Journal of Veterinary Diagnostic Investigation最新文献

Feline oral squamous cell carcinoma (FOSCC) is an aggressive tumor with poor outcomes. Mechanisms of prostaglandin E2 (PGE2)-related inflammation and angiogenesis interact in human OSCC; however, this relationship has not been reported in FOSCC, to our knowledge. We aimed to characterize expression of genes encoding PGE2 synthases (PTGES1-3), PGE2 receptors (EP1-4), hypoxia inducible factor 1α (HIF1A), and vascular and endothelial growth factor A (VEGFA) in FOSCC cell lines (SCCF1-3) in vitro using reverse-transcription quantitative real-time PCR (RT-qPCR). Expression of PTGES1, PTGES3, EP4, and VEGFA were serum-inducible in SCCF2 cells; VEGFA was also inducible in SCCF1 cells (p ≤ 0.05). Compared to other serum-treated cells, SCCF3 cells had the lowest VEGFA expression despite the highest HIF1A (p ≤ 0.05) expression. PGE2 (5 µg/mL and 35 µg/mL) was added to SCCF2 cells for 4 different times (30, 60, 120, 240 min). Both doses of PGE2 stimulated expression of HIF1A and CD147 at 240 min (p ≤ 0.05). PGE2 treatment stimulated cyclooxygenase 2 (COX2) expression at 30 min, followed by suppression at 60 and 120 min and a sharp reduction in EP4 expression at 60 min (p ≤ 0.05). Treatment of SCCF2 with PGE2 and EP4 antagonist L-161,982 increased COX2 expression, and L-161,982 (alone and in combination with PGE2) stimulated EP4 expression (p ≤ 0.05). Genes for PGE2 synthase enzymes, PGE2 receptors, HIF1α and VEGFA were expressed in FOSCC cells in vitro. SCCF2 cells responded to exogenous PGE2 and EP4 antagonism, suggesting that EP4 activity in FOSCC deserves more study.

Since the First International Canine Leukocyte Antigen Workshop in 1994, individual laboratories have worked to expand the repertoire of monoclonal antibodies (mAbs) used in clinical testing and research in dogs. Here, we employed flow cytometry to document the specificity of mAbs submitted to the canine workshop and the animal homologue section of the human HLDA8 international workshop. We also provide an initial characterization of new mAbs derived from hybridomas developed from mice immunized with leukocytes from the blood of a healthy dog and from mice immunized with cells from a dog with B-cell leukemia. These mAbs enhance the tools available for characterizing leukemias, lymphomas, and other hematologic disorders in dogs, as well as for researching the canine immune response to pathogens. Importantly, some of the mAbs submitted to the canine and HLDA8 workshop recognize highly conserved epitopes expressed on orthologues of cluster of differentiation molecules first identified in humans and present an opportunity to develop a cross-species panel for clinical testing and research.

Hypertrophic osteopathy (HO) is a condition in which periosteal bone forms along long bone diaphyses and metaphyses. Lesions generally affect all 4 limbs, and often involve the distal portions. The pathogenesis is incompletely understood, and many, but not all, cases are associated with concurrent neoplasia or space-occupying masses. Among veterinary species, most cases are reported in dogs, cats, and horses, with sporadic cases in other domestic and non-domestic species. In cervids, this condition is often associated with fungal granulomas, typically in the lung. We report HO, with findings consistent to other veterinary species, in a farmed white-tailed deer (WTD) with bacterial pneumonia and in 3 free-ranging WTD, one of which had fungal pneumonia. Recognition of HO in WTD and potentially associated conditions can lead to improved sample collection in the field. Such information can improve characterization of the disease, complement our understanding of comparative pathology of this condition, and raise awareness among individuals or institutions working with cervids.

Polioencephalomalacia (PEM) is a neurologic disorder of ruminants that can affect cattle as a result of high levels of sulfur in the diet. Sulfur can be ingested by an animal through feed or water, and the mineral status of the animal can affect how much sulfur is ultimately absorbed. Broiler (poultry) litter is sometimes used in cattle diets as an economical protein source, although it runs the risk of supplying too much sulfur to the bovine diet. Here we report a case in which 15 cows had neurologic signs before death. Initial testing ruled out other causes of acute death, and histologic findings confirmed a diagnosis of PEM. The sulfur concentration was ~0.70% of the total mixed ration (TMR), which is above the maximum tolerable dietary concentration of sulfur. The broiler litter contributed nearly all of the sulfur to this ration and was the cause of the PEM experienced in this herd. When feeding by- and co-products as part of a TMR, it is important for producers to analyze the feed to mitigate risk and ensure good health.

Neuroglial choristomas are rare in humans and animals. During a routine medical examination, an ovarian mass was found in a 5-y-old female mixed-breed dog. Histologically, the ovarian mass was well-defined, expansile, and composed of large polygonal cells resembling well-differentiated neurons and scattered smaller cells resembling glial cells. Immunohistochemically, the neuron-like cells were positive for neuronal nuclei; the smaller cells were positive for glial fibrillar acidic protein, oligodendrocyte transcription factor 2, and ionized calcium-binding adaptor protein-1. Ovarian neuroglial choristoma was diagnosed.

Bacteria in the Mycobacterium tuberculosis complex and nontuberculous mycobacteria may affect a variety of animal species under human care and pose public health risks as zoonotic pathogens. A case of sudden onset of lethargy and increased respiratory effort in a 5-y-old, intact female reindeer (Rangifer tarandus) under managed care had progressed to severe dyspnea despite aggressive treatment. The animal was euthanized due to poor prognosis. Postmortem findings included: disseminated miliary nodules in the lungs, pleura, small intestine, liver, and spleen; enlarged mesenteric lymph nodes; dilated mesenteric and serosal lymphatic vessels; and renal infarcts. Histologically, granulomatous lymphadenitis and lymphangitis with intrahistiocytic acid-fast bacilli were observed. Mycobacterium sp. DNA was detected in lung via real-time PCR. Mycobacterial culture and sequencing identified Mycobacterium avium subsp. hominissuis (MAH) within pulmonary lesions. Infection with MAH has been reported in humans and many animal species; this nontuberculous mycobacterial infection may be an emerging concern in animals under managed care. To our knowledge, MAH infection has not been reported previously in reindeer.

Compromised gill health is a critical cause of forfeited welfare in Atlantic salmon farming. Detecting and quantifying the early onset of gill disease is important to reveal initial inflicting stimuli. We collected gill samples of 45 Atlantic salmon from 2 commercial recirculating aquaculture systems (RASs) spanning fry-to-market-size fish with no clinical signs of gill disease. Gill samples were assessed histologically by 3 independent raters with different levels of experience. Semiquantitative scoring for 7 types of gill changes was carried out for 10 filaments per gill (450 filaments total) over 3 rounds on anonymized samples. Scores were summarized for each type of gill change. The assumed clinical relevance for each change was transformed into a category score, followed by an assessment of agreement within (intra) and between (inter) raters. A generalized linear model estimated the difference in score levels between raters. For each rater, intra-rater agreement was high for 6 gill changes and moderate for 1 gill change. Inter-rater agreement was moderate to almost-perfect, except for 2 gill changes; generalized linear model regression revealed systematic differences in score usage between the raters. Our scoring protocol worked satisfactorily for mucous cell amount, lamellar clubbing, lamellar hypertrophy and/or hyperplasia, and aneurysms, despite different levels of expertise in histologic evaluation. Intra-rater agreement was consistent, but differences existed for interlamellar hypercellularity, lamellar inflammation, and degeneration. Scoring subclinical gill changes is a challenge, and our scoring system for mild-to-moderate lesions may enable early intervention to limit the detrimental effects of poor gill health in RAS farming.

Equid alphaherpesvirus 4 (EqAHV4; Orthoherpesviridae, Varicellovirus equidalpha4; equine rhinopneumonitis virus) has seldom been associated with complications such as abortion and myeloencephalopathy, given the low tendency of this virus to induce viremia. We investigated the frequency of EqAHV4 viremia in horses with fever and respiratory signs. Case selection included all equids with EqAHV4 quantitative real-time PCR (qPCR)-positive nasal secretions (defined as EqAHV4 qPCR-positive cases) submitted to a diagnostic laboratory. Controls consisted of each case submitted before and after each EqAHV4 qPCR-positive case. Purified nucleic acid from blood samples collected from EqAHV4 qPCR-positive horses and control cases was tested for EqAHV4 by qPCR. We selected 183 EqAHV4 qPCR-positive horses and 376 EqAHV4 qPCR-negative horses. In general, EqAHV4 qPCR-positive horses were younger, and had a lower rate of anorexia and a higher rate of nasal discharge compared to the EqAHV4 qPCR-negative horses. A total of 25 of 183 (13.7%) horses with EqAHV4 qPCR-positive nasal secretions tested qPCR-positive for EqAHV4 in blood. EqAHV4 viremic horses were significantly younger (p < 0.015 for group <1-y-old) and had a significantly higher occurrence of distal limb edema (p < 0.05) than EqAHV4 non-viremic horses. Our data support the observation that EqAHV4 viremia is rarely detected in EqAHV4-infected horses, which explains the low level of reported complications, such as abortion and myeloencephalopathy.

The Serratia marcescens complex contains important opportunistic pathogens of humans and vertebrate animals, as well as insects and other invertebrates. To date, the methods used for the identification of species within the genus Serratia, including PCR assays, have poor discriminatory power and may require further molecular typing or genomic sequence analysis to determine clinical relevance. We developed a duplex TaqMan probe-based quantitative real-time PCR (qPCR) assay targeting the chiP gene, which is involved in chitin degradation and transport, and the ureD gene, which is involved in urease production. This allowed us to simultaneously identify all members of the S. marcescens complex (chiP positive) and differentiate those most likely to act as insect pathogens (chiP and ureD positive). We applied our assay to identify potentially entomopathogenic members of the S. marcescens complex in the context of a conservation program for the critically endangered insect Dryococelus australis and found it to be a useful aid for rapid and accurate detection of infection with S. marcescens complex strains in insects and determination of their clinical relevance. By targeting 2 genes likely to be virulence factors, this assay may also be of use for research investigating the pathogenesis of entomopathogenic Serratia spp.