Journal of Veterinary Diagnostic Investigation最新文献

The rapid geographic spread of chronic wasting disease (CWD) in white-tailed deer (WTD; Odocoileus virginianus) increases the need for the development and validation of new detection tests. Real-time quaking-induced conversion (RT-QuIC) has emerged as a sensitive tool for CWD prion detection, but federal approval in the United States has been challenged by practical constraints on validation and uncertainty surrounding RT-QuIC robustness between laboratories. To evaluate the effect of inter-laboratory variation on CWD prion detection using RT-QuIC, we conducted a multi-institution comparison on a shared anonymized sample set. We hypothesized that RT-QuIC can accurately and reliably detect the prions that cause CWD in postmortem samples from medial retropharyngeal lymph node (RPLN) tissue despite variation in laboratory protocols. Laboratories from 6 U.S. states (Michigan, Minnesota, Missouri, New York, Pennsylvania, Wisconsin) were enlisted to compare the use of RT-QuIC in determining CWD prion status (positive or negative) among 50 anonymized RPLNs of known prion status. Our sample set included animals of 3 codon 96 WTD genotypes known to affect CWD progression and detection (G96G, G96S, S96S). All 6 laboratories successfully identified the true disease status consistently for all 3 tested codon 96 genotypes. Our results indicate that RT-QuIC is a suitable test for the detection of CWD prions in RPLN tissues in several genotypes of WTD.

Prompt diagnosis of equine septic arthritis is crucial for successful treatment. Serum amyloid A (SAA) has been suggested as a reliable biomarker. However, we previously found that synovial fluid SAA increases in nonaffected joints of horses with septic arthritis. We hypothesized that systemic SAA may leak into the nonaffected joints. If this is the case, we also hypothesized that locally produced joint SAA isoforms may be better candidates for septic arthritis biomarkers. Thus, our objectives were 1) to evaluate the temporal kinetics of systemic and synovial fluid SAA in horses with septic arthritis (n = 5), non-septic synovitis (n = 5), and systemic inflammation (n = 5), examining both affected and contralateral joints; and 2) investigate putative locally produced joint SAA isoforms and detect amino-acid differences between them. We confirmed that SAA increases significantly in synovial fluid in nonaffected joints of horses with systemic inflammation (≤352 mg/L), as well as in contralateral nonaffected joins in horses with septic arthritis (≤1,830 mg/L) compared to baseline at time 0 (<0.2 mg/L). We also identified a putative locally produced joint SAA peptide in synovial fluid (FGDSGHGAADSR) that differed in 1 amino acid from 2 systemic peptides found both in plasma and synovial fluid. The putative joint SAA isoform was present in joints of horses with both septic arthritis and systemic inflammation (ion intensities 10⁴-10⁶). Thus, the increase of synovial fluid SAA may be both due to the leakage of SAA from serum into joints and local production of joint SAA isoforms.

South Korea's beekeeping industry has been facing a major crisis due to colony collapse disorder (CCD), manifesting since the winter of 2021. CCD in South Korea is presumed to be caused by a combination of factors, including an abnormal climate, pesticide use, declining source plants, and increased honey bee diseases. We examined the prevalence of 12 major honey bee (Apis mellifera) pathogens by sampling 3,707 colonies with abnormal behavior and suspected pathogen infections from 1,378 apiaries nationwide between 2020 and 2023. Black queen cell virus (BQCV), deformed wing virus (DWV), Israeli acute paralysis virus (IAPV), and Vairimorpha (Nosema) ceranae had the highest infection rates among honey bees in South Korea. BQCV had the highest infection rate (83.3% in 2023) and was highly prevalent throughout the year, regardless of the season. DWV (48.7%) and IAPV (41.3%) had the highest infection rates in October-December, corresponding to the winter season. Among the 12 honey bee pathogens, acute bee paralysis virus and Kashmir bee virus were rarely detected; the remaining 10 honey bee pathogens were detected throughout the year. The differences in honey bee pathogen prevalence among regions were not significant. We suggest that South Korean honey bees are highly exposed to viral pathogens, possibly resulting in the loss of unhealthy honey bees during the winter. Our study is expected to help identify trends in the occurrence of honey bee pathogens in South Korea and predict outbreaks to prepare a prevention system and appropriate control measures for honey bee pathogens.

Pituitary pars intermedia dysfunction (PPID) is a neurodegenerative disease of senior horses. Loss of dopaminergic inhibition of the melanotropes of the pars intermedia leads to increased concentrations of pro-opiomelanocortin (POMC)-derived peptides. Diagnosis is challenging due to pre-analytical variables, such as sample storage, handling, and time to analysis. Our objective was to develop an ELISA for ACTH measurement, which could ultimately form the basis for a stall-side equine ACTH test. We selected 2 ACTH-specific monoclonal antibodies, CBL57 and EPR20361-248, based on the recognition of separate epitopes, strong and rapid color change, and minimal background interference, including no cross-reactivity with themselves, each other, and the test reagents. CBL57 was chosen as the detection antibody (or secondary antibody). EPR20361-248, functionalized on superparamagnetic iron oxide beads, was chosen as the capture antibody (or primary antibody) to bind ACTH in plasma. The incorporation of magnetic beads marks the initial stage in establishing a platform that could potentially be utilized in the field, similar to other stall-side tests. The concentrations of antibodies, magnetic beads, and incubation durations were optimized. Our immunoassay detected unglycosylated rat recombinant ACTH. Further studies are ongoing to optimize and validate our assay using equine plasma and serum samples.

Neoplasia is a common disease in guinea pigs (Cavia porcellus); however, few studies have evaluated the prevalence of neoplasia in all organ systems. We retrospectively analyzed the tumor prevalence in pet guinea pigs and the frequency of metastasis in a multi-institutional study population of 2,474 autopsy cases. Tumors were found in 508 guinea pigs (prevalence: 20.5%), of which 95 cases had >1 tumor, resulting in a total of 627 tumors. The tumor prevalence increased from 1.4% in animals <0.5-y-old to 53.6% for guinea pigs >5-y-old. The most common tumor type was lymphoma or leukemia, affecting 174 guinea pigs (tumor prevalence: 7.0%). Lymphomas or leukemias were disseminated to various organs and/or lymph nodes in 146 (83.9%) cases and localized to 1 organ or 1 lymph node in 28 (16.1%) cases. Primary non-lymphoid tumors were most frequent in the female genital tract (62 of 1,235 cases, mostly uterus), respiratory system (116 of 2,474), skin including mammary gland (81 of 2,474), endocrine system (66 of 2,474, mostly thyroid gland), and alimentary tract (35 of 2,474). Tumors of the alimentary tract were dominated by gastrointestinal stromal tumors. Metastasis was detected in 42 of 453 non-lymphoid tumors (9.3%), with a surprisingly low frequency for pulmonary carcinoma and splenic hemangiosarcoma compared to other species. Our postmortem study demonstrates a high prevalence of disseminated lymphoma or leukemia in pet guinea pigs at the time of death or euthanasia. Additional studies are needed to further characterize these tumors.

In March 2024, highly pathogenic avian influenza A(H5N1) virus, clade 2.3.4.4b, was detected in dairy cows in the United States, and at the same time in resident cats on affected farms. To help guide sample collection and diagnosis in cats, here we report the distribution of lesions and detection of H5N1 clade 2.3.4.4b influenza A virus (IAV) infection by PCR, immunohistochemistry (IHC), and serology in samples from 4 deceased and 2 living cats from 3 separate affected dairy farms. Although gross lesions were not diagnostic, histologically, all 4 deceased cats had nonsuppurative and necrotizing encephalitis and subtle interstitial pneumonia, and some also had significant myocarditis (3 of 4), chorioretinitis (2 of 4), and sialadenitis (1 of 2). The virus was detected by IHC in the aforementioned tissues, and by PCR in each brain (Ct = 9.9-25.1), lung (17.4-32.7), oropharyngeal swab (28.3-30.5), urine (30.3-34.4), and nasal swab (33.5-34.1) collected postmortem; fecal swabs were PCR-negative. In the antemortem samples, the virus was detected by PCR in the oropharyngeal swabs (34.1-36.1), whole-blood samples (30.8-36.6), and one serum sample (31.7). Seroconversion was detected in one cat. Our results support histologic evaluation of brain, lung, eyes, and heart, and PCR testing of brain and lung for postmortem diagnosis, and show that oropharyngeal swabs, urine, serum, and whole blood are suitable samples for antemortem detection of IAV infection in clinically affected cats.

We identified a novel herpesvirus in 2 deceased captive blue penguins (Eudyptula minor). Moderate-to-severe myocardiocyte atrophy and necrosis, and eosinophilic intranuclear inclusion bodies (INIBs), were seen in myocardiocytes in one bird; reticuloendothelial (RE) cell INIBs and multifocal RE cell necrosis were seen in both birds. The histologic findings were suggestive of viral infection. A herpesvirus PCR assay was positive in myocardial tissue from the bird with myocardial degeneration and in splenic tissue from both birds. Sequencing and phylogenetic analysis showed that the virus, accessioned as spheniscid alphaherpesvirus 2 (SpAHV2), groups within the Alphaherpesvirinae subfamily and forms a unique branch point in a subclade containing members of the Mardivirus, Simplexvirus, and Varicellovirus genera. Herpesvirus screening of tissues from 8 additional blue penguin postmortem examination cases (7 spleen, 1 liver) and combined conjunctival-choanal-cloacal swab samples from 13 live penguins revealed 5 additional dead and 7 live penguins that were positive for SpAHV2. The presence of SpAHV2 in healthy live animals and lack of significant herpesvirus-associated lesions as the cause of death in 6 of 7 SpAHV2-positive dead penguins suggests that this virus may be an endemic in blue penguins, and that recrudescence may cause disease and death.

Evaluating stress in shelter and institutionally owned cats is important to help guide improvements in their welfare. Welfare assessments often focus on behavior metrics and physiologic measurements, such as systemic cortisol levels. The gold standard for measuring acute stress is serum cortisol; measuring cortisol in feces and urine gives reliable time-integrated assessments of acute stress. Monitoring chronic stress requires using a matrix that accumulates cortisol over time, such as hair or nails. Hair was collected from 29 cats representing 2 populations: cats from a local shelter and cats owned by a university. Cortisol was extracted from the hair using a method established for extracting cortisol from bovine hair. We measured hair cortisol concentrations with a commercial ELISA that is marketed for human saliva. The mean cortisol concentration was 140 pg/mg for the shelter cats and 98 pg/mg for the university-owned cats. We found no significant difference in hair cortisol concentrations between the 2 groups (p = 0.793). The intra- and inter-assay CVs for the ELISA were 9.3% and 8.4%, respectively. Observed:expected ratios for spiking recovery and dilutional parallelism were 87.7 ± 25.8% and 99.7 ± 37.5%, respectively. Measurement of cortisol in hair samples may provide a noninvasive method to monitor chronic stress and acclimation in cats that live in confinement for prolonged periods.