Pub Date : 2025-01-21DOI: 10.1016/j.vetmic.2025.110398
David Sáez Moreno , Maciej Kuczkowski , Paweł Korzeniowski , Krzysztof Grzymajło , Anna Woźniak-Biel , Paulina Śliwka , Anita Rywińska , Marta Kuźmińska-Bajor
Avian pathogenic Escherichia coli (APEC) is the main causative agent of colibacillosis, causing poultry respiratory infections, mortality and economic loss. APEC poses a serious threat to public health and food safety due to its multi-drug resistance and capacity to form biofilms. Bacteriophages (phages) have emerged as an alternative to antibiotics. To cure APEC-infected chickens, a cocktail consisting of UPWr_E1, UPWr_E2 and UPWr_E4 APEC-specific phages was developed and tested. In this study, we documented the maintenance of their activity in neutralized simulated gastric fluid (SGF) conditions and the efficiency of the UPWr_E124 phage cocktail in inhibiting APEC in biofilm structures on chicken breast meat surfaces. Further, we evaluated the efficacy of the UPWr_E124 phage cocktail against APEC in vivo in murine and chicken infection models. In experimentally infected mice, we evaluated the intraperitoneal and gastric gavage administrations of phages. The study revealed that gastric administration of phages reduced bacterial levels in the respiratory system. Moreover, we tested the UPWr_E124 phage cocktail in a chicken model of infection, where phages effectively reduced the number of APEC in the lungs, bursa of Fabricius and blood. These results suggest that the UPWr_E124 phage cocktail could be a potential treatment for colibacillosis in the poultry industry.
{"title":"Application of UPWr_E124 phage cocktail for effective reduction of avian pathogenic Escherichia coli in mice and broiler chickens","authors":"David Sáez Moreno , Maciej Kuczkowski , Paweł Korzeniowski , Krzysztof Grzymajło , Anna Woźniak-Biel , Paulina Śliwka , Anita Rywińska , Marta Kuźmińska-Bajor","doi":"10.1016/j.vetmic.2025.110398","DOIUrl":"10.1016/j.vetmic.2025.110398","url":null,"abstract":"<div><div>Avian pathogenic <em>Escherichia coli</em> (APEC) is the main causative agent of colibacillosis, causing poultry respiratory infections, mortality and economic loss. APEC poses a serious threat to public health and food safety due to its multi-drug resistance and capacity to form biofilms. Bacteriophages (phages) have emerged as an alternative to antibiotics. To cure APEC-infected chickens, a cocktail consisting of UPWr_E1, UPWr_E2 and UPWr_E4 APEC-specific phages was developed and tested. In this study, we documented the maintenance of their activity in neutralized simulated gastric fluid (SGF) conditions and the efficiency of the UPWr_E124 phage cocktail in inhibiting APEC in biofilm structures on chicken breast meat surfaces. Further, we evaluated the efficacy of the UPWr_E124 phage cocktail against APEC <em>in vivo</em> in murine and chicken infection models. In experimentally infected mice, we evaluated the intraperitoneal and gastric gavage administrations of phages. The study revealed that gastric administration of phages reduced bacterial levels in the respiratory system. Moreover, we tested the UPWr_E124 phage cocktail in a chicken model of infection, where phages effectively reduced the number of APEC in the lungs, bursa of Fabricius and blood. These results suggest that the UPWr_E124 phage cocktail could be a potential treatment for colibacillosis in the poultry industry.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110398"},"PeriodicalIF":2.4,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143068236","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}
Pub Date : 2025-01-21DOI: 10.1016/j.vetmic.2025.110396
Ugo Araújo Souza , Laura Berger , Renata Fagundes-Moreira , Vinícius Baggio-Souza , Adeyldes Reis , Rafaela Mallmann-Bohn , Aline Girotto-Soares , Felipe Bortolotto Peters , Marina Ochoa Favarini , Ana Paula M. Albano , Silvia Resende Terra , Anelise Webster , Bruno Dall’Agnol , Bruno Albuquerque de Almeida , Tatiane Campos Trigo , Stella de Faria Valle , Flávia Pereira Tirelli , José Reck , João Fabio Soares
Hemoplasma infection significantly impacts felines health, yet there is a research gap regarding free-ranging wild small felids. Therefore, this study aimed to investigate the occurrence of hemoplasma in Leopardus geoffroyi and Leopardus wiedii in southern Brazil. For this purpose, we conducted molecular research for Mycoplasma spp. based on the 16S rRNA gene in 79 blood samples from captured wild felids. When positive, the samples were submitted to species-specific reactions for Mycoplasma haemofelis (Mhf), ‘Candidatus Mycoplasma haemominutum’ (CMhm), and ‘Candidatus Mycoplasma turicensis’ (CMt). A total of 39/79 samples were positive for at least one species of hemoplasma. The frequencies found by hemoplasma species were: 39.2 % (31/79) for Mhf, 17.7 % (14/79) for CMhm, and 20.2 % (16/79) for CMt. Factors such as species, sex, age, pelage, capture season, ectoparasite presence, and co-infection with another Mycoplasma were not associated with hemoplasma infection. This study provides the first evidence of circulating hemotropic mycoplasmas in free-living L. geoffroyi and L. wiedii in southern Brazil.
{"title":"Natural infection and diversity of hemotropic mycoplasmas in free-ranging Geoffroy’s cat (Leopardus geoffroyi) and margay cat (Leopardus wiedii) populations in Southern Brazil","authors":"Ugo Araújo Souza , Laura Berger , Renata Fagundes-Moreira , Vinícius Baggio-Souza , Adeyldes Reis , Rafaela Mallmann-Bohn , Aline Girotto-Soares , Felipe Bortolotto Peters , Marina Ochoa Favarini , Ana Paula M. Albano , Silvia Resende Terra , Anelise Webster , Bruno Dall’Agnol , Bruno Albuquerque de Almeida , Tatiane Campos Trigo , Stella de Faria Valle , Flávia Pereira Tirelli , José Reck , João Fabio Soares","doi":"10.1016/j.vetmic.2025.110396","DOIUrl":"10.1016/j.vetmic.2025.110396","url":null,"abstract":"<div><div>Hemoplasma infection significantly impacts felines health, yet there is a research gap regarding free-ranging wild small felids. Therefore, this study aimed to investigate the occurrence of hemoplasma in <em>Leopardus geoffroyi</em> and <em>Leopardus wiedii</em> in southern Brazil. For this purpose, we conducted molecular research for <em>Mycoplasma</em> spp. based on the 16S rRNA gene in 79 blood samples from captured wild felids. When positive, the samples were submitted to species-specific reactions for <em>Mycoplasma haemofelis</em> (<em>Mhf</em>), ‘<em>Candidatus</em> Mycoplasma haemominutum’ (<em>C</em>Mhm), and ‘<em>Candidatus</em> Mycoplasma turicensis’ (<em>C</em>Mt). A total of 39/79 samples were positive for at least one species of hemoplasma. The frequencies found by hemoplasma species were: 39.2 % (31/79) for <em>Mhf</em>, 17.7 % (14/79) for <em>C</em>Mhm, and 20.2 % (16/79) for <em>C</em>Mt. Factors such as species, sex, age, pelage, capture season, ectoparasite presence, and co-infection with another <em>Mycoplasma</em> were not associated with hemoplasma infection. This study provides the first evidence of circulating hemotropic mycoplasmas in free-living <em>L. geoffroyi</em> and <em>L. wiedii</em> in southern Brazil.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110396"},"PeriodicalIF":2.4,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143042062","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 : 2025-01-20DOI: 10.1016/j.vetmic.2025.110403
Catharina E. Exel , Sara M. Tamminga , Sandra Man-Bovenkerk , A. Robin Temming , Astrid Hendriks , Mirlin Spaninks , Nina M. van Sorge , Lindert Benedictus
Staphylococcus aureus (S. aureus) is one of the major causes of bovine mastitis, a disease with detrimental effects on health and wellbeing. Current control measures are costly, laborious and not always effective in eradicating S. aureus. The cell wall-linked polysaccharide wall teichoic acid (WTA) is highly immunogenic in humans and is considered as a prospective vaccine antigen based on promising pre-clinical studies in animals. WTA consist of polymerized ribitol-phosphate backbone that is modified with N-acetylglucosamine (GlcNAc) moieties in different configurations by the glycosyltransferases TarS (β−1,4-GlcNAc), TarM (α−1,4-GlcNAc) and TarP (β−1,3-GlcNAc). This study aimed to characterize the presence and genetic variation in tarS, tarM and tarP in bovine-associated S. aureus strains and how this impacts WTA-glycoprofile. Bioinformatic analyses of a whole genome sequence database consisting of 1047 S. aureus, 10 S. schweitzeri, and 6 S. argenteus strains showed that over 99% of strains contained tarS, 34 % also contained tarM, while 5 % of the strains encoded tarP in addition to tarS. The distribution of WTA-glycosyltransferase genes was similar to what has been reported for human-associated S. aureus strains. Phenotypic analysis of WTA glycosylation by flow cytometry corroborated with tarS/tarM/tarP gene presence. The WTA glycoprofile was variable between bovine-associated strains and the levels and ratios of GlcNAcylation were affected by growth conditions. Interestingly, a divergent tarM allele was present in strains of clonal complexes (CC) 49 and the mastitis-associated CC151, but its function was similar to canonical tarM. In conclusion, we demonstrated that bovine-associated S. aureus strains show similar variation in WTA GlcNAc decoration as human S. aureus strains, despite the presence of a divergent tarM allele.
{"title":"Wall teichoic acid glycosylation of bovine-associated Staphylococcus aureus strains","authors":"Catharina E. Exel , Sara M. Tamminga , Sandra Man-Bovenkerk , A. Robin Temming , Astrid Hendriks , Mirlin Spaninks , Nina M. van Sorge , Lindert Benedictus","doi":"10.1016/j.vetmic.2025.110403","DOIUrl":"10.1016/j.vetmic.2025.110403","url":null,"abstract":"<div><div><em>Staphylococcus aureus</em> (<em>S. aureus</em>) is one of the major causes of bovine mastitis, a disease with detrimental effects on health and wellbeing. Current control measures are costly, laborious and not always effective in eradicating <em>S. aureus</em>. The cell wall-linked polysaccharide wall teichoic acid (WTA) is highly immunogenic in humans and is considered as a prospective vaccine antigen based on promising pre-clinical studies in animals. WTA consist of polymerized ribitol-phosphate backbone that is modified with N-acetylglucosamine (GlcNAc) moieties in different configurations by the glycosyltransferases TarS (β−1,4-GlcNAc), TarM (α−1,4-GlcNAc) and TarP (β−1,3-GlcNAc). This study aimed to characterize the presence and genetic variation in <em>tarS</em>, <em>tarM</em> and <em>tarP</em> in bovine-associated <em>S. aureus</em> strains and how this impacts WTA-glycoprofile. Bioinformatic analyses of a whole genome sequence database consisting of 1047 <em>S. aureus</em>, 10 <em>S. schweitzeri,</em> and 6 <em>S. argenteus</em> strains showed that over 99<em>%</em> of strains contained <em>tarS</em>, 34 % also contained <em>tarM</em>, while 5 % of the strains encoded <em>tarP</em> in addition to <em>tarS.</em> The distribution of WTA-glycosyltransferase genes was similar to what has been reported for human-associated <em>S. aureus</em> strains. Phenotypic analysis of WTA glycosylation by flow cytometry corroborated with <em>tarS/tarM/tarP</em> gene presence. The WTA glycoprofile was variable between bovine-associated strains and the levels and ratios of GlcNAcylation were affected by growth conditions. Interestingly, a divergent <em>tarM</em> allele was present in strains of clonal complexes (CC) 49 and the mastitis-associated CC151, but its function was similar to canonical <em>tarM</em>. In conclusion, we demonstrated that bovine-associated <em>S. aureus</em> strains show similar variation in WTA GlcNAc decoration as human <em>S. aureus</em> strains, despite the presence of a divergent <em>tarM</em> allele.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110403"},"PeriodicalIF":2.4,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143024739","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}
Pub Date : 2025-01-20DOI: 10.1016/j.vetmic.2025.110402
Qingxiao Zhu , Tian Liu , Wenzhen Qin , Xinyu Yang , Wu Tong , Hai Yu , Hao Zheng , Guangzhi Tong , Tongling Shan , Yu Zhang , Xuelan Liu , Ning Kong
BTG3, which belongs to the BTG/Tob gene family, is involved in various physiological processes. Infection with porcine epidemic diarrhea virus (PEDV), an alphacoronavirus, is associated with high mortality rates among piglets, contributing to major economic losses. This study elucidated a novel mechanism through which BTG3 suppresses PEDV replication. Endogenous BTG3 protein expression was upregulated in PEDV-infected host cells. PEDV replication was suppressed upon BTG3 overexpression but enhanced upon BTG3 knockdown. Additionally, BTG3 inhibited viral proliferation by targeting and degrading the S2 subunit of the PEDV spike (S) protein through both autophagy and proteasome pathways. BTG3 interacted and co-localized with the S2 protein, promoting S2 protein degradation through the recruitment of the cargo receptor NDP52 and the E3 ubiquitin ligase MARCHF8. In summary, this study elucidated a novel antiviral mechanism in which the host BTG3 targeted the viral S2 protein to inhibit PEDV proliferation through autophagy and proteasome pathways. These findings indicate that BTG3 is a potential novel target for the prevention and control of PEDV.
{"title":"BTG3 inhibits porcine epidemic diarrhea virus replication by promoting viral S2 protein degradation through the autophagy and proteasome pathways","authors":"Qingxiao Zhu , Tian Liu , Wenzhen Qin , Xinyu Yang , Wu Tong , Hai Yu , Hao Zheng , Guangzhi Tong , Tongling Shan , Yu Zhang , Xuelan Liu , Ning Kong","doi":"10.1016/j.vetmic.2025.110402","DOIUrl":"10.1016/j.vetmic.2025.110402","url":null,"abstract":"<div><div>BTG3, which belongs to the BTG/Tob gene family, is involved in various physiological processes. Infection with porcine epidemic diarrhea virus (PEDV), an alphacoronavirus, is associated with high mortality rates among piglets, contributing to major economic losses. This study elucidated a novel mechanism through which BTG3 suppresses PEDV replication. Endogenous BTG3 protein expression was upregulated in PEDV-infected host cells. PEDV replication was suppressed upon BTG3 overexpression but enhanced upon BTG3 knockdown. Additionally, BTG3 inhibited viral proliferation by targeting and degrading the S2 subunit of the PEDV spike (S) protein through both autophagy and proteasome pathways. BTG3 interacted and co-localized with the S2 protein, promoting S2 protein degradation through the recruitment of the cargo receptor NDP52 and the E3 ubiquitin ligase MARCHF8. In summary, this study elucidated a novel antiviral mechanism in which the host BTG3 targeted the viral S2 protein to inhibit PEDV proliferation through autophagy and proteasome pathways. These findings indicate that BTG3 is a potential novel target for the prevention and control of PEDV.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110402"},"PeriodicalIF":2.4,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143024733","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 : 2025-01-20DOI: 10.1016/j.vetmic.2025.110404
Han Lin , Jingyang Sun , Hua Li , Kesong Chen , Zhendong Qin , Biao Jiang , Wei Li , Qing Wang , Youlu Su , Li Lin , Chun Liu
Acinetobacter johnsonii is a denitrifying bacterium commonly used as an environmental probiotic in wastewater treatment. However, research on its potential pathogenicity to animals is limited. During an epidemiological survey conducted from 2022 to 2024 at bullfrog farms in Guangdong Province, China, multiple strains were isolated from diseased bullfrogs during the low-temperature season. Three representative strains were selected for virulence testing, which showed high virulence to bullfrogs. Molecular identification confirmed these strains as A. johnsonii. One strain was named NW220314S and chosen for further study. Artificial infection through different routes revealed that the mortality rate of bullfrogs infected with this strain was higher at 15°C compared to 28°C. Additionally, the expression of at least 11 virulence genes was significantly higher at 15°C. Pathological examinations of bullfrogs infected with A. johnsonii showed systemic infection with extensive infiltration of inflammatory factors in organs, muscles, and skin. Immune-related gene expression analysis indicated a rapid and intense inflammatory response in bullfrogs post-infection. Our findings uncovered the novel phenomenon of the pathogenicity of A. johnsonii in bullfrogs under low-temperature conditions, warning of the potential threat of A. johnsonii to amphibian populations and the risks associated with its use in various environmental applications.
{"title":"Enhanced virulence of Acinetobacter johnsonii at low temperatures induces acute immune response and systemic infection in American bullfrogs (Aquarana catesbeiana)","authors":"Han Lin , Jingyang Sun , Hua Li , Kesong Chen , Zhendong Qin , Biao Jiang , Wei Li , Qing Wang , Youlu Su , Li Lin , Chun Liu","doi":"10.1016/j.vetmic.2025.110404","DOIUrl":"10.1016/j.vetmic.2025.110404","url":null,"abstract":"<div><div><em>Acinetobacter johnsonii</em> is a denitrifying bacterium commonly used as an environmental probiotic in wastewater treatment. However, research on its potential pathogenicity to animals is limited. During an epidemiological survey conducted from 2022 to 2024 at bullfrog farms in Guangdong Province, China, multiple strains were isolated from diseased bullfrogs during the low-temperature season. Three representative strains were selected for virulence testing, which showed high virulence to bullfrogs. Molecular identification confirmed these strains as <em>A. johnsonii</em>. One strain was named NW220314S and chosen for further study. Artificial infection through different routes revealed that the mortality rate of bullfrogs infected with this strain was higher at 15°C compared to 28°C. Additionally, the expression of at least 11 virulence genes was significantly higher at 15°C. Pathological examinations of bullfrogs infected with <em>A. johnsonii</em> showed systemic infection with extensive infiltration of inflammatory factors in organs, muscles, and skin. Immune-related gene expression analysis indicated a rapid and intense inflammatory response in bullfrogs post-infection. Our findings uncovered the novel phenomenon of the pathogenicity of <em>A. johnsonii</em> in bullfrogs under low-temperature conditions, warning of the potential threat of A. johnsonii to amphibian populations and the risks associated with its use in various environmental applications.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110404"},"PeriodicalIF":2.4,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143060884","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 : 2025-01-20DOI: 10.1016/j.vetmic.2025.110399
Shaohua Sun , Kaili Zhang , Jiajia Zhang , Ping He , Pingping Zhang , Dafu Deng , Chenglin Chi , Sen Jiang , Wanglong Zheng , Nanhua Chen , Jianzhong Zhu
Porcine reproductive and respiratory syndrome virus (PRRSV) causes reproductive failure in sows and respiratory disease in growing pigs, leading to significant economic losses worldwide. Due to the constant mutation and recombination, PRRSV exhibits significant genetic diversity, the general detection of all PRRSV-2 and PRRSV-1 strains is thus needed. In our study, four monoclonal antibodies (mAbs) against PRRSV nucleocapsid (N) protein were generated and the precise and novel B cell epitopes (52KPHF55 and 109HHTVR113) were identified. The epitope 52KPHF55 is highly conserved across all strains of PRRSV-2 lineages and PRRSV-1 subtypes, and the corresponding two mAbs (6D7, 4D12) were selected to develop a sandwich ELISA that was able to detect all tested PRRSV-2 and PRRSV-1 strains. The developed sandwich ELISA demonstrated high specificity, sensitivity and repeatability. In detection of 133 clinical samples, the sandwich ELISA achieved 84.21 % coincidence with the real-time RT-PCR. In conclusion, the mAb based sandwich ELISA can be suitable for detection of potential all PRRSV-2 lineages and PRRSV-1 subtypes, providing a simple, quick and high content method for diagnosis of PRRS.
{"title":"A nucleocapsid monoclonal antibody based sandwich ELISA for the general detection of both PRRSV-2 and PRRSV-1","authors":"Shaohua Sun , Kaili Zhang , Jiajia Zhang , Ping He , Pingping Zhang , Dafu Deng , Chenglin Chi , Sen Jiang , Wanglong Zheng , Nanhua Chen , Jianzhong Zhu","doi":"10.1016/j.vetmic.2025.110399","DOIUrl":"10.1016/j.vetmic.2025.110399","url":null,"abstract":"<div><div>Porcine reproductive and respiratory syndrome virus (PRRSV) causes reproductive failure in sows and respiratory disease in growing pigs, leading to significant economic losses worldwide. Due to the constant mutation and recombination, PRRSV exhibits significant genetic diversity, the general detection of all PRRSV-2 and PRRSV-1 strains is thus needed. In our study, four monoclonal antibodies (mAbs) against PRRSV nucleocapsid (N) protein were generated and the precise and novel B cell epitopes (<sup>52</sup>KPHF<sup>55</sup> and <sup>109</sup>HHTVR<sup>113</sup>) were identified. The epitope <sup>52</sup>KPHF<sup>55</sup> is highly conserved across all strains of PRRSV-2 lineages and PRRSV-1 subtypes, and the corresponding two mAbs (6D7, 4D12) were selected to develop a sandwich ELISA that was able to detect all tested PRRSV-2 and PRRSV-1 strains. The developed sandwich ELISA demonstrated high specificity, sensitivity and repeatability. In detection of 133 clinical samples, the sandwich ELISA achieved 84.21 % coincidence with the real-time RT-PCR. In conclusion, the mAb based sandwich ELISA can be suitable for detection of potential all PRRSV-2 lineages and PRRSV-1 subtypes, providing a simple, quick and high content method for diagnosis of PRRS.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110399"},"PeriodicalIF":2.4,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143029397","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 : 2025-01-20DOI: 10.1016/j.vetmic.2025.110400
Manyu Zhang , Zixiong Zeng , Xia Chen , Guoqing Wang , Xinxin Cai , Zenglei Hu , Min Gu , Shunlin Hu , Xiaowen Liu , Xiaoquan Wang , Daxin Peng , Jiao Hu , Xiufan Liu
Currently, there is increasing spillover of highly pathogenic H5N1 avian influenza virus (AIV) to mammals, raising a concern of pandemic threat about this virus. Although the function of PA protein of the influenza virus is well understood, the understanding of how phosphorylation regulates this protein and influenza viral life cycle is still limited. We previously identified PA S225 as the phosphorylation site in the highly pathogenic H5N1 AIV. In this study, we investigated the role of phosphorylation in regulating PA function and viral fitness through dephosphorylation (PA S225A) or continuous phosphorylation (PA S225E)-mimetic mutation of PA S225. Structure analysis revealed that PA S225A or PA S225E mutation had no obvious effect on the structure of PA protein. Replication assay in vitro showed that PA S225A phosphorylation-ablative mutation significantly inhibited virus replication both in mammalian and avian-derived cells, while PA S225E enhanced viral replication in these cells. Correspondingly, PA S225A dephosphorylation significantly attenuated viral replication and virulence in mice, while PA S225E enhanced these aspects in mice. Mechanistically, PA S225A mutation significantly decreased viral polymerase activity, disabled viral ribonucleoprotein complex (vRNP) assembly and attenuated PA nuclear accumulation. Altogether, our study directly suggested that phosphorylation of PA protein at site S225 enhances viral fitness of the highly pathogenic H5N1 virus in mammals by assuring effective vRNP activity, providing a framework for further study of phosphorylation events in influenza virus life cycle.
{"title":"Phosphorylation of PA at serine 225 enhances viral fitness of the highly pathogenic H5N1 avian influenza virus in mice","authors":"Manyu Zhang , Zixiong Zeng , Xia Chen , Guoqing Wang , Xinxin Cai , Zenglei Hu , Min Gu , Shunlin Hu , Xiaowen Liu , Xiaoquan Wang , Daxin Peng , Jiao Hu , Xiufan Liu","doi":"10.1016/j.vetmic.2025.110400","DOIUrl":"10.1016/j.vetmic.2025.110400","url":null,"abstract":"<div><div>Currently, there is increasing spillover of highly pathogenic H5N1 avian influenza virus (AIV) to mammals, raising a concern of pandemic threat about this virus. Although the function of PA protein of the influenza virus is well understood, the understanding of how phosphorylation regulates this protein and influenza viral life cycle is still limited. We previously identified PA S225 as the phosphorylation site in the highly pathogenic H5N1 AIV. In this study, we investigated the role of phosphorylation in regulating PA function and viral fitness through dephosphorylation (PA S225A) or continuous phosphorylation (PA S225E)-mimetic mutation of PA S225. Structure analysis revealed that PA S225A or PA S225E mutation had no obvious effect on the structure of PA protein. Replication assay <em>in vitro</em> showed that PA S225A phosphorylation-ablative mutation significantly inhibited virus replication both in mammalian and avian-derived cells, while PA S225E enhanced viral replication in these cells. Correspondingly, PA S225A dephosphorylation significantly attenuated viral replication and virulence in mice, while PA S225E enhanced these aspects in mice. Mechanistically, PA S225A mutation significantly decreased viral polymerase activity, disabled viral ribonucleoprotein complex (vRNP) assembly and attenuated PA nuclear accumulation. Altogether, our study directly suggested that phosphorylation of PA protein at site S225 enhances viral fitness of the highly pathogenic H5N1 virus in mammals by assuring effective vRNP activity, providing a framework for further study of phosphorylation events in influenza virus life cycle.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110400"},"PeriodicalIF":2.4,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143029400","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 : 2025-01-17DOI: 10.1016/j.vetmic.2025.110397
Lun Yao , Xiangfei Ren , Ping Zhou , Junjie Yang , Chi Zeng , Zhe Zeng , Yu Shang , Helong Feng , Mengyun Jin , Qianni Xiao , Huabin Shao , Qingping Luo , Sishun Hu , Guoyuan Wen
Cecropin AD (CAD), a hybrid antimicrobial peptide composed of the first 11 residues of cecropin A and last 26 residues of cecropin D, is a promising antibiotic candidate. Therefore, an efficient and convenient method for producing CAD is necessary for commercial applications. The Newcastle disease virus (NDV) has been widely used as a platform for gene delivery and exogenous protein expression. In this study, we constructed a recombinant NDV that expresses CAD. To obtain high expression of the CAD peptide, tandem repeats of the cad gene were inserted into the genomes of the thermostable NDV strain TS09-C using reverse genetic technology. The thermostable recombinant NDV, namely rTS-CAD3, showed thermostability and growth kinetics similar to those of their parental strain. A bacteriostatic test showed that rTS-CAD3 inhibited Staphylococcus aureus (gram-positive bacteria) and Escherichia coli (gram-negative bacteria) in vitro. We further determined the bacteriostatic effects of rTS-CAD3 expressed CAD against S. aureus in skin wound infections. The results showed that rTS-CAD3 subcutaneously injection improved wound healing and reduced S. aureus decolonization. In summary, our results indicate that the rTS-CAD3 expressing CAD peptide is a potent antimicrobial agent against S. aureus and E. coli and may be applied to accelerate wound healing in farm animals.
{"title":"Construction and bacteriostatic effect analyses of a recombinant thermostable Newcastle disease virus expressing cecropin AD","authors":"Lun Yao , Xiangfei Ren , Ping Zhou , Junjie Yang , Chi Zeng , Zhe Zeng , Yu Shang , Helong Feng , Mengyun Jin , Qianni Xiao , Huabin Shao , Qingping Luo , Sishun Hu , Guoyuan Wen","doi":"10.1016/j.vetmic.2025.110397","DOIUrl":"10.1016/j.vetmic.2025.110397","url":null,"abstract":"<div><div>Cecropin AD (CAD), a hybrid antimicrobial peptide composed of the first 11 residues of cecropin A and last 26 residues of cecropin D, is a promising antibiotic candidate. Therefore, an efficient and convenient method for producing CAD is necessary for commercial applications. The Newcastle disease virus (NDV) has been widely used as a platform for gene delivery and exogenous protein expression. In this study, we constructed a recombinant NDV that expresses CAD. To obtain high expression of the CAD peptide, tandem repeats of the <em>cad</em> gene were inserted into the genomes of the thermostable NDV strain TS09-C using reverse genetic technology. The thermostable recombinant NDV, namely rTS-CAD<sub>3</sub>, showed thermostability and growth kinetics similar to those of their parental strain. A bacteriostatic test showed that rTS-CAD<sub>3</sub> inhibited <em>Staphylococcus aureus</em> (gram-positive bacteria) and <em>Escherichia coli</em> (gram-negative bacteria) <em>in vitro</em>. We further determined the bacteriostatic effects of rTS-CAD<sub>3</sub> expressed CAD against <em>S. aureus</em> in skin wound infections. The results showed that rTS-CAD<sub>3</sub> subcutaneously injection improved wound healing and reduced <em>S. aureus</em> decolonization. In summary, our results indicate that the rTS-CAD<sub>3</sub> expressing CAD peptide is a potent antimicrobial agent against <em>S. aureus</em> and <em>E. coli</em> and may be applied to accelerate wound healing in farm animals.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110397"},"PeriodicalIF":2.4,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143042061","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 : 2025-01-15DOI: 10.1016/j.vetmic.2025.110389
Camila N. Foster , Ursula A. Rossi , Carlos A. Rossetti
Enzyme-linked immunosorbent assay (ELISA) is a widely used and effective tool for detection of anti-Brucella antibodies in serum, easy to perform with high sensitivity and specificity. In this study, we validated an in-house indirect ELISA using B. melitensis whole cell lysate as antigen (Bm-WCL iELISA) for the serodiagnosis of caprine brucellosis and evaluated the use of BSL-2 B. neotomae in replacement of BSL-3 Brucella species as an antigen for the detection of Brucella-specific antibodies in ruminant sera. Using 724 serum samples from female crossbred goats classified as brucellosis-positive or -negative by both the buffered plate antigen (BPA) and the complement fixation (CF) tests, the Bm-WCL iELISA was successfully validated with a sensitivity (Se) of 91.83 % (88.51–94.25 %) and a specificity (Sp) of 97.41 % (95.41–98.70 %). In addition, the Bm-WCL iELISA showed a great concordance with a commercial iELISA kit (k = 0.94) in a subset of 217 serum samples. To avoid working with a BSL-3 Brucella for antigen preparation, we replaced it with a less virulent Brucella species such as B. neotomae. A total of 214 goat and 220 cow serum samples were evaluated for the diagnosis of brucellosis using the B. neotomae whole cell homogenate (Bn-WCL) iELISA. The analysis of the ROC curves suggested cut-off values of 63.83 PP for goats and 24.04 PP for cattle, with associated Se and Sp of 98.18 % (93.61–99.68 %) and 90.38 % (83.20–94.69 %) respectively in goat sera, and 95.45 % (89.80–98.04 %) and 96.36 % (91.02–98.58 %) of Se and Sp, respectively in cattle. These results confirm the utility of the in house Bm-WCL iELISA and encourage validation of the Bn-WCL iELISA for the serodiagnosis of ruminant brucellosis in resource-limited areas where the disease is endemic.
{"title":"Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants","authors":"Camila N. Foster , Ursula A. Rossi , Carlos A. Rossetti","doi":"10.1016/j.vetmic.2025.110389","DOIUrl":"10.1016/j.vetmic.2025.110389","url":null,"abstract":"<div><div>Enzyme-linked immunosorbent assay (ELISA) is a widely used and effective tool for detection of anti-<em>Brucella</em> antibodies in serum, easy to perform with high sensitivity and specificity. In this study, we validated an in-house indirect ELISA using <em>B. melitensis</em> whole cell lysate as antigen (Bm-WCL iELISA) for the serodiagnosis of caprine brucellosis and evaluated the use of BSL-2 <em>B. neotomae</em> in replacement of BSL-3 <em>Brucella</em> species as an antigen for the detection of <em>Brucella</em>-specific antibodies in ruminant sera. Using 724 serum samples from female crossbred goats classified as brucellosis-positive or -negative by both the buffered plate antigen (BPA) and the complement fixation (CF) tests, the Bm-WCL iELISA was successfully validated with a sensitivity (Se) of 91.83 % (88.51–94.25 %) and a specificity (Sp) of 97.41 % (95.41–98.70 %). In addition, the Bm-WCL iELISA showed a great concordance with a commercial iELISA kit (<em>k</em> = 0.94) in a subset of 217 serum samples. To avoid working with a BSL-3 <em>Brucella</em> for antigen preparation, we replaced it with a less virulent <em>Brucella</em> species such as <em>B. neotomae</em>. A total of 214 goat and 220 cow serum samples were evaluated for the diagnosis of brucellosis using the <em>B. neotomae</em> whole cell homogenate (Bn-WCL) iELISA. The analysis of the ROC curves suggested cut-off values of 63.83 PP for goats and 24.04 PP for cattle, with associated Se and Sp of 98.18 % (93.61–99.68 %) and 90.38 % (83.20–94.69 %) respectively in goat sera, and 95.45 % (89.80–98.04 %) and 96.36 % (91.02–98.58 %) of Se and Sp, respectively in cattle. These results confirm the utility of the in house Bm-WCL iELISA and encourage validation of the Bn-WCL iELISA for the serodiagnosis of ruminant brucellosis in resource-limited areas where the disease is endemic.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110389"},"PeriodicalIF":2.4,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143012668","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 : 2025-01-14DOI: 10.1016/j.vetmic.2025.110378
Angelica Petersen Dias, Karin Orsel, Corienne Sarah Gammariello, Jeroen De Buck
Digital dermatitis (DD) is a skin infection of cattle’s feet with multiple bacteria suspected to be involved, yet its precise etiopathogenesis remains unclear. This longitudinal study explored the temporal changes of seven DD-associated bacteria in feet developing lesions or remaining healthy, while simultaneously investigating their persistence in potential reservoirs as sources of infection. Weekly swabs were collected from feet skin and saliva of 53 Holstein cows without DD lesions sequentially enrolled at calving in a commercial dairy herd. At the end of the study, samples from all cases and a subset of matched controls were analyzed (1:2 ratio) at five-time points (weeks −3, −2, −1, 0 - when early signs of DD were observed - and +1) and subjected to qPCR targeting Treponema phagedenis, T. medium, T. pedis, Porphyromonas levii, Bacteroides pyogenes, Fusobacterium necrophorum, and F. mortiferum. Linear mixed-effect models assessed the bacterial number changes within cows (cases) and between cows (cases vs controls). Throughout the study, 8 cows developed signs of DD. P. levii, F. necrophorum, and B. pyogenes numbers increased two weeks before the first visible lesion. T. phagedenis and T. pedis numbers increased one week before, suggesting a sequential colonization and potential synergism in triggering DD. Only P. levii and F. necrophorum were persistently present in saliva and skin, while Treponema spp. persisted solely in lesions. Our results inform specific bacterial dynamics associated with DD pathogenesis and might advise future attempts to effectively treat and control DD.
{"title":"Sequential emergence and quantitative dynamics of key bacterial species preceding digital dermatitis lesion onset in dairy cattle","authors":"Angelica Petersen Dias, Karin Orsel, Corienne Sarah Gammariello, Jeroen De Buck","doi":"10.1016/j.vetmic.2025.110378","DOIUrl":"10.1016/j.vetmic.2025.110378","url":null,"abstract":"<div><div>Digital dermatitis (DD) is a skin infection of cattle’s feet with multiple bacteria suspected to be involved, yet its precise etiopathogenesis remains unclear. This longitudinal study explored the temporal changes of seven DD-associated bacteria in feet developing lesions or remaining healthy, while simultaneously investigating their persistence in potential reservoirs as sources of infection. Weekly swabs were collected from feet skin and saliva of 53 Holstein cows without DD lesions sequentially enrolled at calving in a commercial dairy herd. At the end of the study, samples from all cases and a subset of matched controls were analyzed (1:2 ratio) at five-time points (weeks −3, −2, −1, 0 - when early signs of DD were observed - and +1) and subjected to qPCR targeting <em>Treponema phagedenis, T. medium, T. pedis, Porphyromonas levii, Bacteroides pyogenes, Fusobacterium necrophorum</em>, and <em>F. mortiferum</em>. Linear mixed-effect models assessed the bacterial number changes within cows (cases) and between cows (cases vs controls). Throughout the study, 8 cows developed signs of DD. <em>P. levii, F. necrophorum,</em> and <em>B. pyogenes</em> numbers increased two weeks before the first visible lesion. <em>T. phagedenis</em> and <em>T. pedis</em> numbers increased one week before, suggesting a sequential colonization and potential synergism in triggering DD. Only <em>P. levii</em> and <em>F. necrophorum</em> were persistently present in saliva and skin, while <em>Treponema</em> spp. persisted solely in lesions. Our results inform specific bacterial dynamics associated with DD pathogenesis and might advise future attempts to effectively treat and control DD.</div></div>","PeriodicalId":23551,"journal":{"name":"Veterinary microbiology","volume":"302 ","pages":"Article 110378"},"PeriodicalIF":2.4,"publicationDate":"2025-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143024736","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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