Veterinary microbiology最新文献

{"title":"Cholesterol-dependent Nsp5-endosomes co-trafficking to lysosomes facilitates porcine reproductive and respiratory syndrome virus replication by activating autophagy","authors":"Haoxin Dong ,&nbsp;Qiming Pei ,&nbsp;Jiahui Ren ,&nbsp;Yaci Zhang ,&nbsp;Xuedan Wei ,&nbsp;Aijuan Shen ,&nbsp;Yunshuo Lu ,&nbsp;Ziheng Zhang ,&nbsp;Yongkun Du ,&nbsp;Guoqing Zhuang ,&nbsp;Angke Zhang ,&nbsp;Hong Duan","doi":"10.1016/j.vetmic.2025.110507","DOIUrl":"10.1016/j.vetmic.2025.110507","url":null,"abstract":"<div><div>Our previous studies showed that intracellular endosomal vesicles participated in PRRS virions trafficking in the early stage of viral infection, and cholesterol retention in endosomal vesicles disturbed viral replication via blocking PRRSV-endosomal vesicles membrane fusion. However, whether endosomal vesicles were associated with PRRSV protein(s) trafficking and the role of cholesterol in this process was still unclarity. In this study, we sought to elucidate the mechanism of cholesterol in endosomal vesicles-mediated viral protein transportation. The results showed that endosomal vesicles participated in trafficking of PRRSV Nsp5 protein. After being synthesized in endoplasmic reticulum (ER) and Golgi apparatus, Nsp5 was trafficked to early endosomes (EEs), but not endocytic recycling compartments (ERCs), then to late endosomes (LEs), and eventually reached lysosomal compartments, whereas disruption of cholesterol flux or LEs function led to the inability of Nsp5 arriving at lysosomes, where Nsp5 activated cellular autophagy to promote PRRSV replication. Molecular docking predictions revealed that cholesterol could form two hydrogen bonds with 74 alanine and 78 asparagine of Nsp5. After mutating the aforementioned binding sites, the replication efficiency of PRRSV decreased. Subsequently, the role of cholesterol in PRRSV replication was explored. Blocking of cholesterol flux significantly inhibited PRRSV replication. Single virus infection cycle analysis showed that cholesterol flux disorder did not affect virus adsorption, but could inhibit virus entry into host cells and block EEs-LEs-lysosomes mediated trafficking of virions, leading to virions retention in endosomal compartments. The present studies suggest that cholesterol and endosomal vesicles synergistically participate in Nsp5 trafficking to promote PRRSV replication, which may provide new insights for the development of novel antiviral drugs targeting cholesterol metabolism pathways or the improvement of commercial vaccines.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"305 ","pages":"Article 110507"},"PeriodicalIF":2.4,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143815783","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}

{"title":"Single-B-cell cloning and recombinant antibodies generation to analyze the antigenicity of porcine reproductive and respiratory syndrome virus nonstructural protein 12","authors":"Hongyong Duan ,&nbsp;Chaozhi Tang ,&nbsp;Song Han ,&nbsp;Nan Yang ,&nbsp;Shumao Wang ,&nbsp;Fei Gao ,&nbsp;Yanjun Zhou ,&nbsp;Guangzhi Tong ,&nbsp;Kuan Zhao ,&nbsp;Liwei Li","doi":"10.1016/j.vetmic.2025.110506","DOIUrl":"10.1016/j.vetmic.2025.110506","url":null,"abstract":"<div><div>The prevalence and variation of porcine reproductive and respiratory syndrome virus (PRRSV) in China are increasing. The rapid preparation of essential antibodies will effectively reveal the antigenicity, epitopes, and intracellular distribution of viral proteins. Single-B-cell antibody technology is a novel method for screening diverse functional monoclonal antibodies (mAbs). Herein, we successfully expressed PRRSV nonstructural protein 12 (Nsp12) in suspension-cultured Chinese hamster ovary (CHO) cells. Using single-B-cell antibody technology, we utilized fluorescence-activated cell sorting to collect individual immune B cells and prepared single-cell reverse transcription-polymerase chain reaction to clone the variable region of immunoglobulin heavy chain (<em>IgH</em>) and immunoglobulin light chain (<em>IgK</em>). Two recombinant mAbs were generated via transient transfection of CHO cells with the corresponding expression plasmids of IgH and IgK. A novel linear epitope (¹⁰⁴YEFTGNGEDW¹¹³) of Nsp12 was identified using mAb_1N14. This epitope was conserved in lineages 1, 5, and 8 of PRRSV-2 and was located on the surface of the Nsp12 spatial structure. The amino acid mutation in Nsp12 of lineage 3 PRRSV-2 affected the antigenicity of this linear epitope. A conserved conformational epitope was identified using mAb_2S18, and the spatial structure of Nsp12 showed high similarity between PRRSV-1 and different lineages of PRRSV-2. During PRRSV infection, Nsp12 was distributed in the cytoplasm and accumulated in the nucleus. Overall, antigenicity analysis and novel epitope identification contributed to the in-depth exploration of the biological function of Nsp12 and will facilitate the development of detection assays and antiviral strategies.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110506"},"PeriodicalIF":2.4,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791855","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}

{"title":"Potential of recombinant CAV1-Fc in the treatment of ApxI toxin-induced damage by Actinobacillus pleuropneumoniae","authors":"Yaofang Hu ,&nbsp;Mengdi Zhang ,&nbsp;Gan Yang ,&nbsp;Haoran Guo ,&nbsp;Changsheng Jiang ,&nbsp;Pei Zhou ,&nbsp;Yanhong Chen ,&nbsp;Mengjia Zhang ,&nbsp;Ahmed H. Ghonaim ,&nbsp;Wentao Li ,&nbsp;Qigai He","doi":"10.1016/j.vetmic.2025.110504","DOIUrl":"10.1016/j.vetmic.2025.110504","url":null,"abstract":"<div><div>Currently, porcine contagious pleuropneumonia (PCP) caused by <em>Actinobacillus pleuropneumoniae</em> (APP), poses a significant threat to the pig breeding industry. There is an urgent need for effective therapeutic and prophylactic treatments, especially those that can overcome the limitations associated with vaccines and antibiotics. This includes the development of novel antitoxin agents, immunomodulatory therapies, and alternative strategies like phage therapy and herbal extracts. Our previous study has demonstrated membrane protein caveolin-1 (CAV1) is a key protein that acts as a functional receptor of APP ApxI toxin by binding to its acylated region. Here, we developed recombinant human N-CAV1-Fc fusion protein and C-CAV1-Fc fusion protein. Both fusion proteins could tightly bind to ApxI toxin. N-CAV1-Fc and C-CAV1-Fc fusion proteins efficiently blocked the interaction between ApxI toxin and immortalized porcine alveolar macrophages (iPAMs), thereby inhibiting cell apoptosis caused by APP ApxI toxin. Furthermore, prophylactic and therapeutic CAV1-Fc treatments effectively protected mice from ApxI toxin-induced damage, as determined by reduced weight loss, apoptosis factor transcription, and pathological changes in the lungs. The protective effects of N-CAV1-Fc and C-CAV1-Fc showed clear dose-dependent efficacy <em>in vivo</em>. Protein kinetics data indicated that N-CAV1-Fc has a relatively longer half-life <em>in vivo</em> compared to C-CAV1-Fc, making it an excellent candidate for prevention and treatment of APP infections.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"305 ","pages":"Article 110504"},"PeriodicalIF":2.4,"publicationDate":"2025-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143815784","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}

{"title":"Transcriptome profiling reveals that the host BRD4 protein facilitates African swine fever virus infection and suppresses inflammatory cytokine expression by downregulating transcriptional regulatory signaling pathways","authors":"Mengli Wu ,&nbsp;Jifei Yang ,&nbsp;Zhancheng Tian ,&nbsp;Hualin Sun ,&nbsp;Zhonghui Zhang ,&nbsp;Jianxun Luo ,&nbsp;Guiquan Guan ,&nbsp;Hong Yin ,&nbsp;Qingli Niu ,&nbsp;Rongzeng Hao","doi":"10.1016/j.vetmic.2025.110498","DOIUrl":"10.1016/j.vetmic.2025.110498","url":null,"abstract":"<div><div>The African swine fever virus (ASFV), a complex DNA virus belonging to the <em>Asfarviridae</em> family, is a significant threat to the global swine industry because of its high mortality rates and impact on international trade. The establishment of a stable and efficient cell culture model of ASFV <em>in vitro</em> is helpful for the development of effective vaccines. Several passaged cell lines supporting ASFV replication have been reported to meet the scientific purpose of serial passage of ASFV to a certain extent, but it remains to be determined whether gene expression is lost or whether immunogenicity changes after serial passage of the virus. It is also unclear these edited cell lines how to affect ASFV replication. In our previous study, 3D4/21 cells were transduced with a lentivirus packaging system to express the BD1/2 domain of bromodomain-containing protein 4 (BRD4-BD1/2) and establish a 3D4/21-BD1/2 cell line, which efficiently increased ASFV replication. In this study, the role of bromodomain-containing protein 4 (BRD4), particularly its BD1/2 domains,in enhancing ASFV replication was investigated using an engineered 3D4/21 cell line. Through RNA-Seq transcriptomic analysis, we revealed that the host BRD4 protein facilitates ASFV infection and suppresses key transcription factors (CDK9 and p-CDK9) and inflammatory cytokine expression by downregulating transcriptional regulatory signaling pathways and suppressing innate immune responses. This dual mechanism of BRD4-BD1/2 in promoting ASFV immune evasion and adaptation underscores the virus’s strategic exploitation of host epigenetic factors. These findings provide valuable insights into viral pathogenesis and identify potential therapeutic targets, paving the way for future antiviral strategies.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"305 ","pages":"Article 110498"},"PeriodicalIF":2.4,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143815782","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}

{"title":"Tracking of single virus: Dual fluorescent labeling of pseudorabies virus for observing entry and replication in the N2a cells","authors":"Mingzhi Li ,&nbsp;Li Pan ,&nbsp;Caoyuan Ma ,&nbsp;Hongxia Wu ,&nbsp;Guangtao Xiang ,&nbsp;Lian-Feng Li ,&nbsp;Tao Wang ,&nbsp;Rui Luo ,&nbsp;Yongfeng Li ,&nbsp;Di Liu ,&nbsp;Huanjie Zhai ,&nbsp;Moon Assad ,&nbsp;Xin Song ,&nbsp;Yanjin Wang ,&nbsp;Franck Gallardo ,&nbsp;Hua-Ji Qiu ,&nbsp;Yuan Sun","doi":"10.1016/j.vetmic.2025.110503","DOIUrl":"10.1016/j.vetmic.2025.110503","url":null,"abstract":"<div><div>Pseudorabies virus (PRV) is a neurotropic herpesvirus. It is not easy to be track the whole replication progress of PRV, especially the nascent viral genome in the host cells. In this study, we developed a dual-fluorescence-labeled PRV (rPRV-Anchor3-mCherry) with the viral genome and the envelope protein gM labeled by ANCHOR DNA labeling system and mCherry, respectively. Through single-virus tracking of rPRV-Anchor3-mCherry, we observed that PRV invaded mouse neuroblastoma Neuro-2a cells via both endocytosis and plasma membrane fusion pathway. During the replication stage, parental and progeny viral genome of rPRV-Anchor3-mCherry in the cell nuclei could be visible, and viral nucleocapsid appeared more specifically than traditional capsid protein labeled PRV particles (rPRV-VP26-EGFP). We found that numerous progeny viral particles were produced in the nuclear, causing the nucleus membrane to break using three-dimensional (3D) live-cell imaging and electron microscopy. Moreover, our findings confirmed that simultaneously targeting of the <em>UL9</em> and <em>UL54</em> genes using a CRISPR-Cas9 system led to the complete inhibition PRV replication. rPRV-Anchor3-mCherry can be used to research multiple steps of the viral cycle.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110503"},"PeriodicalIF":2.4,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143785663","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}

{"title":"VP4-Specific IgA level as a correlate of neutralizing antibody and fecal shedding of porcine rotavirus infection","authors":"Sufen Li ,&nbsp;Xianyu Bian ,&nbsp;Jianxin Wang ,&nbsp;Dandan Wang ,&nbsp;Jinzhu Zhou ,&nbsp;Jiapeng Song ,&nbsp;Wei Wang ,&nbsp;Nan Han ,&nbsp;Junming Zhou ,&nbsp;Yunchuan Li ,&nbsp;Ran Tao ,&nbsp;Xuejiao Zhu ,&nbsp;Baochao Fan ,&nbsp;Hailong Dong ,&nbsp;Xuehan Zhang ,&nbsp;Bin Li","doi":"10.1016/j.vetmic.2025.110501","DOIUrl":"10.1016/j.vetmic.2025.110501","url":null,"abstract":"<div><div>Rotavirus (RV) causes diarrhea in children, infants, and young animals globally, with public health implications. Porcine rotavirus (PoRV) leads to economic losses in swine farming. Neutralizing antibodies (NAb) are vital for protecting piglets from intestinal infections. However, which serum and mucosal markers correlate with NAbs against PoRV and relate to post-infection fecal shedding remains unclear, crucial for pathogen-specific detection. We used indirect ELISA to measure IgG/IgA in sera, sIgA in colostrum from recovered pigs, and feces from diarrheal piglets against VP4*, VP7*, VP6, and NSP4*. Analyses showed specific IgA/sIgA levels correlated better with NAb titers than IgG. Among them, VP4*-specific IgA/sIgA had the highest positive correlation with NAb titers in sera (R = 0.848, P &lt; 0.0001) and colostrum (R = 0.865, P &lt; 0.0001). Also, VP4*-specific IgA/sIgA in sera (R= −0.446, P &lt; 0.001) and feces (R= −0.497, P &lt; 0.0001) had the strongest inverse relationship with viral RNA load. Piglet passive protection tests confirmed VP4*-specific IgA's high neutralizing capacity, highly correlated with NAb titers (R = 0.858, P &lt; 0.0001), reducing viral shedding. In conclusion, mucosal IgA/sIgA responses to VP4 are important for PoRV diagnosis assays and vaccine efficacy evaluation.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110501"},"PeriodicalIF":2.4,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143759941","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}

{"title":"What’s up ducks? – Antimicrobial resistance of Escherichia coli isolated from duck farm environment in Poland extended with genomic characteristics of cephalosporin-resistant strains","authors":"Magdalena Skarżyńska ,&nbsp;Magdalena Zając ,&nbsp;Renata Kwit ,&nbsp;Anna Lalak ,&nbsp;Aleksandra Śmiałowska-Węglińska ,&nbsp;Paulina Pasim ,&nbsp;Ewelina Skrzypiec ,&nbsp;Weronika Koza ,&nbsp;Dominika Wojdat ,&nbsp;Emilia Mikos-Wojewoda ,&nbsp;Dominika Pastuszka ,&nbsp;Arkadiusz Bomba ,&nbsp;Dariusz Wasyl","doi":"10.1016/j.vetmic.2025.110492","DOIUrl":"10.1016/j.vetmic.2025.110492","url":null,"abstract":"<div><div>Antimicrobial resistance (AMR) in food-producing animals is a source of concern as it may pose a risk to public health. Studies of ducks in this area seem to be scarce. Thus, we aimed to evaluate the AMR occurrence in <em>Escherichia coli</em> from a duck farm environment in Poland. We applied official AMR monitoring methods to investigate AMR in <em>E</em>. <em>coli</em> isolated from boot swabs collected at 306 duck farms in Poland. The samples were screened for indicator, cephalosporin-, carbapenem- and colistin- resistant <em>E</em>. <em>coli</em>. Isolates were tested for antimicrobial susceptibility using the microbroth dilution method and interpreted with epidemiological cut-off values. Whole Genome Sequencing (WGS) of cephalosporin-resistant strains enabled an in-depth insight into specific resistance mechanisms, mobile genetic elements and phylogeny of strains. A total of 340 strains were isolated. The percentage of indicator <em>E</em>. <em>coli</em> equaled 89.9 %, while 19.3 % were obtained through selective screening for <em>E</em>. <em>coli</em> resistant to cephalosporins. Six were recovered on colistin-supplemented MacConkey agar. Among indicator <em>E</em>. <em>coli</em> 81.1 % were resistant and ciprofloxacin, ampicillin, tetracycline and nalidixic acid resistance were the most frequent, followed by folate-path inhibitors. Within the group of strains from cephalosporin resistance screening: 76.3 % exhibited ESBL-, 20.3 % AmpC-, and 3.4 % showed both ESBL- and AmpC- phenotypes. WGS of those strains revealed numerous AMR determinants, not only genes corresponding to mentioned phenotypes but also determinants encoding resistance to other medically important antimicrobials. Our study reveals that <em>E</em>. <em>coli</em> from duck farm environment constitute a reservoir of AMR determinants including those of public health concern.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110492"},"PeriodicalIF":2.4,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143783650","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}

{"title":"The antioxidant protein PntA coordinates with OmpW to resist oxidant stress in Actinobacillus pleuropneumoniae","authors":"Weiyao Han ,&nbsp;Qiuhong Zhang ,&nbsp;Zhen Luo ,&nbsp;Hao Tang ,&nbsp;Xiabing Chen ,&nbsp;Qi Huang ,&nbsp;Rui Zhou ,&nbsp;Lu Li","doi":"10.1016/j.vetmic.2025.110500","DOIUrl":"10.1016/j.vetmic.2025.110500","url":null,"abstract":"<div><div>Bacteria have evolved various strategies to combat oxidative stress caused by reactive oxygen species (ROS). Outer membrane proteins including OmpW play multiple roles in bacterial physiology, stress responses and virulence. In this study, the OmpW protein of <em>Actinobacillus pleuropneumoniae</em>, an important porcine respiratory tract pathogen, was found to contribute to virulence but concurrently to reduce resistance to oxidative stress. An <em>ompW</em> deletion (Δ<em>ompW</em>) showed attenuation in mice, and decreased adherence to pig tracheal epithelial cells and resistance to hyperosmotic stress, compared to the wild-type (WT) strain. However, the Δ<em>ompW</em> strain exhibited increased resistance to H₂O₂, enhanced survival ability within macrophages, and lower intracellular ROS level. OmpW may serve as a H₂O₂ channel. Further study showed that exposure to H₂O₂ significantly suppressed <em>ompW</em> transcription in the WT strain. Overexpression of these two proteins in WT and Δ<em>ompW</em> increased the antioxidative properties of the bacteria. Furthermore, by construction of the double gene mutant Δ<em>ompW</em>Δ<em>pntA</em>, it was found that PntA could reverse the effects of OmpW on the bacterial survivability and intracellular ROS level after H₂O₂ treatment. Therefore, by interacting with OmpW, PntA alleviated the increased oxidative stress sensitivity caused by OmpW. These results suggest a mechanism whereby antioxidant proteins collaborate with OMPs to protect bacteria from oxidative stress.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110500"},"PeriodicalIF":2.4,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143747794","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}

{"title":"Proteome profiling of the lung tissues of the mice co-infected with H9N2 influenza A virus and Pseudomonas aeruginosa","authors":"Sun Ying-Li ,&nbsp;Fan Hua ,&nbsp;Li Wen-qian ,&nbsp;Li Dan ,&nbsp;Bai Ke-fei ,&nbsp;Dong Lu-jiao ,&nbsp;Zhang Hui-ning ,&nbsp;Li Jian-liang ,&nbsp;Xie Zhi-jing","doi":"10.1016/j.vetmic.2025.110499","DOIUrl":"10.1016/j.vetmic.2025.110499","url":null,"abstract":"<div><div>H9N2 Influenza A virus (IAV) can contribute to <em>Pseudomonas aeruginosa</em> (<em>P. aeruginosa</em>) superinfection, causing severe pneumonia. But the underlying mechanisms remain unclear. In this study, to investigate molecular ecology of the lung tissues co-infected with H9N2 IAV and <em>P. aeruginosa</em>, the mouse models were developed to analyze proteome of the lung tissues. As a result, the differential proteins of the lungs sampled from the mice with single H9N2 IAV infection, single <em>P. aeruginosa</em> infection and co-infection with H9N2 IAV and <em>P. aeruginosa</em> were related to immune responses, cell proliferation and apoptosis, which were involved in NF-κB pathway, Toll-like signaling pathway, RIG-I signaling pathway and JAK-STAT signaling pathway. It implied that the different infection combinations of H9N2 IAV and <em>P. aeruginosa</em> influenced the protein expression levels. Based on the proteomics assays, the three proteins, NLRX1, ISG15 and IRF9, were screened to be further characterized using the in vitro MH-S cell models with H9N2 IAV and <em>P. aeruginosa</em> co-infection. The findings demonstrated that NLRX1, IRF9 and ISG15 might play the important roles in immune response against H9N2 IAV and <em>P. aeruginosa</em> co-infection and are potential targets for the development of drugs to prevent and treat the diseases.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110499"},"PeriodicalIF":2.4,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143776834","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}

{"title":"Misidentification of Raoultella spp. (R. terrigena, R. planticola) and Klebsiella spp. (K. variicola, K. grimontii) as Klebsiella pneumoniae: Retrospective study of a necropsy-associated bacterial collection from horses","authors":"Francois Gravey ,&nbsp;Corinne Sévin ,&nbsp;Bénédicte Langlois ,&nbsp;Karine Maillard ,&nbsp;Nathalie Foucher ,&nbsp;Fabien Duquesne ,&nbsp;Albertine Léon ,&nbsp;Simon Le Hello ,&nbsp;Sandrine Petry","doi":"10.1016/j.vetmic.2025.110497","DOIUrl":"10.1016/j.vetmic.2025.110497","url":null,"abstract":"<div><div>Misidentifications as <em>Klebsiella pneumoniae</em> were observed during a French retrospective study of a necropsy-associated <em>K. pneumoniae</em> bacterial collection from horses. Accordingly, the present study aimed to further characterise the 12 <em>Raoultella</em> spp. and <em>Klebsiella</em> spp. strains involved in these misidentifications. The strains were identified and characterised using the Api 20E system, <em>K. pneumoniae</em> PCR detection, matrix-assisted laser desorption/ionisation time-of-flight mass spectroscopy and whole-genome sequencing. Antimicrobial susceptibilities were tested by the disc diffusion method with a panel of 40 antibiotics. Thus, misidentifications as <em>K. pneumoniae</em> mainly concerned <em>Raoultella</em> spp. (<em>R. terrigena</em>, <em>R. planticola</em>) rather than <em>Klebsiella</em> spp. (<em>K. variicola</em>, <em>K. grimontii</em>), with a dominance of <em>R. terrigena</em>. Among the 12 strains, only <em>K. grimontii</em> was multi-drug resistant and none were considered hypervirulent. MALDI-TOF was sufficient to avoid misidentification but commercial spectra databases should be expanded with <em>K. grimontii</em> and <em>R. terrigena</em> reference spectra to improve identification accuracy. This is probably (i) the first report of <em>K. grimontii</em> and <em>R. planticola</em> isolations from horses and (ii) the second report of <em>R. terrigena</em> and <em>K. variicola</em> isolations from horses.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"304 ","pages":"Article 110497"},"PeriodicalIF":2.4,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143725653","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}