Pub Date : 2026-01-05DOI: 10.1016/j.virol.2026.110786
Kun Xu , Chen Yao , Yueying Wang , Shuang Guo , Jiajia Pan , Guoyu Yang
Bovine enterovirus (BEV) is an important pathogen causing calf diarrhea and has been detected in the diarrheic calf feces, although its pathogenicity remains unclear. This study aims to investigate the epidemiological profile and genetic diversity of BEV. A total of 356 clinical samples were analyzed by RT-PCR, including feces and rectal swabs from diarrheic calves in 11 cities across four provinces of China during 2024, the results showed a BEV-positive rate of 5.9 % (21/356, CI: 3.89–8.85). The four-province survey revealed differential BEV prevalence, with the highest rate observed in Qinghai (14.29 %). Five subtypes (E2, F1, F2, F3, F8) were found to co-circulate, notably E2, F3 and F8 in Inner Mongolia. Genomic analyses indicated that purifying selection dominates BEV evolution. Following sequencing of ten P1 nucleotide fragments, sequence comparison and phylogenetic analysis were performed based on P1 and VP1. The results showed that the obtained sequences clustered within a branch containing BEV subtypes E2, F3, F2, F1, and F8, and were genetically more closely related to strains from Australia, Japan, and China. Additionally, through the comparison and analysis of the majority of amino acid residues entropy of VP1, VP2 and VP3, it was determined that the BEV VP1 protein showed a high level of genetic diversity. This study provides powerful insight for us to further understand the epidemic status and evolution of BEV in China.
{"title":"Molecular epidemiological investigation and genetic diversity of bovine enterovirus in China","authors":"Kun Xu , Chen Yao , Yueying Wang , Shuang Guo , Jiajia Pan , Guoyu Yang","doi":"10.1016/j.virol.2026.110786","DOIUrl":"10.1016/j.virol.2026.110786","url":null,"abstract":"<div><div>Bovine enterovirus (BEV) is an important pathogen causing calf diarrhea and has been detected in the diarrheic calf feces, although its pathogenicity remains unclear. This study aims to investigate the epidemiological profile and genetic diversity of BEV. A total of 356 clinical samples were analyzed by RT-PCR, including feces and rectal swabs from diarrheic calves in 11 cities across four provinces of China during 2024, the results showed a BEV-positive rate of 5.9 % (21/356, CI: 3.89–8.85). The four-province survey revealed differential BEV prevalence, with the highest rate observed in Qinghai (14.29 %). Five subtypes (E2, F1, F2, F3, F8) were found to co-circulate, notably E2, F3 and F8 in Inner Mongolia. Genomic analyses indicated that purifying selection dominates BEV evolution. Following sequencing of ten P1 nucleotide fragments, sequence comparison and phylogenetic analysis were performed based on P1 and VP1. The results showed that the obtained sequences clustered within a branch containing BEV subtypes E2, F3, F2, F1, and F8, and were genetically more closely related to strains from Australia, Japan, and China. Additionally, through the comparison and analysis of the majority of amino acid residues entropy of VP1, VP2 and VP3, it was determined that the BEV VP1 protein showed a high level of genetic diversity. This study provides powerful insight for us to further understand the epidemic status and evolution of BEV in China.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"616 ","pages":"Article 110786"},"PeriodicalIF":2.4,"publicationDate":"2026-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145978156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-22DOI: 10.1016/j.virol.2025.110777
Shivani Raj, Minnah Irfan, C.T. Ranjith-Kumar
Chikungunya virus (CHIKV) is a single-stranded positive-sense RNA virus that employs various strategies to evade the host immune response. CHIKV non-structural protein 2 (nsP2) is one of the viral-encoded proteins essential for viral replication as well as modulation of the immune response. In this study, we demonstrated that CHIKV nsP2 suppresses both Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) and Toll-like receptors (TLRs) signaling pathways. Domain mapping of nsP2 identified that the N-terminal region encompassing the N-terminal domain and the helicase domain (NH) is responsible for the inhibition. Furthermore, site-directed mutagenesis experiments showed that a functional helicase is necessary for inhibiting interferon production, but the C-terminal VLoop region, previously implicated in host transcriptional shutoff is not. Lastly, we demonstrated that nsP2 disrupts two key immune RLRs and TLRs mediated signaling pathways by interfering with the common proteins TANK-binding kinase 1 (TBK1) and interferon regulatory factor 3 (IRF3), which are involved in both signaling pathways. These findings enhance our understanding of CHIKV immune evasion strategies and offer potential targets for the development of antiviral therapeutics.
{"title":"Chikungunya virus non-structural protein 2 (nsP2) inhibits RIG-I and TLR-mediated immune response","authors":"Shivani Raj, Minnah Irfan, C.T. Ranjith-Kumar","doi":"10.1016/j.virol.2025.110777","DOIUrl":"10.1016/j.virol.2025.110777","url":null,"abstract":"<div><div>Chikungunya virus (CHIKV) is a single-stranded positive-sense RNA virus that employs various strategies to evade the host immune response. CHIKV non-structural protein 2 (nsP2) is one of the viral-encoded proteins essential for viral replication as well as modulation of the immune response. In this study, we demonstrated that CHIKV nsP2 suppresses both Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) and Toll-like receptors (TLRs) signaling pathways. Domain mapping of nsP2 identified that the N-terminal region encompassing the N-terminal domain and the helicase domain (NH) is responsible for the inhibition. Furthermore, site-directed mutagenesis experiments showed that a functional helicase is necessary for inhibiting interferon production, but the C-terminal VLoop region, previously implicated in host transcriptional shutoff is not. Lastly, we demonstrated that nsP2 disrupts two key immune RLRs and TLRs mediated signaling pathways by interfering with the common proteins TANK-binding kinase 1 (TBK1) and interferon regulatory factor 3 (IRF3), which are involved in both signaling pathways. These findings enhance our understanding of CHIKV immune evasion strategies and offer potential targets for the development of antiviral therapeutics.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"616 ","pages":"Article 110777"},"PeriodicalIF":2.4,"publicationDate":"2025-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145835809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
We have developed a recombinant vaccinia vaccine (rDIs-S) against SARS-CoV-2 from an attenuated vaccinia virus (Dairen-I minute-pock variant: DIs strain). The rDIs-S vaccine containing DNA of an S gene of SARS-CoV-2 early-pandemic Pango B strain protected cynomolgus macaques from severe pneumonia caused by a SARS-CoV-2 Pango A strain and induced a broad reactive neutralization antibody against SARS-CoV-2 variants (B.1.1.7, B.1.351, and P.1) in our previous study. In the present study, to further confirm the safety and efficacy of rDIs-S, the vaccine (1 × 108 PFU) was inoculated intradermally into rhesus macaques, which showed antibody-dependent enhancement (ADE) after vaccination with another vaccinia virus vector, MVA carrying a SARS-CoV S gene, and challenge infection with SARS-CoV. The rDIs-S vaccination induced neutralization activity against multiple SARS-CoV-2 variants in plasma of rhesus macaques [100 % (6/6 macaques)] and reduced viral replication of the B.1.1.7 and B.1.351 variants and infiltration of T cells and macrophages into the lungs of rhesus macaques [100 % (5/5 macaques)] after challenge infection with 1 × 107 TCID50 of the two SARS-CoV-2 variants. In the presence of plasma of vaccinated macaques, viral RNA levels in human FcγR-positive cells increased slightly by up to 1.8-folds, but they were much lower than those previously reported in sera of mRNA-vaccinated individuals, in which ADE was anticipated (ratios approximately 10–1000). Thus, rDIs-S induced protective immune responses against SARS-CoV-2 B.1.1.7 and B.1.351 and vaccination with rDIs-S did not enhance pneumonia or replication of SARS-CoV-2, indicating the efficacy and safety of rDIs-S in the rhesus macaque model.
{"title":"Vaccinia virus-based SARS-CoV-2 vaccine prevents lung immunopathology without antibody-dependent enhancement in female rhesus macaques","authors":"Cong Thanh Nguyen , Misako Nakayama , Fumihiko Yasui , Hirohito Ishigaki , Kenichi Otaki , Naoki Yamamoto , Takahiro Sanada , Kenzaburo Yamaji , Tomoko Honda , Yusuke Matsumoto , Naoko Kitagawa , Koji Ishii , Tomoe Kusayanagi , Yoshiki Yagi , Michinori Kohara , Yasushi Itoh","doi":"10.1016/j.virol.2025.110778","DOIUrl":"10.1016/j.virol.2025.110778","url":null,"abstract":"<div><div>We have developed a recombinant vaccinia vaccine (rDIs-S) against SARS-CoV-2 from an attenuated vaccinia virus (Dairen-I minute-pock variant: DIs strain). The rDIs-S vaccine containing DNA of an S gene of SARS-CoV-2 early-pandemic Pango B strain protected cynomolgus macaques from severe pneumonia caused by a SARS-CoV-2 Pango A strain and induced a broad reactive neutralization antibody against SARS-CoV-2 variants (B.1.1.7, B.1.351, and P.1) in our previous study. In the present study, to further confirm the safety and efficacy of rDIs-S, the vaccine (1 × 10<sup>8</sup> PFU) was inoculated intradermally into rhesus macaques, which showed antibody-dependent enhancement (ADE) after vaccination with another vaccinia virus vector, MVA carrying a SARS-CoV S gene, and challenge infection with SARS-CoV. The rDIs-S vaccination induced neutralization activity against multiple SARS-CoV-2 variants in plasma of rhesus macaques [100 % (6/6 macaques)] and reduced viral replication of the B.1.1.7 and B.1.351 variants and infiltration of T cells and macrophages into the lungs of rhesus macaques [100 % (5/5 macaques)] after challenge infection with 1 × 10<sup>7</sup> TCID<sub>50</sub> of the two SARS-CoV-2 variants. In the presence of plasma of vaccinated macaques, viral RNA levels in human FcγR-positive cells increased slightly by up to 1.8-folds, but they were much lower than those previously reported in sera of mRNA-vaccinated individuals, in which ADE was anticipated (ratios approximately 10–1000). Thus, rDIs-S induced protective immune responses against SARS-CoV-2 B.1.1.7 and B.1.351 and vaccination with rDIs-S did not enhance pneumonia or replication of SARS-CoV-2, indicating the efficacy and safety of rDIs-S in the rhesus macaque model.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"616 ","pages":"Article 110778"},"PeriodicalIF":2.4,"publicationDate":"2025-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145799913","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","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.virol.2025.110776
Saisai Zhao , Dongwei Wang , Yafei Qin , Mingtian Mao , Yiman Li , Yizhang Hao , Yaru Fan , Dalin He , Yi Tang , Youxiang Diao
To combat the emerging challenge of co-circulating hepatotropic pathogens, duck hepatitis A virus type 1 (DHAV-1), duck hepatitis A virus type 3 (DHAV-3), and Duck astrovirus type 1 (DAstV-1), complicating duck viral hepatitis (DVH) control in China, we engineered a novel trivalent egg yolk immunoglobulin (IgY) preparation designed to transcend serotype-specific limitations and enhance passive immunization efficacy. The polyvalent antibody was produced through multistage immunization protocols in high-yielding laying hens, followed by octanoic acid fractionation purification. The preparation exhibited a potent in vitro neutralization index of up to 1:143 against DHAV-1 and 1:256 against DHAV-3 while demonstrating robust immunoreactivity against the DAstV-1 antibody titer of 1:512 via ELISA. Prophylactic administration conferred complete protection (100 %) in challenge models. Therapeutic intervention yielded survival rates of 70 % for DHAV-1, 90 % for DHAV-3, and 80 % for DAstV-1. To our knowledge, this is the first report of a trivalent IgY agent that concurrently neutralizes the three predominant hepatotropic viruses (DHAV-1, DHAV-3, and DAstV-1) complicating DVH, establishing a broad-spectrum immunoprophylactic and therapeutic strategy.
{"title":"Decoding cross-serotype interference in duck hepatitis viruses: Trivalent IgY targeting DHAV-1, DHAV-3 and DAstV-1","authors":"Saisai Zhao , Dongwei Wang , Yafei Qin , Mingtian Mao , Yiman Li , Yizhang Hao , Yaru Fan , Dalin He , Yi Tang , Youxiang Diao","doi":"10.1016/j.virol.2025.110776","DOIUrl":"10.1016/j.virol.2025.110776","url":null,"abstract":"<div><div>To combat the emerging challenge of co-circulating hepatotropic pathogens, duck hepatitis A virus type 1 (DHAV-1), duck hepatitis A virus type 3 (DHAV-3), and Duck astrovirus type 1 (DAstV-1), complicating duck viral hepatitis (DVH) control in China, we engineered a novel trivalent egg yolk immunoglobulin (IgY) preparation designed to transcend serotype-specific limitations and enhance passive immunization efficacy. The polyvalent antibody was produced through multistage immunization protocols in high-yielding laying hens, followed by octanoic acid fractionation purification. The preparation exhibited a potent <em>in vitro</em> neutralization index of up to 1:143 against DHAV-1 and 1:256 against DHAV-3 while demonstrating robust immunoreactivity against the DAstV-1 antibody titer of 1:512 via ELISA. Prophylactic administration conferred complete protection (100 %) in challenge models. Therapeutic intervention yielded survival rates of 70 % for DHAV-1, 90 % for DHAV-3, and 80 % for DAstV-1. To our knowledge, this is the first report of a trivalent IgY agent that concurrently neutralizes the three predominant hepatotropic viruses (DHAV-1, DHAV-3, and DAstV-1) complicating DVH, establishing a broad-spectrum immunoprophylactic and therapeutic strategy.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"615 ","pages":"Article 110776"},"PeriodicalIF":2.4,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145790501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-11DOI: 10.1016/j.virol.2025.110774
Rui Xue , Rang Wang , Haodong Zhang , Weijie Xu , Yiting Liu , Donglingzi Cao , Jiamo Huang , Jinrui Chen , Shixin Liu , Huabo Zhou , Yifeng Qin , Kang Ouyang , Yeshi Yin , Zuzhang Wei , Weijian Huang , Jianming Long , Ying Chen
Feline astrovirus (FAstV), as an intestinal pathogen, has been widely detected worldwide, but little is known about its genetic evolutionary diversity and isolation. This study presents the comprehensive molecular epidemiological investigation of FAstV in Guangxi, China. Screening 775 feline intestinal samples revealed an FAstV prevalence of 4.25 % (33/775), with 39.4 % (13/33) of positive cases co-infected with six other major feline enteric viruses. Notably, FAstV-feline panleukopenia virus (FPV) co-infection emerged as the predominant viral combination (27.27 %, 9/33), suggesting potential synergistic pathogenesis in feline gastroenteritis cases. Phylogenetic analysis of 13 ORF2 genes and 2 complete genomes identified three distinct FAstV genogroups (G1-G3) with significant amino acid divergence (0.445–0.765 p-distance), supporting their classification as separate genotypes. Additionally, we successfully isolated a novel FAstV strain (GXNNF10-2024) in trypsin-supplemented feline kidney-derived FK81 cell cultures, which exhibited characteristic cytopathic effects (CPEs) and was confirmed by RT-PCR and immunofluorescence. These findings significantly enhance our understanding of FAstV genetic diversity, provide essential tools for viral isolation, and establish a foundation for developing diagnostic and control strategies against this emerging feline enteric pathogen.
{"title":"Genetic characterization and isolation of feline astrovirus in domestic cats in Guangxi, China","authors":"Rui Xue , Rang Wang , Haodong Zhang , Weijie Xu , Yiting Liu , Donglingzi Cao , Jiamo Huang , Jinrui Chen , Shixin Liu , Huabo Zhou , Yifeng Qin , Kang Ouyang , Yeshi Yin , Zuzhang Wei , Weijian Huang , Jianming Long , Ying Chen","doi":"10.1016/j.virol.2025.110774","DOIUrl":"10.1016/j.virol.2025.110774","url":null,"abstract":"<div><div>Feline astrovirus (FAstV), as an intestinal pathogen, has been widely detected worldwide, but little is known about its genetic evolutionary diversity and isolation. This study presents the comprehensive molecular epidemiological investigation of FAstV in Guangxi, China. Screening 775 feline intestinal samples revealed an FAstV prevalence of 4.25 % (33/775), with 39.4 % (13/33) of positive cases co-infected with six other major feline enteric viruses. Notably, FAstV-feline panleukopenia virus (FPV) co-infection emerged as the predominant viral combination (27.27 %, 9/33), suggesting potential synergistic pathogenesis in feline gastroenteritis cases. Phylogenetic analysis of 13 ORF2 genes and 2 complete genomes identified three distinct FAstV genogroups (G1-G3) with significant amino acid divergence (0.445–0.765 <em>p</em>-distance), supporting their classification as separate genotypes. Additionally, we successfully isolated a novel FAstV strain (GXNNF10-2024) in trypsin-supplemented feline kidney-derived FK81 cell cultures, which exhibited characteristic cytopathic effects (CPEs) and was confirmed by RT-PCR and immunofluorescence. These findings significantly enhance our understanding of FAstV genetic diversity, provide essential tools for viral isolation, and establish a foundation for developing diagnostic and control strategies against this emerging feline enteric pathogen.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"615 ","pages":"Article 110774"},"PeriodicalIF":2.4,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145737384","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-09DOI: 10.1016/j.virol.2025.110773
Ying Zhai , Hao Peng , Prabu Gnanasekaran , Anirban Roy , Hanu R. Pappu
Croton yellow vein mosaic virus (CYVMV) is a Begomovirus infecting a wide range of vegetables, ornamentals, and weeds. The monopartite CYVMV genome comprises a single-stranded circular DNA (DNA-A). It is also associated with a DNA-betasatellite (DNA-β) called croton yellow vein mosaic betasatellite (CroYVMB). While DNA-A encodes three RNA silencing suppressors viz. V2/C2/C4, DNA-β encodes βC1, which also acts as silencing suppressor. The V2 protein physically interacts with both itself and V1, which is critical for cell-to-cell movement of CYVMV. However, little is known about the molecular mechanism of how V2 suppresses plant RNA silencing machinery and the defense system. Here, we identified twenty-three putative V2-interacting proteins in Nicotiana benthamiana via affinity purification-mass spectrometry (AP-MS) and performed gene ontology (GO) analysis on these V2-interacting candidates. Particularly, two N. benthamiana (Nb) proteins, Ankyrin repeat domain containing protein (NbANK) and Receptor for Activated C Kinase 1 (NbRACK1), were verified to physically interact with V2 in both yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC). Compared to NbRACK1, NbANK exhibited much stronger binding capacity with V2 in Y2H assays, which was consistent with structural predictions of V2-NbANK and V2-NbRACK1 complexes using AlphaFold 3. Virus-induced gene silencing (VIGS) assays using the pTRV vectors demonstrated that the silencing of NbANK significantly increased CYVMV abundance in N. benthamiana, but did not lead to enhanced pathogenicity. In conclusion, our proteomic analysis identifies specific CYVMV V2 interactors in plants, and NbANK is used as an example to show their critical roles in CYVMV propagation.
Croton yellow vein mosaic virus （CYVMV）是一种感染多种蔬菜、观赏植物和杂草的begomavirus。单分子CYVMV基因组由单链环状DNA （DNA- a）组成。它还与一种称为CroYVMB的DNA-β卫星（DNA-β）有关。DNA- a编码3种RNA沉默抑制因子，即V2/C2/C4， DNA-β编码同样具有沉默抑制作用的βC1。V2蛋白与自身和V1相互作用，这对CYVMV的细胞间运动至关重要。然而，关于V2如何抑制植物RNA沉默机制和防御系统的分子机制知之甚少。在这里，我们通过亲和纯化-质谱（AP-MS）鉴定了23个可能与v2相互作用的烟叶（Nicotiana benthamiana）蛋白，并对这些与v2相互作用的候选蛋白进行了基因本体（GO）分析。特别是，两个N. benthamiana （Nb）蛋白，Ankyrin repeat domain containing protein （NbANK）和Receptor for Activated C Kinase 1 (NbRACK1)，被证实在酵母双杂交（Y2H）和双分子荧光互补（BiFC）中与V2有物理相互作用。与NbRACK1相比，在Y2H实验中，NbANK与V2的结合能力更强，这与使用AlphaFold 3对V2-NbANK和V2-NbRACK1复合物的结构预测一致。利用pTRV载体进行的病毒诱导基因沉默（VIGS）实验表明，NbANK的沉默显著增加了benthamiana中CYVMV的丰度，但并未导致致病性增强。总之，我们的蛋白质组学分析确定了植物中特定的CYVMV V2相互作用因子，并以NbANK为例展示了它们在CYVMV繁殖中的关键作用。
{"title":"Proteomic analysis of plant interactors with the RNA silencing suppressor V2 encoded by croton yellow vein mosaic virus","authors":"Ying Zhai , Hao Peng , Prabu Gnanasekaran , Anirban Roy , Hanu R. Pappu","doi":"10.1016/j.virol.2025.110773","DOIUrl":"10.1016/j.virol.2025.110773","url":null,"abstract":"<div><div>Croton yellow vein mosaic virus (CYVMV) is a <em>Begomovirus</em> infecting a wide range of vegetables, ornamentals, and weeds. The monopartite CYVMV genome comprises a single-stranded circular DNA (DNA-A). It is also associated with a DNA-betasatellite (DNA-β) called croton yellow vein mosaic betasatellite (CroYVMB). While DNA-A encodes three RNA silencing suppressors <em>viz.</em> V2/C2/C4, DNA-β encodes βC1, which also acts as silencing suppressor. The V2 protein physically interacts with both itself and V1, which is critical for cell-to-cell movement of CYVMV. However, little is known about the molecular mechanism of how V2 suppresses plant RNA silencing machinery and the defense system. Here, we identified twenty-three putative V2-interacting proteins in <em>Nicotiana benthamiana</em> via affinity purification-mass spectrometry (AP-MS) and performed gene ontology (GO) analysis on these V2-interacting candidates. Particularly, two <em>N. benthamiana</em> (Nb) proteins, Ankyrin repeat domain containing protein (NbANK) and Receptor for Activated C Kinase 1 (NbRACK1), were verified to physically interact with V2 in both yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC). Compared to NbRACK1, NbANK exhibited much stronger binding capacity with V2 in Y2H assays, which was consistent with structural predictions of V2-NbANK and V2-NbRACK1 complexes using AlphaFold 3. Virus-induced gene silencing (VIGS) assays using the pTRV vectors demonstrated that the silencing of <em>NbANK</em> significantly increased CYVMV abundance in <em>N. benthamiana</em>, but did not lead to enhanced pathogenicity. In conclusion, our proteomic analysis identifies specific CYVMV V2 interactors in plants, and NbANK is used as an example to show their critical roles in CYVMV propagation.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"615 ","pages":"Article 110773"},"PeriodicalIF":2.4,"publicationDate":"2025-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145737385","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","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.virol.2025.110768
Matthew D. De Koch , Simona Kraberger , Russell Fielding , Kendal Smith , Kelsie Schiavone , Katharine R. Hall , Vincent S. Reid , Diallo Boyea , Emma L. Smith , Kara Schmidlin , Rafaela S. Fontenele , Darren P. Martin , Mart Krupovic , Arvind Varsani
The family Circoviridae comprises viruses with small single-stranded DNA genomes that are known to infect various animals, resulting in considerable morbidity and mortality in some hosts. Circoviruses have been recently identified through metagenomic sequencing in diverse terrestrial vertebrate species, but their distribution and diversity in marine vertebrates remains underexplored. Here, we use high-throughput sequencing (HTS) to identify circoviruses from archived tissue samples of delphinids (order Artiodactyla, infraorder Cetacea, family Delphinidae). Based on the HTS data, we designed specific abutting primer pairs to recover seven complete circovirus genomes from individual delphinid hosts, namely, the short-finned pilot whale (Globicephala macrorhynchus, n = 5) and the orca (Orcinus orca, n = 2). The circoviruses from the two delphinid species share <65.4 % genome-wide pairwise nucleotide identity with all classified circovirus representative sequences and 66 % amongst themselves. Accordingly, these viruses, which we have named shofin circovirus and orcin circovirus, respectively, represent two novel species. This report also marks the first detection of cetacean circoviruses in the North Atlantic Ocean (near St. Vincent, Caribbean). Notably, analysis of the capsid protein sequences and structures of the delphinid circoviruses revealed notable elaborations within the surface exposed loops that have been previously shown to be a major antigenic epitope in porcine circovirus 2. Collectively, the delphinid circovirus genomes expand the known diversity of circoviruses of marine vertebrates and suggest similar evolutionary pressures exerted by the immune systems of cetacean and suina hosts, both members of the order Artiodactyla.
圆环病毒科包括具有小单链DNA基因组的病毒，已知可感染各种动物，在某些宿主中导致相当高的发病率和死亡率。最近，人们通过宏基因组测序在多种陆生脊椎动物中发现了圆环病毒，但它们在海洋脊椎动物中的分布和多样性仍未得到充分探索。本研究采用高通量测序（HTS）技术，从存档的海豚科（蹄甲目、鲸目、海豚科）组织样本中鉴定环状病毒。基于HTS数据，我们设计了特异性的连接引物对，从短鳍领港鲸（Globicephala macrorhynchus, n = 5）和虎鲸（Orcinus orca, n = 2）个体海豚宿主中恢复7个完整的环状病毒基因组。来自这两种海豚的圆环病毒是相同的
{"title":"Novel circoviruses identified in short-finned pilot whale and orca from the North Atlantic Ocean","authors":"Matthew D. De Koch , Simona Kraberger , Russell Fielding , Kendal Smith , Kelsie Schiavone , Katharine R. Hall , Vincent S. Reid , Diallo Boyea , Emma L. Smith , Kara Schmidlin , Rafaela S. Fontenele , Darren P. Martin , Mart Krupovic , Arvind Varsani","doi":"10.1016/j.virol.2025.110768","DOIUrl":"10.1016/j.virol.2025.110768","url":null,"abstract":"<div><div>The family <em>Circoviridae</em> comprises viruses with small single-stranded DNA genomes that are known to infect various animals, resulting in considerable morbidity and mortality in some hosts. Circoviruses have been recently identified through metagenomic sequencing in diverse terrestrial vertebrate species, but their distribution and diversity in marine vertebrates remains underexplored. Here, we use high-throughput sequencing (HTS) to identify circoviruses from archived tissue samples of delphinids (order Artiodactyla, infraorder Cetacea, family Delphinidae). Based on the HTS data, we designed specific abutting primer pairs to recover seven complete circovirus genomes from individual delphinid hosts, namely, the short-finned pilot whale (<em>Globicephala macrorhynchus</em>, n = 5) and the orca (<em>Orcinus orca</em>, n = 2). The circoviruses from the two delphinid species share <65.4 % genome-wide pairwise nucleotide identity with all classified circovirus representative sequences and 66 % amongst themselves. Accordingly, these viruses, which we have named shofin circovirus and orcin circovirus, respectively, represent two novel species. This report also marks the first detection of cetacean circoviruses in the North Atlantic Ocean (near St. Vincent, Caribbean). Notably, analysis of the capsid protein sequences and structures of the delphinid circoviruses revealed notable elaborations within the surface exposed loops that have been previously shown to be a major antigenic epitope in porcine circovirus 2. Collectively, the delphinid circovirus genomes expand the known diversity of circoviruses of marine vertebrates and suggest similar evolutionary pressures exerted by the immune systems of cetacean and suina hosts, both members of the order Artiodactyla.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"615 ","pages":"Article 110768"},"PeriodicalIF":2.4,"publicationDate":"2025-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145717233","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-04DOI: 10.1016/j.virol.2025.110771
Sanna Tervo , Andrzej Zalewski , Hanna Vauhkonen , Marek Nieoczym , Marta Kołodziej-Sobocińska , Saana Lehikoinen , Kirsi Aaltonen , Teemu Smura , Tarja Sironen , Jenni Pettersson
American mink (Neogale vison) are known carriers of various viruses, including many zoonoses. Their susceptibility to a wide range of pathogens and anatomical similarities to human lungs raise concerns about their potential role in facilitating interspecies transmission. Here we analysed viruses detected in tissue samples of native mustelids, and feral and farmed mink in Poland (2019–2022). The species included American mink, pine and stone marten (Martes martes, Martes foina), weasel (Mustela nivalis), European badger (Meles meles), European polecat (Mustela putorius), and Eurasian otter (Lutra lutra). Blood samples were tested for antibodies against SARS-CoV-2 (n = 270), and lung samples were subjected to genetic studies by PCR (n = 129) or metagenomics (n = 37). No evidence of SARS-CoV-2 was found in feral or wild populations, although two farmed mink tested positive for SARS-CoV-2 by PCR. Aleutian mink disease virus (AMDV) and mink Torque teno neovison virus (mTTV) were identified by metagenomics, and PCR analyses revealed frequent co-infection. AMDV was most prevalent in mink, while other mustelids showed serological evidence but were PCR-negative suggesting viral clearance. AMDV-DNA was found in 26.4 % and mTTV-DNA in 87.8 % of tissue samples. Two animals were co-infected with AMDV, mTTV, and SARS-CoV-2. Phylogenetic analysis revealed both local persistence and inter-farm transmission of AMDV, including transmission between feral and farmed mink. mTTV showed mixing of strains between Poland and China. The findings stress the need for improved surveillance and biosafety to monitor zoonotic threats and cross-species transmission.
{"title":"Virus circulation in native, introduced, and farmed mustelids in Poland","authors":"Sanna Tervo , Andrzej Zalewski , Hanna Vauhkonen , Marek Nieoczym , Marta Kołodziej-Sobocińska , Saana Lehikoinen , Kirsi Aaltonen , Teemu Smura , Tarja Sironen , Jenni Pettersson","doi":"10.1016/j.virol.2025.110771","DOIUrl":"10.1016/j.virol.2025.110771","url":null,"abstract":"<div><div>American mink <em>(Neogale vison</em>) are known carriers of various viruses, including many zoonoses. Their susceptibility to a wide range of pathogens and anatomical similarities to human lungs raise concerns about their potential role in facilitating interspecies transmission. Here we analysed viruses detected in tissue samples of native mustelids, and feral and farmed mink in Poland (2019–2022). The species included American mink, pine and stone marten (<em>Martes martes, Martes foina</em>), weasel (<em>Mustela nivalis</em>), European badger (<em>Meles meles</em>), European polecat (<em>Mustela putorius</em>), and Eurasian otter <em>(Lutra lutra</em>). Blood samples were tested for antibodies against SARS-CoV-2 (n = 270), and lung samples were subjected to genetic studies by PCR (n = 129) or metagenomics (n = 37). No evidence of SARS-CoV-2 was found in feral or wild populations, although two farmed mink tested positive for SARS-CoV-2 by PCR. Aleutian mink disease virus (AMDV) and mink Torque teno neovison virus (mTTV) were identified by metagenomics, and PCR analyses revealed frequent co-infection. AMDV was most prevalent in mink, while other mustelids showed serological evidence but were PCR-negative suggesting viral clearance. AMDV-DNA was found in 26.4 % and mTTV-DNA in 87.8 % of tissue samples. Two animals were co-infected with AMDV, mTTV, and SARS-CoV-2. Phylogenetic analysis revealed both local persistence and inter-farm transmission of AMDV, including transmission between feral and farmed mink. mTTV showed mixing of strains between Poland and China. The findings stress the need for improved surveillance and biosafety to monitor zoonotic threats and cross-species transmission.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"615 ","pages":"Article 110771"},"PeriodicalIF":2.4,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145728050","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study was aimed to have a better understanding of the functional domains of the product of AC2, a protein having a trans-activation function on the late promoters of begomoviruses. The bipartite begomovirus Sri Lankan cassava mosaic virus (SLCMV) was chosen for this study. Infectious SLCMV clones carrying mutations in AC2 were analysed for infectivity, accumulation of viral DNA and trans-activation of the CP promoter in the experimental host Nicotiana benthamiana. The mutations were designed to express truncated AC2 proteins (containing N-terminal 36, N-terminal 101 or C-terminal 22 out of the 135 amino acid residues of the complete protein) or single amino acid replacements targeting the conserved “cysteine-rich” and the “Zn-finger” domains, implicated in trans-activation by AC2 of other begomoviruses. In the inoculated leaves, the DNA of all the above mutants, except the one expressing the C-terminal 22 amino acid residues accumulated at wild type levels. In newly-emerged leaves, only the DNA of the mutant expressing N-terminal 101 residues showed a delayed accumulation compared to the wild type and produced mild symptoms in the inoculated plants. The capability of trans-activation of CP promoter was abolished in all AC2 mutants except in the one expressing the N-terminal 101 residues, lacking the C-terminal trans-activation domain. The results suggest novel functional domains of the AC2 product in SLCMV, not described before.
{"title":"Mutation and trans-activation analyses suggest novel functional motifs for AC2 of the begomovirus Sri Lankan cassava mosaic virus","authors":"Nabanita Gogoi , Kanika Gupta , Rohit Kumar , Fauzia Zarreen, Indranil Dasgupta","doi":"10.1016/j.virol.2025.110772","DOIUrl":"10.1016/j.virol.2025.110772","url":null,"abstract":"<div><div>This study was aimed to have a better understanding of the functional domains of the product of <em>AC2</em>, a protein having a <em>trans</em>-activation function on the late promoters of begomoviruses. The bipartite begomovirus Sri Lankan cassava mosaic virus (SLCMV) was chosen for this study. Infectious SLCMV clones carrying mutations in <em>AC2</em> were analysed for infectivity, accumulation of viral DNA and <em>trans</em>-activation of the CP promoter in the experimental host <em>Nicotiana benthamiana</em>. The mutations were designed to express truncated AC2 proteins (containing N-terminal 36, N-terminal 101 or C-terminal 22 out of the 135 amino acid residues of the complete protein) or single amino acid replacements targeting the conserved “cysteine-rich” and the “Zn-finger” domains, implicated in <em>trans</em>-activation by AC2 of other begomoviruses. In the inoculated leaves, the DNA of all the above mutants, except the one expressing the C-terminal 22 amino acid residues accumulated at wild type levels. In newly-emerged leaves, only the DNA of the mutant expressing N-terminal 101 residues showed a delayed accumulation compared to the wild type and produced mild symptoms in the inoculated plants. The capability of <em>trans</em>-activation of CP promoter was abolished in all <em>AC2</em> mutants except in the one expressing the N-terminal 101 residues, lacking the C-terminal <em>trans</em>-activation domain. The results suggest novel functional domains of the <em>AC2</em> product in SLCMV, not described before.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"615 ","pages":"Article 110772"},"PeriodicalIF":2.4,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145717217","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","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.virol.2025.110770
Zhanzhong Zhao, Gangbin Tang, Xinghui Zhao, Gang Li
The lipid profile of rabies virus infection in hosts remains unknown. Based on the histopathology and neuroinvasiveness, one Flury-LEP of three rabies virus strains was selected and utilized in vitro and in vivo studies. Samples (tissue, plasma and cell pellet) were collected from a mouse model and cell model infected with Flury-LEP strain, and analyzed using a semi-quantitative, untargeted method based on liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS-MS). The resulting lipidomics data were then subjected to statistical analysis. The lipid profile for specific tissues (brain, heart, liver, spleen, lungs, kidneys, intestines, spinal cord, testes and ovaries), plasma (male and female) and cell lines (BHK21 and N2a) were obtained. In mice, the histopathological changes in the tissues, might be associated with changes of the local and circulating lipid profile. Overall, lipid profiling of mice and cells infected with Flury-LEP strain has been achieved, contributing to the understanding of host lipid metabolism and rabies virus-host interactions.
{"title":"Lipid profiling of rabies virus infection in mice and cell lines","authors":"Zhanzhong Zhao, Gangbin Tang, Xinghui Zhao, Gang Li","doi":"10.1016/j.virol.2025.110770","DOIUrl":"10.1016/j.virol.2025.110770","url":null,"abstract":"<div><div>The lipid profile of rabies virus infection in hosts remains unknown. Based on the histopathology and neuroinvasiveness, one Flury-LEP of three rabies virus strains was selected and utilized in vitro and in vivo studies. Samples (tissue, plasma and cell pellet) were collected from a mouse model and cell model infected with Flury-LEP strain, and analyzed using a semi-quantitative, untargeted method based on liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS-MS). The resulting lipidomics data were then subjected to statistical analysis. The lipid profile for specific tissues (brain, heart, liver, spleen, lungs, kidneys, intestines, spinal cord, testes and ovaries), plasma (male and female) and cell lines (BHK21 and N2a) were obtained. In mice, the histopathological changes in the tissues, might be associated with changes of the local and circulating lipid profile. Overall, lipid profiling of mice and cells infected with Flury-LEP strain has been achieved, contributing to the understanding of host lipid metabolism and rabies virus-host interactions.</div></div>","PeriodicalId":23666,"journal":{"name":"Virology","volume":"615 ","pages":"Article 110770"},"PeriodicalIF":2.4,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145682669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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