cyanobacteria interactions, information on zooplankton feeding on toxin-producing cyanobacteria in natural water bodies remains scarce. In this study, we quantified Microcystis-specific mcyE synthase genes from the gut contents of various cladoceran and copepod taxa to assess the in situ crustacean community and taxon-specific ingestion of potentially toxic Microcystis in Lake Peipsi, a large eutrophic lake in Estonia, Northern Europe. Microcystis cells with mcyE genes were found in all crustaceans examined. However, some species, such as the cyclopoid copepod Mesocyclops leuckarti, were more efficient in ingesting potentially toxic Microcystis than other co-occurring cladocerans (Daphnia spp., Bosmina spp., Chydorus sphaericus) and copepods (Eudiaptomus gracilis). The amount of toxigenic Microcystis cells grazed by crustacean population changed temporarily, and copepods were the predominant consumers of toxigenic Microcystis during several months of the 5-month study period. Crustacean ingestion of toxigenic Microcystis was not related to Microcystis biomass or mcyE gene copy numbers in the environment but was instead related to the abundance of major crustacean grazers. Our findings emphasize the close interaction between crustacean zooplankton and toxigenic Microcystis, indicating that some species may play a more significant role in linking toxic cells within the food web than others. Full article
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<p>Location of sampling sites in Peipsi <span class="html-italic">sensu lato</span> (<span class="html-italic">s.l.</span>): P11 and P38 in Peipsi <span class="html-italic">sensu stricto</span> (<span class="html-italic">s.s.</span>) and P17 in Lämmijärv.</p>
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<p>Phytoplankton composition and biomass (<b>A</b>); seasonal dynamics of biomasses of <span class="html-italic">Microcystis</span> species and <span class="html-italic">Microcystis mcyE</span> copy numbers (±SD) (<b>B</b>) in sampling sites P11, P38, and P17 in Lake Peipsi in 2021.</p>
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<p>Seasonal dynamics of abundance (<b>A</b>) and biomass (<b>B</b>) of major crustacean taxa in sampling sites P11, P38, and P17 in Lake Peipsi in 2021.</p>
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<p>Seasonal dynamics of crustacean population feeding on potentially toxic <span class="html-italic">Microcystis</span> cells (based on the detection of <span class="html-italic">mcyE</span>-containing cells in consumer’ guts) in sampling sites P11, P38, and P17 in Lake Peipsi in 2021; ingestion by various cladoceran and copepod taxa (<b>A</b>); proportional contribution (%) of cladoceran and copepod ingestion (<b>B</b>).</p>
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<p>Seasonal dynamics of crustacean population feeding on potentially toxic <span class="html-italic">Microcystis</span> cells (based on the detection of <span class="html-italic">mcyE</span>-containing cells in consumer’ guts) in sampling sites P11, P38, and P17 in Lake Peipsi in 2021; ingestion by various cladoceran and copepod taxa (<b>A</b>); proportional contribution (%) of cladoceran and copepod ingestion (<b>B</b>).</p>
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<p>Principal component analysis plot displaying the association between cladoceran, copepod, and total crustacean population ingestion (<span class="html-italic">mcyE</span> cell/L), <span class="html-italic">Microcystis</span> biomass, <span class="html-italic">McyE</span> copy numbers, and environmental variables in Lake Peipsi in 2021 grouped by month.</p>
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26 pages, 1169 KiB  
Review
Recent Advances of Processing and Detection Techniques on Crustacean Allergens: A Review
by Xin Qu, Zekun Ma, Xuli Wu and Liangtao Lv
Foods 2025, 14(2), 285; https://doi.org/10.3390/foods14020285 - 16 Jan 2025
Viewed by 284
Abstract
Crustaceans are delicious and highly nutritional food. However, crustaceans are one of the main food allergens, causing severe public health issues. Thus, it is important to increase the knowledge on crustacean allergens and protect the health of sensitized individuals. This review systematically summarizes [...] Read more.
Crustaceans are delicious and highly nutritional food. However, crustaceans are one of the main food allergens, causing severe public health issues. Thus, it is important to increase the knowledge on crustacean allergens and protect the health of sensitized individuals. This review systematically summarizes the basic information on major crustacean allergens’ characteristics, structures, and function. It also summarizes the latest evaluation and detection methods of crustacean allergens. In addition, various processing techniques to alleviate crustacean’s allergenicity are discussed and compared. A host of multiplex approaches as innovative research is attractive to decrease crustacean allergenicity. In addition, the strategies to address the risk of crustacean allergens are also reviewed and discussed in detail. This review provides updates and new findings on crustacean allergens, which helps better understand crustacean allergy and provide novel strategies for its prevention and management. Full article
(This article belongs to the Special Issue Food Allergenic Risk Assessment: State of the Art and Challenges)
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<p>A schematic presentation of common crustaceans.</p>
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<p>Three-dimensional structural models based on existing crystal structures (<a href="https://swissmodel.expasy.org/" target="_blank">https://swissmodel.expasy.org/</a>) for major crustacean allergens.</p>
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<p>The detection of crustacean allergens by enzyme-linked immunosorbent assay. (<b>A</b>) Biosensor technology; (<b>B</b>) SPR: Surface plasmon resonance; SELEX: Systematic evolution of ligands by exponential enrichment; AuNP: Gold nanoparticle; NB: Nanomagnetic bead; FMB: Fluorescent magnetic bead.</p>
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<p>(<b>A</b>) The mechanism of food allergy and the changes in the allergenicity by different methods on crustacean allergens; (<b>B</b>) Classification of reducing crustacean allergenicity using physical, chemical, and microbial methods.</p>
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<p>(<b>A</b>) The mechanism of food allergy and the changes in the allergenicity by different methods on crustacean allergens; (<b>B</b>) Classification of reducing crustacean allergenicity using physical, chemical, and microbial methods.</p>
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17 pages, 3883 KiB  
Article
Molecular Mechanisms Underlying Substance Transport, Signal Transduction, and Anti-Stress Regulation, as Well as Anti-Alkaline Regulation via Bursicon in the Cerebral Ganglion of Chinese Mitten Crab Eriocheir sinensis Under Alkaline Stress
by Meiyao Wang, Jun Zhou, Jiachun Ge, Gangchun Xu and Yongkai Tang
Biology 2025, 14(1), 84; https://doi.org/10.3390/biology14010084 - 16 Jan 2025
Viewed by 515
Abstract
(1) Background: Global climate change is intensifying, and the vigorous development and utilization of saline–alkali land is of great significance. As an important economic aquatic species in the context of saline–alkali aquaculture, it is highly significant to explore the regulatory mechanisms of Eriocheir [...] Read more.
(1) Background: Global climate change is intensifying, and the vigorous development and utilization of saline–alkali land is of great significance. As an important economic aquatic species in the context of saline–alkali aquaculture, it is highly significant to explore the regulatory mechanisms of Eriocheir sinensis under alkaline conditions. In particular, the brain (cerebral ganglion for crustaceans) serves as a vital regulatory organ in response to environmental stress; (2) Methods: In this study, a comparative transcriptome approach was employed to investigate the key regulatory genes and molecular regulatory mechanisms in the cerebral ganglion of E. sinensis under alkaline stress. (3) Results: The results demonstrated that the cerebral ganglion of E. sinensis exhibited a positive response to acute alkaline stress. Pathways associated with signal transduction and substance transportation, such as “phagosome” and “regulation of actin cytoskeleton”, along with regulatory genes involved in antioxidation, were upregulated synergistically to maintain homeostasis under alkaline stress. Furthermore, it was discovered for the first time that bursicon plays a positive regulatory role in the adaptation of E. sinensis to alkalinity. (4) Conclusions: The present study elucidates the molecular regulatory pattern of the cerebral ganglion in E. sinensis under acute alkaline stress as well as revealing a novel role of bursicon in facilitating adaptation to alkalinity in E. sinensis, providing valuable theoretical insights into the molecular regulatory mechanisms underlying the responses of cerebral ganglia to saline–alkali environments. These findings also offer a theoretical reference for promoting the sustainable development of the E. sinensis breeding industry under saline–alkali conditions. Full article
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<p>BUSCO assessment.</p>
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<p>Differential expression enrichment analysis on the top 30 GO terms.</p>
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<p>Top 10 KEGG analysis. The results were shown in four circles from outside to inside. First circle: Classifications of top 20 pathways. Coordinatometer indicated numbers of DMs. Second circle: Number of DMs and <span class="html-italic">p</span>-value of pathways. Longer bars indicate more DMs. Third circle: the proportion of upregulated and downregulated DMs; red represents upregulation and blue represents downregulation. Fourth circle: Rich factor for the top 20 pathways. Each gray auxiliary line represents an increment of 0.2.</p>
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<p>qPCR validation. Letters “A” and “B” represent two functional gene clusters of DEGs. “A” represents “substance transport and signal transduction” and “B” represents “anti-stress and antioxidant response”. “C” represents the control group and “E” represents the experimental group. “*” indicated significant difference, <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Pearson correlation analysis. The dots represented the detected DEGs.</p>
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<p>Sequence and phylogenetic analysis of <span class="html-italic">bursicon</span>-α in <span class="html-italic">E. sinensis</span>. (<b>A</b>) The multiple alignment of <span class="html-italic">bursicon</span>-α sequences. The conserved amino acid residues were highlighted in colored shadow. Species names were abbreviated as follows: EsBursA, <span class="html-italic">Eriocheir sinensis bursicon</span>-α; Cm, <span class="html-italic">Carcinus maenas</span>; Ca, <span class="html-italic">Callinectes arcuatus</span>; Cs, <span class="html-italic">Callinectes sapidus</span>; Pc, <span class="html-italic">Procambarus clarkii</span>; Hg, <span class="html-italic">Homarus gammarus</span>; Dm, <span class="html-italic">Drosophila mojavensis</span>; Da, <span class="html-italic">Drosophila arizonae</span>; Sp, <span class="html-italic">Schistocerca piceifrons</span>. (<b>B</b>) Analysis of motifs of reported <span class="html-italic">bursicon</span>-α sequences. Different motifs were identified in distinct colors. (<b>C</b>) phylogenetic tree of <span class="html-italic">bursicon</span>-α. The phylogenetic tree was constructed with the NJ (neighbor-joining) method. The values at the nodes indicate the bootstrap percent when bootstrap replications reach 1000.</p>
Full article ">Figure 6 Cont.
<p>Sequence and phylogenetic analysis of <span class="html-italic">bursicon</span>-α in <span class="html-italic">E. sinensis</span>. (<b>A</b>) The multiple alignment of <span class="html-italic">bursicon</span>-α sequences. The conserved amino acid residues were highlighted in colored shadow. Species names were abbreviated as follows: EsBursA, <span class="html-italic">Eriocheir sinensis bursicon</span>-α; Cm, <span class="html-italic">Carcinus maenas</span>; Ca, <span class="html-italic">Callinectes arcuatus</span>; Cs, <span class="html-italic">Callinectes sapidus</span>; Pc, <span class="html-italic">Procambarus clarkii</span>; Hg, <span class="html-italic">Homarus gammarus</span>; Dm, <span class="html-italic">Drosophila mojavensis</span>; Da, <span class="html-italic">Drosophila arizonae</span>; Sp, <span class="html-italic">Schistocerca piceifrons</span>. (<b>B</b>) Analysis of motifs of reported <span class="html-italic">bursicon</span>-α sequences. Different motifs were identified in distinct colors. (<b>C</b>) phylogenetic tree of <span class="html-italic">bursicon</span>-α. The phylogenetic tree was constructed with the NJ (neighbor-joining) method. The values at the nodes indicate the bootstrap percent when bootstrap replications reach 1000.</p>
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<p>Sequence and phylogenetic analysis of <span class="html-italic">bursicon</span>-β in <span class="html-italic">E. sinensis</span>. (<b>A</b>) The multiple alignment of <span class="html-italic">bursicon</span>-β sequences. The conserved amino acid residues were highlighted in colored shadow. Species names were abbreviated as follows: EsBursA, <span class="html-italic">Eriocheir sinensis bursicon</span>-α; Cm, Carcinus maenas; Ca, <span class="html-italic">Callinectes arcuatus</span>; Cs, <span class="html-italic">Callinectes sapidus</span>; Pc, <span class="html-italic">Procambarus clarkii</span>; Hg, <span class="html-italic">Homarus gammarus</span>; Dm, <span class="html-italic">Drosophila mojavensis</span>; Da, <span class="html-italic">Drosophila arizonae</span>; Sp, <span class="html-italic">Schistocerca piceifrons</span>. (<b>B</b>) Analysis of motifs of reported <span class="html-italic">bursicon</span>-β sequences. Different motifs were represented in distinct colors. (<b>C</b>) Phylogenetic tree of <span class="html-italic">bursicon</span>-β. The phylogenetic tree was constructed with the NJ (neighbor-joining) method. The values at the nodes indicate the bootstrap percent when bootstrap replications reach 1000.</p>
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<p>Sequence and phylogenetic analysis of <span class="html-italic">bursicon</span>-β in <span class="html-italic">E. sinensis</span>. (<b>A</b>) The multiple alignment of <span class="html-italic">bursicon</span>-β sequences. The conserved amino acid residues were highlighted in colored shadow. Species names were abbreviated as follows: EsBursA, <span class="html-italic">Eriocheir sinensis bursicon</span>-α; Cm, Carcinus maenas; Ca, <span class="html-italic">Callinectes arcuatus</span>; Cs, <span class="html-italic">Callinectes sapidus</span>; Pc, <span class="html-italic">Procambarus clarkii</span>; Hg, <span class="html-italic">Homarus gammarus</span>; Dm, <span class="html-italic">Drosophila mojavensis</span>; Da, <span class="html-italic">Drosophila arizonae</span>; Sp, <span class="html-italic">Schistocerca piceifrons</span>. (<b>B</b>) Analysis of motifs of reported <span class="html-italic">bursicon</span>-β sequences. Different motifs were represented in distinct colors. (<b>C</b>) Phylogenetic tree of <span class="html-italic">bursicon</span>-β. The phylogenetic tree was constructed with the NJ (neighbor-joining) method. The values at the nodes indicate the bootstrap percent when bootstrap replications reach 1000.</p>
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<p>Schematic diagram of the regulatory pattern on the cerebral ganglion of <span class="html-italic">E. sinensis</span> under acute alkalinity stress.</p>
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16 pages, 8029 KiB  
Article
A Vermetid Bioconstruction at the Adriatic Coast of Apulia (Italy)
by Maria Mercurio, Isabella Coccia, Manuel Marra, Tamara Lazic, Giuseppe Corriero and Maria Flavia Gravina
Diversity 2025, 17(1), 49; https://doi.org/10.3390/d17010049 - 14 Jan 2025
Viewed by 381
Abstract
This study presents the first comprehensive data on a vermetid formation along the Apulian coast of the Adriatic Sea, representing one of the northernmost records in the Mediterranean. Surveys along the Brindisi coastline employed visual inspection to map the bioconstruction’s distribution and extension. [...] Read more.
This study presents the first comprehensive data on a vermetid formation along the Apulian coast of the Adriatic Sea, representing one of the northernmost records in the Mediterranean. Surveys along the Brindisi coastline employed visual inspection to map the bioconstruction’s distribution and extension. Detailed data on the bioconstruction inner and the outer edge length, thickness, width, slope and topographic complexity were collected at three selected sites. Moreover, photographic replicates were used to assess shell aperture density and diameters of Dendropoma sp. Associated fauna was studied using two quantitative sampling squares in each transect. The results showed that the vermetid bioconstruction consisted of a thin, encrusted monolayer (thickness < 1.5 cm) that extended for 3.273 linear kilometers, covering 17.23% of the investigated area; it had an average width of 0.5 m, with a mean density of Dendropoma sp. at 2.52 ind/cm2. The associated fauna was composed of 47 taxa dominated by crustaceans, mollusks and annelids. Species richness was correlated with the bioconstruction’s thickness and complexity. These findings underline the ecological importance of vermetid bioconstructions as biodiversity hotspots. The lack of massive mortality events along the Apulian coast, in contrast to other Mediterranean vermetid bioconstructions, underscores the necessity for targeted conservation measures. Full article
(This article belongs to the Special Issue Biodiversity and Ecology in the Mediterranean Sea)
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<p>The study area divided in three sites: (i) “RSM site 1” located between Rosa Marina and Villanova; (ii) “TPZ site 2” located between Villanova and Torre Pozzelle; and (iii) “SPC site 3” located between Torre Pozzelle and Specchiolla. The location of the survey transects is highlighted.</p>
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<p>(<b>a</b>) Coast’s profile in the investigated area; (<b>b</b>) detail of the vermetid bioconstruction in the studied area.</p>
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<p>Example of the monitoring sheet at the Villanova–Torre Pozzelle site (TPZ site 2).</p>
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<p>Surface area occupied by the vermetid bioconstruction in each quadrant, in both the first (<b>a</b>) and second (<b>b</b>) parts of the investigated area. The colors represent different ranges of coverage values as indicated in the legend.</p>
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<p>Length of the inner and outer edge (m) and average thickness (mm ± SD) of the vermetid bioconstruction in each transect.</p>
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<p>Average density values (±SD) of <span class="html-italic">Dendropoma</span> sp. (N<sub>ind</sub>/cm<sup>2</sup>) and topographic complexity (Xn/Xi) of the vermetid bioconstruction in each transect.</p>
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<p>Frequency distribution (%) of <span class="html-italic">Dendropoma</span> sp. shell diameters.</p>
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<p>Percentage composition of taxa (number of individuals) at the three sampling sites.</p>
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<p>Dendrogram from hierarchical cluster analysis based on abundance data (Bray–Curtis similarity index).</p>
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<p>Taxonomic richness (N.sp. = number of spieces) (<b>a</b>) and abundance (<b>b</b>), Shannon diversity (<b>c</b>) and Pielou evenness (<b>d</b>) (mean values ± standard deviation, SD) and at each sampling site.</p>
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16 pages, 13773 KiB  
Article
Clinical and Immunological Features in Limpet (Patella sp.) Allergy in Subtropical Areas: A New Trigger for Food Anaphylaxis
by Elena Mederos-Luis, Tania Galán, María J. Martínez, Ruperto González-Pérez, Inmaculada Sánchez-Machín, Fernando Pineda and Paloma Poza-Guedes
Allergies 2025, 5(1), 2; https://doi.org/10.3390/allergies5010002 - 10 Jan 2025
Viewed by 361
Abstract
Seafood is a crucial source of nutrients, with global consumption steadily increasing. Among seafood-related allergies, shellfish are a significant cause of food allergy and anaphylaxis worldwide, affecting approximately 0.5–2.5% of the general population. While the majority of existing research has focused on crustaceans, [...] Read more.
Seafood is a crucial source of nutrients, with global consumption steadily increasing. Among seafood-related allergies, shellfish are a significant cause of food allergy and anaphylaxis worldwide, affecting approximately 0.5–2.5% of the general population. While the majority of existing research has focused on crustaceans, allergic reactions to mollusks, including their clinical characteristics, remain poorly understood. In the Canary Islands, limpets (a type of marine gastropod) are widely consumed as part of the traditional cuisine. Despite isolated reports of limpet allergy, no large-scale studies or comprehensive clinical analyses have been published on this topic. A cohort of patients sensitized to limpets was analyzed: 66 patients were monosensitized to limpets (Group A), while 64 patients demonstrated additional sensitization to other shellfish (Group B). Limpet ingestion was associated with delayed and severe symptoms, including anaphylaxis and severe asthma. Notably, only 11.5% of patients in Group A tested positive for shellfish allergens using ALEX testing compared to 67.9% in Group B. The identification of protein bands in the 25–40 and 50–200 kDa molecular weight ranges in monosensitized patients provides a novel finding that differentiates this study from prior research. Our study represents the largest reported series of patients with documented limpet allergy to date. Full article
(This article belongs to the Section Food Allergy)
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<p>Summary of respiratory symptoms following ingestion of limpets in patients without previous diagnoses of asthma. SBE: severe bronchospasm; others: urticaria, angioedema, rinoconjuncthivitis, etc.</p>
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<p>SDS-PAGE (<b>1</b>) and IgE Western blot (<b>2</b>) under reducing conditions with the sera pool of all patients from Group A and Group B relative to limpet extracts and with the individual sera (<b>3</b>) of 16 random patients from Group A and Group B relative to allergens of raw and cooked black limpets (<span class="html-italic">Patella crenata</span>), raw and cooked white limpets (<span class="html-italic">Patella aspera</span>), and cooked shrimp extract.</p>
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15 pages, 1557 KiB  
Article
Whole-Genome Sequencing Reveals the Progress of Genetic Breeding in Eriocheir sinensis
by Xuanpeng Wang, Gaowei Zhang, Dandan Gao, Yongchun Ge, Yongxu Cheng and Xinhai Wang
Animals 2025, 15(1), 77; https://doi.org/10.3390/ani15010077 - 1 Jan 2025
Viewed by 675
Abstract
Eriocheir sinensis is an important and popular crustacean species in China, producing huge economic benefits. Large individuals of E. sinensis are preferred due to market demand. The long-term goal of our research group is to produce a new variety of E. sinensis with [...] Read more.
Eriocheir sinensis is an important and popular crustacean species in China, producing huge economic benefits. Large individuals of E. sinensis are preferred due to market demand. The long-term goal of our research group is to produce a new variety of E. sinensis with better growth performance and stronger abilities to resist environmental changes through mass selection. The present study aimed to evaluate the progress of the genetic breeding of E. sinensis by analyzing the genetic diversity and genetic distance between the basic breeding population (pooled population of Suqian and Yixing, G0) and generation 1 (G1) using whole-genome sequencing (WGS). The growth traits, including body weight, shell length, shell width, and third appendage length, in the G1 generation increased by 8.3%, 7.9%, 9.6%, and 9.3%, respectively, compared with those of the G0 generation, indicating that the G1 generation showed better growth performance. A total of 372,448,393 high-quality single nucleotide polymorphisms (SNPs) were detected in 40 E. sinensis individuals, with an average of 9,331,209.83 SNPs. The fixation index values were 0.007 between the Yixing and Suqian populations and 0.015 between the G0 generation and G1 generation, indicating a close genetic background between these groups, especially when considered in combination with the phylogenetic tree and principal component analysis. All of these data suggest that genetic information was stably inherited by the G1 generation, with no introduction of foreign genetic information during the genetic breeding process. In addition, the genetic diversity analysis revealed that the G0 and G1 generations showed a high level of genetic diversity and a relatively stable genetic structure. The present study evaluated the recent progress of the genetic improvement of E. sinensis by our research group, providing valuable evidence for further genetic improvement in this species. Future studies will be performed to select growth-related SNPs and genes through genome-wide association studies. Full article
(This article belongs to the Section Aquatic Animals)
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<p>Sampling map of <span class="html-italic">E. sinensis</span> in the present study. SQ indicates the Suqian population. YX indicates the Yixing population.</p>
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<p>Linkage disequilibrium and population structure of 40 individuals. (<b>A</b>) Linkage disequilibrium analysis for the G0 and G1 generations. (<b>B</b>) Phylogenetic tree analysis for 40 individuals. (<b>C</b>) Principal component analysis for 40 individuals.</p>
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21 pages, 5596 KiB  
Article
Impact of Soybean Bioactive Peptides on Growth, Lipid Metabolism, Antioxidant Ability, Molecular Responses, and Gut Microbiota of Oriental River Prawn (Macrobrachium nipponense) Fed with a Low-Fishmeal Diet
by Chang Yang, Bo Liu, Liangkun Pan, Dong Xia, Cunxin Sun, Xiaochuan Zheng, Peng Chen, He Hu and Qunlan Zhou
Biology 2025, 14(1), 11; https://doi.org/10.3390/biology14010011 - 26 Dec 2024
Viewed by 499
Abstract
The substitution of fishmeal with high-level soybean meal in the diet of crustaceans usually induces lipid accumulation and oxidative stress in the hepatopancreas. Therefore, it is essential to alleviate these adverse effects. In the present study, SBPs were used to alleviate the negative [...] Read more.
The substitution of fishmeal with high-level soybean meal in the diet of crustaceans usually induces lipid accumulation and oxidative stress in the hepatopancreas. Therefore, it is essential to alleviate these adverse effects. In the present study, SBPs were used to alleviate the negative effects of a fishmeal decrease on the growth performance, lipid metabolism, antioxidant capacity, and gut microbiota of oriental river prawn (Macrobrachium nipponense) in an 8-week feeding trial. Three isonitrogenic and isolipidic diets were prepared as follows: R (reference diet with 32% fishmeal), CT (control diet with 22% fishmeal), and SBP (22% fishmeal with 1.25 g/kg soybean bioactive peptides). The prawns (initial biomass per tank 17 g) were randomly divided into three groups with four replicates. The results showed that the low-fishmeal diet induced the following: (1) the inhibition of growth performance and survival of prawns; (2) an increase in triglyceride content in the hepatopancreas and hemolymph and downregulation of carnitine palmitoyl transferase 1 (cpt1) gene expression; (3) a reduction in antioxidant enzymes’ activities and their genes expression levels and an increase malondialdehyde (MDA) content; and (4) an increase in the abundance of the conditional pathogen Pseudomonas in the gut. SBPs supplementation in the CT diet effectively alleviated most of the above adverse effects. SBPs enhanced inducible nitric oxide synthase (iNOS) activity to synthesize nitric oxide (NO) by activating the imd-relish pathway. Most importantly, SBPs increased the potential probiotic Rikenellaceae_RC9_gut_group abundance and decreased the abundance of the conditional pathogen Pseudomonas in the gut. In conclusion, SBPs supplementation can improve low-fishmeal-diet-induced growth inhibition by regulating the gut microbiota composition to ameliorate lipid deposition and oxidative stress and strengthen immune status in oriental river prawn. Full article
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Graphical abstract
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<p>Effects of SBPs on growth performance of oriental river prawn: (<b>a</b>) survival; (<b>b</b>) average final body weight; (<b>c</b>) final biomass; (<b>d</b>) biomass increase; (<b>e</b>) feed conversion ratio. <span class="html-italic">n</span> = 4. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>SBPs reduced lipid accumulation in the hepatopancreas of oriental river prawn fed with the low-fishmeal diet. (<b>a</b>–<b>d</b>) Assay of the hepatopancreatic and hemolymph triglyceride (TG) and total cholesterol (TC) (<span class="html-italic">n</span> = 4) (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001).</p>
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<p>SBPs improved antioxidative capacity in prawns’ hepatopancreas: (<b>a</b>) malondialdehyde (MDA) content in the hepatopancreas (<span class="html-italic">n</span> = 4). (<b>b</b>–<b>d</b>) The hepatopancreatic superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) activities (<span class="html-italic">n</span> = 4). (<b>e</b>,<b>f</b>) The inducible nitric oxide synthase (iNOS) activity and nitric oxide (NO) content in hepatopancreas (<span class="html-italic">n</span> = 4) (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001).</p>
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<p>SBPs activated lipid catabolism genes’ expressions in hepatopancreas. (<b>a</b>) Relative expression of lipid synthesis genes in hepatopancreas (<span class="html-italic">n</span> = 4). <span class="html-italic">fas</span>, fatty acid synthase; <span class="html-italic">acc</span>, acetyl-CoA carboxylase; <span class="html-italic">scd</span>, acyl-CoA delta-9 desaturase; (<b>b</b>) Relative expression of lipid catabolism genes in hepatopancreas (<span class="html-italic">n</span> = 4) <span class="html-italic">cpt1</span>, carnitine palmitoyltransferase 1; <span class="html-italic">sr-b I</span>, scavenger receptor class B type I; <span class="html-italic">hsl</span>, hormone-sensitive triglayceride lipase. (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01).</p>
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<p>SBPs upregulated genes’ expressions on antioxidant enzymes and <span class="html-italic">imd</span>/<span class="html-italic">relish</span> in hepatopancreas. (<b>a</b>) Relative expression of genes on antioxidant enzymes in hepatopancreas (<span class="html-italic">n</span> = 4) (<b>b</b>) Relative expression of genes like <span class="html-italic">tlr</span>, <span class="html-italic">dorsal</span>, <span class="html-italic">imd</span>, and <span class="html-italic">relish</span> in hepatopancreas (<span class="html-italic">n</span> = 4) (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001).</p>
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<p>SBPs ameliorated the gut microbiota composition. (<b>a</b>–<b>c</b>) Observed species, Chao1 and Shannon index (<span class="html-italic">n</span> = 4). (<b>d</b>) Principal coordinates analysis between three groups. (<b>e</b>,<b>f</b>) Venn diagram comparing gut microbiota distribution at phylum and genus level. (<b>g</b>,<b>h</b>) Relative abundances of the major taxonomic lineages at phylum and genus level.</p>
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<p>Analysis of microbiota differences between groups: (<b>a</b>) LEfSe analysis between R and CT as well as CT and SBP. Bar chart indicating the log-transformed LDA scores between groups, with a threshold of LDA &gt; 3.0. (<b>b</b>) Bar diagram of altered microbes at genus level in the CT group compared with R group or SBP group; <span class="html-italic">p</span>-value &lt; 0.05 represents a significant difference.</p>
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<p>Correlation analyses between genera of gut microbiota and biochemical and molecular indexes. Red or green circles suggest positive or negative relations, respectively. Asterisks within the different circles represent significance, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Ecological interaction network analysis of gut microbiota at genus level. (<b>a</b>–<b>c</b>) Interspecies interaction network of gut microbiota for oriental river prawn fed with different diet. Each node corresponds to a genus. Node colors represent genus belonged to different classes. A red or green edge suggests positive or negative correlations between two individual nodes.</p>
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23 pages, 3333 KiB  
Article
Identification of Allergenic Tropomyosins with Known and Unknown Sequences in Seafood Products
by Marta Godlewska, Piotr Minkiewicz, Małgorzata Darewicz and Anna Iwaniak
Appl. Sci. 2025, 15(1), 63; https://doi.org/10.3390/app15010063 - 25 Dec 2024
Viewed by 379
Abstract
Invertebrate tropomyosins belong to the key food allergens. Several peptides likely to be released during proteolysis can be found in many sequences of proteins from this family. The aim of the present study was to evaluate the possibility of identifying tropomyosins with known [...] Read more.
Invertebrate tropomyosins belong to the key food allergens. Several peptides likely to be released during proteolysis can be found in many sequences of proteins from this family. The aim of the present study was to evaluate the possibility of identifying tropomyosins with known and unknown amino acid sequences in unheated, boiled and fried seafoods. The workflow included in silico proteolysis simulation of tropomyosin sequences and analysis of the distribution of resulting peptides among proteins. The experiment entailed the proteolysis of unheated, boiled and fried products, containing crustaceans or mollusks, and the identification of resulting peptides using LC-MS/MS. Finally, taxonomic lineages of identified peptides were determined. Predicted peptides were identified in unheated samples. The boiling of seafoods resulted in an increase in the length of peptides containing predicted sequences. Some peptides from the boiled samples contained entire linear epitopes. The prediction of tropomyosin cleavage sites failed in the case of fried products. Peptides from the unheated and boiled samples were attributed to crustacean, arthropod or molluscan tropomyosins. In turn, peptides from the fried samples possessed inconclusive taxonomic lineages. Our results show that bioinformatics analysis (especially using Unipept program) may be a viable tool supporting LC-MS/MS experiments aimed at the detection of allergens. Full article
(This article belongs to the Special Issue Proteins and Peptides from Foods in 2024)
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<p>The workflow used in the study: blue—bioinformatics analysis; red—laboratory experiment.</p>
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<p>Percentage of proteins belonging to the tropomyosin family (ID: IPR000533 in the InterPro database), containing predicted products of simulated hydrolysis by trypsin. Peptides present in at least two tropomyosins are included. Data visualized in this figure are shown also in <a href="#app1-applsci-15-00063" class="html-app">Table S2 in the Supplementary Materials</a>.</p>
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<p>Example of the MS/MS spectrum of a tropomyosin marker, with the MDALENQLK sequence, generated by the PEAKS program. Nomenclature of fragment ions according to Roepstorff and Fohlman [<a href="#B62-applsci-15-00063" class="html-bibr">62</a>].</p>
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<p>Heatmap presenting peptides detected in in individual samples. Yellow—detected peptides, containing entire tropomyosin epitopes; green—detected peptides without epitopes; undetected peptides; sequences written using red font—peptides predicted (See <a href="#applsci-15-00063-f002" class="html-fig">Figure 2</a>), green font—longer peptides containing predicted peptides as fragments; black front—unpredicted peptides. Samples numbered as in <a href="#applsci-15-00063-t001" class="html-table">Table 1</a>. Raw data is shown in <a href="#app1-applsci-15-00063" class="html-app">Tables S3 and S5 in the Supplementary Materials</a>. Peptides containing entire epitopes annotated in the IEDB database are listed in <a href="#app1-applsci-15-00063" class="html-app">Table S4 in the Supplementary Materials</a>. Characteristics of sets of peptides obtained from particular samples is provided in <a href="#app1-applsci-15-00063" class="html-app">Table S6 in the Supplementary Materials</a>.</p>
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<p>Correlation matrix between sets of peptides identified in the hydrolysates of individual samples. Only peptides detected in at least one sample were considered (108 peptides). Yellow—strong or moderate correlation, green—weak correlation, blue—no correlation. Correlation coefficients are provided in <a href="#app1-applsci-15-00063" class="html-app">Table S7 in the Supplementary Materials</a>. Correlation matrix was generated using HeatMapper program.</p>
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<p>Distribution of peptides identified in seafood samples and present in at least 10 parent proteins determined using the Unipept program. The number of all proteins containing a given peptide is taken as 100%. (<b>a</b>) Number of proteins containing a given peptide; (<b>b</b>) number of InterPro families containing a given peptide; (<b>c</b>) percentage of tropomyosins among proteins containing a given peptide; (<b>d</b>) percentage of proteins of organisms belonging to various taxa among the ones containing a given peptide. Heatmaps generated using HeatMapper program. Raw data are annotated in <a href="#app1-applsci-15-00063" class="html-app">Table S8 in the Supplementary Materials</a>. Meaning of font colors is the same as in <a href="#applsci-15-00063-f004" class="html-fig">Figure 4</a>.</p>
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<p>Taxonomic lineage trees of peptides: (<b>a</b>) LAEASQAADESERMR as an example of peptide characteristic for <span class="html-italic">Crustacea</span>; (<b>b</b>) IQLLEEDLER as an example of peptide characteristic for <span class="html-italic">Arthropoda</span>; (<b>c</b>) IVELEEELKVVGNNMK as an example of peptide characteristic for <span class="html-italic">Mollusca</span>; (<b>d</b>) LSNQRQR as an example of peptide with inconclusive taxonomic lineage. Trees were generated using the Unipept program.</p>
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17 pages, 3994 KiB  
Article
Transcriptomic Analysis of Neocaridina denticulata sinensis Gills Following FPPS Knockdown Reveals Its Regulatory Role in Immune Response
by Hongrui Li, Dandan Feng, Chunyu Zhang, Mengfei Liu, Zixuan Wu, Yuke Bu, Jiquan Zhang and Yuying Sun
Int. J. Mol. Sci. 2025, 26(1), 65; https://doi.org/10.3390/ijms26010065 - 25 Dec 2024
Viewed by 316
Abstract
Farnesyl pyrophosphate synthase (FPPS) is a key enzyme in the terpenoid biosynthesis pathway, responsible for converting isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) into farnesyl pyrophosphate (FPP). In crustaceans, FPPS plays an important role in various physiological processes, particularly in synthesizing the crustacean-specific [...] Read more.
Farnesyl pyrophosphate synthase (FPPS) is a key enzyme in the terpenoid biosynthesis pathway, responsible for converting isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) into farnesyl pyrophosphate (FPP). In crustaceans, FPPS plays an important role in various physiological processes, particularly in synthesizing the crustacean-specific hormone methyl farnesoate (MF). This study analyzed the evolutionary differences in the physicochemical properties, subcellular localization, gene structure, and motif composition of FPPS in Neocaridina denticulata sinensis (named NdFPPS) compared to other species. The significant evolutionary divergence of FPPS was observed in crustaceans, likely linked to its role in MF synthesis. After the RNA interference (RNAi)-mediated knockdown of NdFPPS, transcriptomic analysis of gills revealed the significant enrichment of differentially expressed genes (DEGs) in pathways related to metabolism and immunity. Gene set enrichment analysis (GSEA) showed that most of these immune-related pathways were significantly suppressed, suggesting that NdFPPS may indirectly regulate the immune response by modulating metabolic levels. During the early stages of Vibrio parahaemolyticus infection, the expression of NdFPPS in the gills was significantly downregulated and subsequently returned to its original levels. Overall, our results provide new perspectives on the role of FPPS in immune regulation and enrich the functional information of FPPS. Full article
(This article belongs to the Section Molecular Immunology)
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<p>The structure and motif analysis of FPPS. (<b>A</b>) The phylogenetic relationships of FPPS across different species, represented as a dendrogram. The FPPS proteins of different species include <span class="html-italic">B.mori</span> (BmFPPS-1, BmFPPS-2), <span class="html-italic">C. elegans</span> (CeFPPS), <span class="html-italic">Cherax quadricarinatus</span> (CqFPPS), <span class="html-italic">D. melanogaster</span> (DmFPPS), <span class="html-italic">D. rerio</span> (DrFPPS), <span class="html-italic">E. sinensis</span> (EsFPPS-1, EsFPPS-2), <span class="html-italic">Homarus americanus</span> (HaFPPS), <span class="html-italic">Macrobrachium rosenbergii</span> (MrFPPS), <span class="html-italic">P. chinensis</span> (PchFPPS), <span class="html-italic">P. japonicus</span> (PjFPPS), <span class="html-italic">P. monodon</span> (PmFPPS), <span class="html-italic">P. trituberculatus</span> (PtFPPS), <span class="html-italic">P. vannamei</span> (PvFPPS), <span class="html-italic">Procambarus clarkii</span> (PclFPPS), and <span class="html-italic">Scylla paramamosain</span> (SpFPPS). (<b>B</b>) The distribution of conserved motifs within FPPS proteins, with different colors representing distinct motifs (motif 1 to motif 10). (<b>C</b>) A representation of the polyprenyl_synt superfamily domain, with green indicating conserved catalytic domains across species. (<b>D</b>) The structure of FPPS genes, with green boxes representing coding sequences (CDS). The GenBank accession numbers for the sequences mentioned in the phylogenetic tree are detailed in <a href="#app1-ijms-26-00065" class="html-app">Supplementary Materials Table S1</a>.</p>
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<p>A phylogenetic time tree of crustacean species. This time tree illustrates the evolutionary divergence times among various crustacean species. Blue circles indicate calibration points, and horizontal blue lines represent 95% confidence intervals. The timeline, marked in millions of years ago (Mya), spans the evolutionary divergence from the Carboniferous period to the Neogene of the Cenozoic era. The GenBank accession numbers for the genome data mentioned in the phylogenetic tree are detailed in <a href="#app1-ijms-26-00065" class="html-app">Supplementary Materials Table S2</a>.</p>
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<p>A volcano plot showing the distribution of DEGs. This volcano plot illustrates the distribution of gene expression changes. The <span class="html-italic">x</span>-axis represents the fold change in gene expression (log2(FC)), and the <span class="html-italic">y</span>-axis shows the significance level (−log10(FDR)). Red dots indicate significantly upregulated genes (Up), green dots represent significantly downregulated genes (Down), and blue dots denote genes without significant differential expression (Normal).</p>
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<p>Functional and pathway enrichment analysis of DEGs. (<b>A</b>) The GO classification shows the top 10 enriched GO terms for upregulated and downregulated DEGs across the BP, CC, and MF categories. Blue bars represent upregulated genes, while pink bars represent downregulated genes. (<b>B</b>) GO enrichment analysis of DEGs, with each dot size indicating the number of genes and the color intensity representing the <span class="html-italic">p</span>-value (darker colors correspond to smaller <span class="html-italic">p</span>-values). The gene ratio represents the ratio of DEGs in each GO term to the total DEGs analyzed. (<b>C</b>) KEGG pathway enrichment analysis for all DEGs, displaying significantly enriched pathways. The dot size indicates the gene count, while the color intensity indicates the <span class="html-italic">p</span>-value significance.</p>
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<p>Functional and pathway enrichment analysis of DEGs. (<b>A</b>) The GO classification shows the top 10 enriched GO terms for upregulated and downregulated DEGs across the BP, CC, and MF categories. Blue bars represent upregulated genes, while pink bars represent downregulated genes. (<b>B</b>) GO enrichment analysis of DEGs, with each dot size indicating the number of genes and the color intensity representing the <span class="html-italic">p</span>-value (darker colors correspond to smaller <span class="html-italic">p</span>-values). The gene ratio represents the ratio of DEGs in each GO term to the total DEGs analyzed. (<b>C</b>) KEGG pathway enrichment analysis for all DEGs, displaying significantly enriched pathways. The dot size indicates the gene count, while the color intensity indicates the <span class="html-italic">p</span>-value significance.</p>
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<p>GSEA enrichment analysis of KEGG pathways. This figure presents the GSEA enrichment curves for five immune-related KEGG pathways. The top curves illustrate the changes in enrichment scores for each pathway across gene rankings, with the <span class="html-italic">x</span>-axis representing the ranked gene positions and the <span class="html-italic">y</span>-axis showing the running enrichment scores. The middle bar chart displays the gene distribution for each pathway within the ranked dataset.</p>
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<p>COG functional classification. This figure displays the distribution of downregulated DEGs across various COG functional categories. The <span class="html-italic">x</span>-axis shows COG category labels, while the <span class="html-italic">y</span>-axis indicates the gene count in each category. Different colors represent distinct COG functions.</p>
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<p>Relative expression of seven DEGs. Asterisks indicate significant differences in seven selected genes’ expression compared to control group, respectively (*: <span class="html-italic">p</span> &lt; 0.05; ***: <span class="html-italic">p</span> &lt; 0.001; and ****: <span class="html-italic">p</span> &lt; 0.0001).</p>
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<p>The semi-quantitative validation of the <span class="html-italic">NdFPPS</span> expression pattern in the early stage of <span class="html-italic">V. parahaemolyticus</span> challenge. (<b>A</b>) Gel maps of <span class="html-italic">NdFPPS</span> and <span class="html-italic">18S</span> rRNA genes challenged by <span class="html-italic">V. parahaemolyticus</span> at different times. (<b>B</b>) Significance analysis of grayscale values in gel results. The transcript expression levels of <span class="html-italic">NdFPPS</span> were normalized to the <span class="html-italic">18S</span> rRNA gene. The different letters on the column represent statistical significance (<span class="html-italic">p</span> &lt; 0.05), and the same letters indicate no significance among groups (<span class="html-italic">p</span> &gt; 0.05).</p>
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13 pages, 2619 KiB  
Article
Ecological Indicators and Estimators of Fish and Crustacean Diversity in the Yucatan Peninsula
by Martha Angélica Gutiérrez-Aguirre, Adrián Cervantes-Martínez and Eduardo Suárez-Morales
Diversity 2025, 17(1), 5; https://doi.org/10.3390/d17010005 - 25 Dec 2024
Viewed by 594
Abstract
Biodiversity patterns provide insights into the conservation value of ecosystems and also aid in an understanding of some research priorities. This research paper presents an exhaustive inventory of crustaceans and fish species from the Yucatan Peninsula’s epicontinental, underground, and anchialine aquatic systems. Using [...] Read more.
Biodiversity patterns provide insights into the conservation value of ecosystems and also aid in an understanding of some research priorities. This research paper presents an exhaustive inventory of crustaceans and fish species from the Yucatan Peninsula’s epicontinental, underground, and anchialine aquatic systems. Using records spanning from 1936 to 2024, this study analyzed over 2600 entries across 670 aquatic systems, including cenotes, flooded cave lakes, springs, and wetlands. A total of 329 species were recorded, comprising 117 fish and 212 crustacean species. The richness and endemism of these species were assessed through a completeness analysis, utilizing non-parametric models (Chao 2, Jackknife 1, Bootstrap) to estimate potential species richness. Notably, the region exhibits high endemism, especially among crustaceans, with up to 50 species and 2 endemic families being unique to the Yucatan Peninsula. This study underscores the importance of the eastern YP for crustaceans and the central YP for fish, highlighting areas critical for conservation. These findings emphasize the need for further exploration, as the diversity of aquatic species remains incompletely understood. This comprehensive synthesis aims to inform future conservation strategies, environmental management, and regional planning efforts, particularly highlighting the vulnerability of these unique aquatic systems to environmental degradation. Full article
(This article belongs to the Section Freshwater Biodiversity)
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<p>Yucatan Peninsula—records of fish or crustaceans.</p>
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<p>Ranges of species richness of Crustacea (red) and fish (blue) for each site.</p>
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<p>Species richness estimators (Chao 2, Jackknife 1, and Bootstrap): (<b>a</b>) fish and (<b>b</b>) crustaceans of the Yucatan Peninsula, Mexico. Actual species richness per site (sampled units) is represented (=S Obs, blue line).</p>
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19 pages, 7157 KiB  
Article
The Bile Acid Metabolism of Intestinal Microorganisms Mediates the Effect of Different Protein Sources on Muscle Protein Deposition in Procambarus clarkii
by Xiaodi Xu, Xiaochuan Zheng, Qunlan Zhou, Cunxin Sun, Aimin Wang, Aimin Zhu, Yuanyuan Zhang and Bo Liu
Microorganisms 2025, 13(1), 11; https://doi.org/10.3390/microorganisms13010011 - 24 Dec 2024
Viewed by 497
Abstract
The most economically important trait of the Procambarus clarkii is meat quality. Protein deposition is essential in muscle growth and nutritional quality formation. The effects and potential mechanisms of feed protein sources on crustaceans’ muscle protein deposition have not been elucidated. This study [...] Read more.
The most economically important trait of the Procambarus clarkii is meat quality. Protein deposition is essential in muscle growth and nutritional quality formation. The effects and potential mechanisms of feed protein sources on crustaceans’ muscle protein deposition have not been elucidated. This study established an all-animal protein source (AP) and an all-plant protein source group (PP), with a feeding period of 8 weeks (four replicates per group, 45 individuals per replicate). The results demonstrated that muscle protein deposition, muscle fiber diameter, and hardness were significantly higher in the PP group (p < 0.05). The transcript levels of genes involved in protein synthesis were notably upregulated, while those of protein hydrolysis and negative regulators of myogenesis notably downregulated in PP group (p < 0.05). Furthermore, protein sources shaped differential intestinal microbiota composition and microbial metabolites profiles, as evidenced by a significant decrease in g_Bacteroides (p = 0.030), and a significant increase in taurochenodeoxycholic acid (TCDCA) in PP group (p = 0.027). A significant correlation was further established by Pearson correlation analysis between the g_Bacteroides, TCDCA, and genes involved in the MSTN-mediated protein deposition pathway (p < 0.05). In vitro anaerobic fermentation confirmed the ability of the two groups of intestinal flora to metabolically produce differential TCDCA (p = 0.038). Our results demonstrated that the ‘Bacteroides-TCDCA-MSTN’ axis may mediate the effects of different protein sources on muscle development and protein deposition in P. clarkii, which was anticipated to represent a novel target for the muscle quality modulation in crustaceans. Full article
(This article belongs to the Special Issue Aquatic Microorganisms and Their Application in Aquaculture)
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<p>The effects of animal and plant protein sources on the amino acid composition in the muscle of <span class="html-italic">P. clarkii</span>. Note: (<b>A</b>) Amino acid composition in the AP and PP groups; (<b>B</b>) Differentiated amino acids and classification; AP, all-animal protein source group; PP, all-plant protein source group. Red data mean the <span class="html-italic">p</span> value &lt; 0.05.</p>
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<p>The effects of animal and plant protein sources on the histological morphology of muscle in <span class="html-italic">P. clarkii</span>. Note: AP, all-animal protein source group; PP, all-plant protein source group. Red data with asterisk mean the <span class="html-italic">p</span> value &lt; 0.05, and ns means no significant difference.</p>
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<p>The effects of animal and plant protein sources on the muscle texture characteristics of <span class="html-italic">P. clarkii.</span> Note: Red means the texture parameters of AP group, blue means the texture parameters of PP group. * means <span class="html-italic">p</span> &lt; 0.05. AP, all-animal protein source group; PP, all-plant protein source group.</p>
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<p>Effects of animal and plant protein sources on the transcription levels of genes related to muscle development in <span class="html-italic">P. clarkii</span>. Note: (<b>A</b>) Relative expression of genes related to protein synthesis signaling molecules and transcription factors; (<b>B</b>) Relative expression of genes related to myosins and muscle regulatory factors; (<b>C</b>) Relative expression of genes related to autophagy factors; (<b>D</b>) Relative expression of genes related to ubiquitination factors. AP, all-animal protein source group; PP, all-plant protein source group; Red data means the <span class="html-italic">p</span> value &lt; 0.05.</p>
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<p>Effects of animal and plant protein sources on the intestinal microbiota structure and intestinal content composition of <span class="html-italic">P. clarkii</span>. Note: (<b>A</b>,<b>B</b>), The relative abundance of the top seven predominant phyla (with a mean relative abundance &gt; 1%). (<b>C</b>,<b>D</b>), The relative abundance of the top seven predominant phyla (with a mean relative abundance &gt; 0.01%). (<b>E</b>), Heatmap analysis of 20 differential metabolites. AP, all-animal protein source group; PP, all-plant protein source group. Red data means the <span class="html-italic">p</span> value &lt; 0.05.</p>
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<p>Correlation analysis. Note: (<b>A</b>) Intestinal microbiome and differential metabolites correlation analysis; (<b>B</b>) Correlation analysis of selected differential metabolites and muscle development-related genes. Red ball means positive correlation, while blue ball means negative correlation, * means <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Bile acid profile of in vitro intestinal anaerobic fermentation broth. Note: Independent Samples <span class="html-italic">t</span>-test was used to detect the significant differences between the AP and PP group. * means <span class="html-italic">p</span> &lt; 0.05. AP, all-animal protein source group; PP, all-plant protein source group.</p>
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14 pages, 576 KiB  
Article
Enhanced Bioaccessibility of Carotenoids, Antioxidants, and Minerals from Red Lobster By-Products Through High-Hydrostatic Pressure and Ultrasound Extraction
by Vilbett Briones-Labarca and Claudia Giovagnoli-Vicuña
Processes 2025, 13(1), 10; https://doi.org/10.3390/pr13010010 - 24 Dec 2024
Viewed by 457
Abstract
Efficient extraction of bioactive compounds from red lobster by-products is crucial for maximizing their nutritional and economic value. This study compared high-hydrostatic pressure extraction (HHPE), ultrasound-assisted extraction (UAE), and conventional extraction (CE), assessing the yield and bioaccessibility of carotenoids (astaxanthin, β-carotene, lycopene), antioxidants [...] Read more.
Efficient extraction of bioactive compounds from red lobster by-products is crucial for maximizing their nutritional and economic value. This study compared high-hydrostatic pressure extraction (HHPE), ultrasound-assisted extraction (UAE), and conventional extraction (CE), assessing the yield and bioaccessibility of carotenoids (astaxanthin, β-carotene, lycopene), antioxidants (DPPH, FRAP assays), and minerals (sodium, magnesium, potassium, calcium). HHPE and UAE significantly enhanced carotenoid extraction (p < 0.05), with UAE yielding the highest astaxanthin (3.61 mg/100 g FW) and HHPE producing the most β-carotene (0.64 mg/100 g FW). HHPE also significantly increased antioxidant capacity (38.04% increase over CE, p < 0.05). Despite reduced bioaccessibility after in vitro digestion, HHPE and UAE represent sustainable methods for recovering valuable bioactive compounds, improving the nutritional profile of red lobster by-products. Full article
(This article belongs to the Special Issue Feature Papers in the "Food Process Engineering" Section)
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<p>Chemical structures: (<b>A</b>) lycopene, (<b>B</b>) β-carotene, and (<b>C</b>) astaxanthin.</p>
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