Journal of Veterinary Diagnostic Investigation最新文献

{"title":"In silico-derived <i>Actinobacillus equuli</i>-specific DNA markers and development of associated PCR assays.","authors":"Warangkhana Songsungthong, Wichai Pornthanakasem, Ubolsree Leartsakulpanich, Gun Srijuntongsiri","doi":"10.1177/10406387251382186","DOIUrl":"10.1177/10406387251382186","url":null,"abstract":"<p><p>The ability to accurately and rapidly identify causative agents of infectious diseases facilitates precise treatment, improves clinical outcomes, and augments epidemiology studies. For many veterinary and zoonotic pathogens, however, simple molecular tests for species identification are not available. <i>Actinobacillus equuli</i> causes severe diseases, such as sleepy foal disease, septicemia, and meningitis in horses and pigs. <i>A. equuli</i> can also cause severe diseases in humans bitten by infected animals. Existing <i>A. equuli</i> identification methods are biochemical tests, 16S rRNA gene amplification followed by DNA sequencing, and MALDI-TOF MS. Nonetheless, differentiating among <i>Actinobacillus</i> spp. by these methods is still challenging. We identified novel DNA markers specific to <i>A. equuli</i> by computational genome analysis. We then designed PCR primers specific to <i>A. equuli</i> based on <i>A. equuli</i> marker sequences. We validated 2 <i>A. equuli</i>-specific PCR assays using genomic DNA from 10 strains of <i>A. equuli</i>, 15 strains of other <i>Actinobacillus</i> species, and 5 other bacterial species. Both assays gave the PCR products of expected sizes for genomic DNA of all 10 strains of <i>A. equuli</i> but not for those of other <i>Actinobacillus</i> and other bacterial species. Our novel PCR assays can accelerate <i>A. equuli</i> identification and disease diagnosis, leading to timely and appropriate antimicrobial treatment, and enable high-resolution epidemiologic studies.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251382186"},"PeriodicalIF":1.1,"publicationDate":"2025-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12511006/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251597","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simultaneous detection of canine astrovirus and canine kobuvirus by duplex qPCR: validation and evidence of extraintestinal viral RNA in naturally infected dogs.","authors":"Tin Van Nguyen, Tanit Kasantikul, Chutchai Piewbang, Somporn Techangamsuwan","doi":"10.1177/10406387251382915","DOIUrl":"10.1177/10406387251382915","url":null,"abstract":"<p><p>Canine astrovirus (mamastrovirus 5 [MAstV5], formerly CaAstV; family <i>Astroviridae</i>, taxon species <i>Mamastrovirus canis</i>) and canine kobuvirus (aichivirus A2 [AiV-A2], formerly CaKoV; <i>Picornaviridae</i>, <i>Kobuvirus aichi</i>) are emerging enteric viruses increasingly detected in both diarrheic and subclinical dogs. Although primarily associated with gastrointestinal illness, recent evidence suggests a potential for systemic dissemination, which remains insufficiently explored. To improve detection, we developed and validated a duplex quantitative real-time PCR (dqPCR) assay for simultaneous identification of MAstV5 and AiV-A2. Primers and TaqMan probes were designed based on conserved regions of the MAstV5 <i>ORF1b</i> and AiV-A2 <i>3D</i> polymerase genes. Assay optimization included primer-probe concentration titration and thermal gradient analysis. Analytical performance was assessed using synthetic plasmid standards and RNA from non-target viruses, including canine parvovirus (CPV), canine morbillivirus (CDV), and canine enteric coronavirus (CCoV). Our dqPCR assay had high linearity (<i>R</i>² > 0.99) and sensitivity, with limits of detection of 10 copies/μL for MAstV5 and 100 copies/μL for AiV-A2. No cross-reactivity or interference was observed in coinfection simulations across various target ratios. Intra- and inter-assay CV values were <2.2%, indicating excellent reproducibility. Validation using 50 clinical rectal swabs and tissue samples from 25 autopsied dogs revealed 1 additional AiV-A2-positive case undetected by RT-PCR but confirmed by sequencing. Importantly, viral RNA was also found in extraintestinal tissues-spleen, liver, trachea, and mesenteric lymph nodes-suggesting systemic distribution in naturally infected dogs. Our dqPCR assay provides a sensitive, specific, and efficient tool for the detection of MAstV5 and AiV-A2, supporting both clinical testing and epidemiologic studies.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251382915"},"PeriodicalIF":1.1,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12507789/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145244638","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Coat color and other factors influencing hair cortisol concentration in domestic cats.","authors":"Kirsten L Nutter, Andrew S Cooke","doi":"10.1177/10406387251384320","DOIUrl":"10.1177/10406387251384320","url":null,"abstract":"<p><p>Hair cortisol quantification can be used to understand long-term stress in cats and other animals. The technique is becoming increasingly common; however, there is uncertainty as to the factors that may affect or confound hair cortisol quantification, in particular, hair color. Although some studies show that hair of different colors has different abilities to store cortisol, others do not. We collected hair samples from 27 domestic cats with either black-and-white or ginger-and-white haircoat coloring. From each cat, 2 samples were taken, 1 of white hair and 1 of the other color (black or ginger). Samples underwent cortisol quantification by ELISA, and pairwise analysis was conducted. Hair cortisol was also compared against information provided by the cat owners regarding their cat (e.g., sex, age) and behavioral issues. Black hair contained significantly greater concentrations of cortisol than white hair (<i>p</i> = 0.016). Although ginger hair tended to have higher mean cortisol concentrations than white hair, the difference was not statistically significant (<i>p</i> = 0.613). A significant positive correlation was also found between hair cortisol and behavioral issues reported by owners (<i>p</i> = 0.010). To our knowledge, the impact of the color of the hair on feline hair cortisol concentrations has not been reported previously.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251384320"},"PeriodicalIF":1.1,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12507796/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145244419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Letter to the editor: Celomitis revisited.","authors":"Wes Baumgartner, Brian Speer","doi":"10.1177/10406387251382891","DOIUrl":"10.1177/10406387251382891","url":null,"abstract":"","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251382891"},"PeriodicalIF":1.1,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12507780/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145244460","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of 3 qPCR primer-probe sets for the detection of equid alphaherpesvirus 1.","authors":"Yoshinori Kambayashi, Hiroshi Bannai, Manabu Nemoto, Nanako Kawanishi, Hidekazu Niwa, Koji Tsujimura","doi":"10.1177/10406387251379857","DOIUrl":"10.1177/10406387251379857","url":null,"abstract":"<p><p>With the revision of the World Organisation for Animal Health (WOAH) Terrestrial Manual on equine rhinopneumonitis in 2024, 3 recommended qPCR primer-probe sets were added for the detection of equid alphaherpesvirus 1 (EqAHV1; formerly equine herpesvirus 1 [EHV1]; family <i>Orthoherpesviridae</i>, taxon species <i>Varicellovirus equidalpha1</i>), also known as equine abortion virus. We compared the sensitivity and specificity of the 3 qPCR primer-probe sets to determine the most reliable set. Sets gB1H and gB1P, which target the glycoprotein B (<i>gB</i>) gene of EqAHV1, detected all 10 copies and even lower copy numbers. In contrast, set gC1 (ISO 17025-accredited method used at the WOAH reference laboratory), which targets the glycoprotein C (<i>gC</i>) gene, failed to detect ≤10 copies of EqAHV1. Our results showed the lower sensitivity of gC1, which was not improved by modification of primer and probe concentrations. gB1P detected not only EqAHV1 but also equid alphaherpesvirus 4 (EqAHV4; <i>Orthoherpesviridae</i>, <i>Varicellovirus equidalpha4</i>), likely owing to an erroneous amplification of the homologous EqAHV4 <i>gB</i> gene, indicating that gB1P is not suitable for the detection of EqAHV1 with high specificity. We then compared gB1H with gB1D, a set recommended in the previous version of the Manual, using 120 nasal swabs collected from febrile horses. gB1H had slightly higher sensitivity than gB1D. gB1H proved to be the most reliable primer-probe set for detecting EqAHV1, with high sensitivity and specificity. Nevertheless, individual laboratories are encouraged to validate these methods under their own conditions before implementation.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251379857"},"PeriodicalIF":1.1,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12504209/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145238875","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selected microRNAs as biomarkers in sarcoid-affected horses under immunotherapy with a mistletoe extract.","authors":"Anke Beermann, Eman Hamza, Sonja Reinhard, Christoph Koch, Thomas Oberhänsli, Lucia Unger","doi":"10.1177/10406387251362820","DOIUrl":"10.1177/10406387251362820","url":null,"abstract":"<p><p>We investigated microRNAs (miRNAs) as potential prognostic biomarkers for equine sarcoid (ES) disease. In a breed-, age-, and sex-matched case-controlled study involving 45 ES-affected and 15 control horses, we assessed the diagnostic, prognostic, and theragnostic value of 3 miRNAs (eca-miR-127, eca-miR-379, eca-miR-432) in horses treated with European mistletoe (<i>Viscum album</i>) extract versus placebo. Whole-blood miRNA concentrations were measured using reverse-transcription quantitative real-time PCR (RT-qPCR) at 3 different times. We found that eca-miR-432 expression was lower in ES-affected (median = -1.93; 95% CI: -2.03 to -.86) compared to control (median = -1.71; 95% CI: -1.92 to -1.6) horses (<i>p</i> = 0.03, <i>r</i> = 0.3; 95% CI: 0.024-0.57) with a median difference of -1.93 versus -1.71, respectively. The ROC curve analysis indicated an area under the curve of 0.71 (95% CI: 0.51-0.84; <i>p</i> = 0.005) with a sensitivity of 74% (95% CI: 61-88%) and a specificity of 73% (95% CI: 39-94%) to diagnose ES. However, none of the miRNAs evaluated had prognostic potential or significant changes in expression following treatment. Additionally, miRNA expression was not influenced by breed, sex, or season. Although whole-blood eca-miR-432 had moderate diagnostic potential for ES, identifying prognostic miRNA biomarkers for ES remains a challenge.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251362820"},"PeriodicalIF":1.1,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12494582/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145213145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ectopic splenic tissue in 46 dogs, 2000-2024.","authors":"C Robert Stilz, Ricardo E Mendes, Claudio S L Barros, Daniel R Rissi","doi":"10.1177/10406387251379922","DOIUrl":"10.1177/10406387251379922","url":null,"abstract":"<p><p>Ectopic splenic tissue (accessory spleen or splenosis) occurs as dark-red-to-brown or purple nodules outside the spleen. Accessory spleens are congenital lesions histologically identical to a normal spleen. Splenosis results from implantation of splenic tissue following splenic rupture and lacks features of normal spleen. However, these distinctions have been largely applied to human cases, and the terms are often used interchangeably in domestic animals. Here we describe ectopic splenic tissue in 46 canine surgical biopsy specimens examined at the Athens Veterinary Diagnostic Laboratory, 2000-2024. The omentum (39 cases) and mesentery (5) were the most commonly affected sites. Original diagnoses were accessory spleen (28 cases), splenosis (14), accessory spleen or splenosis (2), and ectopic splenic tissue and normal splenic tissue (1 each). Updated diagnoses, modified after histologic assessment for a fibrous capsule, smooth muscle trabeculae, and white and red pulp, were accessory spleen (37 cases) and splenosis (9). Concurrent splenic lesions were reported in 12 cases in which accessory spleens were diagnosed and only 2 splenosis cases, confirming that the histologic diagnosis of accessory spleen and splenosis is not always correlated with the clinical history and gross findings (no splenic lesions vs. splenic lesions with rupture). For that reason, <i>ectopic splenic tissue</i> may be a more inclusive and better term for these lesions. Hemangiosarcoma was diagnosed in the spleen in 4 of the 12 cases with splenic masses, which underscores the importance of the differentiation between ectopic splenic tissue and hemangiosarcoma.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251379922"},"PeriodicalIF":1.1,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12494584/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145213090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Insights from 17 years of culture and PCR detection of animal mollicutes in a Canadian provincial laboratory.","authors":"Lisa Ledger, Fernando Munevar, Pauline Nelson-Smikle, Calvin Kellendonk, Qiumei You, Lois Parker, Patricia McRaild, Rebeccah McDowall, Jason Eidt, Nathan Benoit, Pat Bell-Rogers, Grant Maxie, Hugh Y Cai","doi":"10.1177/10406387251377526","DOIUrl":"10.1177/10406387251377526","url":null,"abstract":"<p><p>Since ~1980, the Animal Health Laboratory (AHL) in Ontario, Canada, has isolated animal mollicute species by culture. Data for the most recent 17 y (2007-2024) captures over 90,000 test results. Advancements in PCR, qPCR, and DNA sequencing have shifted the percentage of testing by PCR from 18.7% in 2007 to 91.1% in 2024. The bulk of this shift is due to the uptake of molecular testing as a screening tool for clinically normal animals, but this shift has not been universal, particularly for ureaplasma testing. Culture remains the gold standard for the detection and identification of rare pathogens and plays a key role in research through our mycoplasma cryobank, which includes 40+ y of isolates. Synergizing the microbiologic and molecular techniques developed over the AHL's multi-decade history has presented novel opportunities for detection, characterization, and local eradication of animal mollicutes, including the development of new assays, tracking of historical trends for antimicrobial resistance (AMR), and identifying AMR-associated mutations in <i>Mycoplasmopsis</i> (<i>Mycoplasma</i>) <i>bovis</i>.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251377526"},"PeriodicalIF":1.1,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12494587/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145213166","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High mortality in a commercial turkey flock associated with coinfection by <i>Pasteurella multocida</i> and <i>Mycoplasmoides</i> (<i>Mycoplasma</i>) <i>gallisepticum</i>.","authors":"Carlos Daniel Gornatti-Churria, Carmen Jerry, Shayne Ramsubeik, Mark Bland, Francisco A Uzal, Tiffany Santoro, Simone T Stoute","doi":"10.1177/10406387251378687","DOIUrl":"10.1177/10406387251378687","url":null,"abstract":"<p><p>Six 11-wk-old, commercial, Broad-Breasted White, meat turkeys were submitted to the Turlock branch of the California Animal Health & Food Safety (CAHFS) laboratory for autopsy and diagnostic work-up. Clinical signs in the turkeys of the affected flock included depression, ruffled feathers, swollen periorbital areas, rales, and sneezing. A mortality of 50% (5,000 of 10,000) was reported at the time of case submission. Flock morbidity was 100% by 12 wk of age, and mortality eventually exceeded 90%. Fibrinous pleuropneumonia, airsacculitis, increased luminal mucoid exudate in the nasal cavities and tracheas, mottled and enlarged spleens, and hepatomegaly were the most remarkable gross findings. Microscopically, fibrinoheterophilic pneumonia and epicarditis with intralesional bacterial colonies, and necrotizing hepatitis and splenitis, were noted. <i>Mycoplasmoides</i> (<i>Mycoplasma</i>) <i>gallisepticum</i> (MG) was detected in tracheal and sinus pools by quantitative real-time PCR. Multilocus sequence analysis of the <i>mgc2</i> gene and IGSR segment of MG differentiated our strain from MG vaccine strains, but were similar to MG isolates detected previously in other commercial turkey operations in California. <i>Pasteurella multocida</i> was isolated from air sacs, lungs, tracheas, hearts, and livers, and classified as profile H<i>ha</i>I 0001, strain X-73, by restriction enzyme analysis DNA fingerprinting. Coinfection with <i>P. multocida</i> and MG in a susceptible flock resulted in rapid elevation of mortality and significant economic losses in this commercial meat turkey operation.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251378687"},"PeriodicalIF":1.1,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12463915/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145149774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pineal parenchymal tumor in a domestic rabbit.","authors":"Katti R Crakes, Charles G Eberhart, John A Flanders, John G Trupkiewicz","doi":"10.1177/10406387251371014","DOIUrl":"10.1177/10406387251371014","url":null,"abstract":"<p><p>An 11-y-old male lionhead rabbit (<i>Oryctolagus cuniculus</i>) was presented with progressive hindlimb weakness and right-sided neurologic deficits, and was subsequently euthanized due to poor prognosis. Autopsy revealed a 1.6 × 1.1 × 1.0-cm, well-circumscribed, extra-axial mass compressing the occipital lobe and affecting both telencephalic hemispheres. Histologic and immunohistochemical analyses demonstrating positivity for synaptophysin and neuron-specific enolase, along with a Ki67 proliferative index of ~20%, were highly suggestive of a high-grade pineal parenchymal tumor (PPT). The tumor was densely cellular with marked atypia and frequent binucleation, and lacked pineocytomatous rosettes-features most consistent with a pineal parenchymal tumor of intermediate differentiation in humans. No evidence of metastasis was observed. Pineal tumors are exceptionally rare in domestic animals, with limited documentation in species such as dogs, horses, goats, cattle, and birds. To our knowledge, PPT has not been reported previously in a rabbit, underscoring the diagnostic challenges associated with intracranial neoplasms in this species.</p>","PeriodicalId":17579,"journal":{"name":"Journal of Veterinary Diagnostic Investigation","volume":" ","pages":"10406387251371014"},"PeriodicalIF":1.1,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12463860/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145149820","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}