Gills are the main respiratory organs of fish and bear important physiological and immunological functions, but the functional heterogeneity of interlamellar cell mass (ILCM) at the single-cell level has rarely been reported. Here, we identified 19 cell types from the gills of crucian carp (Carassius auratus) by single-cell RNA sequencing (scRNA-seq) in combination with histological analysis. We annotated ILCM and analyzed its functional heterogeneity at the single-cell level for the first time. Functional enrichment analysis and cell cycle analysis identified ILCM as a type of metabolically active cells in a state of constant proliferation, and identified the major pathways responsible for ILCM immunoregulation. Histological analysis revealed the morphology and positional relationships of 6 cell types. Meanwhile, the gene regulatory network of ILCM was established through weighted gene co-expression network analysis (WGCNA), and one transcription factor and five hub genes related to immunoregulation were identified. We found that pyroptosis might be an important pathway responsible for the immune response of ILCM. Our findings provide an insight into the physiological and immune functions of gills and ILCM at the single-cell level and lay a solid foundation for further exploration of the molecular mechanism of ILCM immunity functions.
{"title":"Single-cell transcriptome sequencing analysis of physiological and immune profiling of crucian carp (Carassius auratus) gills","authors":"Zhi-Guang Hou , Meng-Chao Xing , Jia-Xing Luo , Yi-Huan Xu , Li-Han Zhang , Xiao-Wei Gao , Jiang-Jiang Wang , Fazhan Hanafiah , Waiho Khor , Xin Hao , Xin Zhao , Cheng-Bin Wu","doi":"10.1016/j.fsi.2024.110087","DOIUrl":"10.1016/j.fsi.2024.110087","url":null,"abstract":"<div><div>Gills are the main respiratory organs of fish and bear important physiological and immunological functions, but the functional heterogeneity of interlamellar cell mass (ILCM) at the single-cell level has rarely been reported. Here, we identified 19 cell types from the gills of crucian carp (<em>Carassius auratus</em>) by single-cell RNA sequencing (scRNA-seq) in combination with histological analysis. We annotated ILCM and analyzed its functional heterogeneity at the single-cell level for the first time. Functional enrichment analysis and cell cycle analysis identified ILCM as a type of metabolically active cells in a state of constant proliferation, and identified the major pathways responsible for ILCM immunoregulation. Histological analysis revealed the morphology and positional relationships of 6 cell types. Meanwhile, the gene regulatory network of ILCM was established through weighted gene co-expression network analysis (WGCNA), and one transcription factor and five hub genes related to immunoregulation were identified. We found that pyroptosis might be an important pathway responsible for the immune response of ILCM. Our findings provide an insight into the physiological and immune functions of gills and ILCM at the single-cell level and lay a solid foundation for further exploration of the molecular mechanism of ILCM immunity functions.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110087"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142812425","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 : 2025-02-01DOI: 10.1016/j.fsi.2024.110096
Ottavia Benedicenti, Maria K. Dahle, Shokouh Makvandi-Nejad , Adriana Magalhaes Santos Andresen, Torfinn Moldal, Hilde Sindre, Johanna Hol Fosse
Infectious Salmon Anaemia virus (ISAV) is an orthomyxovirus that causes large economic losses in Atlantic salmon (Salmo salar L.) aquaculture. All virulent ISAV variants originally emerged from a non-virulent subtype, ISAV-HPR0. Transient ISAV-HPR0 infections are common in both freshwater and marine environments. ISAV-HPR0 infects juveniles, marine salmon at on-growing sites, and broodstock salmon. The shift in virulence from ISAV-HPR0 to the virulent HPRΔ is suggested to be a stochastic event that depends on the virus's replication frequency. Therefore, reducing the capacity to maintain ISAV-HPR0 infection within individual farms may limit the risk of emerging pathogenic ISAV variants and ISA disease. The absence of infection-related clinical signs and the lack of experimental models limit our understanding of ISAV-HPR0-host interactions. We characterise the host transcriptional response to natural ISAV-HPR0 infection, using Atlantic salmon gill tissues collected on three Norwegian smolt farms. The comparison of all infected (qPCR-positive) and non-infected (qPCR-negative) individuals revealed a classic antiviral response in the gills of ISAV-HPR0 infected fish in a site-independent transcriptomic analysis. Complementary analyses showed that the response to infection varied considerably between sites. Site-specific differences could be associated with a range of factors that are challenging to control in field studies, such as fish size, the stage of infection, and the presence of additional microorganisms.
Our findings enhance our understanding of how Atlantic salmon respond to ISAV-HPR0 infection, pinpointing common HPR0-induced antiviral response genes. Future studies should investigate whether these candidate genes limit virus replication in the gill for risk of novel transitions to virulence.
{"title":"The Atlantic salmon gill transcriptional response to natural infection with HPR0-ISAV (Isavirus salaris) in three Norwegian smolt farms","authors":"Ottavia Benedicenti, Maria K. Dahle, Shokouh Makvandi-Nejad , Adriana Magalhaes Santos Andresen, Torfinn Moldal, Hilde Sindre, Johanna Hol Fosse","doi":"10.1016/j.fsi.2024.110096","DOIUrl":"10.1016/j.fsi.2024.110096","url":null,"abstract":"<div><div>Infectious Salmon Anaemia virus (ISAV) is an orthomyxovirus that causes large economic losses in Atlantic salmon (<em>Salmo salar</em> L.) aquaculture. All virulent ISAV variants originally emerged from a non-virulent subtype, ISAV-HPR0. Transient ISAV-HPR0 infections are common in both freshwater and marine environments. ISAV-HPR0 infects juveniles, marine salmon at on-growing sites, and broodstock salmon. The shift in virulence from ISAV-HPR0 to the virulent HPRΔ is suggested to be a stochastic event that depends on the virus's replication frequency. Therefore, reducing the capacity to maintain ISAV-HPR0 infection within individual farms may limit the risk of emerging pathogenic ISAV variants and ISA disease. The absence of infection-related clinical signs and the lack of experimental models limit our understanding of ISAV-HPR0-host interactions. We characterise the host transcriptional response to natural ISAV-HPR0 infection, using Atlantic salmon gill tissues collected on three Norwegian smolt farms. The comparison of all infected (qPCR-positive) and non-infected (qPCR-negative) individuals revealed a classic antiviral response in the gills of ISAV-HPR0 infected fish in a site-independent transcriptomic analysis. Complementary analyses showed that the response to infection varied considerably between sites. Site-specific differences could be associated with a range of factors that are challenging to control in field studies, such as fish size, the stage of infection, and the presence of additional microorganisms.</div><div>Our findings enhance our understanding of how Atlantic salmon respond to ISAV-HPR0 infection, pinpointing common HPR0-induced antiviral response genes. Future studies should investigate whether these candidate genes limit virus replication in the gill for risk of novel transitions to virulence.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110096"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142893193","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}
Pub Date : 2025-02-01DOI: 10.1016/j.fsi.2024.110093
Meng Zhang , Xiao Ma , Zerui Wang , Yuqing Han , Zhilin Jia , Dongcai Chen , Yue Xu , Zhigang Qiao , Xinyu Jiang , Lei Wang , Hongxia Jiang , Miao Yu , Yongjing Li , Yawei Shen
Largemouth bass (Micropterus salmoides) has become one of the most important freshwater economic fish farmed almost all over China in recent years. At the same time, the increasing outbreaks of diseases in its aquaculture process have caused substantial economic losses to this industry. However, at present, the genetic basis of disease resistance, including resistance against Aeromonas veronii infection, in largemouth bass is very limited. Therefore, a genome-wide association study (GWAS) on host resistance against the A. veronii of largemouth bass was conducted in the present study. A total of 627 largemouth bass were artificially challenged by A. veronii, among which 160 of the earliest deaths and 173 of the final survivals were genotyped. A total of 3076 high-quality SNPs were used for further analysis employing two analysis models, of which six shared SNPs were finally identified as significant molecular markers with the explaining phenotypic variance ranging from 2.28 % to 8.95 %. Furthermore, seven candidate genes were identified, including one gene, T-cell surface antigen CD2, which is directly involved in T cell activation and the cellular immune response. Additionally, the other identified genes play roles in critical processes such as cell survival, inflammatory responses, and signal transduction. This study lays a genetic foundation for research on largemouth bass disease resistance and studies related to A. veronii. It also contributes significantly to the future development of the commercial production of largemouth bass.
{"title":"Genome-wide association analysis study on host resistance against the Aeromonas veronii of largemouth bass Micropterus salmoides","authors":"Meng Zhang , Xiao Ma , Zerui Wang , Yuqing Han , Zhilin Jia , Dongcai Chen , Yue Xu , Zhigang Qiao , Xinyu Jiang , Lei Wang , Hongxia Jiang , Miao Yu , Yongjing Li , Yawei Shen","doi":"10.1016/j.fsi.2024.110093","DOIUrl":"10.1016/j.fsi.2024.110093","url":null,"abstract":"<div><div>Largemouth bass (<em>Micropterus salmoides</em>) has become one of the most important freshwater economic fish farmed almost all over China in recent years. At the same time, the increasing outbreaks of diseases in its aquaculture process have caused substantial economic losses to this industry. However, at present, the genetic basis of disease resistance, including resistance against <em>Aeromonas veronii</em> infection, in largemouth bass is very limited. Therefore, a genome-wide association study (GWAS) on host resistance against the <em>A. veronii</em> of largemouth bass was conducted in the present study. A total of 627 largemouth bass were artificially challenged by <em>A. veronii</em>, among which 160 of the earliest deaths and 173 of the final survivals were genotyped. A total of 3076 high-quality SNPs were used for further analysis employing two analysis models, of which six shared SNPs were finally identified as significant molecular markers with the explaining phenotypic variance ranging from 2.28 % to 8.95 %. Furthermore, seven candidate genes were identified, including one gene, T-cell surface antigen <em>CD2</em>, which is directly involved in T cell activation and the cellular immune response. Additionally, the other identified genes play roles in critical processes such as cell survival, inflammatory responses, and signal transduction. This study lays a genetic foundation for research on largemouth bass disease resistance and studies related to <em>A. veronii</em>. It also contributes significantly to the future development of the commercial production of largemouth bass.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110093"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142906810","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 : 2025-02-01DOI: 10.1016/j.fsi.2025.110113
Qiang Hao , Xiuyan Gao , Mingzhe Sun , Yuan Liu
Fibrinogen-related domain (FReD) containing proteins are an evolutionarily conserved immune gene family characterized by the C-terminal fibrinogen (FBG) and diverse N-terminal domains. To understand the complexity of this family in crustaceans, we performed genome screening and identified 43 full-length FReDs encoding genes in Litopenaeus vannamei. Structural classification analysis revealed these putative FReDs could be divided into six types, including two reported types (LvFReDI and II) and four new types (LvFReDIII-VI). Sequence and phylogenetic analysis showed that FBG domains were highly conserved throughout and phylogeny clusters correlated strongly with gene type. We analyzed the temporal and spatial expression patterns of LvFReD genes based on the transcriptomes of developmental stages, adult tissues or pathogen infected tissues of L. vannamei. Most LvFReDs were expressed from larval in membrane stage, and exhibited tissue-specific expression patterns and immune-responsive transcription after challenge with bacteria or virus. Further time-course expression analysis suggested that LvFReDII genes with additional coiled-coil region were more sensitive to pathogens than LvFReDI genes. Our findings provided comprehensive gene sequence resources and expression profiles of FReD genes in shrimp, which give insights into clarifying the diversity and function of these genes in crustaceans.
{"title":"Genomic insights into fibrinogen-related proteins and expression analysis in the Pacific white shrimp, Litopenaeus vannamei","authors":"Qiang Hao , Xiuyan Gao , Mingzhe Sun , Yuan Liu","doi":"10.1016/j.fsi.2025.110113","DOIUrl":"10.1016/j.fsi.2025.110113","url":null,"abstract":"<div><div>Fibrinogen-related domain (FReD) containing proteins are an evolutionarily conserved immune gene family characterized by the C-terminal fibrinogen (FBG) and diverse N-terminal domains. To understand the complexity of this family in crustaceans, we performed genome screening and identified 43 full-length FReDs encoding genes in <em>Litopenaeus vannamei</em>. Structural classification analysis revealed these putative FReDs could be divided into six types, including two reported types (LvFReDI and II) and four new types (LvFReDIII-VI). Sequence and phylogenetic analysis showed that FBG domains were highly conserved throughout and phylogeny clusters correlated strongly with gene type. We analyzed the temporal and spatial expression patterns of LvFReD genes based on the transcriptomes of developmental stages, adult tissues or pathogen infected tissues of <em>L. vannamei</em>. Most LvFReDs were expressed from larval in membrane stage, and exhibited tissue-specific expression patterns and immune-responsive transcription after challenge with bacteria or virus. Further time-course expression analysis suggested that LvFReDII genes with additional coiled-coil region were more sensitive to pathogens than LvFReDI genes. Our findings provided comprehensive gene sequence resources and expression profiles of FReD genes in shrimp, which give insights into clarifying the diversity and function of these genes in crustaceans.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110113"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142946750","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 : 2025-02-01DOI: 10.1016/j.fsi.2025.110112
Seung-Hyeon Kim , Hyoun Joong Kim , S.D.N.K. Bathige , Soohwan Kim , Kyung-Il Park
In this study, we investigated the variability in virulence among different strains of Perkinsus marinus and other Perkinsus species in Eastern oysters (Crassostrea virginica), examining the immune responses and mortality rates of oysters exposed to different Perkinsus isolates. Compared with the other assessed strains, P. marinus strain ATCC 50787 was found to induce significantly (P < 0.05) higher levels of reactive oxygen species, nitric oxide, and necrosis in oyster hemocytes. Perkinsus chesapeaki (ATCC PRA-65) elicited strong immune responses and high mortality in Eastern oysters at rates similar to those induced by the most virulent P. marinus strain. In contrast, P. olseni and P. honshuensis induced low levels of immune response and mortality. In vivo survival assays confirmed that strains inducing high immune responses in vitro also caused high mortalities in oysters. Our findings in this study highlight the importance of considering strain-specific virulence when studying Perkinsus infections in oysters. These findings have implications for understanding host–parasite interactions and managing Perkinsus-related diseases in oyster populations, particularly from the perspectives of aquaculture and conservation.
{"title":"Strain-specific virulence of Perkinsus marinus and related species in Eastern oysters: A comprehensive analysis of immune responses and mortality","authors":"Seung-Hyeon Kim , Hyoun Joong Kim , S.D.N.K. Bathige , Soohwan Kim , Kyung-Il Park","doi":"10.1016/j.fsi.2025.110112","DOIUrl":"10.1016/j.fsi.2025.110112","url":null,"abstract":"<div><div>In this study, we investigated the variability in virulence among different strains of <em>Perkinsus marinus</em> and other <em>Perkinsus</em> species in Eastern oysters (<em>Crassostrea virginica</em>), examining the immune responses and mortality rates of oysters exposed to different <em>Perkinsus</em> isolates. Compared with the other assessed strains, <em>P. marinus</em> strain ATCC 50787 was found to induce significantly (<em>P</em> < 0.05) higher levels of reactive oxygen species, nitric oxide, and necrosis in oyster hemocytes. <em>Perkinsus chesapeaki</em> (ATCC PRA-65) elicited strong immune responses and high mortality in Eastern oysters at rates similar to those induced by the most virulent <em>P. marinus</em> strain. In contrast, <em>P. olseni</em> and <em>P. honshuensis</em> induced low levels of immune response and mortality. <em>In vivo</em> survival assays confirmed that strains inducing high immune responses <em>in vitro</em> also caused high mortalities in oysters. Our findings in this study highlight the importance of considering strain-specific virulence when studying <em>Perkinsus</em> infections in oysters. These findings have implications for understanding host–parasite interactions and managing <em>Perkinsus</em>-related diseases in oyster populations, particularly from the perspectives of aquaculture and conservation.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110112"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142946807","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 : 2025-02-01DOI: 10.1016/j.fsi.2025.110169
Shunli Che , Jiawei Chen , Hai Zhang, Weizhen Xu, Yanwei Li, Xueming Dan, Zequan Mo
The mandarin fish (Siniperca chuatsi) is a significant freshwater carnivorous species in Chinese aquaculture industry, and its farming scale is continuously expanding. The use of feed in aquaculture has become an increasingly common practice. However, the impacts of substituting artificial feed for live bait on fish's biochemical and immune responses are poorly understood. In this study, two hundred forty mandarin fish (weight: 5.60 ± 0.41 g) were divided into two groups and fed live bait or artificial feed (LB and AF groups) over a 63-day aquaculture experiment. We compared the differences between the two groups in terms of histology, biochemical indicators, gene expression, and bacterial resistance. The results showed that artificial feed promoted enhanced growth, evidenced by higher weight (p < 0.05). The AF group exhibited higher liver and intestinal somatic indices (p < 0.05), and histological examination revealed denser cytoplasmic content in liver cells, less fragmentation of renal tubular epithelial cells, and less detachment of intestinal epithelial cells in the AF group. Regarding biochemical indicators and gene expression, the AF group showed better performance in glucose regulation and lipid metabolism. The AF group maintained glucose balance (p < 0.05) and effectively regulated cholesterol transport (p < 0.05), promoting lipolysis (p < 0.05) while inhibiting lipogenesis (p < 0.05). In contrast, live bait consumption resulted in reduced lipolysis (p < 0.05), increased lipogenesis (p < 0.05), impaired endoplasmic reticulum function (p < 0.05), heightened inflammation (p < 0.05), and diminished antioxidant capacity (p < 0.05). Additionally, the LB group exhibited lower survival rates and lysozyme levels during bacterial challenges. Overall, artificial feed was more beneficial for the growth, regulate physiology and enhance disease resistance of S. chuatsi, highlighting its potential to improve fish health and increase aquaculture yield.
{"title":"Impacts of live and artificial feed on histology, biochemical indicators, gene expression, and bacterial resistance in mandarin fish (Siniperca chuatsi)","authors":"Shunli Che , Jiawei Chen , Hai Zhang, Weizhen Xu, Yanwei Li, Xueming Dan, Zequan Mo","doi":"10.1016/j.fsi.2025.110169","DOIUrl":"10.1016/j.fsi.2025.110169","url":null,"abstract":"<div><div>The mandarin fish (<em>Siniperca chuatsi</em>) is a significant freshwater carnivorous species in Chinese aquaculture industry, and its farming scale is continuously expanding. The use of feed in aquaculture has become an increasingly common practice. However, the impacts of substituting artificial feed for live bait on fish's biochemical and immune responses are poorly understood. In this study, two hundred forty mandarin fish (weight: 5.60 ± 0.41 g) were divided into two groups and fed live bait or artificial feed (LB and AF groups) over a 63-day aquaculture experiment. We compared the differences between the two groups in terms of histology, biochemical indicators, gene expression, and bacterial resistance. The results showed that artificial feed promoted enhanced growth, evidenced by higher weight (<em>p</em> < 0.05). The AF group exhibited higher liver and intestinal somatic indices (<em>p</em> < 0.05), and histological examination revealed denser cytoplasmic content in liver cells, less fragmentation of renal tubular epithelial cells, and less detachment of intestinal epithelial cells in the AF group. Regarding biochemical indicators and gene expression, the AF group showed better performance in glucose regulation and lipid metabolism. The AF group maintained glucose balance (<em>p</em> < 0.05) and effectively regulated cholesterol transport (<em>p</em> < 0.05), promoting lipolysis (<em>p</em> < 0.05) while inhibiting lipogenesis (<em>p</em> < 0.05). In contrast, live bait consumption resulted in reduced lipolysis (<em>p</em> < 0.05), increased lipogenesis (<em>p</em> < 0.05), impaired endoplasmic reticulum function (<em>p</em> < 0.05), heightened inflammation (<em>p</em> < 0.05), and diminished antioxidant capacity (<em>p</em> < 0.05). Additionally, the LB group exhibited lower survival rates and lysozyme levels during bacterial challenges. Overall, artificial feed was more beneficial for the growth, regulate physiology and enhance disease resistance of <em>S. chuatsi</em>, highlighting its potential to improve fish health and increase aquaculture yield.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"158 ","pages":"Article 110169"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143122448","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 : 2025-02-01DOI: 10.1016/j.fsi.2024.110082
U.P.E. Arachchi , K.P. Madushani , K.A.S.N. Shanaka , Gaeun Kim , Chaehyeon Lim , Hyerim Yang , B.P.M. Vileka Jayamali , Yasara Kavindi Kodagoda , W.A.D.L.R. Warnakula , Sumi Jung , Qiang Wan , Jehee Lee
The tripartite motif-containing (TRIM) superfamily is the largest family of RING-type E3 ubiquitin ligases that is conserved across the metazoan kingdom. Previous studies in mammals have demonstrated that TRIM59 possesses ubiquitin-protein ligase activity and acts as a negative regulator of NF-κB signaling. However, TRIM59 has rarely been characterized in fish. This study aimed to characterize TRIM59 from Epinephelus akaara (Eatrim59) and elucidate its structural features, expression patterns, and functional properties in innate immune responses and in the regulation of apoptosis. Eatrim59 is composed of 406 amino acids with a molecular weight of 45.84 kDa and a theoretical isoelectric point of 5.25. It comprises a conserved RING domain, a B-box motif, and a coiled-coil region. Subcellular localization analysis revealed that Eatrim59 was localized in the endoplasmic reticulum. Eatrim59 was ubiquitously expressed in all tissues examined, with the highest relative expression detected in the blood, followed by the brain and spleen. Temporal expression of Eatrim59 was dynamically regulated in response to in vivo immune stimulation by Toll-like receptor ligands and nervous necrosis virus infection. In FHM cells overexpressing Eatrim59, an increase in viral replication was observed upon infection with the Viral hemorrhagic septicemia virus. This phenomenon is attributed to Eatrim59-mediated downregulation of interferon, pro-inflammatory cytokines, and other antiviral pathways. Moreover, macrophages stably overexpressing Eatrim59 exhibited a decrease in nitric oxide production and the formation of a filamentous actin structure upon lipopolysaccharide stimulation, indicating dampened M1 polarization. Furthermore, a decrease in apoptosis was observed in Eatrim59-overexpressing FHM cells under oxidative stress induced by H2O2. In conclusion, these findings demonstrate the multifaceted role of Eatrim59 as a regulator of innate immune response and apoptosis in E. akaara.
{"title":"Characterization of tripartite motif containing 59 (TRIM59) in Epinephelus akaara: Insights into its immune involvement and functional properties in viral pathogenesis, macrophage polarization, and apoptosis regulation","authors":"U.P.E. Arachchi , K.P. Madushani , K.A.S.N. Shanaka , Gaeun Kim , Chaehyeon Lim , Hyerim Yang , B.P.M. Vileka Jayamali , Yasara Kavindi Kodagoda , W.A.D.L.R. Warnakula , Sumi Jung , Qiang Wan , Jehee Lee","doi":"10.1016/j.fsi.2024.110082","DOIUrl":"10.1016/j.fsi.2024.110082","url":null,"abstract":"<div><div>The tripartite motif-containing (TRIM) superfamily is the largest family of RING-type E3 ubiquitin ligases that is conserved across the metazoan kingdom. Previous studies in mammals have demonstrated that TRIM59 possesses ubiquitin-protein ligase activity and acts as a negative regulator of NF-κB signaling. However, TRIM59 has rarely been characterized in fish. This study aimed to characterize TRIM59 from <em>Epinephelus akaara</em> (<em>Eatrim59</em>) and elucidate its structural features, expression patterns, and functional properties in innate immune responses and in the regulation of apoptosis. Eatrim59 is composed of 406 amino acids with a molecular weight of 45.84 kDa and a theoretical isoelectric point of 5.25. It comprises a conserved RING domain, a B-box motif, and a coiled-coil region. Subcellular localization analysis revealed that Eatrim59 was localized in the endoplasmic reticulum. Eatrim59 was ubiquitously expressed in all tissues examined, with the highest relative expression detected in the blood, followed by the brain and spleen. Temporal expression of Eatrim59 was dynamically regulated in response to <em>in vivo</em> immune stimulation by Toll-like receptor ligands and nervous necrosis virus infection. In FHM cells overexpressing Eatrim59, an increase in viral replication was observed upon infection with the Viral hemorrhagic septicemia virus. This phenomenon is attributed to Eatrim59-mediated downregulation of interferon, pro-inflammatory cytokines, and other antiviral pathways. Moreover, macrophages stably overexpressing Eatrim59 exhibited a decrease in nitric oxide production and the formation of a filamentous actin structure upon lipopolysaccharide stimulation, indicating dampened M1 polarization. Furthermore, a decrease in apoptosis was observed in Eatrim59-overexpressing FHM cells under oxidative stress induced by H<sub>2</sub>O<sub>2</sub>. In conclusion, these findings demonstrate the multifaceted role of Eatrim59 as a regulator of innate immune response and apoptosis in <em>E. akaara</em>.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110082"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791404","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 : 2025-02-01DOI: 10.1016/j.fsi.2024.110098
T. Sathish Kumar , B. Naveen Rajeshwar , T. Sivaramakrishnan , Sujeet Kumar , Vidya Rajendran , Shiva Kumar , Lane Pineda , Morvarid Rezvani , S. Saravanan
Vibriosis caused by Vibrio spp. is imposing severe havoc and adverse effects on shrimp culture. Antibiotics are the most widely used therapeutic measures against vibriosis. However, the uncontrolled use of antibiotics may spread the antibiotic residues and increase antibiotic-resistant bacteria. In this study, a product based on a synergistic blend of short- and medium-chain fatty acids (Selacid® GG, herein referred as Sel) was evaluated against vibriosis in shrimp, P. vannamei. The shrimps (n = 30 in triplicate per treatment) were fed with Sel (0.0 %, 0.1 %, 0.2 % or 0.3 %) for 14 days. The shrimp were challenged on 8th day by immersion, and the samples were collected on the 6th day post-challenge and the 14th day post-feeding. The shrimp fed with Sel (0.1 %, 0.2 %, and 0.3 %) diet showed significantly higher total haemocyte count, while the phenoloxidase activity was significantly increased in the Sel 0.2 % diet fed group. Histological analysis unveiled inflammatory responses with haemocytic infiltration, encapsulation, and granuloma in Sel 0.2 % diet-fed shrimps with less degeneration and necrosis of hepatopancreas tubules and epithelial cells. qPCR analysis of vibrio loads in hepatopancreas revealed significantly lower vibrio count in all Sel diet-fed groups and further confirmed by culture-dependent method which showed significantly lower total vibrio count in the hepatopancreas and haemolymph of Sel 0.2 % and Sel 0.3 % diet fed shrimp. Growth parameters showed significantly higher values in weight gain, specific growth rate, and survival rate in Sel 0.2 % diet-fed shrimps. Thus, the current study confirms the efficacy of Sel (0.2 %) against vibriosis by enhancing the immune and inflammatory responses, reducing vibrio load and improving the growth and survival. Hence Sel, can be used as an alternative therapeutic for managing vibriosis in shrimp aquaculture.
{"title":"Evaluation of a synergistic blend of short- and medium-chain fatty acids as a dietary intervention for the effective management of Vibriosis in shrimp culture","authors":"T. Sathish Kumar , B. Naveen Rajeshwar , T. Sivaramakrishnan , Sujeet Kumar , Vidya Rajendran , Shiva Kumar , Lane Pineda , Morvarid Rezvani , S. Saravanan","doi":"10.1016/j.fsi.2024.110098","DOIUrl":"10.1016/j.fsi.2024.110098","url":null,"abstract":"<div><div>Vibriosis caused by <em>Vibrio</em> spp. is imposing severe havoc and adverse effects on shrimp culture. Antibiotics are the most widely used therapeutic measures against vibriosis. However, the uncontrolled use of antibiotics may spread the antibiotic residues and increase antibiotic-resistant bacteria. In this study, a product based on a synergistic blend of short- and medium-chain fatty acids (Selacid® GG, herein referred as Sel) was evaluated against vibriosis in shrimp, <em>P. vannamei</em>. The shrimps (n = 30 in triplicate per treatment) were fed with Sel (0.0 %, 0.1 %, 0.2 % or 0.3 %) for 14 days. The shrimp were challenged on 8th day by immersion, and the samples were collected on the 6th day post-challenge and the 14th day post-feeding. The shrimp fed with Sel (0.1 %, 0.2 %, and 0.3 %) diet showed significantly higher total haemocyte count, while the phenoloxidase activity was significantly increased in the Sel 0.2 % diet fed group. Histological analysis unveiled inflammatory responses with haemocytic infiltration, encapsulation, and granuloma in Sel 0.2 % diet-fed shrimps with less degeneration and necrosis of hepatopancreas tubules and epithelial cells. qPCR analysis of vibrio loads in hepatopancreas revealed significantly lower vibrio count in all Sel diet-fed groups and further confirmed by culture-dependent method which showed significantly lower total vibrio count in the hepatopancreas and haemolymph of Sel 0.2 % and Sel 0.3 % diet fed shrimp. Growth parameters showed significantly higher values in weight gain, specific growth rate, and survival rate in Sel 0.2 % diet-fed shrimps. Thus, the current study confirms the efficacy of Sel (0.2 %) against vibriosis by enhancing the immune and inflammatory responses, reducing vibrio load and improving the growth and survival. Hence Sel, can be used as an alternative therapeutic for managing vibriosis in shrimp aquaculture.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110098"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142893096","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 : 2025-02-01DOI: 10.1016/j.fsi.2024.110071
Li He , Meiqin Mao , Hui Ge , Jiaonan Zhang , Jiaolin Zhang , Qingpi Yan
Flagella-mediated swarming motility plays a crucial role in facilitating the rapid colonization and dissemination of bacterial within the host. The swarming motility of Pseudomonas plecoglossicida is intricately associated with its lateral flagella, and notably, the lateral flagella system of P. plecoglossicida encompasses a transcriptional regulator known as LafK. However, the regulatory role of LafK and its impact on bacteria-host interactions remain to be elucidated. In this study, we systematically investigated the regulatory role of LafK by constructing lafK deletion strain on the biological characteristics, virulence, and pathogenic process of P. plecoglossicida, as well as its impact on the host immune response. Our findings demonstrated that the deletion of lafK led to a significant down-regulation in the expression of type III secretion system-associated genes within the lateral flagella of P. plecoglossicida, consequently impairing bacterial swarming motility, biofilm formation, adhesion, and chemotaxis ability. Furthermore, in vitro infection experiments demonstrated that the deletion of lafK resulted in a diminished pathogenicity of P. plecoglossicida through down-regulation of flagella-related genes, thereby triggering an expedited immune response for bacterial clearance, and subsequently leading to reduced bacterial load within the host and attenuated tissue damage during infection. In summary, this study presents a novel theoretical framework for elucidating the regulatory mechanism of virulence in P. plecoglossicida.
{"title":"lafK contributes the regulation of swarming motility of Pseudomonas plecoglossicida and bacterial-host interaction","authors":"Li He , Meiqin Mao , Hui Ge , Jiaonan Zhang , Jiaolin Zhang , Qingpi Yan","doi":"10.1016/j.fsi.2024.110071","DOIUrl":"10.1016/j.fsi.2024.110071","url":null,"abstract":"<div><div>Flagella-mediated swarming motility plays a crucial role in facilitating the rapid colonization and dissemination of bacterial within the host. The swarming motility of <em>Pseudomonas plecoglossicida</em> is intricately associated with its lateral flagella, and notably, the lateral flagella system of <em>P</em>. <em>plecoglossicida</em> encompasses a transcriptional regulator known as LafK. However, the regulatory role of LafK and its impact on bacteria-host interactions remain to be elucidated. In this study, we systematically investigated the regulatory role of LafK by constructing <em>lafK</em> deletion strain on the biological characteristics, virulence, and pathogenic process of <em>P</em>. <em>plecoglossicida</em>, as well as its impact on the host immune response. Our findings demonstrated that the deletion of <em>lafK</em> led to a significant down-regulation in the expression of type III secretion system-associated genes within the lateral flagella of <em>P</em>. <em>plecoglossicida</em>, consequently impairing bacterial swarming motility, biofilm formation, adhesion, and chemotaxis ability. Furthermore, in vitro infection experiments demonstrated that the deletion of <em>lafK</em> resulted in a diminished pathogenicity of <em>P</em>. <em>plecoglossicida</em> through down-regulation of flagella-related genes, thereby triggering an expedited immune response for bacterial clearance, and subsequently leading to reduced bacterial load within the host and attenuated tissue damage during infection. In summary, this study presents a novel theoretical framework for elucidating the regulatory mechanism of virulence in <em>P</em>. <em>plecoglossicida</em>.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110071"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142784634","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 : 2025-02-01DOI: 10.1016/j.fsi.2024.110088
Thomais Tsoulia , Arvind YM. Sundaram , Marit M. Amundsen , Espen Rimstad , Øystein Wessel , Jorunn B. Jørgensen , Maria K. Dahle
Piscine orthoreovirus (PRV) infection is common in aquaculture of salmonids. The three known PRV genotypes (PRV-1-3) have host species specificity and cause different diseases, but all infect and replicate in red blood cells (RBCs) in early infection phase. PRV-1 is the causative agent of heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon (Salmo salar), PRV-2 causes erythrocytic inclusion body syndrome (EIBS) in coho salmon (Oncorhynchus kisutch), while PRV-3 induces HSMI-like disease in farmed rainbow trout (Oncorhynchus mykiss). PRV-3 can also infect A. salmon without causing clinical disease and has been shown to cross-protect against PRV-1 infection and HSMI, while PRV-2 or inactivated adjuvanted PRV-1 vaccine only partially reduced HSMI pathologic changes. In the present work, we studied the transcriptional responses in blood cells of A. salmon two- and five-weeks post infection with PRV-1, PRV-2, PRV-3, or post injection with inactivated PRV-1 vaccine. PRV-1 and PRV-3 replicated well in A. salmon blood cells, and both induced the typical innate antiviral responses triggered by dsRNA viruses. Two weeks post infection, PRV-3 triggered stronger antiviral responses than PRV-1, despite their similar viral RNA replication levels, but after five weeks the induced responses were close to equal. PRV-2 and the InPRV-1 vaccine did not trigger the same typical antiviral responses as the replicating PRV-1 and PRV-3 genotypes, but induced genes involved in membrane trafficking and signaling pathways that may regulate physiological functions. These findings propose that the protection mediated by PRV-3 against a secondary infection by PRV-1 occur due to a potent and early activation of the same type of innate immune responses. The difference in the timing of antiviral responses may give PRV-1 an evolutionary edge, facilitating its dissemination to A. salmon heart, a critical step for HSMI development.
{"title":"Comparison of transcriptome responses in blood cells of Atlantic salmon infected by three genotypes of Piscine orthoreovirus","authors":"Thomais Tsoulia , Arvind YM. Sundaram , Marit M. Amundsen , Espen Rimstad , Øystein Wessel , Jorunn B. Jørgensen , Maria K. Dahle","doi":"10.1016/j.fsi.2024.110088","DOIUrl":"10.1016/j.fsi.2024.110088","url":null,"abstract":"<div><div><em>Piscine orthoreovirus</em> (PRV) infection is common in aquaculture of salmonids. The three known PRV genotypes (PRV-1-3) have host species specificity and cause different diseases, but all infect and replicate in red blood cells (RBCs) in early infection phase. PRV-1 is the causative agent of heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon (<em>Salmo salar</em>), PRV-2 causes erythrocytic inclusion body syndrome (EIBS) in coho salmon (<em>Oncorhynchus kisutch</em>), while PRV-3 induces HSMI-like disease in farmed rainbow trout (<em>Oncorhynchus mykiss</em>). PRV-3 can also infect A. salmon without causing clinical disease and has been shown to cross-protect against PRV-1 infection and HSMI, while PRV-2 or inactivated adjuvanted PRV-1 vaccine only partially reduced HSMI pathologic changes. In the present work, we studied the transcriptional responses in blood cells of A. salmon two- and five-weeks post infection with PRV-1, PRV-2, PRV-3, or post injection with inactivated PRV-1 vaccine. PRV-1 and PRV-3 replicated well in A. salmon blood cells, and both induced the typical innate antiviral responses triggered by dsRNA viruses. Two weeks post infection, PRV-3 triggered stronger antiviral responses than PRV-1, despite their similar viral RNA replication levels, but after five weeks the induced responses were close to equal. PRV-2 and the InPRV-1 vaccine did not trigger the same typical antiviral responses as the replicating PRV-1 and PRV-3 genotypes, but induced genes involved in membrane trafficking and signaling pathways that may regulate physiological functions. These findings propose that the protection mediated by PRV-3 against a secondary infection by PRV-1 occur due to a potent and early activation of the same type of innate immune responses. The difference in the timing of antiviral responses may give PRV-1 an evolutionary edge, facilitating its dissemination to A. salmon heart, a critical step for HSMI development.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"157 ","pages":"Article 110088"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142812338","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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