Veterinary immunology and immunopathology最新文献

{"title":"In vitro characterization of the E. ruminantium pLAMP multi-epitope DNA poly (lactic-co-glycolic acid) nanoparticle vaccine in sheep peripheral blood mononuclear cells","authors":"M. Nesane ,&nbsp;A. Pretorius ,&nbsp;W. van Wyngaardt ,&nbsp;S.I. Tshilwane ,&nbsp;F.E. Faber ,&nbsp;H. Steyn ,&nbsp;Y. Lemmer ,&nbsp;M. van Kleef ,&nbsp;N. Thema","doi":"10.1016/j.vetimm.2025.110977","DOIUrl":"10.1016/j.vetimm.2025.110977","url":null,"abstract":"<div><div>Heartwater is a tick-borne disease caused by <em>Ehrlichia ruminantium</em>, that has a major economic impact on smallholder farmers. This study investigates the potential of Poly Lactic-co-Glycolic Acid (PLGA) nanoparticles (NPs) as a delivery system for the pLAMP multi-epitope DNA vaccine against heartwater. The vaccine was designed to enhance antigen presentation and activation of specific immune responses, including CD4 + and CD8 + T cell activation. Spherical microspheres with smooth surfaces ranging from 180 nm to 5 µm in diameter were produced, with an adsorption efficiency of 83 %. The <em>in vitro</em> release kinetics demonstrated an initial release of adsorbed pLAMP DNA from PLGA NPs peaking at day 7 and again at day 35. Cellular uptake and gene expression were confirmed using the Vitality hrGFP II plasmid that was adsorbed onto PLGA NPs. High throughput transcriptome sequencing was utilized to determine the immune response activated by the vaccine <em>in vitro</em> in immune sheep peripheral blood mononuclear cells (PBMCs). The pLAMP plasmid transcripts were shown to be present, and key immune pathways, including DNA sensing pathways, MHC presentation and CD4 + T cell and CD8 + T cell pathways were activated that corresponded to those identified and used for the vaccine design previously. This is an indication of the capability of the pLAMP-NP vaccine to induce the desired immune responses, demonstrating potential for <em>in vivo</em> studies.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"286 ","pages":"Article 110977"},"PeriodicalIF":1.4,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144702621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mucoadhesive chitosan-based nano vaccine as promising immersion vaccine against Edwardsiella tarda challenge in Nile tilapia (Oreochromis niloticus)","authors":"Nandhakumar ,&nbsp;Ishwarya Ramachandran ,&nbsp;Preetham Elumalai","doi":"10.1016/j.vetimm.2025.110976","DOIUrl":"10.1016/j.vetimm.2025.110976","url":null,"abstract":"<div><div>Immersion vaccination with a biomimetic mucoadhesive nanovaccine has been shown to induce a strong mucosal immune response against Edwardsiellosis, a serious bacterial disease in Nile tilapia caused by <em>Edwardsiella tarda</em>. This study aims to develop and investigate the efficacy of immersion delivery using a chitosan nano-vaccine (CS) with β-glucan (BG) as an adjuvant to enhance the mucoadhesive properties of the vaccine. The prepared vaccines were nano-sized and spherical as confirmed by scanning electron microscope (SEM), and the images show that nano vaccine greatly increased the binding and penetrating ability into gills when compared with formalin-killed vaccine. Fingerling-sized Nile tilapia (12 ± 2 g) were randomly assigned to four groups: control, Formalin Killed vaccine (Positive control), Chitosan/β-glucan alone (CS/BG), and Chitosan/β-glucan Vaccine (CS/BG V). Fish received immersion baths on days 1 and 21 with a 30-min booster dose. Samples were collected at two time intervals (14, and 28 days post-vaccination (<em>Dpv</em>)) to evaluate innate immune responses through lysozyme, myeloperoxidase, NBT, and superoxide dismutase activity were significantly elevated in vaccinated fish compared to the control group (<em>p </em>&lt; 0.05). IgM antibody titers, measured by ELISA, peaked at 14 and 28<em>Dpv</em> compared to the non-vaccinated group. Furthermore, after vaccination, gene expression analysis using qRT-PCR showed a significant increase <em>in IgM, TNF-α, IL-1β, TCR- β, MHC-I, and IL-8</em> in the spleen of CS/BG V fishes, with similar antibody responses observed. The efficacy of the vaccine was further assessed by challenging the fish with virulent <em>E. tarda</em> after 36 <em>Dpv</em>, and observed 15 days for cumulative mortality. The results demonstrate that the vaccine showed significant protection of 24.44 %, 37.78 %, 48.89 % and 68.89 % respectively, in groups and high relative percentage survival (RPS) in the C/BG V group compared to the control group. Histopathological examinations of head kidney, spleen, and gills were performed for all four groups showed mild infiltrations. This chitosan nano formulation, adjuvanted with a β-glucan immersion vaccine delivery method, will prove effective for Nile tilapia (<em>Oreochromis niloticus</em>), significantly impacting aquaculture and potentially being applicable against other pathogens in global aquaculture systems.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"286 ","pages":"Article 110976"},"PeriodicalIF":1.4,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144614085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Progress and challenges in developing swine immune reagents","authors":"Joan K. Lunney","doi":"10.1016/j.vetimm.2025.110975","DOIUrl":"10.1016/j.vetimm.2025.110975","url":null,"abstract":"<div><div>Swine immune reagents are required for scientists to reveal the mechanisms underlying responses to infectious disease and vaccines. This review highlights availability of antibodies and immune proteins to assess complex cellular and tissue interactions and the issues involved in assuring the best panel of reagents continue to be available for the research community. Continuing issues are discussed for the production, characterization and availability of expressed swine immune proteins and monoclonal antibodies (mAbs) reactive with those proteins and immune cell subset CD (cluster of differentiation) markers. Finally, suggestions are presented for future investments in veterinary immune toolkit efforts.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"286 ","pages":"Article 110975"},"PeriodicalIF":1.4,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144588074","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimisation of bronchoalveolar lavage fluid preparation for mononuclear cell isolation and cytologic evaluation in free-ranging African elephants (Loxodonta africana)","authors":"Stacey C. Engel ,&nbsp;Tanya J. Kerr ,&nbsp;Gian D. van der Spuy ,&nbsp;Tracey Jooste ,&nbsp;Peter E. Buss ,&nbsp;Jennifer L. Johns ,&nbsp;Michele A. Miller ,&nbsp;Léanie Kleynhans","doi":"10.1016/j.vetimm.2025.110974","DOIUrl":"10.1016/j.vetimm.2025.110974","url":null,"abstract":"<div><div>Understanding immune responses to infectious diseases, such as tuberculosis (TB), is essential for developing diagnostic tests and studying disease progression. Although TB affects African savanna elephants (<em>Loxodonta africana)</em>, few studies have investigated immune cells and function in this species, especially in the respiratory tract. Techniques for isolating immune cells from elephant bronchoalveolar lavage (BAL) samples have not been previously reported. Therefore, this study aimed to develop and optimise a protocol to isolate and characterise alveolar cell types in BAL fluid collected from free-ranging, African savanna elephants. The optimised protocol incorporated a mucin digestion step, filtration, Ficoll gradient separation and wash steps to remove contaminants and successfully isolate viable populations of alveolar mononuclear cells. The isolated cells were stained with Rapi-Diff and microscopically examined to differentiate and characterise each cell type present. Cells isolated from healthy African elephant BAL samples, using this method, were predominantly alveolar macrophages (92.5 – 100.0 %) followed by lymphocytes (0.0 – 6.0 %), neutrophils (0.0 – 3.0 %) and eosinophils (0.0 – 1.0 %). This study provides the first optimised protocol for the isolation of alveolar mononuclear cells for future investigations into local immune responses to respiratory diseases such as tuberculosis.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"286 ","pages":"Article 110974"},"PeriodicalIF":1.4,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144588073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"BVDV non-structural protein 4B promoted autophagy by regulating p62/LC3-II molecule","authors":"Beiyan Wang ,&nbsp;Kaiyue Liu ,&nbsp;Shuyu Wei ,&nbsp;Min Wang ,&nbsp;Jiacheng Zhou ,&nbsp;Shuangshuang Wu ,&nbsp;Yazun Dong ,&nbsp;Yixuan Ma ,&nbsp;Hongyan Liu ,&nbsp;Liquan Yu ,&nbsp;Jinzhu Ma ,&nbsp;Baifen Song","doi":"10.1016/j.vetimm.2025.110973","DOIUrl":"10.1016/j.vetimm.2025.110973","url":null,"abstract":"<div><div>Bovine viral diarrhea virus (BVDV) causes bovine viral diarrhea-mucosal disease that is characterized by diarrhea, fever, leukopenia, and propagation disorders in host animals. Autophagy acts an important regulatory role during virial infection. However, the molecular mechanisms that BVDV induces autophagy remain poorly defined. Here, we report that BVDV non-structural protein 4B (NS4B) promotes autophagy in MDBK cells and HEK-293T cells by reducing the level of p62 protein and increasing the production of LC3-II protein, and NS4B proteins were able to improve autophagy in MDBK cells by inhibiting the phosphorylation of mTOR protein. In addition, NS4B (90–260aa) acts a main role in promoting autophagy. The results reveal a new mechanism of BVDV to induce autophagy in host cells and also provide a new theoretical basis for the interaction between BVDV and host cells, and an important target for the prevention and control of BVDV.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"286 ","pages":"Article 110973"},"PeriodicalIF":1.4,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144491030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Monoclonal antibodies targeting the Erns protein of classical swine fever virus: application for virus detection","authors":"Shreya Gopinath ,&nbsp;Madhusudan Hosamani ,&nbsp;Suresh Basagoudanavar ,&nbsp;B.P. Sreenivasa ,&nbsp;Biljo V. Joseph ,&nbsp;Sharanagouda S. Patil","doi":"10.1016/j.vetimm.2025.110965","DOIUrl":"10.1016/j.vetimm.2025.110965","url":null,"abstract":"<div><div>Classical swine fever (CSF), caused by the classical swine fever virus (CSFV; genus <em>Pestivirus</em>), is a highly transmissible disease in pigs that leads to severe economic losses for pig farmers worldwide. Early and accurate diagnosis of CSFV infections is crucial for effective containment of the disease, especially in endemic areas. In this study, the full-length viral protein E^rns was successfully expressed in <em>E. coli</em>. The recombinant protein was purified using Ni-NTA affinity chromatography, and its antigenicity confirmed by western blot and indirect ELISA (iELISA) using CSFV-specific antiserum. The purified E^rns protein was then used as an immunogen to generate murine hybridomas producing monoclonal antibodies (mAbs). A panel of 15 mAbs targeting this protein was characterized using iELISA and western blotting, for their ability to react with native antigens in cultures infected with CSFV. These mAbs have the potential to be used in the development of diagnostic tools, particularly for the detection and titration of virus-infected cultures through endpoint dilution assays.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"286 ","pages":"Article 110965"},"PeriodicalIF":1.4,"publicationDate":"2025-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144322926","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Negative influence of suboptimal quality of drinking water on avian coronavirus pathogenesis and immune response: A Controlled Study","authors":"Muhammad Farooq ,&nbsp;Awais Ghaffar ,&nbsp;Ahmed Ali ,&nbsp;Ryan Rahimi ,&nbsp;Muhammad Azhar ,&nbsp;Ishara M. Isham ,&nbsp;Heshanthi Herath-Mudiyanselage ,&nbsp;Sufna M. Suhail ,&nbsp;Mohamed Faizal Abdul-Careem","doi":"10.1016/j.vetimm.2025.110964","DOIUrl":"10.1016/j.vetimm.2025.110964","url":null,"abstract":"<div><div>This study investigated the impact of poor drinking water quality on infectious bronchitis virus (IBV) pathogenesis. Drinking water samples from Alberta layer farms were assessed based on physical, chemical, and microbiological properties. The highest-scoring field water (FW), which is suboptimal with higher pH, hardness and bicarbonate concentration was selected, transported in clean containers, and used in this control experiment. Forty-eight specific pathogen free White Leghorn chicks were divided into four groups: Tap water non-infected (TW-control), field water non-infected (FW-control), tap water infected (TW-infected), and field water infected (FW-infected). They were maintained on their respective water types for 7 weeks. The IBV genome load was significantly higher in the lungs of the FW-infected when compared to TW-infected group at 4 days post-infection (dpi). The histopathological lesion scores in the trachea and lungs were higher in the FW-infected birds when compared to the uninfected controls at observed time points. However, the histopathological lesion scores in the trachea and lungs of the TW-infected birds were not different when compared to that of FW-infected group. In the lungs, the CD4 + and CD8 + T cell populations were significantly higher in the TW-infected group at observed time points when compared to uninfected controls. However, the CD4 + and CD8 + T cell populations in lungs of the FW-infected birds were not different when compared to that of TW-infected group. In the spleen, CD4 + and CD8 + T cell populations were significantly higher in TW-infected and FW-infected birds when compared to uninfected controls depending on the observed time point and we did not observe differences in CD4 + and CD8 + T cell populations in spleen between TW-infected and FW-infected birds. These findings suggest that sub-optimal drinking water can exacerbate IBV infection by weakening immune responses and increasing disease severity. Further studies are necessary to observe the effect of suboptimal water quality on the development of vaccine-mediated immune response. Understanding these interactions is key for improving water management strategies for maintaining poultry health and productivity.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110964"},"PeriodicalIF":1.4,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144223624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of 1,25-dihydroxy vitamin D3 on inflammation, antimicrobial peptide, and D-dimer levels in Escherichia coli-induced sepsis in neonatal calves","authors":"Emre Eren ,&nbsp;Mustafa Sinan Aktaş","doi":"10.1016/j.vetimm.2025.110963","DOIUrl":"10.1016/j.vetimm.2025.110963","url":null,"abstract":"<div><div>This study evaluated the immunomodulatory, antimicrobial, and anticoagulant effects of 1.25-dihydroxy vitamin D3 in neonatal calves with Escherichia coli-induced sepsis. Thirty neonatal Simmental calves were assigned to three groups: Control (<em>n</em> = 10), Medical Treatment (MT; <em>n</em> = 10), and Medical Treatment plus vitamin D₃ (MT + D₃; <em>n</em> = 10). The MT group received standard sepsis therapy, while the MT + D₃ group was additionally administered 20,000 IU/kg intramuscular vitamin D₃. Blood samples were collected on days 0, 1, 3, and 5 to analyze inflammatory cytokines (NF-κB, TNF-α, IL-1β, IL-10), cathelicidin, D-dimer, iron levels, and hematological parameters. Biochemical indicators including liver, kidney, and heart function, as well as calcium and vitamin D₃ levels, were assessed on days 0 and 5. The MT + D₃ group showed significant clinical and laboratory improvements compared to the MT group. Notably, SpO₂ levels increased, and metabolic acidosis resolved earlier. Hematological findings indicated reduced sepsis-associated anemia, with better preservation of RBC, HGB, and HCT levels. Inflammatory cytokines (NF-κB, TNF-α, IL-1β) significantly decreased, and IL-10 levels were more effectively regulated. Lower D-dimer levels indicated improved coagulation balance. Although cathelicidin levels initially increased, their subsequent decline by day 5 suggested controlled innate immune activation. In conclusion, vitamin D₃ supplementation in combination with standard treatment effectively reduced systemic inflammation, supported innate immunity, and improved coagulation in neonatal calves with sepsis. These results suggest that vitamin D₃ may serve as a beneficial adjunct therapy in bovine neonatal sepsis. Further research is needed to determine optimal dosing strategies and long-term clinical benefits.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110963"},"PeriodicalIF":1.4,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144195112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Serological diagnosis and post-treatment monitoring of Echinococcus granulosus in experimentally infected dogs using crude and recombinant fibronectin antigens","authors":"Figen Celik ,&nbsp;Muhammed Ahmed Selcuk ,&nbsp;Muhammet Uslug ,&nbsp;Sami Simsek","doi":"10.1016/j.vetimm.2025.110962","DOIUrl":"10.1016/j.vetimm.2025.110962","url":null,"abstract":"<div><div><em>Echinococcus granulosus</em> is a zoonotic helminth responsible for cystic echinococcosis, a significant public health concern. The diagnosis of <em>E. granulosus</em> infections in definitive hosts, such as dogs, is challenging due to the absence of clinical signs. This study aimed to evaluate the diagnostic performance of crude (EgSCA) and recombinant (rEgFN162) antigens for the detection of <em>E. granulosus</em> infection in dogs using ELISA and Western blot assays. Additionally, it sought to identify the most suitable antigen and method for population-based screening and post-treatment monitoring. Adult <em>E. granulosus</em> parasites were collected from experimentally infected dogs using arecoline hydrobromide purgation. Soluble crude antigen (EgSCA) was prepared through freeze-thaw cycles, sonication, and filtration, while recombinant fibronectin protein (rEgFN162) was obtained via gene cloning, expression, and purification in <em>E. coli</em>. The antigenic properties of EgSCA and rEgFN162 were analyzed by SDS-PAGE and Western blot. ELISA assays were performed to assess IgG and IgM responses in experimentally infected and treated dogs. Based on IgG ELISA results, EgSCA showed a sensitivity of 96.66 % and specificity of 66.66 %, while rEgFN162 demonstrated a sensitivity of 76.66 % and specificity of 46.66 %. In Western blot analysis, EgSCA achieved a sensitivity of 90 % and specificity of 83.33 %, whereas rEgFN162 showed 66.66 % sensitivity and 73.33 % specificity. The recombinant antigen showed a higher ability to differentiate <em>E. granulosus</em> infections from other helminth infections. The findings suggest that rEgFN162 is a promising candidate for the serodiagnosis of <em>E. granulosus</em> in dogs, with potential applications in epidemiological studies and post-treatment follow-up. Further validation with larger sample sizes is needed to confirm its diagnostic accuracy in natural infections.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110962"},"PeriodicalIF":1.4,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144195113","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Does gradient isolation preserve equine neutrophil viability and oxidative function for up to six hours?","authors":"Laís Sodré Santana Alves ,&nbsp;Amanda Lopes Hasuda ,&nbsp;Lucienne Garcia Pretto Giordano ,&nbsp;Isabela Frederico ,&nbsp;Isabelle Hadid dos Santos ,&nbsp;Priscilla Fajardo Valente Pereira ,&nbsp;Júlio Augusto Naylor Lisbôa ,&nbsp;Karina Keller Marques da Costa Flaiban","doi":"10.1016/j.vetimm.2025.110961","DOIUrl":"10.1016/j.vetimm.2025.110961","url":null,"abstract":"<div><div>This study investigated isolation of equine neutrophils and the assessment of their antioxidant function using the nitroblue tetrazolium (NBT) reduction test, an indicator of reactive oxygen species (ROS) activation. The objective was to test the viability of neutrophils isolated from equine whole blood and their ability to undergo respiratory burst, comparing the results with whole blood. The hypothesis tested was that isolation of equine neutrophils allows for effective functional evaluation, even after isolation process, with no significant differences between whole blood and isolated cells. The study showed that neutrophils maintained their ability to reduce NBT dye over a 6-hour period, with no significant differences between time points analyzed (0 h, 3 h, and 6 h). Isolation was performed using Histopaque®, and total neutrophil concentration and oxidative function were evaluated at three different time points. Cell counts and viability assessments were conducted using both automated and manual techniques, confirming the prolonged viability of neutrophils and the accuracy of antioxidant function testing.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110961"},"PeriodicalIF":1.4,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144189565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}