Pub Date : 2025-02-27DOI: 10.1016/j.vetmic.2025.110441
Ruijiao Jiang , Qiuyan Huang , Ruiting Shen, Yongning Zhang, Lei Zhou, Xinna Ge, Jun Han, Xin Guo, Hanchun Yang
Porcine circovirus type 2 (PCV2) is the pathogen that causes porcine circovirus disease, characterized by severe immunosuppression and significant economic losses in the swine industry. The replicase (Rep), one of the most critical non-structural proteins of PCV2, plays a pivotal role in viral replication. However, the mechanism by which Rep regulates the replication of PCV2 still requires further investigation. Our study demonstrated that PCV2 can infect regulatory T cells (Tregs), and within the nucleus, Rep interacted with Foxp3, while the structural protein capsid protein (Cap) did not exhibit this interaction. Further investigations revealed that the Forkhead domain of Foxp3 was crucial for mediating its interaction with the C-terminal region of Rep, which had an ATPase activity-regulating domain. The interaction between Foxp3 and Rep reduced the ATPase activity of Rep, thereby inhibiting PCV2 replication. This study provided a theoretical foundation for elucidating the role of Rep in PCV2 pathogenesis and contributed to a deeper understanding of the molecular mechanisms underlying PCV2 immune evasion.
{"title":"Foxp3 inhibits PCV2 replication by reducing the ATPase activity of Rep","authors":"Ruijiao Jiang , Qiuyan Huang , Ruiting Shen, Yongning Zhang, Lei Zhou, Xinna Ge, Jun Han, Xin Guo, Hanchun Yang","doi":"10.1016/j.vetmic.2025.110441","DOIUrl":"10.1016/j.vetmic.2025.110441","url":null,"abstract":"<div><div>Porcine circovirus type 2 (PCV2) is the pathogen that causes porcine circovirus disease, characterized by severe immunosuppression and significant economic losses in the swine industry. The replicase (Rep), one of the most critical non-structural proteins of PCV2, plays a pivotal role in viral replication. However, the mechanism by which Rep regulates the replication of PCV2 still requires further investigation. Our study demonstrated that PCV2 can infect regulatory T cells (Tregs), and within the nucleus, Rep interacted with Foxp3, while the structural protein capsid protein (Cap) did not exhibit this interaction. Further investigations revealed that the Forkhead domain of Foxp3 was crucial for mediating its interaction with the C-terminal region of Rep, which had an ATPase activity-regulating domain. The interaction between Foxp3 and Rep reduced the ATPase activity of Rep, thereby inhibiting PCV2 replication. This study provided a theoretical foundation for elucidating the role of Rep in PCV2 pathogenesis and contributed to a deeper understanding of the molecular mechanisms underlying PCV2 immune evasion.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110441"},"PeriodicalIF":2.4,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143643205","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 : 2025-02-26DOI: 10.1016/j.vetmic.2025.110446
Chenbo Yan , Tianning Dong , Yiyi Shan , Bingru Zhao , Hua Yang , Yu Cai , Shanglai Li , Qiuyue Liu , Yuefeng Chu , Huafang Hao , Zilong Cheng , Maojun Liu , Yanli Zhang
Mycoplasma pneumonia is a chronic respiratory disease that seriously affects the health of sheep. To date, little information is available about the damage caused by Mycoplasma ovipneumoniae (MO) pneumonia to host lungs. Here, after sheep were infected with MO for 28 days, severe inflammatory reactions and pathological damage occurred. By using single-cell RNA sequencing (scRNA-seq), all the transcriptome changes in 11 cell types in sheep lung tissue were systematically analyzed, and the key biological processes regulating inflammation and immunity were identified. Moreover, we constructed both intercellular communication models and differential expression maps of key regulatory genes for each cell subgroup. We also specifically focused on the response of T cell subpopulations and neutrophils to MO infection. Long-term infection may affect an organism's immune response, inhibit intercellular communication, and highlight the important role of the cyclophilin A (CypA) and macrophage migration inhibitory factor (MIF) pathways in intercellular communication. Notably, MO infection decreased the toxicity of CD8 effector T cells and depleted regulatory T cells, thus inhibiting normal cell function. Subsequently, emphasis was placed on the important role of the neutrophil marker gene S100A9 in promoting neutrophil clearance of MO through activation of the ERK signaling pathway and reactive oxygen species (ROS) burst in vitro. These results contribute to understanding the progression of MO infection in the lungs and provide a rich database on the molecular basis of the response to different cell types in MO infection.
{"title":"Mycoplasma ovipnuemoniae impairs the immune response of sheep and suppresses neutrophil function by inhibiting S100A9","authors":"Chenbo Yan , Tianning Dong , Yiyi Shan , Bingru Zhao , Hua Yang , Yu Cai , Shanglai Li , Qiuyue Liu , Yuefeng Chu , Huafang Hao , Zilong Cheng , Maojun Liu , Yanli Zhang","doi":"10.1016/j.vetmic.2025.110446","DOIUrl":"10.1016/j.vetmic.2025.110446","url":null,"abstract":"<div><div><em>Mycoplasma pneumonia</em> is a chronic respiratory disease that seriously affects the health of sheep. To date, little information is available about the damage caused by <em>Mycoplasma ovipneumoniae</em> (MO) pneumonia to host lungs. Here, after sheep were infected with MO for 28 days, severe inflammatory reactions and pathological damage occurred. By using single-cell RNA sequencing (scRNA-seq), all the transcriptome changes in 11 cell types in sheep lung tissue were systematically analyzed, and the key biological processes regulating inflammation and immunity were identified. Moreover, we constructed both intercellular communication models and differential expression maps of key regulatory genes for each cell subgroup. We also specifically focused on the response of T cell subpopulations and neutrophils to MO infection. Long-term infection may affect an organism's immune response, inhibit intercellular communication, and highlight the important role of the cyclophilin A (CypA) and macrophage migration inhibitory factor (MIF) pathways in intercellular communication. Notably, MO infection decreased the toxicity of CD8 effector T cells and depleted regulatory T cells, thus inhibiting normal cell function. Subsequently, emphasis was placed on the important role of the neutrophil marker gene <em>S100A9</em> in promoting neutrophil clearance of MO through activation of the ERK signaling pathway and reactive oxygen species (ROS) burst <em>in vitro</em>. These results contribute to understanding the progression of MO infection in the lungs and provide a rich database on the molecular basis of the response to different cell types in MO infection.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"303 ","pages":"Article 110446"},"PeriodicalIF":2.4,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143512622","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}
Between June 2017 and October 2023, five cases of epididymitis caused by Pasteurella multocida were confirmed in four Japanese Black calves and one Japanese Black-Holstein crossbreed calf across five farms in three prefectures in Japan. Although P. multocida is primarily known to cause respiratory diseases in cattle, its role in reproductive disorders, particularly in epididymitis, has not been extensively studied. Histopathological examination showed pyogranulomatous lesions in the epididymal duct and surrounding areas, consisting of ductal extension, immune cell infiltration, epithelial degeneration, abscess formation, and fibrosis with angiogenesis. In the immunohistochemical analysis, positive reactions to anti-P. multocida serotype A antibodies were observed in macrophages and neutrophils in the epididymal ducts and surrounding areas, suggesting an ascending infection rather than hematogenous spread. This report is the first to directly confirm P. multocida involvement in bovine epididymitis through immunohistochemical examination. P. multocida was isolated in pure culture from the scrotal contents in all cases. The isolates were grouped into sequence types ST79 and ST13, commonly found in the respiratory tract and oral cavity of cattle, using the Rural Industries Research and Development Corporation multilocus sequence typing scheme; pulsed-field gel electrophoresis revealed unique band patterns in the isolates from each case. These results suggest that multiple genetically distinct strains, commonly found in the nasal and oral cavities, incidentally infected the epididymis via an ascending route. This bacterium warrants further investigation as a potential cause of reproductive disorders, particularly in Japanese Black cattle.
{"title":"Pathological and bacteriological investigations of Pasteurella multocida-induced epididymitis in calves","authors":"Kaori Namba , Aya Terao , Yuichi Ueno , Chie Teratani , Ikumi Yamamoto , Risa Kaji , Sachi Tamaboko , Chiaki Mizukami , Akihiko Hashida , Mitsutaka Ikezawa , Shogo Tanaka , Kumiko Kimura","doi":"10.1016/j.vetmic.2025.110445","DOIUrl":"10.1016/j.vetmic.2025.110445","url":null,"abstract":"<div><div>Between June 2017 and October 2023, five cases of epididymitis caused by <em>Pasteurella multocida</em> were confirmed in four Japanese Black calves and one Japanese Black-Holstein crossbreed calf across five farms in three prefectures in Japan. Although <em>P. multocida</em> is primarily known to cause respiratory diseases in cattle, its role in reproductive disorders, particularly in epididymitis, has not been extensively studied. Histopathological examination showed pyogranulomatous lesions in the epididymal duct and surrounding areas, consisting of ductal extension, immune cell infiltration, epithelial degeneration, abscess formation, and fibrosis with angiogenesis. In the immunohistochemical analysis, positive reactions to anti-<em>P. multocida</em> serotype A antibodies were observed in macrophages and neutrophils in the epididymal ducts and surrounding areas, suggesting an ascending infection rather than hematogenous spread. This report is the first to directly confirm <em>P. multocida</em> involvement in bovine epididymitis through immunohistochemical examination. <em>P. multocida</em> was isolated in pure culture from the scrotal contents in all cases. The isolates were grouped into sequence types ST79 and ST13, commonly found in the respiratory tract and oral cavity of cattle, using the Rural Industries Research and Development Corporation multilocus sequence typing scheme; pulsed-field gel electrophoresis revealed unique band patterns in the isolates from each case. These results suggest that multiple genetically distinct strains, commonly found in the nasal and oral cavities, incidentally infected the epididymis via an ascending route. This bacterium warrants further investigation as a potential cause of reproductive disorders, particularly in Japanese Black cattle.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110445"},"PeriodicalIF":2.4,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143609916","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 : 2025-02-26DOI: 10.1016/j.vetmic.2025.110444
Xiuhua Yu , Ning Su , Jinna Luo , Daining Zhang , Hansi Zhang , Ming Duan , Ning Shi
Long noncoding RNAs (lncRNAs) are important regulators of gene expression. Although evidence accumulated over the past decade shows that lncRNAs have key roles in the interaction between viruses and hosts, the functions of the majority of differentially expressed lncRNAs in response to viral infections remain uncharacterized so far. In this study, we have identified that USP30 antisense RNA 1 (USP30-AS1), a host antisense lncRNA, is hijacked by influenza A virus (IAV) to assist its replication. We show that USP30-AS1 is IAV-induced via the Janus protein tyrosine kinase-signal transducer and the activator of transcription (JAK-STAT) signaling pathway. Functionally, ectopic expression of USP30-AS1 significantly promotes IAV replication. Conversely, silencing USP30-AS1 suppresses IAV replication. Mechanistically, USP30-AS1 directly binds prohibitin 1 (PHB1) and modulates its protein stability and function. On the one hand, the binding of USP30-AS1 sequesters PHB1 away from the E3 ubiquitin ligase, tripartite motif containing 21 (TRIM21), thereby protecting the protein stability of PHB1. On the other hand, USP30-AS1 serves as a molecular scaffold for enhancing the interaction between PHB1 and interferon regulatory factor 3 (IRF3), which in turn impedes the nuclear import of IRF3. Therefore, our data unveil an important role of USP30-AS1 in promoting viral replication by modulating PHB1 stability and functions, providing a new insight into the role of lncRNAs in the interplay between IAV and host.
{"title":"Long noncoding RNA USP30-AS1 promotes influenza A virus replication by enhancing PHB1 function","authors":"Xiuhua Yu , Ning Su , Jinna Luo , Daining Zhang , Hansi Zhang , Ming Duan , Ning Shi","doi":"10.1016/j.vetmic.2025.110444","DOIUrl":"10.1016/j.vetmic.2025.110444","url":null,"abstract":"<div><div>Long noncoding RNAs (lncRNAs) are important regulators of gene expression. Although evidence accumulated over the past decade shows that lncRNAs have key roles in the interaction between viruses and hosts, the functions of the majority of differentially expressed lncRNAs in response to viral infections remain uncharacterized so far. In this study, we have identified that USP30 antisense RNA 1 (USP30-AS1), a host antisense lncRNA, is hijacked by influenza A virus (IAV) to assist its replication. We show that USP30-AS1 is IAV-induced via the Janus protein tyrosine kinase-signal transducer and the activator of transcription (JAK-STAT) signaling pathway. Functionally, ectopic expression of USP30-AS1 significantly promotes IAV replication. Conversely, silencing USP30-AS1 suppresses IAV replication. Mechanistically, USP30-AS1 directly binds prohibitin 1 (PHB1) and modulates its protein stability and function. On the one hand, the binding of USP30-AS1 sequesters PHB1 away from the E3 ubiquitin ligase, tripartite motif containing 21 (TRIM21), thereby protecting the protein stability of PHB1. On the other hand, USP30-AS1 serves as a molecular scaffold for enhancing the interaction between PHB1 and interferon regulatory factor 3 (IRF3), which in turn impedes the nuclear import of IRF3. Therefore, our data unveil an important role of USP30-AS1 in promoting viral replication by modulating PHB1 stability and functions, providing a new insight into the role of lncRNAs in the interplay between IAV and host.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"303 ","pages":"Article 110444"},"PeriodicalIF":2.4,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143512621","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 : 2025-02-25DOI: 10.1016/j.vetmic.2025.110443
Mengtao Wang , Mengjuan Ma , Lijie Yu, Kun He, Tengli Zhang, Yiming Feng, Gongzheng Hu, Dandan He, Yushan Pan, Yajun Zhai
Antimicrobial resistance (AMR) poses a significant global health threat, particularly due to increasing bacterial resistance to β-lactam and aminoglycoside antibiotics, primarily mediated by extended-spectrum β-lactamases (ESBLs) and 16S rRNA methylases in Enterobacteriaceae. In this study, a multidrug resistant (MDR) E. coli strain HN257 isolated from chicken belonging to ST224 and serotype O88:H23 was characterized. SNP-based phylogenetic analysis revealed two distinct clades among poultry-associated E. coli ST224 in this study and others from Genbank, with strain HN257 closely related to chicken-derived E. coli YH17148 (serotype O78:H23), from China. The E. coli HN257 harbored four plasmids with 16 resistance determinants. Two blaCTX-M-65 genes were located on different plasmids with an IS26-bracketed resistance module IS26-traI-fip-∆ISEcp1-blaCTX-M-65-IS903D-iroN-IS26. The plasmid pHN257–2 belonged to the IncI1 ST71 epidemic lineage and carried blaCTX-M-65, blaTEM-1b, rmtB, fosA3, floR, aac(3)-IV and oqxAB, while plasmid pHN257–4 belonged to the non-conjugative IncX1 and carried blaCTX-M-65 and fosA3. Under experimental conditions, a rmtB-positive conjugative helper IncI1 ST136 plasmid could fuse with the non-conjugative pHN257–4 carrying blaCTX-M-65, resulting in the formation of a cointegrate pHN257–F mediated by IS26. Importantly, both single and fused plasmids in transconjugants showed minimal impact on bacterial growth. This study highlights the first identification of a non-conjugative IncX1 plasmid carrying blaCTX-M-65 and fosA3 in MDR E. coli ST224 from poultry, offering critical insights into the presence and transmission dynamics of blaCTX-M-65.
{"title":"Characterization of IS26-bracketed blaCTX-M-65 resistance module on IncI1 and IncX1 plasmids in Escherichia coli ST224 isolated from a chicken in China","authors":"Mengtao Wang , Mengjuan Ma , Lijie Yu, Kun He, Tengli Zhang, Yiming Feng, Gongzheng Hu, Dandan He, Yushan Pan, Yajun Zhai","doi":"10.1016/j.vetmic.2025.110443","DOIUrl":"10.1016/j.vetmic.2025.110443","url":null,"abstract":"<div><div>Antimicrobial resistance (AMR) poses a significant global health threat, particularly due to increasing bacterial resistance to β-lactam and aminoglycoside antibiotics, primarily mediated by extended-spectrum β-lactamases (ESBLs) and 16S rRNA methylases in <em>Enterobacteriaceae</em>. In this study, a multidrug resistant (MDR) <em>E. coli</em> strain HN257 isolated from chicken belonging to ST224 and serotype O88:H23 was characterized. SNP-based phylogenetic analysis revealed two distinct clades among poultry-associated <em>E. coli</em> ST224 in this study and others from Genbank, with strain HN257 closely related to chicken-derived <em>E. coli</em> YH17148 (serotype O78:H23), from China. The <em>E. coli</em> HN257 harbored four plasmids with 16 resistance determinants. Two <em>bla</em><sub>CTX-M-65</sub> genes were located on different plasmids with an IS<em>26</em>-bracketed resistance module IS<em>26-traI-fip</em>-∆IS<em>Ecp1</em>-<em>bla</em><sub>CTX-M-65</sub>-IS<em>903D</em>-<em>iroN</em>-IS<em>26</em>. The plasmid pHN257–2 belonged to the IncI1 ST71 epidemic lineage and carried <em>bla</em><sub>CTX-M-65</sub>, <em>bla</em><sub>TEM-1b</sub>, <em>rmtB</em>, <em>fosA3</em>, <em>floR</em>, <em>aac</em>(3)<em>-IV</em> and <em>oqxAB</em>, while plasmid pHN257–4 belonged to the non-conjugative IncX1 and carried <em>bla</em><sub>CTX-M-65</sub> and <em>fosA3</em>. Under experimental conditions, a <em>rmtB</em>-positive conjugative helper IncI1 ST136 plasmid could fuse with the non-conjugative pHN257–4 carrying <em>bla</em><sub>CTX-M-65</sub>, resulting in the formation of a cointegrate pHN257–F mediated by IS<em>26</em>. Importantly, both single and fused plasmids in transconjugants showed minimal impact on bacterial growth. This study highlights the first identification of a non-conjugative IncX1 plasmid carrying <em>bla</em><sub>CTX-M-65</sub> and <em>fosA3</em> in MDR <em>E. coli</em> ST224 from poultry, offering critical insights into the presence and transmission dynamics of <em>bla</em><sub>CTX-M-65</sub>.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"303 ","pages":"Article 110443"},"PeriodicalIF":2.4,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143512629","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 : 2025-02-25DOI: 10.1016/j.vetmic.2025.110442
Xiang-hui Ling , Biao Zhang , Hao-jie Ren , Ming-yang Li , Shun-da Liu , Meng-ru Luo , Ke-wei Guo , Shi-chong Han , Wen-rui He , Gai-ping Zhang , Yu-hang Zhang , Bo Wan
Senecavirus A (SVA) infection causes vesicular disease in pigs and acute death of newborn piglets. Frequent gene mutation and recombination events lead to difficulties for SVA prevention and eradication, especially impeding the development of effective vaccine or treatment drug. SVA nonstructural 3 A protein plays an important role in viral replication and various immune evasion pathways, which makes it a potential therapeutic target. In this study, prokaryotic 3 A protein was successfully expressed in Escherichia.coli BL21 (DE3) system and purified with Ni-affinity chromatography. Western blotting results showed 3 A protein specifically reacted with serum of SVA infected pig, indicating that nonstructural 3 A protein was a potential diagnostic maker for SVA serological testing, especially for differentiating infected from vaccinated animals. In addition, specific monoclonal antibody (mAb) 5A7 against 3 A protein was also prepared. Indirect immunofluorescence assay and Western blotting assay showed mAb 5A7 specifically reacted with SVA cultured in IBRS-2 cells. To characterize the epitope of mAb 5A7, serial truncated peptides of 3 A protein were prepared. Western blotting assay showed the epitope of mAb 5A7 was 5NDDTPVDEALGR16. Bioinformatic studies revealed that the epitopes are located on the extrinsic membrane domain of 3 A protein with good antigenicity. To sum up, SVA 3 A protein and its specific mAb 5A7 were successfully prepared in this study, which will contribute to biological function study of 3 A protein and the pathogenic mechanism of SVA, as well as the diagnosis and prevention of this disease.
{"title":"Preparation and epitope identification of a novel monoclonal antibody against 3A protein of Senecavirus A","authors":"Xiang-hui Ling , Biao Zhang , Hao-jie Ren , Ming-yang Li , Shun-da Liu , Meng-ru Luo , Ke-wei Guo , Shi-chong Han , Wen-rui He , Gai-ping Zhang , Yu-hang Zhang , Bo Wan","doi":"10.1016/j.vetmic.2025.110442","DOIUrl":"10.1016/j.vetmic.2025.110442","url":null,"abstract":"<div><div>Senecavirus A (SVA) infection causes vesicular disease in pigs and acute death of newborn piglets. Frequent gene mutation and recombination events lead to difficulties for SVA prevention and eradication, especially impeding the development of effective vaccine or treatment drug. SVA nonstructural 3 A protein plays an important role in viral replication and various immune evasion pathways, which makes it a potential therapeutic target. In this study, prokaryotic 3 A protein was successfully expressed in <em>Escherichia.coli</em> BL21 (DE3) system and purified with Ni-affinity chromatography. Western blotting results showed 3 A protein specifically reacted with serum of SVA infected pig, indicating that nonstructural 3 A protein was a potential diagnostic maker for SVA serological testing, especially for differentiating infected from vaccinated animals. In addition, specific monoclonal antibody (mAb) 5A7 against 3 A protein was also prepared. Indirect immunofluorescence assay and Western blotting assay showed mAb 5A7 specifically reacted with SVA cultured in IBRS-2 cells. To characterize the epitope of mAb 5A7, serial truncated peptides of 3 A protein were prepared. Western blotting assay showed the epitope of mAb 5A7 was <sup>5</sup>NDDTPVDEALGR<sup>16</sup>. Bioinformatic studies revealed that the epitopes are located on the extrinsic membrane domain of 3 A protein with good antigenicity. To sum up, SVA 3 A protein and its specific mAb 5A7 were successfully prepared in this study, which will contribute to biological function study of 3 A protein and the pathogenic mechanism of SVA, as well as the diagnosis and prevention of this disease.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"303 ","pages":"Article 110442"},"PeriodicalIF":2.4,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527471","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 : 2025-02-24DOI: 10.1016/j.vetmic.2025.110440
Ying Sun , Shenglin Huang , Fengjuan Li , Shulun Huang , Pinghua Li , Qiongqiong Zhao , Tao Wang , Huifang Bao , Yuanfang Fu , Pu Sun , Xingwen Bai , Hong Yuan , Xueqing Ma , Zhixun Zhao , Jing Zhang , Jian Wang , Dong Li , Qiang Zhang , Yimei Cao , Kun Li , Huiying Fan
Foot-and-mouth disease virus (FMDV) is highly infectious and lacks cross-protection among serotypes, with antibodies playing a key role in antiviral immunity. To map conserved epitopes on the FMDV surface that exhibit cross-serotype reactivity, we constructed a pig-specific B-cell receptor (BCR) library through single B-cell sorting and high-throughput sequencing. This led to the identification of 16 broadly reactive, non-neutralizing monoclonal antibodies (mAbs), with 10 targeting VP2 (pOTB-1, pOTB-10, pOTB-13, pOTB-33, pOTB-37, pONY-14, pONY-17, pONY-23, pONY-30, pONY-60) and 6 targeting VP3 (pOTB-6, pOTB-11, pOTB-22, pOTB-23, pONY-3, pONY-59). Among these, a novel free linear epitope was identified at the C-terminus of VP2, recognized by pOTB-1, with the minimal recognition motif "KE." Key residues, T53 and W101, within the complementarity-determining region (CDR) of the pOTB-1 heavy chain, interact with the carboxyl group of the C-terminal glutamate through hydrogen bonding, contributing to the free-form nature of the epitope. Competitive enzyme-linked immunosorbent assays (cELISA) showed that most non-neutralizing antibodies (nNAbs) interfered with the binding of neutralizing antibodies B82 (site 2) and C4 (site 4), confirming the overlap between non-neutralizing and neutralizing epitopes. It has been confirmed that nNAbs mediate antiviral activity in vivo through various mechanisms, such as the formation of immune complexes. These findings reveal new epitopes on VP2 and VP3 and their spatial overlap with neutralizing sites, enhancing our understanding of FMDV immunogenicity and providing novel targets for vaccine and therapeutic development.
{"title":"Porcine antibodies reveal novel non-neutralizing universal epitopes on FMDV and their overlaps with neutralization sites","authors":"Ying Sun , Shenglin Huang , Fengjuan Li , Shulun Huang , Pinghua Li , Qiongqiong Zhao , Tao Wang , Huifang Bao , Yuanfang Fu , Pu Sun , Xingwen Bai , Hong Yuan , Xueqing Ma , Zhixun Zhao , Jing Zhang , Jian Wang , Dong Li , Qiang Zhang , Yimei Cao , Kun Li , Huiying Fan","doi":"10.1016/j.vetmic.2025.110440","DOIUrl":"10.1016/j.vetmic.2025.110440","url":null,"abstract":"<div><div>Foot-and-mouth disease virus (FMDV) is highly infectious and lacks cross-protection among serotypes, with antibodies playing a key role in antiviral immunity. To map conserved epitopes on the FMDV surface that exhibit cross-serotype reactivity, we constructed a pig-specific B-cell receptor (BCR) library through single B-cell sorting and high-throughput sequencing. This led to the identification of 16 broadly reactive, non-neutralizing monoclonal antibodies (mAbs), with 10 targeting VP2 (pOTB-1, pOTB-10, pOTB-13, pOTB-33, pOTB-37, pONY-14, pONY-17, pONY-23, pONY-30, pONY-60) and 6 targeting VP3 (pOTB-6, pOTB-11, pOTB-22, pOTB-23, pONY-3, pONY-59). Among these, a novel free linear epitope was identified at the C-terminus of VP2, recognized by pOTB-1, with the minimal recognition motif \"KE.\" Key residues, T53 and W101, within the complementarity-determining region (CDR) of the pOTB-1 heavy chain, interact with the carboxyl group of the C-terminal glutamate through hydrogen bonding, contributing to the free-form nature of the epitope. Competitive enzyme-linked immunosorbent assays (cELISA) showed that most non-neutralizing antibodies (nNAbs) interfered with the binding of neutralizing antibodies B82 (site 2) and C4 (site 4), confirming the overlap between non-neutralizing and neutralizing epitopes. It has been confirmed that nNAbs mediate antiviral activity in vivo through various mechanisms, such as the formation of immune complexes. These findings reveal new epitopes on VP2 and VP3 and their spatial overlap with neutralizing sites, enhancing our understanding of FMDV immunogenicity and providing novel targets for vaccine and therapeutic development.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"303 ","pages":"Article 110440"},"PeriodicalIF":2.4,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143529211","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 : 2025-02-20DOI: 10.1016/j.vetmic.2025.110439
Maureen Sijtsema , Thijs Stringer , Nonke E.M. Hopman , Linda van der Graaf – van Bloois , Birgitta Duim , Astrid van den Brom-Spierenburg , Mathijs Theelen , Aldert Zomer , Els M. Broens
Healthcare-associated infections (HAIs) associated with ESKAPEE pathogens are an emerging concern in equine hospitals, especially in the intensive care unit (ICU). To gain insight into the occurrence of HAIs and to identify potential reservoirs and transmission routes of ESKAPEE pathogens in an equine ICU, a pilot study of two periods (December 2022-January 2023 and April-June 2023) was performed, where patient and environmental samples were obtained upon ICU admission and discharge. A sampling and laboratory protocol for the detection and identification of different ESKAPEE pathogens and Salmonella spp. was developed. Samples consisted of patient samples and patient-bound environmental samples, including nose and environmental swabs, feces, twitch ropes and catheter tips. Non-patient bound samples were collected from high-contact surfaces for staff members. In total, 271 patient-bound samples (n = 122 in sampling period 1 and n = 149 in sampling period 2) from 30 patients and 28 non-patient bound environmental samples were collected resulting in the isolation of 207 ESKAPEE isolates. A selection of isolates (n = 115) was sequenced to identify clusters and antimicrobial resistance genes. ESKAPEE pathogens were present in 90 % of the horses hospitalized at the ICU or their immediate environment. Different genetic clusters of MRSA, A. baumannii and E. cloacae isolates were identified over time suggesting transmission and persistence within the ICU. During both sampling periods no HAIs were observed, but the abundance of these pathogens might pose a risk for HAIs. This study shows the importance for every (veterinary) hospital to implement surveillance and to further optimize hygiene protocols.
{"title":"Active surveillance in an equine intensive care unit identifies clusters of ESKAPEE pathogens in the veterinary hospital environment","authors":"Maureen Sijtsema , Thijs Stringer , Nonke E.M. Hopman , Linda van der Graaf – van Bloois , Birgitta Duim , Astrid van den Brom-Spierenburg , Mathijs Theelen , Aldert Zomer , Els M. Broens","doi":"10.1016/j.vetmic.2025.110439","DOIUrl":"10.1016/j.vetmic.2025.110439","url":null,"abstract":"<div><div>Healthcare-associated infections (HAIs) associated with ESKAPEE pathogens are an emerging concern in equine hospitals, especially in the intensive care unit (ICU). To gain insight into the occurrence of HAIs and to identify potential reservoirs and transmission routes of ESKAPEE pathogens in an equine ICU, a pilot study of two periods (December 2022-January 2023 and April-June 2023) was performed, where patient and environmental samples were obtained upon ICU admission and discharge. A sampling and laboratory protocol for the detection and identification of different ESKAPEE pathogens and <em>Salmonella</em> spp. was developed. Samples consisted of patient samples and patient-bound environmental samples, including nose and environmental swabs, feces, twitch ropes and catheter tips. Non-patient bound samples were collected from high-contact surfaces for staff members. In total, 271 patient-bound samples (n = 122 in sampling period 1 and n = 149 in sampling period 2) from 30 patients and 28 non-patient bound environmental samples were collected resulting in the isolation of 207 ESKAPEE isolates. A selection of isolates (n = 115) was sequenced to identify clusters and antimicrobial resistance genes. ESKAPEE pathogens were present in 90 % of the horses hospitalized at the ICU or their immediate environment. Different genetic clusters of MRSA, <em>A. baumannii</em> and <em>E. cloacae</em> isolates were identified over time suggesting transmission and persistence within the ICU. During both sampling periods no HAIs were observed, but the abundance of these pathogens might pose a risk for HAIs. This study shows the importance for every (veterinary) hospital to implement surveillance and to further optimize hygiene protocols.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"303 ","pages":"Article 110439"},"PeriodicalIF":2.4,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143464779","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}
A classical swine fever (CSF) vaccine combining high potency and an immunological marker for differentiating infected animals from vaccinated animals facilitates disease control and provides proof of eradication to promote international pig trade. Previously, CSF virus (CSFV) recombinant live vaccine strains, guinea-pig exaltation of Newcastle disease virus-negative strain vaccine [vGPE– (genotype 1.1)], were developed with the Erns gene replaced by non-CSF pestiviruses (Pronghorn antelope or Phocoena pestiviruses). We evaluated the potency of these marker vaccines against a Japanese circulating CSFV strain (genotype 2.1), which is genetically distant from the vaccine strain. Pigs were experimentally vaccinated with the vGPE– and two marker vaccine strains. All vaccinated and unvaccinated pigs were challenged with CSFV JPN/1/2018 at 26 days post-vaccination. The clinical signs and viral titers in blood and oral swabs were monitored for three weeks post-challenge, and antibodies against CSFV E2 and Erns were detected using commercial enzyme-linked immunosorbent assay kits. Unvaccinated pigs showed typical CSF clinical signs and viremia, and one pig died at 19 days post-challenge. Meanwhile, none of the vaccinated pigs showed any clinical signs, and the replication of infectious virus was substantially suppressed. Both vGPE–-vaccinated and unvaccinated pigs had CSFV E2 and Erns antibodies after vaccination and virus challenge; meanwhile, notably, marker-vaccinated pigs had only E2 antibodies, while both E2 and Erns antibodies detected only after the challenge. In conclusion, the marker vaccine strains provided protective immunity to suppress clinical signs, viremia, and virus excretion, comparable to the GPE– live vaccine, and successfully differentiated infection from vaccination.
{"title":"Potency of two chimeric vaccine candidates derived from the classical swine fever GPE– vaccine strain against a circulating virus strain isolated in Japan","authors":"Tatsuya Nishi , Loc Tan Huynh , Tomoko Kato , Mitsutaka Ikezawa , Takehisa Yamamoto , Yoshihiro Sakoda , Katsuhiko Fukai","doi":"10.1016/j.vetmic.2025.110438","DOIUrl":"10.1016/j.vetmic.2025.110438","url":null,"abstract":"<div><div>A classical swine fever (CSF) vaccine combining high potency and an immunological marker for differentiating infected animals from vaccinated animals facilitates disease control and provides proof of eradication to promote international pig trade. Previously, CSF virus (CSFV) recombinant live vaccine strains, guinea-pig exaltation of Newcastle disease virus-negative strain vaccine [vGPE<sup>–</sup> (genotype 1.1)], were developed with the E<sup>rns</sup> gene replaced by non-CSF pestiviruses (Pronghorn antelope or Phocoena pestiviruses). We evaluated the potency of these marker vaccines against a Japanese circulating CSFV strain (genotype 2.1), which is genetically distant from the vaccine strain. Pigs were experimentally vaccinated with the vGPE<sup>–</sup> and two marker vaccine strains. All vaccinated and unvaccinated pigs were challenged with CSFV JPN/1/2018 at 26 days post-vaccination. The clinical signs and viral titers in blood and oral swabs were monitored for three weeks post-challenge, and antibodies against CSFV E2 and E<sup>rns</sup> were detected using commercial enzyme-linked immunosorbent assay kits. Unvaccinated pigs showed typical CSF clinical signs and viremia, and one pig died at 19 days post-challenge. Meanwhile, none of the vaccinated pigs showed any clinical signs, and the replication of infectious virus was substantially suppressed. Both vGPE<sup>–</sup>-vaccinated and unvaccinated pigs had CSFV E2 and E<sup>rns</sup> antibodies after vaccination and virus challenge; meanwhile, notably, marker-vaccinated pigs had only E2 antibodies, while both E2 and E<sup>rns</sup> antibodies detected only after the challenge. In conclusion, the marker vaccine strains provided protective immunity to suppress clinical signs, viremia, and virus excretion, comparable to the GPE<sup>–</sup> live vaccine, and successfully differentiated infection from vaccination.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"303 ","pages":"Article 110438"},"PeriodicalIF":2.4,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143510629","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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