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A nanobody-based blocking enzyme-linked immunosorbent assay for detecting antibodies against pseudorabies virus glycoprotein E

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Journal of Integrative Agriculture 2024年第4期23卷 1354-1368页

作者： Huanhuan Lü Pinpin Ji Siyu Liu Ziwei Zhang Lei Wang Yani Sun Baoyuan Liu Lizhen Wang Qin Zhao Department of preventive veterinary medicine College of Veterinary MedicineNorthwest A&F University/Shaanxi Scientific Observing and Experimental Station of Veterinary Pharmacology and Diagnostic TechnologyMinistry of Agriculture and Rural AffairsYangling 712100China

Pseudorabies(PR)is an acute infectious disease of pigs caused by the PR virus(PRV)and results in great economic losses to the pig industry *** glycoprotein E(gE)-based enzyme-linked immunosorbent assay(ELISA)has been used to distinguish gE-deleted vaccine-immunized pigs from wild-type virus-infected pigs to eradicate PR in some *** has the advantages of small size and easy genetic engineering and has been a promising diagnostic ***,there were few reports about developing nanobody-based ELISA for detecting anti-PRV-gE *** the present study,the recombinant PRV-gE was expressed with a bacterial system and used to immunize the Bactrian ***,two nanobodies against PRV-gE were screened from the immunized camel by phage display ***,two nanobody-HRP fusion proteins were expressed with HEK293T *** PRV-gE-Nb36-HRP fusion protein was selected as the probe for developing the blocking ELISA(bELISA)to detect anti-PRV-gE *** optimizing the conditions of bELISA,the amount of coated antigen was 200 ng per well,and dilutions of the fusion protein and tested pig sera were separately 1:320 and 1:*** cut-off value of bELISA was 24.20%,and the sensitivity and specificity were 96.43 and 92.63%,*** detecting 233 clinical pig sera with the developed bELISA and a commercial kit,the results showed that the coincidence rate of two assays was 93.99%.Additionallly,epitope mapping showed that PRV-gE-Nb36 recognized a conserved conformational epitope in different reference PRV ***,great stability and low-cost nanobody-based bELISA for detecting anti-PRV-gE antibodies were *** bELISA could be used for monitoring and eradicating PR.

关键词： nanobody nanobody-HRP blocking ELISA PRV-gE antibody

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Nanobodies against African swine fever virus p72 and CD2v proteins as reagents for developing two cELISAs to detect viral antibodies

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Virologica Sinica 2024年第3期39卷 478-489页

作者： Jiahong Zhu Qingyuan Liu Liuya Li Runyu Zhang Yueting Chang Jiakai Zhao Siyu Liu Xinyu Zhao Xu Chen Yani Sun Qin Zhao Department of preventive veterinary medicine College of Veterinary MedicineNorthwest A&F UniversityYangling Observing and Experimental Station of National Data Center of Animal HealthMinistry of AgricultureYangling712100China

African swine fever virus(ASFV)poses a significant threat to the global swine ***,there are no effective vaccines or treatments available to combat ASFV infection in *** primary means of controlling the spread of the disease is through rapid detection and subsequent elimination of infected ***,a lower virulent ASFV isolate with a deleted EP402R gene(CD2v-deleted)has been reported in China,which further complicates the control of ASFV infection in pig ***,an EP402R-deleted ASFV variant has been developed as a potential live attenuated vaccine candidate ***,it is crucial to develop detection methods that can distinguish wild-type and EP402R-deleted ASFV *** this study,two recombinant ASFV-p72 and-CD2v proteins were expressed using a prokaryotic system and used to immunize Bactrian ***,eight nanobodies against ASFV-p72 and ten nanobodies against ASFV-CD2v were *** the production of these nanobodies with horse radish peroxidase(HRP)fusion proteins,the ASFV-p72-Nb2-HRP and ASFV-CD2v-Nb22-HRP fusions were selected for the development of two competitive ELISAs(cELISAs)to detect anti-ASFV *** two cELISAs exhibited high sensitivity,good specificity,repeatability,and *** coincidence rate between the two cELISAs and commercial ELISA kits was 98.6%and 97.6%,***,the two cELISA for detecting antibodies against ASFV demonstrated ease of operation,a low cost,and a simple production *** two cELISAs could determine whether pigs were infected with wild-type or CD2v-deleted ASFV,and could play an important role in monitoring ASFV infections in pig farms.

关键词： African swine fever virus(ASFV) ASFV-p72 ASFV-CD2v Nanobody-HRP Competitive ELISA

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Urbanization does not affect red foxes’interest in anthropogenic food,but increases their initial cautiousness

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Current Zoology 2024年第3期70卷 394-405页

作者： Martina Lazzaroni Rudy Brogi Valentina Napolitano Marco Apollonio Friederike Range Sarah Marshall-Pescini Domestication laboratory Konrad Lorenz Institute of EthologyUniversity of Veterinary MedicineVienna A-1160Austria Department of veterinary medicine University of Sassarivia Vienna 2I-07100 SassariItaly

Human presence and activities have profoundly altered animals’habitats,exposing them to greater risks but also providing new opportunities and *** animals’capacity to effectively navigate and strike a balance between risks and benefts is crucial for their survival in the Anthropocene *** foxes(Vulpes vulpes),adept urban dwellers,exhibit behavioral plasticity in human-altered *** investigated variations in detection frequency on trail cameras and the behavioral responses(explorative,bold,and fearful)of wild red foxes living along an urbanization gradient when exposed to a metal bin initially presented clean and then flled with anthropogenic *** fox populations displayed an increased interest and similar explorative behavioral responses toward the anthropogenic food source,irrespective of the urbanization *** no impact on explorative behaviors,foxes in more urbanized areas initially showed heightened fear toward the empty bin,indicating increased apprehension toward novel ***,this fear diminished over time,and in the presence of food,urban foxes displayed slightly reduced fear compared with their less urban *** results highlight foxes’potential for adaptability to human landscapes,additionally underscoring the nuanced interplay of fear and explorative behavioral response of populations living along the urbanization gradient.

关键词： anthropization bold explorative fearful human footprint Vulpes vulpes

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Emerging tropane alkaloids:Global development and potential health threats

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Food Quality and Safety 2024年第1期8卷 12-26页

作者： Zile Wang Shiqi Chen Pimiao Zheng Zhenhui Ren Huixia Zhang Jing Zhang Bing Shao Congming Wu Haiyang Jiang Department of veterinary Pharmacology and Toxicology Beijing Key Laboratory of Detection Technology for Animal-Derived Food SafetyBeijing Laboratory for Food Quality and SafetyNational Key Laboratory of Veterinary Public Health and SafetyCollege of Veterinary MedicineChina Agricultural UniversityBeijingChina. College of veterinary medicine Shandong Agricultural UniversityTaianChina. Tianjin Key laboratory of Agricultural Animal Breeding and Healthy Husbandry College of Animal Science and Veterinary MedicineTianjin Agricultural UniversityTianjinChina Beijing Key laboratory of Diagnostic and Traceability Technologies for Food Poisoning Beijing Center for Disease Prevention and ControlBeijingChina.

Tropane alkaloids(TAs)are a kind of plant secondary metabolite that mainly originate from *** have potent anticholinergic activity,and are well-known anticholinergic drugs,but have also been reported as plant *** studies have been conducted on TAs,but no scientometric research has been *** study aimed to expound the knowledge network and development of the feld of TAs and predict the emerging development topics based on *** particular,this research combined VOSviewer and CiteSpace for visualization and covered 1,298 related scientifc publications(1953–2022).The research on TAs is rapidly developing,with the participation of 86 countries,1,129 research institutions,and 4,087 ***,the University of Geneva,and *** are the most productive country,institution,and researcher,***,the main research topics on TAs include biosynthesis,in vitro synthesis,chemical synthesis,and determination *** 2018,the monitoring of TAs in food related to human health has shown citation burst characteristics,which is considered an emerging development trend in the *** fndings of this study will enable researchers to quickly tap into the knowledge background/structure of TAs from massive data to provide a reference for further research.

关键词： Tropane alkaloids bibliometrics biosynthesis in vitro synthesis chemical synthesis determination contamination food safety

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New insights into developmental biology of Eimeria tenella revealed by comparative analysis of mRNA N6-methyladenosine modification between unsporulated oocysts and sporulated oocysts

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Journal of Integrative Agriculture 2024年第1期23卷 239-250页

作者： Qing Liu Bingjin Mu Yijing Meng Linmei Yu Zirui Wang Tao Jia Wenbin Zheng Wenwei Gao Shichen Xie Xingquan Zhu laboratory of Parasitic Diseases College of Veterinary MedicineShanxi Agricultural UniversityTaigu 030801China Research Center for Parasites&Vectors College of Veterinary MedicineHunan Agricultural UniversityChangsha 410128China

Evidence showed that N6-methyladenosine(m^(6)A)modification plays a pivotal role in influencing RNA fate and is strongly associated with cell growth and developmental processes in many ***,no information regarding m^(6)A modification in Eimeria tenella is currently *** the present study,we surveyed the transcriptome-wide prevalence of m^(6)A in sporulated oocysts and unsporulated oocysts of *** RNA immunoprecipitation sequencing(MeRIP-seq)analysis showed that m^(6)A modification was most abundant in the coding sequences,followed by stop *** were 3,903 hypermethylated and 3,178 hypomethylated mRNAs in sporulated oocysts compared with unsporulated *** joint analysis suggested that m^(6)A modification of the majority of genes was positively correlated with mRNA *** mRNA relative expression and m^(6)A level of the selected genes were confirmed by quantitative reverse transcription PCR(RT-qPCR)and *** and KEGG analysis indicated that differentially m^(6)A methylated genes(DMMGs)with significant differences in mRNA expression were closely related to processes such as regulation of gene expression,epigenetic,microtubule,autophagy-other and TOR ***,a total of 96 DMMGs without significant differences in mRNA expression showed significant differences at protein *** and pathway enrichment analysis of the 96 genes showed that RNA methylation may be involved in cell biosynthesis and metabolism of *** firstly present a map of RNA m^(6)A modification in ***,which provides significant insights into developmental biology of ***.

关键词： Eimeria tenella m^(6)A RNA methylation MeRIP-seq RNA-seq proteomic analysis

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Enterogenic Stenotrophomonas maltophilia migrates to the mammary gland to induce mastitis by activating the calcium-ROS-AMPK-mTOR-autophagy pathway

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Journal of Animal Science and Biotechnology 2024年第1期15卷 236-252页

作者： Zhaoqi He Caijun Zhao Yuhong He Zhuoyu Liu Guyue Fan Kun Zhu Yiqi Wang Naisheng Zhang Yunhe Fu Xiaoyu Hu Department of Clinical veterinary medicine College of Veterinary MedicineJilin UniversityChangchun 130062Jilin ProvinceChina

Background Mastitis is an inflammatory disease of the mammary gland that has serious economic impacts on the dairy industry and endangers food *** previous study found that the body has a gut/rumen-mammary gland axis and that disturbance of the gut/rumen microbiota could result in‘gastroenterogenic mastitis'.However,the mechanism has not been fully ***,we found that long-term feeding of a high-concentrate diet induced mastitis in dairy cows,and the abundance of Stenotrophomonas maltophilia(***)was significantly increased in both the rumen and milk ***,we hypothesized that‘gastroenterogenic mastitis'can be induced by the migration of endogenous gut bacteria to the mammary ***,this study investigated the mechanism by which enterogenic *** induces *** First,*** was labelled with superfolder GFP and administered to mice via *** results showed that treatment with *** promoted the occurrence of mastitis and increased the permeability of the blood-milk barrier,leading to intestinal inflammation and intestinal ***,tracking of ingested *** revealed that *** could migrate from the gut to the mammary gland and induce ***,mammary gland transcriptome analysis showed that the calcium and AMPK signalling pathways were significantly upregulated in mice treated with ***,using mouse mammary epithelial cells(MMECs),we verified that *** induces mastitis through activation of the calcium-ROS-AMPK-mTOR-autophagy *** In conclusion,the results showed that enterogenic *** could migrate from the gut to the mammary gland via the gut-mammary axis and activate the calcium-ROS-AMPK-mTOR-autophagy pathway to induce *** the gut-mammary gland axis may also be an effective method to treat mastitis.

关键词： Calcium-ROS-AMPK-mTOR-autophagy pathway Gut-mammary axis Mastitis S.maltophilia

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Two nanobody‑based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus

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Animal Diseases 2024年第2期4卷 98-114页

作者： Xu Chen Yueting Chang Lu Zhang Xinyu Zhao Zhihan Li Zhijie Zhang Pinpin Ji Qingyuan Liu Jiakai Zhao Jiahong Zhu Baoyuan Liu Xinjie Wang Yani Sun Qin Zhao Department of preventive veterinary medicine College of Veterinary MedicineNorthwest A&F UniversityYanglingShaanxi 712100China Shenzhen Branch Guangdong Laboratory of Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of Agriculture and Rural AffairsAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518100China

Porcine reproductive and respiratory syndrome virus(PRRSV)infection causes significant economic loss to the global pig *** 1 and 2 PRRSV(PRRSV-1 and-2)infections have been reported in China,Europe and *** accurate prevention,nanobodies were first used as diagnostic reagents for PRRSV *** this study three nanobodies targeting both PRRSV-1 and-2,two targeting PRRSV-1 and three targeting PRRSV-2,were screened and *** develop two competitive ELISAs(cELISAs),the g1-2-PRRSV-Nb3-HRP nanobody was chosen for the g1-2-cELISA,to detect common antibodies against PRRSV-1 and-2,and the g1-PRRSV-Nb136-HRP nanobody was chosen for the g1-cELISA,to detect anti-PRRSV-1 *** two cELISAs were developed using PRRSV-1-N protein as coating antigen,and the amounts for both were 100 ng/*** optimized dilution of testing pig sera was 1:20,the optimized reaction times were 30 min,and the colorimetric reaction times were 15 ***,the cut-off values of the g1-2-cELISA and g1-cELISA were 26.6%and 35.6%,*** of them have high sensitivity,strong specificity,good repeatability,and *** addition,for the 1534 clinical pig sera,an agreement rate of 99.02%(Kappa values=0.97)was determined between the g1-2-cELISA and the commercial IDEXX ELISA *** the g1-cELSIA,it can specifically detect anti-PRRSV-1 antibodies in the clinical pig ***,combining two nanobody-based cELISAs can differentially detect antibodies against PRRSV-1 and-2.

关键词： PRRSV Competitive ELISA Nanobody Antigen epitope

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Toward innovative veterinary nanoparticle vaccines

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Animal Diseases 2024年第2期4卷 78-97页

作者： Meiqi Sun Aldryan Cristianto Pratama He Qiu Zehui Liu Fang He Institute of preventive veterinary medicine College of Animal Sciences of Zhejiang University866 Yuhangtang RoadHangzhou 310058China Zhejiang Provincial Key laboratory of preventive veterinary medicine Hangzhou 310058China ZJU-Xinchang Joint Innovation Centre(TianMu laboratory) Gaochuang Hi‑Tech ParkXinchang 312500ZhejiangChina Department of Cardiology Cardiovascular Key Lab of Zhejiang ProvinceThe Second Affiliated HospitalZhejiang University School of MedicineZhejiang UniversityHangzhouChina

Nanoparticles are significant for veterinary vaccine development because they are safer and more effective than conventional *** promising area of research involves self-assembled protein nanoparticles(SAPNs),which have shown potential for enhancing antigen-presenting cell uptake,B-cell activation,and lymph node *** nanovaccines have been utilized in veterinary medicine,including natural self-assembled protein nanoparticles,rationally designed self-assembled protein nanoparticles,animal virus-derived nanoparticles,bacteriophagederived nanoparticles,and plant-derived nanoparticles,which will be discussed in this *** vaccines can produce robust cellular and humoral immune responses and have been shown to protect against various animal infectious *** article attempts to summarize these diverse nanovaccine types and their recent research progress in the field of veterinary ***,this paper highlights their disadvantages and methods for improving their immunogenicity.

关键词： Nanoparticles veterinary vaccine Self-assembling protein nanoparticles(SAPNs) Virus-like nanoparticles(VLPs) Immune responses Animal infectious diseases Optimization strategies

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Establishment and application of a surrogate model for human Ebola virus disease in BSL-2 laboratory

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Virologica Sinica 2024年第3期39卷 434-446页

作者： Wanying Yang Wujian Li Wujie Zhou Shen Wang Weiqi Wang Zhenshan Wang Na Feng Tiecheng Wang Ying Xie Yongkun Zhao Feihu Yan Xianzhu Xia Hebei Key Lab of laboratory Animal Science Department of Laboratory Animal ScienceHebei Medical UniversityShijiazhuang050017China Key laboratory of Jilin Province for Zoonosis Prevention and Control Changchun Veterinary Research InstituteChinese Academy of Agricultural SciencesChangchun130122China College of veterinary medicine Jilin UniversityChangchun130062China College of veterinary medicine Jilin Agricultural UniversityChangchun130118China

The Ebola virus(EBOV)is a member of the Orthoebolavirus genus,Filoviridae family,which causes severe hemorrhagic diseases in humans and non-human primates(NHPs),with a case fatality rate of up to 90%.The development of countermeasures against EBOV has been hindered by the lack of ideal animal models,as EBOV requires handling in biosafety level(BSL)-4 ***,accessible and convenient animal models are urgently needed to promote prophylactic and therapeutic approaches against *** this study,a recombinant vesicular stomatitis virus expressing Ebola virus glycoprotein(VSV-EBOV/GP)was constructed and applied as a surrogate virus,establishing a lethal infection in *** infection with VSV-EBOV/GP,3-week-old female Syrian hamsters exhibited disease signs such as weight loss,multi-organ failure,severe uveitis,high viral loads,and developed severe systemic diseases similar to those observed in human EBOV *** animals succumbed at 2–3 days post-infection(dpi).Histopathological changes indicated that VSV-EBOV/GP targeted liver cells,suggesting that the tissue tropism of VSV-EBOV/GP was comparable to wild-type EBOV(WT EBOV).Notably,the pathogenicity of the VSV-EBOV/GP was found to be species-specific,age-related,gender-associated,and challenge ***,equine anti-EBOV immunoglobulins and a subunit vaccine were validated using this ***,this surrogate model represents a safe,effective,and economical tool for rapid preclinical evaluation of medical countermeasures against EBOV under BSL-2 conditions,which would accelerate technological advances and breakthroughs in confronting Ebola virus disease.

关键词： Ebola virus(EBOV) Recombinant vesicular stomatitis virus Pathogenicity Syrian hamster Surrogate models Vaccine evaluation and drug screening

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Prevention and treatment of gerbil hepatitis E using the programmable CRISPR-Cas13d system

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Genes & Diseases 2024年第4期11卷 94-97页

作者： Chengcheng Zhao Chong Lia Sa Li Hanyu Wu Pengyao Ren Tianlong Liu Xiaoxiang Hu Ran Zhang State Key laboratory of Animal Biotech Breeding College of Biological SciencesChina Agricultural UniversityBeijing 100193China laboratory of veterinary Pathology and Nanopathology College of Veterinary MedicineChina Agricultural UniversityBeijing 100193China

In recent years,the global incidence of hepatitis E virus(HEV)has been rising,leading to increased morbidity and mortality associated with ***13,a CRISPR effector,shows promise as an antiviral agent against singlestranded RNA ***13d,a type VI-D effector,exhibits higher efficiency in suppressing RNA viruses compared to other type VI ***,its in vivo activity against RNA viruses in mammals remains unknown.

关键词： mortality hepatitis treatment

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