Geormery Belén Mera-Loor, Ana María Santana-Piñeros, Yanis Cruz-Quintana, Limber José Alcívar-Mendoza, Isabel Valles-Vega, Víctor Hugo Caña-Bozada
Cestode infestations in cultured fish have resulted in significant economic problems associated with international trade. Pharmacological treatments containing albendazole (ABZ) represent a promising alternative for controlling these infestations. This study evaluated the in vitro efficacy of ABZ against the metacestode Parvitaenia cochlearii, a parasite of the Pacific fat sleeper (Dormitator latifrons), by exposing the parasites to different concentrations (200, 600, 800, and 1000 mg/L). The median lethal concentration (LC50) was determined at 10 and 15 min of exposure. Results showed that ABZ was 100% effective in killing P. cochlearii, achieving complete mortality at 15 min with a concentration of 600 mg/L, and at 30 min with 200 mg/L. The LC50 value decreased over time, with 618.91 mg/L at 10 min and 260.32 mg/L at 15 min, indicating a cumulative effect of ABZ on metacestode tissues. In conclusion, ABZ is an effective treatment against cestode infestations, and further evaluation under experimental and commercial scales is recommended to establish control protocols in aquaculture systems.
{"title":"In Vitro Efficacy of Albendazole Against the Metacestode Parvitaenia cochlearii (Cestoda), a Parasite of the Pacific Fat Sleeper (Dormitator latifrons).","authors":"Geormery Belén Mera-Loor, Ana María Santana-Piñeros, Yanis Cruz-Quintana, Limber José Alcívar-Mendoza, Isabel Valles-Vega, Víctor Hugo Caña-Bozada","doi":"10.1111/jfd.70077","DOIUrl":"https://doi.org/10.1111/jfd.70077","url":null,"abstract":"<p><p>Cestode infestations in cultured fish have resulted in significant economic problems associated with international trade. Pharmacological treatments containing albendazole (ABZ) represent a promising alternative for controlling these infestations. This study evaluated the in vitro efficacy of ABZ against the metacestode Parvitaenia cochlearii, a parasite of the Pacific fat sleeper (Dormitator latifrons), by exposing the parasites to different concentrations (200, 600, 800, and 1000 mg/L). The median lethal concentration (LC50) was determined at 10 and 15 min of exposure. Results showed that ABZ was 100% effective in killing P. cochlearii, achieving complete mortality at 15 min with a concentration of 600 mg/L, and at 30 min with 200 mg/L. The LC50 value decreased over time, with 618.91 mg/L at 10 min and 260.32 mg/L at 15 min, indicating a cumulative effect of ABZ on metacestode tissues. In conclusion, ABZ is an effective treatment against cestode infestations, and further evaluation under experimental and commercial scales is recommended to establish control protocols in aquaculture systems.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70077"},"PeriodicalIF":2.2,"publicationDate":"2025-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145458637","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}
Bo Li, Penghui Yu, Lei Su, Jianguo Su, Yu Gao, Gailing Yuan
Aeromonas hydrophila-induced bacterial sepsis is a major threat to aquaculture, characterised by excessive inflammation, ferroptosis (an iron-dependent lipid peroxidation-driven cell death) and oxidative damage, which collectively lead to high mortality. Taurine (TAU) and glutathione (GSH) have been demonstrated to have potential therapeutic efficacy against ferroptosis-associated pathologies through redox modulation. However, their combined effects and underlying mechanisms in alleviating A. hydrophila infection remain unclear. In this study, we investigated the protective roles of TAU, GSH and their combination in vitro using yellow catfish (Pelteobagrus fulvidraco) macrophages and grass carp (Ctenopharyngodon idella) kidney (CIK) cells and in vivo using yellow catfish challenged with A. hydrophila. The results showed that TAU and GSH, either alone or in combination, alleviated oxidative stress by significantly reducing reactive oxygen species (ROS) accumulation and inhibited nuclear factor-κB (NF-κB) pathway activation. This led to downregulated expression of pro-inflammatory cytokines (IL-1β and TNF-α) and upregulated anti-inflammatory cytokines (IL-10 and TGF-β), accompanied by alleviated ferroptosis. In vivo, dietary supplementation with TAU (10 g/kg) or GSH (350 mg/kg) alone improved survival rates (51.7% and 38.3%, respectively), reduced tissue bacterial loads and protected intestinal and hepatic tissues by preserving mucosal barrier function. However, the TAU + GSH combination unexpectedly decreased survival (28.3%) due to excessive immunosuppression (overproduction of IL-10 and TGF-β) and impaired mucosal barrier, which exacerbated pathogen colonisation. Together, our findings demonstrate that TAU and GSH alleviate A. hydrophila-induced oxidative stress by reducing mitochondrial ROS overproduction and disrupting the ROS/NF-κB signalling pathway, thereby attenuating inflammatory cytokine storms and ferroptosis. These results provide novel insights into the pathological mechanisms of bacterial sepsis in fish and develop sustainable strategies to improve disease resistance in aquaculture.
{"title":"Taurine and Glutathione Alleviate Aeromonas hydrophila Infection by Inhibiting Inflammation and Ferroptosis via ROS/NF-κB Pathway in Fishes.","authors":"Bo Li, Penghui Yu, Lei Su, Jianguo Su, Yu Gao, Gailing Yuan","doi":"10.1111/jfd.70070","DOIUrl":"https://doi.org/10.1111/jfd.70070","url":null,"abstract":"<p><p>Aeromonas hydrophila-induced bacterial sepsis is a major threat to aquaculture, characterised by excessive inflammation, ferroptosis (an iron-dependent lipid peroxidation-driven cell death) and oxidative damage, which collectively lead to high mortality. Taurine (TAU) and glutathione (GSH) have been demonstrated to have potential therapeutic efficacy against ferroptosis-associated pathologies through redox modulation. However, their combined effects and underlying mechanisms in alleviating A. hydrophila infection remain unclear. In this study, we investigated the protective roles of TAU, GSH and their combination in vitro using yellow catfish (Pelteobagrus fulvidraco) macrophages and grass carp (Ctenopharyngodon idella) kidney (CIK) cells and in vivo using yellow catfish challenged with A. hydrophila. The results showed that TAU and GSH, either alone or in combination, alleviated oxidative stress by significantly reducing reactive oxygen species (ROS) accumulation and inhibited nuclear factor-κB (NF-κB) pathway activation. This led to downregulated expression of pro-inflammatory cytokines (IL-1β and TNF-α) and upregulated anti-inflammatory cytokines (IL-10 and TGF-β), accompanied by alleviated ferroptosis. In vivo, dietary supplementation with TAU (10 g/kg) or GSH (350 mg/kg) alone improved survival rates (51.7% and 38.3%, respectively), reduced tissue bacterial loads and protected intestinal and hepatic tissues by preserving mucosal barrier function. However, the TAU + GSH combination unexpectedly decreased survival (28.3%) due to excessive immunosuppression (overproduction of IL-10 and TGF-β) and impaired mucosal barrier, which exacerbated pathogen colonisation. Together, our findings demonstrate that TAU and GSH alleviate A. hydrophila-induced oxidative stress by reducing mitochondrial ROS overproduction and disrupting the ROS/NF-κB signalling pathway, thereby attenuating inflammatory cytokine storms and ferroptosis. These results provide novel insights into the pathological mechanisms of bacterial sepsis in fish and develop sustainable strategies to improve disease resistance in aquaculture.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70070"},"PeriodicalIF":2.2,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145438170","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}
Koi herpesvirus disease (KHVD), caused by Cyprinid herpesvirus-3 (CyHV-3), poses a significant threat to global aquaculture due to its high mortality rates and economic impact. Current diagnostic methods, such as PCR, are limited by equipment dependency and procedural complexity, hindering point-of-care (POC) applications. To address this, we developed an integrated assay combining recombinase-aided amplification (RAA) with CRISPR-Cas13a-mediated SHERLOCK technology and lateral flow detection (LFD) for rapid and visual detection of CyHV-3 in clinical samples. The KHV-SHERLOCK assay targets a conserved region of the CyHV-3 thymidine kinase (TK) gene, demonstrating exceptional specificity with no cross-reactivity to related pathogens or host DNA. Sensitivity evaluations revealed a detection limit of 100 ag/μL for CyHV-3 plasmid DNA, tenfold more sensitive than the conventional PCR (1 fg/μL) assay, even in the presence of 100 ng of carp genomic DNA as background interference. Clinical validation using 50 archived samples showed 100% concordance with reference PCR results, confirming diagnostic reliability. The assay's isothermal RAA step (37°C, 40 min) and CRISPR-Cas13a detection (37°C, 1 h) enable equipment-free operation, while LFD provides unambiguous visual results within minutes. This platform merges high sensitivity with POC practicality, offering a transformative tool for field-based KHVD surveillance.
{"title":"CRISPR-Enhanced RAA-SHERLOCK Assay for Point-of-Care Detection of Cyprinid Herpesvirus-3: Development, Validation and Clinical Application.","authors":"Hatem Soliman, Naveed Akram, Mona Saleh","doi":"10.1111/jfd.70079","DOIUrl":"https://doi.org/10.1111/jfd.70079","url":null,"abstract":"<p><p>Koi herpesvirus disease (KHVD), caused by Cyprinid herpesvirus-3 (CyHV-3), poses a significant threat to global aquaculture due to its high mortality rates and economic impact. Current diagnostic methods, such as PCR, are limited by equipment dependency and procedural complexity, hindering point-of-care (POC) applications. To address this, we developed an integrated assay combining recombinase-aided amplification (RAA) with CRISPR-Cas13a-mediated SHERLOCK technology and lateral flow detection (LFD) for rapid and visual detection of CyHV-3 in clinical samples. The KHV-SHERLOCK assay targets a conserved region of the CyHV-3 thymidine kinase (TK) gene, demonstrating exceptional specificity with no cross-reactivity to related pathogens or host DNA. Sensitivity evaluations revealed a detection limit of 100 ag/μL for CyHV-3 plasmid DNA, tenfold more sensitive than the conventional PCR (1 fg/μL) assay, even in the presence of 100 ng of carp genomic DNA as background interference. Clinical validation using 50 archived samples showed 100% concordance with reference PCR results, confirming diagnostic reliability. The assay's isothermal RAA step (37°C, 40 min) and CRISPR-Cas13a detection (37°C, 1 h) enable equipment-free operation, while LFD provides unambiguous visual results within minutes. This platform merges high sensitivity with POC practicality, offering a transformative tool for field-based KHVD surveillance.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70079"},"PeriodicalIF":2.2,"publicationDate":"2025-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145421825","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}
Isolating leukocytes from small-bodied fish presents significant technical challenges, especially in species such as grouper, which exhibited rapid blood coagulation. Attaining high purity and quality leukocyte recovery in such species is often hindered by clot formation and erythrocyte contamination. Conventional methods using anticoagulants, buffy coat extraction, and density gradient centrifugation are typically labor-intensive and often yielded inconsistent results. In this study, a streamlined and efficient whole blood lysis protocol for isolating leukocytes from grouper fingerling is described. This protocol offers improvements in blood handling, cell recovery, and cryopreservation. The optimized method significantly reduces processing time, preserves cellular integrity, and facilitates downstream applications in fish immunological studies. Overall, this protocol presented a cost-effective and scalable solution for leukocyte isolation in teleost fish, particularly grouper species.
{"title":"Streamlined Blood Handling and Leukocyte Isolation Procedure for Immunological Applications in Hybrid Grouper Fingerlings, Epinephelus fuscoguttatus × Epinephelus lanceolatus.","authors":"Nurhikmah Abu Aziz, Chou-Min Chong, Chen-Fei Low","doi":"10.1111/jfd.70080","DOIUrl":"https://doi.org/10.1111/jfd.70080","url":null,"abstract":"<p><p>Isolating leukocytes from small-bodied fish presents significant technical challenges, especially in species such as grouper, which exhibited rapid blood coagulation. Attaining high purity and quality leukocyte recovery in such species is often hindered by clot formation and erythrocyte contamination. Conventional methods using anticoagulants, buffy coat extraction, and density gradient centrifugation are typically labor-intensive and often yielded inconsistent results. In this study, a streamlined and efficient whole blood lysis protocol for isolating leukocytes from grouper fingerling is described. This protocol offers improvements in blood handling, cell recovery, and cryopreservation. The optimized method significantly reduces processing time, preserves cellular integrity, and facilitates downstream applications in fish immunological studies. Overall, this protocol presented a cost-effective and scalable solution for leukocyte isolation in teleost fish, particularly grouper species.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70080"},"PeriodicalIF":2.2,"publicationDate":"2025-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145377563","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}
Danielle Souza Vieira, Caio Francisco Santana Farias, Marco Shizuo Owatari, Mauricio Laterça Martins, Aline Brum, Caio Magnotti
While nephrocalcinosis (kidney stones) is uncommon in wild teleost fish, various environmental and nutritional factors could lead to its occurrence in aquacultured fish. This study presents the first documented case of kidney stones in aquacultured Brazilian sardine (Sardinella brasiliensis). During necropsy, eighteen hard, white kidney stones were found in the posterior kidney, with an average diameter of 3.71 mm and a total length of 16.8 mm. Morphological analysis revealed stones of different sizes and shapes, including elongated and irregular structures. This discovery enhances our understanding of pathological conditions in S. brasiliensis and underscores the importance of further research into the causes, prevalence and potential implications for fish health and fisheries sustainability.
{"title":"Short Communication: First Report of Nephrocalcinosis in Aquacultured Brazilian Sardine (Sardinella brasiliensis Steindachner, 1879).","authors":"Danielle Souza Vieira, Caio Francisco Santana Farias, Marco Shizuo Owatari, Mauricio Laterça Martins, Aline Brum, Caio Magnotti","doi":"10.1111/jfd.70081","DOIUrl":"https://doi.org/10.1111/jfd.70081","url":null,"abstract":"<p><p>While nephrocalcinosis (kidney stones) is uncommon in wild teleost fish, various environmental and nutritional factors could lead to its occurrence in aquacultured fish. This study presents the first documented case of kidney stones in aquacultured Brazilian sardine (Sardinella brasiliensis). During necropsy, eighteen hard, white kidney stones were found in the posterior kidney, with an average diameter of 3.71 mm and a total length of 16.8 mm. Morphological analysis revealed stones of different sizes and shapes, including elongated and irregular structures. This discovery enhances our understanding of pathological conditions in S. brasiliensis and underscores the importance of further research into the causes, prevalence and potential implications for fish health and fisheries sustainability.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70081"},"PeriodicalIF":2.2,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145354951","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}
Myxoid neurothekeoma, also known as nerve sheath myxoma, is a rare benign peripheral nerve sheath tumour primarily reported in mammals, with scarce documentation in teleost fishes. This study reports a confirmed case of myxoid neurothekeoma in the Indian oil sardine (Sardinella longiceps ), a commercially significant marine species along India's southeast coast. An adult female specimen exhibiting a fleshy, dome-shaped, slightly pale brownish mass on the dorsal head was examined grossly and subjected to histopathological analysis using haematoxylin and eosin, Alcian blue and Masson's trichrome staining. Immunohistochemical staining for S100 protein confirmed the neural origin of the tumour cells. Microscopic analysis revealed a well-circumscribed lobular neoplasm composed of spindle to stellate cells within a myxoid stroma rich in acid mucopolysaccharides. Tumour lobules were separated by thin collagenous septa, and widespread S100 positivity supported Schwann cell lineage. The lack of mitotic activity, cytological atypia or invasive features supported a diagnosis of benign myxoid neurothekeoma. This case expands the current understanding of peripheral nerve sheath tumours in marine teleosts and contributes to the broader characterisation of rare neoplasms in aquatic species.
{"title":"Myxoid Neurothekeoma in Indian Oil Sardine (Sardinella longiceps ) From the Southeast Coast of India.","authors":"Vijayapoopathi Singaravel, Ayyaru Gopalakrishnan, Nguyen Dinh-Hung","doi":"10.1111/jfd.70073","DOIUrl":"https://doi.org/10.1111/jfd.70073","url":null,"abstract":"<p><p>Myxoid neurothekeoma, also known as nerve sheath myxoma, is a rare benign peripheral nerve sheath tumour primarily reported in mammals, with scarce documentation in teleost fishes. This study reports a confirmed case of myxoid neurothekeoma in the Indian oil sardine (Sardinella longiceps ), a commercially significant marine species along India's southeast coast. An adult female specimen exhibiting a fleshy, dome-shaped, slightly pale brownish mass on the dorsal head was examined grossly and subjected to histopathological analysis using haematoxylin and eosin, Alcian blue and Masson's trichrome staining. Immunohistochemical staining for S100 protein confirmed the neural origin of the tumour cells. Microscopic analysis revealed a well-circumscribed lobular neoplasm composed of spindle to stellate cells within a myxoid stroma rich in acid mucopolysaccharides. Tumour lobules were separated by thin collagenous septa, and widespread S100 positivity supported Schwann cell lineage. The lack of mitotic activity, cytological atypia or invasive features supported a diagnosis of benign myxoid neurothekeoma. This case expands the current understanding of peripheral nerve sheath tumours in marine teleosts and contributes to the broader characterisation of rare neoplasms in aquatic species.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70073"},"PeriodicalIF":2.2,"publicationDate":"2025-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145345632","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}
Justin M Stilwell, Rachel Sheffler, Lauren Michaels, Whitney Daniel, Jamie Torres, Alvin C Camus
An adult female striped burrfish from a public display aquarium, mixed species recirculating system presented for an exophytic yellow-orange mass on the dorsal head suggestive of neoplasia. The mass was surgically excised twice, adding cryosurgical ablation during the second procedure, but recurred both times post-operatively. After approximately 9 months, euthanasia was elected due to lesion progression. Histopathologic evaluation of surgical biopsy samples and tissues collected at necropsy suggested chromatophoromas. Immunohistochemistry using SOX10, Melan-A and PNL-2 antibodies produced no labelling of tumour cells. Ultrastructurally, neoplastic cells contained poorly formed pterinosomes and vesicles consistent with either xanthophoroma, erythrophoroma or a mixed xanthoerythrophroma. Pigment analysis using absorbance spectrophotometry and gas chromatography mass spectrophotometry identified the primary pigment as xanthopterin with low amounts of the carotenoid tunaxanthin, confirming the diagnosis of xanthophoroma, a rare subtype of chromatophoroma in fish.
{"title":"Xanthophoroma in a Striped Burrfish, Chilomycterus schoepfi.","authors":"Justin M Stilwell, Rachel Sheffler, Lauren Michaels, Whitney Daniel, Jamie Torres, Alvin C Camus","doi":"10.1111/jfd.70075","DOIUrl":"https://doi.org/10.1111/jfd.70075","url":null,"abstract":"<p><p>An adult female striped burrfish from a public display aquarium, mixed species recirculating system presented for an exophytic yellow-orange mass on the dorsal head suggestive of neoplasia. The mass was surgically excised twice, adding cryosurgical ablation during the second procedure, but recurred both times post-operatively. After approximately 9 months, euthanasia was elected due to lesion progression. Histopathologic evaluation of surgical biopsy samples and tissues collected at necropsy suggested chromatophoromas. Immunohistochemistry using SOX10, Melan-A and PNL-2 antibodies produced no labelling of tumour cells. Ultrastructurally, neoplastic cells contained poorly formed pterinosomes and vesicles consistent with either xanthophoroma, erythrophoroma or a mixed xanthoerythrophroma. Pigment analysis using absorbance spectrophotometry and gas chromatography mass spectrophotometry identified the primary pigment as xanthopterin with low amounts of the carotenoid tunaxanthin, confirming the diagnosis of xanthophoroma, a rare subtype of chromatophoroma in fish.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70075"},"PeriodicalIF":2.2,"publicationDate":"2025-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145345591","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}
Anguillid herpesvirus (AngHV) is a highly pathogenic agent that causes "Mucus sloughing and hemorrhagic septicemia disease" in eels, resulting in high morbidity and cumulative mortality rates. Therefore, accurate diagnosis of AngHV infection is essential for effective clinical management and epidemiological control. In this study, three monoclonal antibodies (mAbs) against AngHV were developed, and their specificity and affinity were validated using Western blotting and indirect immunofluorescence assays. Subsequently, a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was established for the detection of AngHV, employing mAb 10G8-9F4 as the capture antibody and rabbit polyclonal antibody 9NA (which targets AngHV-ORF95) as the detection antibody. Specificity testing indicated no cross-reactivity with other common aquatic viruses, including Koi herpesvirus (KHV), Grass carp reovirus (GCRV), Infectious pancreatic necrosis virus (IPNV), and Large yellow croaker iridovirus (LYCI). The developed assay effectively identified AngHV in various tissues of infected eels, including the heart, liver, spleen, kidney, gills, intestines, muscle, skin mucus, and fins. Compared to quantitative real-time PCR (qPCR), the detection limit of this assay was determined to be 10,000 copies of AngHV, with an 88.89% concordance rate observed in the analysis of clinical samples between the results of DAS-ELISA and qPCR, thereby demonstrating the reliability of this immunoassay for detecting AngHV in field samples. Overall, the DAS-ELISA developed in this study demonstrates high reactivity and specificity, serving as a rapid and effective diagnostic tool for the detection and prevention of AngHV in eel populations.
{"title":"Development and Validation of a Double-Antibody Sandwich ELISA (DAS-ELISA) for Detection of Anguillid Herpesvirus.","authors":"Hua Chen, Jin-Xian Yang, Jun-Qing Ge","doi":"10.1111/jfd.70071","DOIUrl":"https://doi.org/10.1111/jfd.70071","url":null,"abstract":"<p><p>Anguillid herpesvirus (AngHV) is a highly pathogenic agent that causes \"Mucus sloughing and hemorrhagic septicemia disease\" in eels, resulting in high morbidity and cumulative mortality rates. Therefore, accurate diagnosis of AngHV infection is essential for effective clinical management and epidemiological control. In this study, three monoclonal antibodies (mAbs) against AngHV were developed, and their specificity and affinity were validated using Western blotting and indirect immunofluorescence assays. Subsequently, a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was established for the detection of AngHV, employing mAb 10G8-9F4 as the capture antibody and rabbit polyclonal antibody 9NA (which targets AngHV-ORF95) as the detection antibody. Specificity testing indicated no cross-reactivity with other common aquatic viruses, including Koi herpesvirus (KHV), Grass carp reovirus (GCRV), Infectious pancreatic necrosis virus (IPNV), and Large yellow croaker iridovirus (LYCI). The developed assay effectively identified AngHV in various tissues of infected eels, including the heart, liver, spleen, kidney, gills, intestines, muscle, skin mucus, and fins. Compared to quantitative real-time PCR (qPCR), the detection limit of this assay was determined to be 10,000 copies of AngHV, with an 88.89% concordance rate observed in the analysis of clinical samples between the results of DAS-ELISA and qPCR, thereby demonstrating the reliability of this immunoassay for detecting AngHV in field samples. Overall, the DAS-ELISA developed in this study demonstrates high reactivity and specificity, serving as a rapid and effective diagnostic tool for the detection and prevention of AngHV in eel populations.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70071"},"PeriodicalIF":2.2,"publicationDate":"2025-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336975","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}
Nocardia seriolae is the primary cause of fish nocardiosis, and it is urgent to develop an effective vaccine for controlling N. seriolae infection in aquaculture. In our previous study, two potential antigenic proteins, glutamate-cysteine ligase EgtA (EgtA) and hypothetical protein 6747 (hp6747), were determined via the in vivo-induced antigen technique (IVIAT). A DNA vaccine was aimed to be developed using these two in vivo-induced antigens as candidates for ligation with the pcDNA3.1-myc-his-A plasmid, and their immunological effects were evaluated in hybrid snakehead (Channa maculata ♀ × Channa argus ♂). Multiple epitopes of the EgtA and hp6747 proteins from N. seriolae were predicted on both T cells and B cells, and the transcription of the EgtA and hp6747 genes was found in the spleen, trunk kidney, muscle and liver in the fish after immunisation with pcDNA-EgtA and pcDNA-hp6747 DNA vaccines, respectively. Notably, the serum specific antibody (IgM) titres, as well as LYZ, ACP, AKP and SOD activities, were elevated in the vaccinated fish. Meanwhile, quantitative real-time PCR analysis demonstrated that the expression levels of several immune-related genes (TNFα, CD4, CD8α, IL-1β, MHCIIα and MHCIα) were significantly elevated across the aforementioned tissues in the vaccinated fish. Furthermore, they could provide the relative percent survival (RPS) at 61.04% and 43.73% against artificial challenge with N. seriolae in the fish after immunisation with pcDNA-EgtA and pcDNA-hp6747 DNA vaccines, respectively. Taken together, using these two in vivo-induced antigens of EgtA and hp6747 for vaccine development was the ideal strategy for controlling fish nocardiosis in aquaculture.
{"title":"Formulation and Immunological Assessment of DNA Vaccines Utilising In Vivo-Induced Antigens Against Nocardia seriolae in Hybrid Snakehead (Channa maculata ♀ × Channa argus ♂).","authors":"Xu Kang, Guoquan Chen, Na Li, Yuhao Li, Liqun Xia, Yishan Lu, Ting Huang, Jianlin Chen","doi":"10.1111/jfd.70074","DOIUrl":"https://doi.org/10.1111/jfd.70074","url":null,"abstract":"<p><p>Nocardia seriolae is the primary cause of fish nocardiosis, and it is urgent to develop an effective vaccine for controlling N. seriolae infection in aquaculture. In our previous study, two potential antigenic proteins, glutamate-cysteine ligase EgtA (EgtA) and hypothetical protein 6747 (hp6747), were determined via the in vivo-induced antigen technique (IVIAT). A DNA vaccine was aimed to be developed using these two in vivo-induced antigens as candidates for ligation with the pcDNA3.1-myc-his-A plasmid, and their immunological effects were evaluated in hybrid snakehead (Channa maculata ♀ × Channa argus ♂). Multiple epitopes of the EgtA and hp6747 proteins from N. seriolae were predicted on both T cells and B cells, and the transcription of the EgtA and hp6747 genes was found in the spleen, trunk kidney, muscle and liver in the fish after immunisation with pcDNA-EgtA and pcDNA-hp6747 DNA vaccines, respectively. Notably, the serum specific antibody (IgM) titres, as well as LYZ, ACP, AKP and SOD activities, were elevated in the vaccinated fish. Meanwhile, quantitative real-time PCR analysis demonstrated that the expression levels of several immune-related genes (TNFα, CD4, CD8α, IL-1β, MHCIIα and MHCIα) were significantly elevated across the aforementioned tissues in the vaccinated fish. Furthermore, they could provide the relative percent survival (RPS) at 61.04% and 43.73% against artificial challenge with N. seriolae in the fish after immunisation with pcDNA-EgtA and pcDNA-hp6747 DNA vaccines, respectively. Taken together, using these two in vivo-induced antigens of EgtA and hp6747 for vaccine development was the ideal strategy for controlling fish nocardiosis in aquaculture.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70074"},"PeriodicalIF":2.2,"publicationDate":"2025-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145337044","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}
Silvia Colussi, Giuseppe Esposito, Khalid Shahin, Pier Luigi Acutis, Lucio Fariano, Claudio Ghittino, Maria Goria, Fabio Bondavalli, Paolo Ajmone Marsan, Giorgia Bignami, Elena Bozzetta, Marino Prearo, Paolo Pastorino
Lactococcus garvieae, Lactococcus petauri and Lactococcus formosensis are etiological agents of piscine lactococcosis, a disease reported in Italy since the early 1990s and linked to significant aquaculture losses. To the best of our knowledge, this study reports the first detection of L. formosensis subsp. formosensis in farmed rainbow trout (Oncorhynchus mykiss) in Europe. In 2024, a total of 70 trout were sampled in the Piedmont region (Northern Italy), as part of a health monitoring programme. Bacteriological and molecular analyses showed the presence of L. garvieae in four fish (5.7%) and the identification of L. formosensis subsp. formosensis in one single fish exhibiting mild clinical signs via 16S-23S rRNA ITS sequencing. Phylogenetic analysis confirmed the clustering of the isolate within the L. formosensis clade, clearly distinct from L. garvieae and L. petauri. Virulotype comparison showed greater similarity between L. petauri and L. formosensis subsp. formosensis, while the latter exhibited a distinct biochemical profile and was highly susceptible to antibiotics commonly used in aquaculture. Nevertheless, monitoring and advanced diagnostics are essential to clarify the role of this bacterium and to develop any necessary preventive and control measures.
{"title":"First Detection of Lactococcus formosensis subsp. formosensis in Rainbow Trout (Oncorhynchus mykiss) in Europe.","authors":"Silvia Colussi, Giuseppe Esposito, Khalid Shahin, Pier Luigi Acutis, Lucio Fariano, Claudio Ghittino, Maria Goria, Fabio Bondavalli, Paolo Ajmone Marsan, Giorgia Bignami, Elena Bozzetta, Marino Prearo, Paolo Pastorino","doi":"10.1111/jfd.70068","DOIUrl":"https://doi.org/10.1111/jfd.70068","url":null,"abstract":"<p><p>Lactococcus garvieae, Lactococcus petauri and Lactococcus formosensis are etiological agents of piscine lactococcosis, a disease reported in Italy since the early 1990s and linked to significant aquaculture losses. To the best of our knowledge, this study reports the first detection of L. formosensis subsp. formosensis in farmed rainbow trout (Oncorhynchus mykiss) in Europe. In 2024, a total of 70 trout were sampled in the Piedmont region (Northern Italy), as part of a health monitoring programme. Bacteriological and molecular analyses showed the presence of L. garvieae in four fish (5.7%) and the identification of L. formosensis subsp. formosensis in one single fish exhibiting mild clinical signs via 16S-23S rRNA ITS sequencing. Phylogenetic analysis confirmed the clustering of the isolate within the L. formosensis clade, clearly distinct from L. garvieae and L. petauri. Virulotype comparison showed greater similarity between L. petauri and L. formosensis subsp. formosensis, while the latter exhibited a distinct biochemical profile and was highly susceptible to antibiotics commonly used in aquaculture. Nevertheless, monitoring and advanced diagnostics are essential to clarify the role of this bacterium and to develop any necessary preventive and control measures.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70068"},"PeriodicalIF":2.2,"publicationDate":"2025-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145274456","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}