Pub Date : 2025-01-24DOI: 10.1016/j.fsi.2025.110155
Xiaoran Liu , Yanyan Cao , Xiucai Hu, Aijun Lv
Rahnella aquatilis is an emerging opportunistic pathogen that usually causes septicemia in fish and poses a potential threat to human health. VgrG gene is an important virulence factor of type VI secretion system in R. aquatilis, but its regulatory mechanism underlying PANoptosis is still unknown. Here, VgrG deletion mutant strain of R. aquatilis (ΔVgrG-RA) and recombinant plasmid pET32a-VgrG were respectively constructed, and immunohistochemistry for VgrG as well as PANoptosis features were evaluated. Moreover, the interaction transcriptome of ΔVgrG-mediated pathogen and host was determined by dual RNA-seq using an in vitro model of the primary macrophage cells from crucian carp Carassius auratus, and a total of 889 and 3765 differentially expressed genes (DEGs) were identified in pathogen-host interaction genes, respectively. Notably, GO and KEGG enrichment analysis were significantly involved in the PANoptosis pathways in ΔVgrG mutant-infected macrophages. The regulatory relationship of potential PANoptosis-related genes (PRGs) were analysed comprehensively, and their binding interaction of several hub proteins (eg., YcgR, Bcl2a, Calr3a, IL-1β) were determined by molecular docking analysis. To our best knowledge, this is first report of R. aquatilis VgrG-mediated interactions between pathogen and host macrophage cells, which will provide a new reference for understanding of molecular mechanism underlying PANoptosis in fish.
{"title":"Rahnella aquatilis VgrG-mediated PANoptosis in macrophages of Carassius auratus by dual RNA-seq analysis","authors":"Xiaoran Liu , Yanyan Cao , Xiucai Hu, Aijun Lv","doi":"10.1016/j.fsi.2025.110155","DOIUrl":"10.1016/j.fsi.2025.110155","url":null,"abstract":"<div><div><em>Rahnella aquatilis</em> is an emerging opportunistic pathogen that usually causes septicemia in fish and poses a potential threat to human health. <em>VgrG</em> gene is an important virulence factor of type VI secretion system in <em>R. aquatilis</em>, but its regulatory mechanism underlying PANoptosis is still unknown. Here, <em>VgrG</em> deletion mutant strain of <em>R. aquatilis</em> (Δ<em>VgrG</em>-RA) and recombinant plasmid pET32a-<em>VgrG</em> were respectively constructed, and immunohistochemistry for VgrG as well as PANoptosis features were evaluated. Moreover, the interaction transcriptome of Δ<em>VgrG</em>-mediated pathogen and host was determined by dual RNA-seq using an in vitro model of the primary macrophage cells from crucian carp <em>Carassius auratus</em>, and a total of 889 and 3765 differentially expressed genes (DEGs) were identified in pathogen-host interaction genes, respectively. Notably, GO and KEGG enrichment analysis were significantly involved in the PANoptosis pathways in Δ<em>VgrG</em> mutant-infected macrophages. The regulatory relationship of potential PANoptosis-related genes (PRGs) were analysed comprehensively, and their binding interaction of several hub proteins (eg., YcgR, Bcl2a, Calr3a, IL-1β) were determined by molecular docking analysis. To our best knowledge, this is first report of <em>R. aquatilis VgrG</em>-mediated interactions between pathogen and host macrophage cells, which will provide a new reference for understanding of molecular mechanism underlying PANoptosis in fish.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110155"},"PeriodicalIF":4.1,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143045358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-24DOI: 10.1016/j.fsi.2025.110157
Tongzhen Sun, Jinqiang Huang, Yongjuan Li, Shenji Wu, Lu Zhao, Yujun Kang
MicroRNAs (miRNAs) are highly conserved endogenous non-coding RNAs that play a crucial role in fish immune response by regulating gene expression at the post-transcriptional level. In recent years, the viral diseases caused by infectious hematopoietic necrosis virus (IHNV) have caused significant economic losses in rainbow trout (Oncorhynchus mykiss) aquaculture, whereas the immune regulatory mechanisms of miRNAs involved in rainbow trout resistance to IHNV infection remains largely undefined. In this study, we analyzed the structural characteristics of Oncorhynchus mykiss tumor necrosis factor receptor-associated factor 3 (OmTRAF3) by bioinformatics software and explored the molecular mechanism of miR-203-3p in rainbow trout resistance to IHNV by regulating OmTRAF3 in vivo and in vitro. The open reading frame (ORF) of OmTRAF3 gene was 1 731 bp and encoded 576 amino acids including an N-terminal RING finger domain, two zinc finger domains, a coiled-coil domain, and a C-terminal MATH domain. The expression pattern analysis showed that the expression of miR-203-3p and OmTRAF3 in immune-related tissues (head kidney, spleen, and liver) and liver cells of rainbow trout infected with IHNV varied with some regularity and had opposite trends at key time points, and a targeting relationship between miR-203-3p and OmTRAF3 was confirmed using a dual luciferase reporter gene assay. Further, we found that in vivo and in vitro overexpression of miR-203-3p significantly reduced the expression of OmTRAF3, downstream immune-related genes (OmTANK, OmIKKε, OmIFN1, and OmISG15) and promoted IHNV copy number replication, while silencing of miR-203-3p yielded opposite results. More importantly, OmTRAF3 and downstream genes as well as IHNV copy number changed accordingly with the silencing of OmTRAF3. The above results revealed that miR-203-3p participates in the immune response against IHNV by targeting OmTRAF3, and provide a theoretical basis for the screening of antiviral drugs in rainbow trout.
{"title":"MicroRNA-203-3p participates in antiviral immune response by negatively regulating TRAF3 in the rainbow trout (Oncorhynchus mykiss).","authors":"Tongzhen Sun, Jinqiang Huang, Yongjuan Li, Shenji Wu, Lu Zhao, Yujun Kang","doi":"10.1016/j.fsi.2025.110157","DOIUrl":"https://doi.org/10.1016/j.fsi.2025.110157","url":null,"abstract":"<p><p>MicroRNAs (miRNAs) are highly conserved endogenous non-coding RNAs that play a crucial role in fish immune response by regulating gene expression at the post-transcriptional level. In recent years, the viral diseases caused by infectious hematopoietic necrosis virus (IHNV) have caused significant economic losses in rainbow trout (Oncorhynchus mykiss) aquaculture, whereas the immune regulatory mechanisms of miRNAs involved in rainbow trout resistance to IHNV infection remains largely undefined. In this study, we analyzed the structural characteristics of Oncorhynchus mykiss tumor necrosis factor receptor-associated factor 3 (OmTRAF3) by bioinformatics software and explored the molecular mechanism of miR-203-3p in rainbow trout resistance to IHNV by regulating OmTRAF3 in vivo and in vitro. The open reading frame (ORF) of OmTRAF3 gene was 1 731 bp and encoded 576 amino acids including an N-terminal RING finger domain, two zinc finger domains, a coiled-coil domain, and a C-terminal MATH domain. The expression pattern analysis showed that the expression of miR-203-3p and OmTRAF3 in immune-related tissues (head kidney, spleen, and liver) and liver cells of rainbow trout infected with IHNV varied with some regularity and had opposite trends at key time points, and a targeting relationship between miR-203-3p and OmTRAF3 was confirmed using a dual luciferase reporter gene assay. Further, we found that in vivo and in vitro overexpression of miR-203-3p significantly reduced the expression of OmTRAF3, downstream immune-related genes (OmTANK, OmIKKε, OmIFN1, and OmISG15) and promoted IHNV copy number replication, while silencing of miR-203-3p yielded opposite results. More importantly, OmTRAF3 and downstream genes as well as IHNV copy number changed accordingly with the silencing of OmTRAF3. The above results revealed that miR-203-3p participates in the immune response against IHNV by targeting OmTRAF3, and provide a theoretical basis for the screening of antiviral drugs in rainbow trout.</p>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":" ","pages":"110157"},"PeriodicalIF":4.1,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143046141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-23DOI: 10.1016/j.fsi.2025.110154
Shishun Gui , Jiang Wu , Lishan Liao , Xiaocong Zheng , Peng Zhu , Lei Zhang , Ziyi Zhang , Dan Xu , Fei Ke , Hong Liu , Jie Sun , Lang Gui
Frog virus 3-like ranaviruses (FV3-like viruses), particularly FV3 (Frog virus 3), represent typical species within the genus Ranavirus, primarily infecting amphibians and reptiles, thereby posing serious threats to aquaculture and biodiversity conservation. We designed a pair of universal primers and a probe targeting the conserved region of the major capsid protein (MCP) genes of FV3-like viruses. By integrating recombinase-aided amplification (RAA) with lateral flow dipstick (LFD) technology and real-time fluorescence (RF) modification, we established RAA-LFD and RF-RAA assays. The limit of detection (LoD) of RAA-LFD was determined to be 35.4 copies/μL under optimized amplification conditions at 35 °C for 15 min. Similarly, the RF-RAA assay exhibited high specificity with a satisfactory LoD of 3.54 × 102 copies/μL at 39 °C over a duration of 17–20 min. In diagnosing 53 clinical samples infected by an isolated strain of FV3-like viruses, both assays demonstrated consistency with results obtained from real-time fluorescence PCR assay. These experiments indicate that both methods serve as promising alternatives for point-of care testing (POST), adaptable to various scenarios. This study represents the first establishment of RAA-LFD and RF-RAA assays for FV3-likes viruses, enabling rapid and intuitive assessment of detection results while fulfilling on-site testing requirements, thus contributing positively to swift diagnosis and long-term monitoring in aquaculture.
{"title":"Development of two recombinase-aided amplification assays combined with lateral flow dipstick (RAA-LFD) and real-time fluorescence (RF-RAA) for the detection of Frog virus 3-like ranaviruses","authors":"Shishun Gui , Jiang Wu , Lishan Liao , Xiaocong Zheng , Peng Zhu , Lei Zhang , Ziyi Zhang , Dan Xu , Fei Ke , Hong Liu , Jie Sun , Lang Gui","doi":"10.1016/j.fsi.2025.110154","DOIUrl":"10.1016/j.fsi.2025.110154","url":null,"abstract":"<div><div>Frog virus 3-like ranaviruses (FV3-like viruses), particularly FV3 (Frog virus 3), represent typical species within the genus <em>Ranavirus</em>, primarily infecting amphibians and reptiles, thereby posing serious threats to aquaculture and biodiversity conservation. We designed a pair of universal primers and a probe targeting the conserved region of the major capsid protein (MCP) genes of FV3-like viruses. By integrating recombinase-aided amplification (RAA) with lateral flow dipstick (LFD) technology and real-time fluorescence (RF) modification, we established RAA-LFD and RF-RAA assays. The limit of detection (LoD) of RAA-LFD was determined to be 35.4 copies/μL under optimized amplification conditions at 35 °C for 15 min. Similarly, the RF-RAA assay exhibited high specificity with a satisfactory LoD of 3.54 × 10<sup>2</sup> copies/μL at 39 °C over a duration of 17–20 min. In diagnosing 53 clinical samples infected by an isolated strain of FV3-like viruses, both assays demonstrated consistency with results obtained from real-time fluorescence PCR assay. These experiments indicate that both methods serve as promising alternatives for point-of care testing (POST), adaptable to various scenarios. This study represents the first establishment of RAA-LFD and RF-RAA assays for FV3-likes viruses, enabling rapid and intuitive assessment of detection results while fulfilling on-site testing requirements, thus contributing positively to swift diagnosis and long-term monitoring in aquaculture.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110154"},"PeriodicalIF":4.1,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143037706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-22DOI: 10.1016/j.fsi.2025.110148
Xinru Wang, Lei Zhu, Zhengyan Du, Hao Li, Libo Hou, Chen Li, Xinyu Jiang, Jie Zhang, Chao Pei, Li Li, Xianghui Kong
Finding effective alternatives to antibiotics is crucial for sustainable aquaculture. Host-derived probiotics have great potential as a promising alternative to antibiotics for immune regulation and disease control in fish farming. However, limited research exists regarding the application of native probiotics in largemouth bass (Micropterus salmoides). This study aims to evaluate the potential of the endogenous strain Pediococcus acidilactici B49 as a probiotic in modulating host immunity and disease control through in vitro and in vivo experiments. The results demonstrated that P. acidilactici B49 exhibited no hemolytic activity and displayed susceptibility to most tested antibiotics. It successfully survived and colonized in the intestinal tract of the largemouth bass. Furthermore, this strain showed remarkable antibacterial activity against common aquatic pathogens, including gram-positive and gram-negative bacteria, and also exhibited resistance against Aeromonas hydrophila on the head kidney leukocytes of largemouth bass in vitro. Following an 8-week feeding trial, P. acidilactici B49 improved host immunity by increasing intestinal lysozyme activity, enhancing IL-8 expression, reducing TGF-β expression, and enhancing IgM levels in both serum and intestinal mucus. It also potentiated the phagocytic activity of peripheral blood lymphocytes. In addition, the B49 feeding group showed a significant increase in intestinal villus height. The challenge test with A. hydrophila demonstrated that the administration of P. acidilactici B49 effectively maintained intestinal barrier integrity, reduced gut inflammation, decreased pathogen load in the spleen, and improved survival rates in largemouth bass. In conclusion, the host-derived strain P. acidilactici B49 exhibited broad-spectrum antibacterial ability, biosafety, and intestinal colonization in largemouth bass. It effectively improved immune function, intestinal health, and resistance against A. hydrophila in the host. Therefore, P. acidilactici B49 is a promising probiotic for immunomodulation and disease control in largemouth bass aquaculture.
{"title":"Host-derived Pediococcus acidilactici B49: A promising probiotic for immunomodulation and disease control in largemouth bass (Micropterus salmoides)","authors":"Xinru Wang, Lei Zhu, Zhengyan Du, Hao Li, Libo Hou, Chen Li, Xinyu Jiang, Jie Zhang, Chao Pei, Li Li, Xianghui Kong","doi":"10.1016/j.fsi.2025.110148","DOIUrl":"10.1016/j.fsi.2025.110148","url":null,"abstract":"<div><div>Finding effective alternatives to antibiotics is crucial for sustainable aquaculture. Host-derived probiotics have great potential as a promising alternative to antibiotics for immune regulation and disease control in fish farming. However, limited research exists regarding the application of native probiotics in largemouth bass (<em>Micropterus salmoides</em>). This study aims to evaluate the potential of the endogenous strain <em>Pediococcus acidilactici</em> B49 as a probiotic in modulating host immunity and disease control through <em>in vitro</em> and <em>in vivo</em> experiments. The results demonstrated that <em>P. acidilactici</em> B49 exhibited no hemolytic activity and displayed susceptibility to most tested antibiotics. It successfully survived and colonized in the intestinal tract of the largemouth bass. Furthermore, this strain showed remarkable antibacterial activity against common aquatic pathogens, including gram-positive and gram-negative bacteria, and also exhibited resistance against <em>Aeromonas hydrophila</em> on the head kidney leukocytes of largemouth bass <em>in vitro</em>. Following an 8-week feeding trial, <em>P. acidilactici</em> B49 improved host immunity by increasing intestinal lysozyme activity, enhancing <em>IL-8</em> expression, reducing <em>TGF-β</em> expression, and enhancing IgM levels in both serum and intestinal mucus. It also potentiated the phagocytic activity of peripheral blood lymphocytes. In addition, the B49 feeding group showed a significant increase in intestinal villus height. The challenge test with <em>A. hydrophila</em> demonstrated that the administration of <em>P. acidilactici</em> B49 effectively maintained intestinal barrier integrity, reduced gut inflammation, decreased pathogen load in the spleen, and improved survival rates in largemouth bass. In conclusion, the host-derived strain <em>P. acidilactici</em> B49 exhibited broad-spectrum antibacterial ability, biosafety, and intestinal colonization in largemouth bass. It effectively improved immune function, intestinal health, and resistance against <em>A. hydrophila</em> in the host<em>.</em> Therefore, <em>P. acidilactici</em> B49 is a promising probiotic for immunomodulation and disease control in largemouth bass aquaculture.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110148"},"PeriodicalIF":4.1,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-21DOI: 10.1016/j.fsi.2025.110149
Min-Young Sohn , Ji-Min Jeong , Gyoungsik Kang , Won-Sik Woo , Kyung-Ho Kim , Ha-Jeong Son , Chan-Il Park
Dendritic cells (DCs) play a pivotal role in activating naïve T-cells and bridging innate and adaptive immunity. This study aimed to identify and characterize CD83 and CD276 as potential markers for DCs in olive flounder (Paralichthys olivaceus). Specific antibodies against these markers were developed and used to analyze their distribution in peripheral blood leukocytes (PBLs) and intestinal tissues. Flow cytometry and immunohistochemistry revealed that CD83 and CD276 are expressed on DCs, with peak expression observed one week after oral administration of an inactivated viral hemorrhagic septicemia virus (VHSV) vaccine. Gene expression analysis further demonstrated significant activation of immune-related genes, including CD3, IgM, and TLRs, indicating that oral vaccine administration effectively activates the intestinal mucosal immune system. These findings provide valuable insights into fish immune responses and establish CD83 and CD276 as promising DC markers, contributing to the development of mucosal vaccine strategies in aquaculture.
{"title":"Identification and characterization of CD83 and CD276 as markers of dendritic cells in olive flounder (Paralichthys olivaceus)","authors":"Min-Young Sohn , Ji-Min Jeong , Gyoungsik Kang , Won-Sik Woo , Kyung-Ho Kim , Ha-Jeong Son , Chan-Il Park","doi":"10.1016/j.fsi.2025.110149","DOIUrl":"10.1016/j.fsi.2025.110149","url":null,"abstract":"<div><div>Dendritic cells (DCs) play a pivotal role in activating naïve T-cells and bridging innate and adaptive immunity. This study aimed to identify and characterize CD83 and CD276 as potential markers for DCs in olive flounder (<em>Paralichthys olivaceus</em>). Specific antibodies against these markers were developed and used to analyze their distribution in peripheral blood leukocytes (PBLs) and intestinal tissues. Flow cytometry and immunohistochemistry revealed that CD83 and CD276 are expressed on DCs, with peak expression observed one week after oral administration of an inactivated viral hemorrhagic septicemia virus (VHSV) vaccine. Gene expression analysis further demonstrated significant activation of immune-related genes, including CD3, IgM, and TLRs, indicating that oral vaccine administration effectively activates the intestinal mucosal immune system. These findings provide valuable insights into fish immune responses and establish CD83 and CD276 as promising DC markers, contributing to the development of mucosal vaccine strategies in aquaculture.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110149"},"PeriodicalIF":4.1,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-20DOI: 10.1016/j.fsi.2025.110143
Wenhui Xiao , Fang Song , Zilin Yang , Xiaoshan Wu , Xiaolin Zhang , Jianyu He , Yue Wang , Isabella Buttino , Xiaojun Yan , Zhi Liao
Mytilus are sessile filter feeders that live in close contact with numerous marine microorganisms. Hemocytes, the immunocompetent cells of Mytilus, participate in the immune response in a very efficient manner. Lipopolysaccharide (LPS) and peptidoglycan (PGN) follow specific microbe/pathogen-associated molecular patterns (MAMPs or PAMPs) and are involved in immune stimulation in host cells. This study evaluated the molecular profiles and reactions at protein level of Mytilus hemocytes after stimulation with LPS and PGN. Mytilus coruscus was challenged in vivo with LPS and PGN. The hemocytes were collected after 48 h and analyzed for quantitative proteomics, cell subpopulations, and the free amino acid composition. 4D-DIA technology-based proteomic analysis revealed different protein profiles, as well as different responses at protein level, under either the LPS or PGN challenge. C-type lectins, collagens, and CD151 protein were highly upregulated in LPS-challenged mussels, while phospholipase A2 and dCMP deaminase were highly upregulated in PGN-challenged mussels. Moreover, LPS challenge disrupted dsRNA-mediated translation and stimulated energy-related metabolism, while PGN challenge stimulated proteins involved in the inflammatory response and suppressed amino acid metabolism. In addition, the LPS and PGN challenges differed in their effects on the free amino acid composition and granulocytes ratio of the hemocytes. These findings highlight the different strategies employed by mussel hemocytes in response to different MAMPs, providing insights into the effects of LPS and PGN on Mytilus.
{"title":"Molecular responses of Mytilus coruscus hemocytes to lipopolysaccharide and peptidoglycan as revealed by 4D-DIA based quantitative proteomics analysis","authors":"Wenhui Xiao , Fang Song , Zilin Yang , Xiaoshan Wu , Xiaolin Zhang , Jianyu He , Yue Wang , Isabella Buttino , Xiaojun Yan , Zhi Liao","doi":"10.1016/j.fsi.2025.110143","DOIUrl":"10.1016/j.fsi.2025.110143","url":null,"abstract":"<div><div><em>Mytilus</em> are sessile filter feeders that live in close contact with numerous marine microorganisms. Hemocytes, the immunocompetent cells of <em>Mytilus</em>, participate in the immune response in a very efficient manner. Lipopolysaccharide (LPS) and peptidoglycan (PGN) follow specific microbe/pathogen-associated molecular patterns (MAMPs or PAMPs) and are involved in immune stimulation in host cells. This study evaluated the molecular profiles and reactions at protein level of <em>Mytilus</em> hemocytes after stimulation with LPS and PGN. <em>Mytilus coruscus</em> was challenged <em>in vivo</em> with LPS and PGN. The hemocytes were collected after 48 h and analyzed for quantitative proteomics, cell subpopulations, and the free amino acid composition. 4D-DIA technology-based proteomic analysis revealed different protein profiles, as well as different responses at protein level, under either the LPS or PGN challenge. C-type lectins, collagens, and CD151 protein were highly upregulated in LPS-challenged mussels, while phospholipase A2 and dCMP deaminase were highly upregulated in PGN-challenged mussels. Moreover, LPS challenge disrupted dsRNA-mediated translation and stimulated energy-related metabolism, while PGN challenge stimulated proteins involved in the inflammatory response and suppressed amino acid metabolism. In addition, the LPS and PGN challenges differed in their effects on the free amino acid composition and granulocytes ratio of the hemocytes. These findings highlight the different strategies employed by mussel hemocytes in response to different MAMPs, providing insights into the effects of LPS and PGN on <em>Mytilus</em>.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110143"},"PeriodicalIF":4.1,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143022674","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-20DOI: 10.1016/j.fsi.2025.110146
Qiongyue Xu , Siting Wu , Xiaoxia Lei , Helong Cao , Zhouling Zhan , Qiwei Qin , Jingguang Wei
During viral infection, RIG-I-like receptors (RLRs) are cytoplasmic pattern recognition receptors that recognize and bind to viral RNA components, initiating the transcription of interferon-related genes, inflammatory cytokines and other factors, thereby triggering the cellular production of an antiviral innate immune response. The protein inhibitor of activated signal transducer and activator of transcription (STAT) (PIAS) protein family has become a hot research topic due to its extensive involvement in the regulation of cytokines, inflammatory factors and innate immune signaling pathways. In the present study, we investigated the role of fish PIASy in Singapore grouper iridovirus (SGIV) and red spotted grouper nervous necrosis virus (RGNNV) infections. The homologous sequence of orange-spotted grouper (Epinephelus coioides) PIASy gene (EcPIASy) was cloned and characterized, which encoded a 498-amino acid protein with 99.20 % homology to Plectropomus leopardus. EcPIASy is expressed mainly in gills, blood, and liver. Subcellular localization showed that EcPIASy was uniformly distributed in the nucleus. Overexpression of EcPIASy promoted SGIV and RGNNV replication, and inhibited the expression of interferon related genes and pro-inflammatory factors induced by viruses. In addition, EcPIASy interacts with RLR signaling pathway-related genes EcMDA5, EcIRF3 and EcIRF7, whereas the interaction between EcPIASy and EcIRF3 does not depend on any specific structural domain of EcPIASy. The results provide a better understanding of the relationship between PIASy and viral infection in fish.
{"title":"PIASy of orange-spotted grouper (Epinephelus coioides) negatively regulates RLRs-mediated innate antiviral immunity","authors":"Qiongyue Xu , Siting Wu , Xiaoxia Lei , Helong Cao , Zhouling Zhan , Qiwei Qin , Jingguang Wei","doi":"10.1016/j.fsi.2025.110146","DOIUrl":"10.1016/j.fsi.2025.110146","url":null,"abstract":"<div><div>During viral infection, RIG-I-like receptors (RLRs) are cytoplasmic pattern recognition receptors that recognize and bind to viral RNA components, initiating the transcription of interferon-related genes, inflammatory cytokines and other factors, thereby triggering the cellular production of an antiviral innate immune response. The protein inhibitor of activated signal transducer and activator of transcription (STAT) (PIAS) protein family has become a hot research topic due to its extensive involvement in the regulation of cytokines, inflammatory factors and innate immune signaling pathways. In the present study, we investigated the role of fish PIASy in Singapore grouper iridovirus (SGIV) and red spotted grouper nervous necrosis virus (RGNNV) infections. The homologous sequence of orange-spotted grouper (<em>Epinephelus coioides</em>) PIASy gene (EcPIASy) was cloned and characterized, which encoded a 498-amino acid protein with 99.20 % homology to <em>Plectropomus leopardus</em>. EcPIASy is expressed mainly in gills, blood, and liver. Subcellular localization showed that EcPIASy was uniformly distributed in the nucleus. Overexpression of EcPIASy promoted SGIV and RGNNV replication, and inhibited the expression of interferon related genes and pro-inflammatory factors induced by viruses. In addition, EcPIASy interacts with RLR signaling pathway-related genes EcMDA5, EcIRF3 and EcIRF7, whereas the interaction between EcPIASy and EcIRF3 does not depend on any specific structural domain of EcPIASy. The results provide a better understanding of the relationship between PIASy and viral infection in fish.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110146"},"PeriodicalIF":4.1,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143022678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-20DOI: 10.1016/j.fsi.2025.110144
Noé Wambreuse , Guillaume Caulier , Igor Eeckhaut , Laura Borrello , Fabrice Bureau , Laurence Fievez , Jérôme Delroisse
Holothuria scabra is one of the most valuable species of sea cucumber owing to its exploitation as a seafood product. This study aims to describe the main molecular and cellular actors in the immunology of this species. First, a detailed description of the immune cells – the cœlomocytes – is provided, highlighting five main cell types including phagocytes, small round cells (SRCs), spherulocytes, fusiform cells, and crystal cells, with a further five subtypes identified using transmission electron microscopy. Cœlomocyte aggregates were also described morphologically, yielding two main types, one comprising three successive maturation stages. A comparison of the concentration and proportion of cell populations was carried out between the two main body fluids, namely the hydrovascular fluid of the Polian vesicle (HF) and the perivisceral fluid of the general cavity (PF), and no clear relation could be highlighted. Next, the cœlomocyte immune response was studied 24 h after lipopolysaccharide (LPS) injection in the two body fluids. Firstly, the fluctuation in cell populations was assessed, and despite a high inter-individual variability, it shows a decrease in the phagocyte proportion and an increase in the SRC proportion. Secondly, the differential gene expression of PF cœlomocytes was studied by de novo RNA-sequencing between LPS-injected and control-injected individuals: 945 genes were differentially expressed, including 673 up-regulated and 272 down-regulated in the LPS-injected individuals. Among these genes, 80 had a presumed function in immunity based on their annotation, covering a wide range of immune mechanisms. Overall, this study reveals a complex immune system at both molecular and cellular levels and constitutes a baseline reference on H. scabra immunity, which may be useful for the development of sustainable aquaculture and provides valuable data for comparative immunology.
{"title":"Morpho-functional characterisation of cœlomocytes in the aquacultivated sea cucumber Holothuria scabra: From cell diversity to transcriptomic immune response","authors":"Noé Wambreuse , Guillaume Caulier , Igor Eeckhaut , Laura Borrello , Fabrice Bureau , Laurence Fievez , Jérôme Delroisse","doi":"10.1016/j.fsi.2025.110144","DOIUrl":"10.1016/j.fsi.2025.110144","url":null,"abstract":"<div><div><em>Holothuria scabra</em> is one of the most valuable species of sea cucumber owing to its exploitation as a seafood product. This study aims to describe the main molecular and cellular actors in the immunology of this species. First, a detailed description of the immune cells – the cœlomocytes – is provided, highlighting five main cell types including phagocytes, small round cells (SRCs), spherulocytes, fusiform cells, and crystal cells, with a further five subtypes identified using transmission electron microscopy. Cœlomocyte aggregates were also described morphologically, yielding two main types, one comprising three successive maturation stages. A comparison of the concentration and proportion of cell populations was carried out between the two main body fluids, namely the hydrovascular fluid of the Polian vesicle (HF) and the perivisceral fluid of the general cavity (PF), and no clear relation could be highlighted. Next, the cœlomocyte immune response was studied 24 h after lipopolysaccharide (LPS) injection in the two body fluids. Firstly, the fluctuation in cell populations was assessed, and despite a high inter-individual variability, it shows a decrease in the phagocyte proportion and an increase in the SRC proportion. Secondly, the differential gene expression of PF cœlomocytes was studied by <em>de novo</em> RNA-sequencing between LPS-injected and control-injected individuals: 945 genes were differentially expressed, including 673 up-regulated and 272 down-regulated in the LPS-injected individuals. Among these genes, 80 had a presumed function in immunity based on their annotation, covering a wide range of immune mechanisms. Overall, this study reveals a complex immune system at both molecular and cellular levels and constitutes a baseline reference on <em>H. scabra</em> immunity<em>,</em> which may be useful for the development of sustainable aquaculture and provides valuable data for comparative immunology.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110144"},"PeriodicalIF":4.1,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143022675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-20DOI: 10.1016/j.fsi.2025.110150
Chang Cai , Peng Yang , Yong Shi, Xinyou Wang, Guanghui Chen, Qianqian Zhang, Gaofeng Cheng, Weiguang Kong, Zhen Xu
Sexual dimorphism is well-documented in aquaculture, particularly regarding growth differences, wherein one sex often grows faster than the other. However, despite the phenomenon being so widely documented, its underlying molecular mechanisms remain poorly understood. As an important digestive and immune organ, the gut plays key roles in the regulation of fish growth. In this study, we conducted RNA-seq and metabolomic analysis on the gut of female and male common carp. We discovered that growth-related pathways, such as “Glycolysis/Gluconeogenesis” and “Riboflavin metabolism” are significantly enriched in the gut of female carp. Conversely, pathways linked to disease resistance, such as “Th17 cell differentiation” and “Autophagy-animal” are predominantly enriched in male carp. Following intraperitoneal injection of spring viraemia of carp virus (SVCV) into both male and female carp, quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis and histopathological staining confirmed that male carp exhibit greater disease resistance compared to females. This study identified the disease resistance dimorphism in common carp and specific mechanisms underlying growth differences. Our findings offer valuable insights for the application of growth dimorphism and disease-resistant breeding in fish.
{"title":"Transcriptomic and metabolomic analysis revealed potential mechanisms of growth and disease resistance dimorphism in male and female common carp (Cyprinus carpio)","authors":"Chang Cai , Peng Yang , Yong Shi, Xinyou Wang, Guanghui Chen, Qianqian Zhang, Gaofeng Cheng, Weiguang Kong, Zhen Xu","doi":"10.1016/j.fsi.2025.110150","DOIUrl":"10.1016/j.fsi.2025.110150","url":null,"abstract":"<div><div>Sexual dimorphism is well-documented in aquaculture, particularly regarding growth differences, wherein one sex often grows faster than the other. However, despite the phenomenon being so widely documented, its underlying molecular mechanisms remain poorly understood. As an important digestive and immune organ, the gut plays key roles in the regulation of fish growth. In this study, we conducted RNA-seq and metabolomic analysis on the gut of female and male common carp. We discovered that growth-related pathways, such as “Glycolysis/Gluconeogenesis” and “Riboflavin metabolism” are significantly enriched in the gut of female carp. Conversely, pathways linked to disease resistance, such as “Th17 cell differentiation” and “Autophagy-animal” are predominantly enriched in male carp. Following intraperitoneal injection of spring viraemia of carp virus (SVCV) into both male and female carp, quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis and histopathological staining confirmed that male carp exhibit greater disease resistance compared to females. This study identified the disease resistance dimorphism in common carp and specific mechanisms underlying growth differences. Our findings offer valuable insights for the application of growth dimorphism and disease-resistant breeding in fish.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110150"},"PeriodicalIF":4.1,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143022680","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-19DOI: 10.1016/j.fsi.2025.110147
Marco Gerdol, Alberto Pallavicini
The Mediterranean mussel (Mytilus galloprovincialis) is a key species in European aquaculture, known for its economic and societal importance, particularly as a primary source of income for local fisheries in European coastal areas. While historically resilient to the mass mortality events that have affected other bivalve species, M. galloprovincialis may face increasing threats from emerging pathogens, including bacteria, viruses, and eukaryotic parasites. These microorganisms, often opportunistic, pose heightened risks in the current climate change scenario, where heatwaves are becoming increasingly frequent and the persistent presence of pollutants is suspected to impair the functional response of hemocytes. Over the past decade, significant advancements in immunological research have provided deeper insights into the cellular and molecular mechanisms underlying the robust defense system of M. galloprovincialis, which allows this species to efficiently cope with a broad range of infections. By analyzing the scientific literature published on mussel immunology over the past ten years, this review consolidates current knowledge on the immune system of the Mediterranean mussel. We place a particular focus on the cellular and molecular components involved in the recognition and elimination of microbial pathogens and discuss how the most recent discoveries may inform improved management and disease mitigation strategies for Mediterranean mussel farming in the in the years to come.
{"title":"Exploring the immune resilience of Mediterranean mussels: Recent advances and future directions","authors":"Marco Gerdol, Alberto Pallavicini","doi":"10.1016/j.fsi.2025.110147","DOIUrl":"10.1016/j.fsi.2025.110147","url":null,"abstract":"<div><div>The Mediterranean mussel (<em>Mytilus galloprovincialis</em>) is a key species in European aquaculture, known for its economic and societal importance, particularly as a primary source of income for local fisheries in European coastal areas. While historically resilient to the mass mortality events that have affected other bivalve species, <em>M. galloprovincialis</em> may face increasing threats from emerging pathogens, including bacteria, viruses, and eukaryotic parasites. These microorganisms, often opportunistic, pose heightened risks in the current climate change scenario, where heatwaves are becoming increasingly frequent and the persistent presence of pollutants is suspected to impair the functional response of hemocytes. Over the past decade, significant advancements in immunological research have provided deeper insights into the cellular and molecular mechanisms underlying the robust defense system of <em>M. galloprovincialis</em>, which allows this species to efficiently cope with a broad range of infections. By analyzing the scientific literature published on mussel immunology over the past ten years, this review consolidates current knowledge on the immune system of the Mediterranean mussel. We place a particular focus on the cellular and molecular components involved in the recognition and elimination of microbial pathogens and discuss how the most recent discoveries may inform improved management and disease mitigation strategies for Mediterranean mussel farming in the in the years to come.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110147"},"PeriodicalIF":4.1,"publicationDate":"2025-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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