Pub Date : 2026-01-01DOI: 10.1016/j.dci.2025.105543
Jiangwei Zhou , Wenwei Huang , Weicheng Yang , Sixun Li , Mengmeng Wu , Min Zhang , Zihang Xie , Lilin Zhan , Qiang Li , Jianrong Huang
Toll-like receptors (TLRs) are a family of pattern recognition receptors (PRRs) central to innate immunity. In teleost fish, TLRs play a pivotal role in defending against diverse microbial invaders, yet their evolutionary adaptation and functional specialization in Spinibarbus hollandi (army fish) remain uncharacterized. This study reports the first molecular cloning and functional analysis of TLR3, TLR4, TLR5M, and TLR18 in S. hollandi. All four TLRs possess conserved intracellular Toll/IL-1 receptor (TIR) domains and extracellular leucine-rich repeat (LRR) domains. Basal expression profiling reveals distinct tissue specificity: TLR3 and TLR5M are highly expressed in the liver, TLR4 dominates in the brain and eye, and TLR18 shows minimal constitutive expression. After Aeromonas veronii challenge, liver TLR5M is most dramatically upregulated and acts as an early-response core gene. Phylogenetic analysis confirms high homology between S. hollandi TLRs and cyprinid orthologs. Molecular evolutionary analysis identifies positive selection sites in the LRR domains of TLR3 and TLR5M, indicative of adaptive optimization for pathogen recognition. These findings clarify TLR-mediated bacterial immunity in S. hollandi and provide candidate targets for aquaculture disease resistance breeding.
{"title":"Molecular characterization and expression analysis of four toll-like receptors genes: TLR3, TLR4, TLR5M and TLR18 in Spinibarbu hollandi under Aeromonas veronii infection","authors":"Jiangwei Zhou , Wenwei Huang , Weicheng Yang , Sixun Li , Mengmeng Wu , Min Zhang , Zihang Xie , Lilin Zhan , Qiang Li , Jianrong Huang","doi":"10.1016/j.dci.2025.105543","DOIUrl":"10.1016/j.dci.2025.105543","url":null,"abstract":"<div><div>Toll-like receptors (TLRs) are a family of pattern recognition receptors (PRRs) central to innate immunity. In teleost fish, TLRs play a pivotal role in defending against diverse microbial invaders, yet their evolutionary adaptation and functional specialization in <em>Spinibarbus hollandi</em> (army fish) remain uncharacterized. This study reports the first molecular cloning and functional analysis of TLR3, TLR4, TLR5M, and TLR18 in <em>S. hollandi</em>. All four TLRs possess conserved intracellular Toll/IL-1 receptor (TIR) domains and extracellular leucine-rich repeat (LRR) domains. Basal expression profiling reveals distinct tissue specificity: TLR3 and TLR5M are highly expressed in the liver, TLR4 dominates in the brain and eye, and TLR18 shows minimal constitutive expression. After <em>Aeromonas veronii</em> challenge, liver TLR5M is most dramatically upregulated and acts as an early-response core gene. Phylogenetic analysis confirms high homology between <em>S. hollandi</em> TLRs and cyprinid orthologs. Molecular evolutionary analysis identifies positive selection sites in the LRR domains of TLR3 and TLR5M, indicative of adaptive optimization for pathogen recognition. These findings clarify TLR-mediated bacterial immunity in <em>S. hollandi</em> and provide candidate targets for aquaculture disease resistance breeding.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"174 ","pages":"Article 105543"},"PeriodicalIF":2.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145849320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2025-12-27DOI: 10.1016/j.dci.2025.105544
Yuwen Luo , Mengyi Dong , Yuxi Shen , Xuelian Xiang , Jiadai Lv , Yi Sun , Yongxin Li , Yamei Huang , Min Cui , Xinfeng Han , Jing Xia , Yong Huang
China has a great demand for poultry, while avian influenza (AI) remains widespread and exhibits high instability, posing a persistent challenge to the poultry industry. Elucidating the genetic stability of avian influenza variants and their implications for pathogenicity is key for pandemic risk mitigation. H9 and H6 AIVs have co-circulated in China due to a specific evolutionary dynamic: H9 AIVs serve as stable genetic reservoirs, whereas H6 AIVs act as platforms for multi-subtype reassortment. This reassortment process continually generates novel variants with unpredictable virulence, posing a persistent biosecurity threat to poultry production. Genetic stability analysis of H9/H6 AI viral Ribonucleoprotein (vRNP) complexes (PB2, PB1, PA, NP) revealed clear subtype-specific divergence. H9 AIVs retained highly conserved gene constellations, whereas H6 AIVs exhibited pronounced plasticity, acquiring 42.5 % of PB2, 20.1 % of PA, and 23.7 % of NP genes from other AIV subtypes. The H9vRNP-H6 reassortant (H6N2 backbone carrying H9N2 vRNP) showed enhanced replicability in both MDCK cells (4.7 → 5.7 log10TCID50/mL) and embryonated eggs (6.7 → 7.7 log10EID50/mL), with significantly promoted pathogenicity in chicks. These findings highlight the urgent need to integrate genetic surveillance of H9 and H6 AIVs into targeted prevention frameworks, thereby forestalling the emergence of pandemic-prone reassortants and mitigating potential losses to the poultry industry and public health. Hence, such research would provide a theoretical basis for the development of novel vaccines and enhance strategies for the control and prevention of avian influenza.
{"title":"Impact of viral ribonucleoprotein complex genetic stability on pathogenicity in H9N2 and H6N2 avian influenza viruses","authors":"Yuwen Luo , Mengyi Dong , Yuxi Shen , Xuelian Xiang , Jiadai Lv , Yi Sun , Yongxin Li , Yamei Huang , Min Cui , Xinfeng Han , Jing Xia , Yong Huang","doi":"10.1016/j.dci.2025.105544","DOIUrl":"10.1016/j.dci.2025.105544","url":null,"abstract":"<div><div>China has a great demand for poultry, while avian influenza (AI) remains widespread and exhibits high instability, posing a persistent challenge to the poultry industry. Elucidating the genetic stability of avian influenza variants and their implications for pathogenicity is key for pandemic risk mitigation. H9 and H6 AIVs have co-circulated in China due to a specific evolutionary dynamic: H9 AIVs serve as stable genetic reservoirs, whereas H6 AIVs act as platforms for multi-subtype reassortment. <strong>This reassortment process continually generates novel variants with unpredictable virulence, posing a persistent biosecurity threat to poultry production.</strong> Genetic stability analysis of H9/H6 AI viral Ribonucleoprotein (vRNP) complexes (<em>PB2, PB1, PA, NP</em>) revealed clear subtype-specific divergence. H9 AIVs retained highly conserved gene constellations, whereas H6 AIVs exhibited pronounced plasticity, acquiring 42.5 % of <em>PB2</em>, 20.1 % of <em>PA</em>, and 23.7 % of <em>NP</em> genes from other AIV subtypes. The H9vRNP-H6 reassortant (H6N2 backbone carrying H9N2 vRNP) showed enhanced replicability in both MDCK cells (4.7 → 5.7 log<sub>10</sub>TCID<sub>50</sub>/mL) and embryonated eggs (6.7 → 7.7 log<sub>10</sub>EID<sub>50</sub>/mL), with significantly promoted pathogenicity in chicks. These findings <strong>highlight the urgent need to integrate genetic surveillance of H9 and H6 AIVs into targeted prevention frameworks</strong>, thereby forestalling the emergence of pandemic-prone reassortants and mitigating potential losses to the poultry industry and public health. Hence, such research would provide a theoretical basis for the development of novel vaccines and enhance strategies for the control and prevention of avian influenza.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"175 ","pages":"Article 105544"},"PeriodicalIF":2.4,"publicationDate":"2025-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145855021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2025-12-16DOI: 10.1016/j.dci.2025.105539
Xiaofeng Liu , Qian Hou , Zejun Zhou
Bacterial pathogens like Aeromonas hydrophila threaten aquaculture by compromising fish immune defenses. The intestinal barrier is a critical frontline defense against pathogenic bacteria in fish, yet its regulation by neuroimmune mechanisms remains poorly understood. In this study, we investigated the role of vasoactive intestinal peptide (VIP) in the immune response of grass carp (Ctenopharyngodon idella) following bacterial challenge. The full-length grass carp VIP gene was 462 bp, encoding a 153-amino acid precursor protein with high sequence conservation across vertebrates. VIP expression was highest in the intestinal tract and significantly upregulated upon A. hydrophila infection. Administration of recombinant grass carp VIP protein enhanced intestinal barrier function by increasing the expression of tight junction molecules (ZO-1, Occludin, Claudin-1) and the immunoregulatory cytokine IL-22, while reducing plasma LPS and D-lactate levels. Furthermore, grass carp VIP treatment promoted the expression of the mucosal barrier component MUC2 and antimicrobial peptides (LEAP-2, Lyz1, Hepcidin-1), reduced bacterial load, and significantly improved survival rates. Mechanistic studies confirmed that VIP interacted specifically with the VIP receptor 2 (VIPR2), and knockdown of VIPR2 abolished the protective effects of VIP on barrier integrity and antimicrobial immunity. These findings demonstrate that the VIP-VIPR2 axis plays a crucial role in modulating intestinal immunity in grass carp, offering new insights into neuroimmune regulation in aquatic animals during pathogenic challenge.
{"title":"The VIP-VIPR2 axis strengthens intestinal barrier integrity and antimicrobial immunity in Ctenopharyngodon idella","authors":"Xiaofeng Liu , Qian Hou , Zejun Zhou","doi":"10.1016/j.dci.2025.105539","DOIUrl":"10.1016/j.dci.2025.105539","url":null,"abstract":"<div><div>Bacterial pathogens like <em>Aeromonas hydrophila</em> threaten aquaculture by compromising fish immune defenses. The intestinal barrier is a critical frontline defense against pathogenic bacteria in fish, yet its regulation by neuroimmune mechanisms remains poorly understood. In this study, we investigated the role of vasoactive intestinal peptide (VIP) in the immune response of grass carp (<em>Ctenopharyngodon idella</em>) following bacterial challenge. The full-length grass carp VIP gene was 462 bp, encoding a 153-amino acid precursor protein with high sequence conservation across vertebrates. VIP expression was highest in the intestinal tract and significantly upregulated upon <em>A. hydrophila</em> infection. Administration of recombinant grass carp VIP protein enhanced intestinal barrier function by increasing the expression of tight junction molecules (<em>ZO-1</em>, <em>Occludin</em>, <em>Claudin-1</em>) and the immunoregulatory cytokine IL-22, while reducing plasma LPS and D-lactate levels. Furthermore, grass carp VIP treatment promoted the expression of the mucosal barrier component <em>MUC2</em> and antimicrobial peptides (<em>LEAP-2</em>, <em>Lyz1</em>, <em>Hepcidin-1</em>), reduced bacterial load, and significantly improved survival rates. Mechanistic studies confirmed that VIP interacted specifically with the VIP receptor 2 (VIPR2), and knockdown of VIPR2 abolished the protective effects of VIP on barrier integrity and antimicrobial immunity. These findings demonstrate that the VIP-VIPR2 axis plays a crucial role in modulating intestinal immunity in grass carp, offering new insights into neuroimmune regulation in aquatic animals during pathogenic challenge.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"174 ","pages":"Article 105539"},"PeriodicalIF":2.4,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145780393","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2025-12-15DOI: 10.1016/j.dci.2025.105538
Wei-Wei Fang , Yi-Xin Wu , Yi-Bin Cao , Xin-Jiang Lu , Jian-Rao Hu
Hypoxia compromises host defense in fish by impairing macrophage function, but the underlying mechanisms remain unclear. This study elucidates the molecular mechanisms by which hypoxia disrupts monocytes/macrophages (MO/MФ) function in Japanese sea bass (Lateolabrax japonicus), with a particular focus on the regulatory role of heme biosynthesis and its key gene, 5′-aminolevulinate synthase 2 (ALAS2). Phylogenetic analysis confirmed high evolutionary conservation of ALAS2 across vertebrates, supporting its fundamental role in heme metabolism. Transcriptomic analysis revealed that hypoxia induced the upregulation of genes involved in heme biosynthesis of MO/MФ, particularly ALAS2. Functional assays demonstrated that heme accumulation under hypoxic conditions impaired MO/MФ phagocytic and bactericidal activities, while also promoted the production of pro-inflammatory cytokines. These dysfunctions were mediated by the ALAS2-heme signaling axis, which orchestrated inflammatory responses and metabolic reprogramming throughout the study. Knockdown of ALAS2 enhanced phagocytic activity and suppressed pro-inflammatory cytokine expression, suggesting its critical role in modulating MO/MФ function under hypoxia. Furthermore, inhibition of heme synthesis improved fish survival rates and reduced bacterial burdens during Vibrio harveyi infection. These findings elucidate the pivotal role of heme in regulating MO/MФ function and host defense under hypoxic conditions, providing mechanistic insights into hypoxia-induced immune suppression in aquaculture.
{"title":"Hypoxia impairs monocyte/macrophage function and host defense via ALAS2-mediated heme biosynthesis in Japanese sea bass (Lateolabrax japonicus)","authors":"Wei-Wei Fang , Yi-Xin Wu , Yi-Bin Cao , Xin-Jiang Lu , Jian-Rao Hu","doi":"10.1016/j.dci.2025.105538","DOIUrl":"10.1016/j.dci.2025.105538","url":null,"abstract":"<div><div>Hypoxia compromises host defense in fish by impairing macrophage function, but the underlying mechanisms remain unclear. This study elucidates the molecular mechanisms by which hypoxia disrupts monocytes/macrophages (MO/MФ) function in Japanese sea bass (<em>Lateolabrax japonicus</em>), with a particular focus on the regulatory role of heme biosynthesis and its key gene, <em>5′-aminolevulinate synthase 2</em> (<em>ALAS2</em>). Phylogenetic analysis confirmed high evolutionary conservation of <em>ALAS2</em> across vertebrates, supporting its fundamental role in heme metabolism. Transcriptomic analysis revealed that hypoxia induced the upregulation of genes involved in heme biosynthesis of MO/MФ, particularly <em>ALAS2</em>. Functional assays demonstrated that heme accumulation under hypoxic conditions impaired MO/MФ phagocytic and bactericidal activities, while also promoted the production of pro-inflammatory cytokines. These dysfunctions were mediated by the <em>ALAS2</em>-heme signaling axis, which orchestrated inflammatory responses and metabolic reprogramming throughout the study. Knockdown of <em>ALAS2</em> enhanced phagocytic activity and suppressed pro-inflammatory cytokine expression, suggesting its critical role in modulating MO/MФ function under hypoxia. Furthermore, inhibition of heme synthesis improved fish survival rates and reduced bacterial burdens during <em>Vibrio harveyi</em> infection. These findings elucidate the pivotal role of heme in regulating MO/MФ function and host defense under hypoxic conditions, providing mechanistic insights into hypoxia-induced immune suppression in aquaculture.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"174 ","pages":"Article 105538"},"PeriodicalIF":2.4,"publicationDate":"2025-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145773879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2025-12-05DOI: 10.1016/j.dci.2025.105534
Xiantong Liu , Ruixue Wu , Ning Ning Wang , Hua Xu , Xiaojun Rong , Chao Li , Min Cao
The intestinal microbiota is a critical mediator of host immune responses and metabolic homeostasis, especially during the pathogenic challenge. This study investigated the dynamic remodeling of the gut microbiota and metabolome in Sebastes schlegelii following infection with the major aquaculture pathogen Edwardsiella piscicida. Using 16S rRNA gene sequencing and untargeted metabolomics, we analyzed the microbial community composition and metabolic profiles of intestinal samples at multiple time points post-infection. Our results revealed significant temporal shifts in bacterial diversity and structure. Notably, the abundances of genera such as Bacteroides, Bacillus, and Lactobacillus increased, while Comamonas and Cutibacterium decreased. Metabolomic analysis identified 1063 metabolites, with lipids and lipid-like molecules being the most abundant. Differential analysis revealed stage-specific metabolic alterations: early infection was marked by the upregulation of pro-inflammatory mediators such as lithocholic acid and palmitoylethanolamide, whereas late infection featured elevated levels of anti-inflammatory metabolites, including cholic acid and agmatine. KEGG pathway analysis indicated an initial enrichment in general metabolic processes, followed by a shift to steroid biosynthesis later in the infection. These findings suggest a coordinated “metabolic switch" mechanism that modulates inflammation and promotes recovery. This study provides novel insights into the microbiota–metabolite–immune network in S. schlegelii and highlights potential biomarkers for monitoring fish health status in aquaculture.
{"title":"Dynamic remodeling of gut microbiota and untargeted etabolomics in Sebastes schlegelii during Edwardsiella piscicida infection","authors":"Xiantong Liu , Ruixue Wu , Ning Ning Wang , Hua Xu , Xiaojun Rong , Chao Li , Min Cao","doi":"10.1016/j.dci.2025.105534","DOIUrl":"10.1016/j.dci.2025.105534","url":null,"abstract":"<div><div>The intestinal microbiota is a critical mediator of host immune responses and metabolic homeostasis, especially during the pathogenic challenge. This study investigated the dynamic remodeling of the gut microbiota and metabolome in <em>Sebastes schlegelii</em> following infection with the major aquaculture pathogen <em>Edwardsiella piscicida</em>. Using 16S rRNA gene sequencing and untargeted metabolomics, we analyzed the microbial community composition and metabolic profiles of intestinal samples at multiple time points post-infection. Our results revealed significant temporal shifts in bacterial diversity and structure. Notably, the abundances of genera such as <em>Bacteroides</em>, <em>Bacillus</em>, and <em>Lactobacillus</em> increased, while <em>Comamonas</em> and <em>Cutibacterium</em> decreased. Metabolomic analysis identified 1063 metabolites, with lipids and lipid-like molecules being the most abundant. Differential analysis revealed stage-specific metabolic alterations: early infection was marked by the upregulation of pro-inflammatory mediators such as lithocholic acid and palmitoylethanolamide, whereas late infection featured elevated levels of anti-inflammatory metabolites, including cholic acid and agmatine. KEGG pathway analysis indicated an initial enrichment in general metabolic processes, followed by a shift to steroid biosynthesis later in the infection. These findings suggest a coordinated “metabolic switch\" mechanism that modulates inflammation and promotes recovery. This study provides novel insights into the microbiota–metabolite–immune network in <em>S. schlegelii</em> and highlights potential biomarkers for monitoring fish health status in aquaculture.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"174 ","pages":"Article 105534"},"PeriodicalIF":2.4,"publicationDate":"2025-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145696210","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2025-12-05DOI: 10.1016/j.dci.2025.105532
Boyu Chen , Zihang Ma , Jinxia Liu , Lei Yan , Zhenyu Lan , Fanxi Gao , Zizhuo Liu , Shimiao Li , Zhenwen Zhang , Yan Zhang , Peng Zhu , Yongyan Liao
Microcystin-LR (MC-LR), a cyanobacterial toxin, poses a considerable threat to aquatic organisms. Herein, we focused on the economically important crustacean Eriocheir hepuensis to examine the acute toxic effects of MC-LR stress and its molecular mechanisms. The half-lethal concentration (LD50) of MC-LR for the E. hepuensis was determined to be 64.78 μg/kg, with a calculated safety concentration of 6.48 μg/kg through a 96-h acute toxicity test (n = 240). Exposure to a sublethal concentration (60 μg/kg) for 48 h resulted in 56.7 % survival rate (n = 60) and caused substantial hepatopancreatic pathological damage, including cell vacuolation, chromatin condensation, and mitochondrial fragmentation. To elucidate the underlying mechanisms, further cloning and identification of the apoptosis-related genes p53 (open reading frame (ORF 1284 bp), Bax (ORF 1482 bp), and Bcl-2 (ORF 732 bp) showed that their expression levels were the highest in the hepatopancreas (2.9, 3.7, and 3.5 times that of the control group, respectively). Time-series analysis further showed that the p53 and Bax gene expression rapidly increased at stress onset but gradually declined, whereas the Bcl-2 gene expression was temporarily downregulated, and a substantial negative correlation was observed with Bax expression. These molecular dynamics reveal that MC-LR-induced hepatopancreatic damage and mortality in E. hepuensis are mediated through coordinated apoptotic gene regulation, with the p53-Bax-Bcl2 pathway orchestrating the cellular stress response. This study providing crucial scientific insights into the molecular responses of crustaceans to algal toxins.
{"title":"Study on the regulatory mechanism of apoptosis and expression patterns of key genes (p53, Bax, and Bcl-2) in the crustacean Eriocheir hepuensis under Microcystin-LR stress","authors":"Boyu Chen , Zihang Ma , Jinxia Liu , Lei Yan , Zhenyu Lan , Fanxi Gao , Zizhuo Liu , Shimiao Li , Zhenwen Zhang , Yan Zhang , Peng Zhu , Yongyan Liao","doi":"10.1016/j.dci.2025.105532","DOIUrl":"10.1016/j.dci.2025.105532","url":null,"abstract":"<div><div>Microcystin-LR (MC-LR), a cyanobacterial toxin, poses a considerable threat to aquatic organisms. Herein, we focused on the economically important crustacean <em>Eriocheir hepuensis</em> to examine the acute toxic effects of MC-LR stress and its molecular mechanisms. The half-lethal concentration (LD<sub>50</sub>) of MC-LR for the <em>E</em>. <em>hepuensis</em> was determined to be 64.78 μg/kg, with a calculated safety concentration of 6.48 μg/kg through a 96-h acute toxicity test (n = 240). Exposure to a sublethal concentration (60 μg/kg) for 48 h resulted in 56.7 % survival rate (n = 60) and caused substantial hepatopancreatic pathological damage, including cell vacuolation, chromatin condensation, and mitochondrial fragmentation. To elucidate the underlying mechanisms, further cloning and identification of the apoptosis-related genes <em>p53</em> (open reading frame (ORF 1284 bp), <em>Bax</em> (ORF 1482 bp), and <em>Bcl-2</em> (ORF 732 bp) showed that their expression levels were the highest in the hepatopancreas (2.9, 3.7, and 3.5 times that of the control group, respectively). Time-series analysis further showed that the <em>p53</em> and <em>Bax</em> gene expression rapidly increased at stress onset but gradually declined, whereas the <em>Bcl-2</em> gene expression was temporarily downregulated, and a substantial negative correlation was observed with <em>Bax</em> expression. These molecular dynamics reveal that MC-LR-induced hepatopancreatic damage and mortality in <em>E. hepuensis</em> are mediated through coordinated apoptotic gene regulation, with the <em>p53</em>-<em>Bax</em>-<em>Bcl2</em> pathway orchestrating the cellular stress response. This study providing crucial scientific insights into the molecular responses of crustaceans to algal toxins.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"174 ","pages":"Article 105532"},"PeriodicalIF":2.4,"publicationDate":"2025-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145699500","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2025-12-03DOI: 10.1016/j.dci.2025.105537
Yanqiu Cui , Yaqi Tao , Tianjun Xu , Yuena Sun
In recent years, the unique circular structure of circular RNAs (circRNAs) has been the focus of much attention and research. RNA molecules without coding function weave a complex regulatory network in mammals, affecting the progress of many biological processes. This discovery opens up a new perspective on the function of the genome. However, the powerful regulatory functions of circular RNAs in fish are still unclear. We identified a novel circular RNA, dubbing it circeIF4G3. Through qPCR and dual luciferase assays, we found that circeIF4G3 reduces the production of antiviral genes and inflammatory factors. We discovered that circeIF4G3 both hindered cell proliferation and diminished cell viability concurrently. These results suggest that circeIF4G3 is not only involved in the innate immunity of miiuy croaker, but also plays an inhibitory role in it. This discovery provides a new research approach for a deeper understanding of the immune regulatory mechanism of miiuy croaker. This study enriches the non-coding RNAs regulatory network in innate immunity in teleost fish, and establishes a foundation for investigating the role of circular RNAs in teleost fish's innate immunity.
{"title":"Identification and functional characterization of a circular RNA circeIF4G3: a negative regulator of innate immunity in miiuy croaker (Miichthys miiuy)","authors":"Yanqiu Cui , Yaqi Tao , Tianjun Xu , Yuena Sun","doi":"10.1016/j.dci.2025.105537","DOIUrl":"10.1016/j.dci.2025.105537","url":null,"abstract":"<div><div>In recent years, the unique circular structure of circular RNAs (circRNAs) has been the focus of much attention and research. RNA molecules without coding function weave a complex regulatory network in mammals, affecting the progress of many biological processes. This discovery opens up a new perspective on the function of the genome. However, the powerful regulatory functions of circular RNAs in fish are still unclear. We identified a novel circular RNA, dubbing it circeIF4G3. Through qPCR and dual luciferase assays, we found that circeIF4G3 reduces the production of antiviral genes and inflammatory factors. We discovered that circeIF4G3 both hindered cell proliferation and diminished cell viability concurrently. These results suggest that circeIF4G3 is not only involved in the innate immunity of miiuy croaker, but also plays an inhibitory role in it. This discovery provides a new research approach for a deeper understanding of the immune regulatory mechanism of miiuy croaker. This study enriches the non-coding RNAs regulatory network in innate immunity in teleost fish, and establishes a foundation for investigating the role of circular RNAs in teleost fish's innate immunity.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"174 ","pages":"Article 105537"},"PeriodicalIF":2.4,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145687313","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2025-12-01DOI: 10.1016/j.dci.2025.105530
Rodrigo Pinto , Muhammad Salman Malik , Hanne Brenne , Fernando Afonso , Erik Burgerhout , Carlo C. Lazado
This study investigated immune responses to Yersinia ruckeri in the olfactory organ and brain of Atlantic salmon parr and the influence of embryonic temperature history on immunity and disease resistance. Atlantic salmon were reared at 4 °C or 8 °C from fertilisation to the eyed stage of embryogenesis and maintained under standard rearing conditions until the parr stage (∼20 g). Fish were bath-exposed to Y. ruckeri, and immune responses were assessed at 1, 3, and 14 days post-infection (dpi). At 14 dpi, cumulative mortality in the 8 °C group (∼35 %) was significantly higher than in the 4 °C group (∼22 %). Low bacteria levels were detected in the olfactory organ and brain. Y. ruckeri antigens were present in the lamina propria of the olfactory lamellae and the optic tectum, including the meninges and nucleus of the vagus nerve. Mild histological changes, such as congestion and leukocyte infiltration were observed in the olfactory organ, although mucosal morphometric parameters were unaffected by infection or embryonic temperature history. Y. ruckeri modulated the expression of cytokines, antibacterial defence genes, and immunoglobulins in the olfactory organ. The most significant changes occurred at 3 and 14 dpi. Embryonic temperature history did not broadly alter immune gene expression, but interleukin-1β (il1b) and tumour necrosis factor-α (tnfa) had higher expression in the 8 °C group. In the brain, microglial markers including aif1 and cd45 were significantly upregulated at 14 dpi, particularly in the 8 °C group. This study provides insights into the nasal immune responses of Atlantic salmon to Y. ruckeri. Our findings suggest that lower embryonic temperature history influenced disease resistance against Y. ruckeri in Atlantic salmon parr, but only minimally shaped immune responses in the olfactory organ and brain following infection.
{"title":"Imprints of the embryonic thermal environment on nasal mucosal immunity and disease resistance to Yersinia ruckeri in Atlantic salmon (Salmo salar) parr","authors":"Rodrigo Pinto , Muhammad Salman Malik , Hanne Brenne , Fernando Afonso , Erik Burgerhout , Carlo C. Lazado","doi":"10.1016/j.dci.2025.105530","DOIUrl":"10.1016/j.dci.2025.105530","url":null,"abstract":"<div><div>This study investigated immune responses to <em>Yersinia ruckeri</em> in the olfactory organ and brain of Atlantic salmon parr and the influence of embryonic temperature history on immunity and disease resistance. Atlantic salmon were reared at 4 °C or 8 °C from fertilisation to the eyed stage of embryogenesis and maintained under standard rearing conditions until the parr stage (∼20 g). Fish were bath-exposed to <em>Y. ruckeri</em>, and immune responses were assessed at 1, 3, and 14 days post-infection (dpi). At 14 dpi, cumulative mortality in the 8 °C group (∼35 %) was significantly higher than in the 4 °C group (∼22 %). Low bacteria levels were detected in the olfactory organ and brain. <em>Y. ruckeri</em> antigens were present in the lamina propria of the olfactory lamellae and the optic tectum, including the meninges and nucleus of the vagus nerve. Mild histological changes, such as congestion and leukocyte infiltration were observed in the olfactory organ, although mucosal morphometric parameters were unaffected by infection or embryonic temperature history. <em>Y. ruckeri</em> modulated the expression of cytokines, antibacterial defence genes, and immunoglobulins in the olfactory organ. The most significant changes occurred at 3 and 14 dpi. Embryonic temperature history did not broadly alter immune gene expression, but <em>interleukin-1β</em> (<em>il1b</em>) and <em>tumour necrosis factor-α</em> (<em>tnfa</em>) had higher expression in the 8 °C group. In the brain, microglial markers includin<em>g aif1</em> and <em>cd45</em> were significantly upregulated at 14 dpi, particularly in the 8 °C group. This study provides insights into the nasal immune responses of Atlantic salmon to <em>Y. ruckeri</em>. Our findings suggest that lower embryonic temperature history influenced disease resistance against <em>Y. ruckeri</em> in Atlantic salmon parr, but only minimally shaped immune responses in the olfactory organ and brain following infection.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105530"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145631117","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Crustaceans, like other invertebrates, lack adaptive immunity and rely primarily on innate immune responses for defense against invading pathogens. Cytokines are signaling molecules that play vital roles in coordinating innate immune responses. Cytokine-like molecules such as astakines, interferon-like proteins (namely Vago), Spätzle, interleukin-like proteins, and tumor necrosis factors have been identified in crustaceans including shrimp, crayfish, and crabs. These molecules, although structurally distinct from vertebrate cytokines, have critical roles in regulating innate immune responses. They mediate processes such as hemocyte proliferation, production of antimicrobial peptides, antiviral response, and anti-inflammation through key immune signaling pathways including Toll, JAK/STAT, and IMD. Astakines are particularly important in crustacean hematopoiesis; Vago is involved in antiviral immunity, and Spätzle functions as a key ligand in Toll signaling. Interleukin-like proteins and tumor necrosis factors, although less well characterized in crustaceans, likely play crucial roles in the innate immune system. Studying the functions of these cytokine-like proteins enables better understanding crustacean immunity and has significant implications for improving disease management in aquaculture.
{"title":"Functional insights into cytokine-like molecules in crustacean innate immunity","authors":"Warumporn Yingsunthonwattana, Zittipong Nanakorn, Anchalee Tassanakajon","doi":"10.1016/j.dci.2025.105528","DOIUrl":"10.1016/j.dci.2025.105528","url":null,"abstract":"<div><div>Crustaceans, like other invertebrates, lack adaptive immunity and rely primarily on innate immune responses for defense against invading pathogens. Cytokines are signaling molecules that play vital roles in coordinating innate immune responses. Cytokine-like molecules such as astakines, interferon-like proteins (namely Vago), Spätzle, interleukin-like proteins, and tumor necrosis factors have been identified in crustaceans including shrimp, crayfish, and crabs. These molecules, although structurally distinct from vertebrate cytokines, have critical roles in regulating innate immune responses. They mediate processes such as hemocyte proliferation, production of antimicrobial peptides, antiviral response, and anti-inflammation through key immune signaling pathways including Toll, JAK/STAT, and IMD. Astakines are particularly important in crustacean hematopoiesis; Vago is involved in antiviral immunity, and Spätzle functions as a key ligand in Toll signaling. Interleukin-like proteins and tumor necrosis factors, although less well characterized in crustaceans, likely play crucial roles in the innate immune system. Studying the functions of these cytokine-like proteins enables better understanding crustacean immunity and has significant implications for improving disease management in aquaculture.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105528"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145582049","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2025-12-01DOI: 10.1016/j.dci.2025.105529
Zhitao Qi , Jinquan He , Chenfei Gu , Yushuai Xie , Mingzhu Pan , Qihuan Zhang , Youchuan Wei
CD28 signaling pathway plays critical roles in the T cell activation. The golden pompano (Trachinotus ovatus, Tro) is a commercially important marine fish that widely cultured in southern China. Frequently pathogenic disease outbreaks greatly hinder the industry of golden pompano. Understanding the T cell activation signaling pathway of golden pompano contributes to prevent pathogenic diseases using immunological strategies. In the present study, three molecules of CD28 signaling pathway (TroCD28, growth factor receptor-bound protein 2 (TroGrb2) and SH2 domain-containing leukocyte phosphoprotein of 76 kDa (TroSLP-76)) were identified from golden pompano. The cDNA of the three genes respectively encoded 248, 217 and 375 amino acids. TroCD28, TroGrb2 and TroSLP-76 contained several conserved functional domains, similar to their mammalian counterparts. qPCR analysis revealed that these three genes were ubiquitously expressed in all selected tissues and peripheral blood lymphocytes (PBLs), with high expressions in immune-related tissues (spleen, head kidney, gill). Further, expressions of these three genes in head kidney were significantly induced at different time points post lipopolysaccharide (LPS) or polyriboinosinic polyribocytidylic acid (polyI:C) stimulation and Vibrio alginolyticus infection. The coimmunoprecipitation (Co-IP) assay verified the TroCD28-TroGrb2 interaction. These results provide basis for understanding the process of T cell activation in golden pompano.
{"title":"Characterization of the CD28 signaling pathway related genes (CD28, Grb2 and SLP-76) of golden pompano (Trachinotus ovatus)","authors":"Zhitao Qi , Jinquan He , Chenfei Gu , Yushuai Xie , Mingzhu Pan , Qihuan Zhang , Youchuan Wei","doi":"10.1016/j.dci.2025.105529","DOIUrl":"10.1016/j.dci.2025.105529","url":null,"abstract":"<div><div>CD28 signaling pathway plays critical roles in the T cell activation. The golden pompano (<em>Trachinotus ovatus</em>, Tro) is a commercially important marine fish that widely cultured in southern China. Frequently pathogenic disease outbreaks greatly hinder the industry of golden pompano. Understanding the T cell activation signaling pathway of golden pompano contributes to prevent pathogenic diseases using immunological strategies. In the present study, three molecules of CD28 signaling pathway (TroCD28, growth factor receptor-bound protein 2 (TroGrb2) and SH2 domain-containing leukocyte phosphoprotein of 76 kDa (TroSLP-76)) were identified from golden pompano. The cDNA of the three genes respectively encoded 248, 217 and 375 amino acids. TroCD28, TroGrb2 and TroSLP-76 contained several conserved functional domains, similar to their mammalian counterparts. qPCR analysis revealed that these three genes were ubiquitously expressed in all selected tissues and peripheral blood lymphocytes (PBLs), with high expressions in immune-related tissues (spleen, head kidney, gill). Further, expressions of these three genes in head kidney were significantly induced at different time points post lipopolysaccharide (LPS) or polyriboinosinic polyribocytidylic acid (polyI:C) stimulation and <em>Vibrio alginolyticus</em> infection. The coimmunoprecipitation (Co-IP) assay verified the TroCD28-TroGrb2 interaction. These results provide basis for understanding the process of T cell activation in golden pompano.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105529"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145615645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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