bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatitis and Enteritis Caused by a Novel Herpesvirus in Two Monitor Lizards (Varanus Spp.)
Authors: Hughes-Hanks Jennifer M. , Schommer Susan K. , Mitchell William J. , Shaw Daniel P.
Published: May 2011

Reported cases of herpesvirus-induced disease are uncommon in most species of reptiles, with the majority of reports in chelonians. Two monitor lizards ( Varanus spp.) presented for postmortem examination at the Veterinary Medical Diagnostic Laboratory at the University of Missouri. Tan, 1–2-mm foci were grossly visible on the mucosal surface of the intestine and in the liver. Microscopically, there was multifocal necrosis in the lamina propria of the small intestine and in the liver. Many of the degenerate cells contained large, eosinophilic intranuclear inclusions. Enveloped icosahedral virions consistent with herpesvirus were detected by electron microscopy. A 180-bp DNA fragment was amplified by polymerase chain reaction from samples of small intestine and liver using primers that targeted a portion of the herpesvirus DNA polymerase gene. The sequence of the fragment was determined to be most closely related to Varanid herpesvirus 2 (80% nucleotide identity, 82% amino acid identity). Based on histological and molecular findings, a novel pathogenic herpesvirus of lizards in the family Varanidae is proposed.
Polyglucosan Inclusions (Lafora Bodies) in a Gray-Headed Flying Fox (Pteropus Poliocephalus)
Authors: Gabor Les J. , Srivastava Mukesh
Published: May 2011

Polyglucosan bodies (Lafora bodies) were identified in a juvenile gray-headed flying fox ( Pteropus poliocephalus) with neurological signs. The structures were only noted in the brain stem, and no associated degenerative changes were present. These structures have not been previously identified in any species in the order Chiroptera.
A Gastric Neuroendocrine Carcinoma Expressing Somatostatin in a Bearded Dragon (Pogona Vitticeps)
Authors: Lyons Jeremiah A. , Newman Shelley J. , Greenacre Cheryl B. , Dunlap John
Published: May 2011

A metastatic gastric neuroendocrine carcinoma in a 2.5-year-old inland bearded dragon ( Pogona vitticeps) with a chronic history of anorexia, weight loss, depression, and acute melena is described. Histologic examination of the gastric mass revealed a densely cellular tumor arranged in nests and occasional rosettes of hyperchromatic cells with oval to spindle-shaped nuclei and minimal cytoplasm; the tumor was supported by a moderate fibrovascular stroma. Similar cells invaded through the gastric mucosa, and there were multiple hepatic metastases. The neoplastic cells were weakly immunopositive for neuron-specific enolase and moderately positive for somatostatin but were negative for chromogranin AB and gastrin. Ultrastructural studies revealed scattered neurosecretory granules in the neoplastic cells, confirming the diagnosis of a neuroendocrine carcinoma.
An Intra-Abdominal Malignant Mesenchymoma Associated with Nonabsorbable Sutures in a Ferret (Mustela Putorius Furo)
Authors: Petterino Claudio , Bedin Marco , Vascellari Marta , Mutinelli Franco , Ratto Alessandra
Published: May 2011

A 6-year-old ferret ( Mustela putorius furo) was presented with abdominal enlargement. Clinical examination revealed an intra-abdominal mass measuring approximately 5 cm in diameter. Abdominal survey radiographs revealed a sharply marginated mass with multifocal radiodense foci, suggestive of pathologic calcification. A complete blood cell count revealed a moderate, normocytic, normochromic, nonregenerative anemia. The mass was surgically removed en bloc, fixed in 10% neutral buffered formalin solution, and routinely processed for histologic and immunohistochemical investigation. The neoplastic tissue consisted of a mixed neoplastic cell population, including osteosarcoma and fibrosarcoma components. Immunohistochem-istry revealed that both neoplastic cell populations were positive for vimentin and negative for actin (smooth and striated), desmin, and myoglobin. Nonabsorbable suture material was admixed with the neoplastic tissue in the histologic section. This material was birefringent when viewed microscopically under polarized light.
Evaluation of Diagnostic Tests for Johne's Disease (Mycobacterium Avium Subspecies Paratuberculosis) in New Zealand Dairy Cows
Authors: Norton Solis , Johnson Wesley O. , Jones Geoff , Heuer Cord
Published: May 2011

To quantify the sensitivity and specificity of a serum enzyme-linked immunosorbent assay (ELISA) and fecal culture (FC) tests and to estimate the prevalence of Johne's disease (JD) in New Zealand dairy herds using Bayesian methods, 4 New Zealand dairy herds were tested simultaneously by ELISA and FC 5 times over 3 lactations. Test results were dichotomized. A Bayesian regression model was developed that considered test sensitivity as a function of the covariates parity, lactation stage, and prevalence of JD, which is expected to vary between herds. It was applied to a cross-sectional subset of the data and the full, repeated measures data set. Results were compared with frequentist pseudo gold standard results of the full data. Using the regression model, sensitivity of the ELISA was higher in older animals, but the sensitivity of the FC test showed no trend across age groups. Both FC and ELISA sensitivity were lower in late lactation. Estimated prevalence was lower and FC sensitivity higher when analyzing the complete data. The regression model enabled a more accurate diagnosis of JD to be made because it incorporated cow-specific information in the diagnosis, such as age and lactation stage. The model also enabled the incorporation of previous test results for an individual when diagnosing disease. The trends in results from the regression model support the current understanding of the disease process. The advantage of repeated testing of individuals in the assessment of test performance is discussed in the current study.
Antigenic Categorization of Contemporary H3N2 Swine Influenza Virus Isolates Using a High-Throughput Serum Neutralization Assay
Authors: Hause Ben M. , Oleson Tracy A. , Bey Russell F. , Stine Douglas L. , Simonson Randy R.
Published: May 2011

In vivo, neutralizing antibodies are critical for viral clearance. A high-throughput serum neutralization (HTSN) assay was developed to antigenically categorize Swine influenza virus (SIV) isolates. Uncategorized viruses were tested using a panel of antisera representing the H3N2 SIV subtypes and the results expressed as a serum neutralization ratio. Antisera were generated against contemporary isolates representing circulating H3N2 SIV subtypes (clusters I, III, IV). Reference viruses and the corresponding antisera were evaluated using traditional hemagglutination inhibition (HI) and the HTSN assays and good correlation ( r = 0.84) was observed between the 2 tests. Categorical clustering of 40 recent (2008-2009) SIV isolates was assessed using the HTSN assay. The H3N2 SIV isolates with amino acid similarity >97% to the commonly used H3N2 cluster IV reference strain A/Swine/Ontario/33853/2005 (ON05) showed strong reactivity with cluster IV antisera. Isolates with <97% amino acid similarity to ON05 sporadically or completely failed to react with any antiserum. A cluster of 3 isolates with weak reaction with cluster III antiserum may be a potential emerging cluster of H3N2 with moderate genetic similarity to cluster II H3N2 (93% similarity). Potential uses of the HTSN assay include identification of broadly cross-reactive or antigenically distinct SIV isolates for use in vaccine virus selection or as part of surveillance efforts monitoring antigenic drift.
Comparison of Genotypic and Phenotypic Characterization Methods for Pasteurella Multocida Isolates from Fatal Cases of Bovine Respiratory Disease
Authors: Taylor Jared D. , Fulton Robert W. , Dabo S. Mady , Lehenbauer Terry W. , Confer Anthony W.
Published: May 2011

Bovine respiratory disease (BRD) is the most costly disease of beef cattle in North America. Because Pasteurella multocida is a commensal of the upper respiratory tract, it is generally considered an opportunistic pathogen. However, studies in swine indicated that there may be a limited number of strains associated with disease, suggesting that some are more virulent than others. Although this may also be true of isolates from cattle, appropriate typing methods must be established before this possibility can be investigated. The purpose of this study was to compare effectiveness of polymerase chain reaction (PCR) fingerprinting to more traditional approaches for typing bovine P. multocida isolates. Isolates were obtained from 41 cases of fatal BRD and subjected to random amplified polymorphic DNA PCR (RAPD-PCR), whole cell protein (WCP) profiles, outer membrane protein (OMP) profiles, and serotyping. The discrimination index was calculated for each typing method and combinations of each using Simpson's index of diversity. Correlation coefficients were calculated to assess concordance between classification results achieved through genotypic (RAPD-PCR) and phenotypic (WCP, OMP, and serotyping) approaches. All characterization methods were capable of discriminating between isolates. However, there was poor concordance between techniques. There were also few significant associations between typing results and epidemiologic data. Random amplified polymorphic DNA PCR was validated as being a repeatable and reliable means of discriminating between P. multocida isolates obtained from cattle. Isolates obtained from fatal cases of BRD in calves in a commercial feedlot demonstrated significant diversity, justifying additional investigation into whether P. multocida is a strictly opportunistic pathogen in cattle.
Jellyfish as Vectors of Bacterial Disease for Farmed Salmon (Salmo Salar)
Authors: Ferguson Hugh W. , Christian M. J. Delannoy , Hay Stephen , Nicolson James , Sutherland David , Crumlish Margaret
Published: May 2011

Swarms or blooms of jellyfish are increasingly problematic and can result in high mortality rates of farmed fish. Small species of jellyfish, such as Phialella quadrata (13 mm in diameter), are capable of passing through the mesh of sea cages and being sucked into the mouth of fish during respiration. Results of the current study show that the initial damage to gills of farmed Atlantic salmon, likely produced by nematocyst-derived toxins from the jellyfish, was compounded by secondary bacterial infection with Tenacibaculum maritimum. Results also demonstrate that these filamentous bacteria were present on the mouth of the jellyfish and that their DNA sequences were almost identical to those of bacteria present on the salmon gills. This suggests that the bacterial lesions were not the result of an opportunistic infection of damaged tissue, as previously thought. Instead, P. quadrata is probably acting as a vector for this particular bacterial pathogen, and it is the first time that evidence to support such a link has been presented. No prior literature describing the presence of bacteria associated with jellyfish, except studies about their decay, could be found. It is not known if all jellyfish of this and other species carry similar bacteria or the relationship to each other. Their source, the role they play under other circumstances, and indeed whether the jellyfish were themselves diseased are also not known. The high proteolytic capabilities of T. maritimum mean that partially digested gill tissues were readily available to the jellyfish, which rely heavily on intracellular digestion for their nutrition.
Immunohistochemical Investigation of PNL2 Reactivity of Canine Melanocytic Neoplasms and Comparison with Melan A
Authors: Giudice Chiara , Ceciliani Fabrizio , Rondena Marco , Stefanello Damiano , Grieco Valeria
Published: May 2011

PNL2 is a recently generated monoclonal antibody (mAb) that recognizes normal and neoplastic melanocytes. Although the antigen recognized by PNL2 remains unknown, recent studies of human and mouse melanomas have confirmed its usefulness as a diagnostic marker. In the current study, the immunoreactivity of PNL2 in canine melanomas was tested and compared with Melan A (A103). Validation of PNL2 was performed by Western blot analysis. PNL2 and Melan A immunoreactivity were tested on frozen samples of canine melanomas and on 69 formalin-fixed, paraffin-embedded melanocytic neoplasms. Normal canine tissues and nonmelanocytic neoplasms were included as negative controls. Western blot confirmed the presence of a protein recognized by the PNL2 antibody in canine melanomas. Immunohistochemically, PNL2 stained the melanocytic neoplastic cells with an intracytoplasmic, granular pattern. Among the melanocytic neoplasms tested, 62% stained positively with PNL2 and 59% with Melan A; 50.7% stained positively with both mAbs. The overall percentage of neoplasms that stained positively with at least 1 of these 2 antibodies was 68%. The extent of staining (i.e., the percentage of cells stained per specimen) was greater with PNL2 than with Melan A. With both mAbs, staining was most intense and diffuse in the epithelioid cell phenotype. Neither nonspecific staining nor staining in cells other than melanocytes was detected with either mAb. In contrast to human granulocytes, canine granulocytes were negative by both Western blot and immunohistochemical analyses. PNL2 mAb proved to be highly specific for the identification of formalin-fixed canine melanocytic neoplasms and should be a valuable diagnostic reagent.
C-Reactive Protein as a Prognostic Indicator in Dogs with Acute Abdomen Syndrome
Authors: Galezowski Angelica M. , Snead Elisabeth C. R. , Kidney Beverly A. , Jackson Marion L.
Published: May 2011

The objective of the present study was to measure serum C-reactive protein (CRP) concentrations in 32 dogs with acute abdomen syndrome at presentation and after 48–72 hr. Data were evaluated to determine if there was an association between CRP concentration and outcome, and if CRP concentration correlated with the white blood cell (WBC) count at both time points. An immunoturbidimetric assay for human CRP, previously validated for use in dogs, was used for serum CRP analysis. Increased serum CRP concentrations were found in 21 dogs at presentation. Fifteen of these dogs had declining serum CRP concentrations by 48–72 hr, but 3 of the 15 dogs were later euthanized. Serum CRP concentrations increased by 48–72 hr in 4 dogs. Of the 32 dogs, 4 were dead or were euthanized prior to the 48–72 hr time point. No significant difference between initial CRP concentration and outcome was found ( P = 0.054). Initial and 48–72 hr CRP values taken together were significantly different between outcome groups ( P < 0.001). Serum CRP concentrations that were elevated at both time points were associated with a poor prognosis. No correlation was found between CRP concentrations and WBC counts at presentation ( P = 0.83); however, a significant correlation was noted at 48–72 hr ( P = 0.03). Evaluation of sequential CRP concentrations in dogs with acute abdomen syndrome may be helpful in assessing clinical response to treatment and predicting outcome. Also, serum CRP may be better in detecting tissue injury and/or inflammation at presentation than WBC counts in select cases.
Single-Step Multiplex Conventional and Real-Time Reverse Transcription Polymerase Chain Reaction Assays for Simultaneous Detection and Subtype Differentiation of Influenza A Virus in Swine
Authors: Nagarajan Malliga M. , Simard Geneviève , Longtin Diane , Simard Carole
Published: May 2011

Because pigs are considered intermediate hosts for the emergence of novel influenza virus reassortants with associated zoonotic potential, monitoring and characterization of circulating influenza viruses in pigs are important for adequate control of infection. For this, rapid molecular diagnostic methods other than immunoassays are needed. Three novel single-step multiplex reverse transcription polymerase chain reaction (RT-PCR) assays were developed in the current study for simultaneous detection and subtype differentiation of Influenza A virus in pigs. A conventional single-step pentaplex RT-PCR was designed for concomitant detection of the generic matrix (M) gene, hemagglutinin H1 and H3, and neuraminidase N1 and N2 genes of Swine influenza virus (SIV). In the other 2 single-step tetraplex real-time RT-PCR assays, the primers and fluorescent probes were targeted for the simultaneous detection of common M, H1, H3, and N2 SIV genes (first assay), and for M, H1, and H3 SIV genes and the H5 gene of highly pathogenic avian influenza virus of Eurasian lineage (second assay). The real-time RT-PCR assays had detection sensitivity limits ranging from 101 to 103 copies of respective in vitro RNA transcripts of M, H1, H3, H5, and N2 genes. The multiplex assays were evaluated by using SIV isolates, clinical specimens, and the appropriate synthetic template. The recent H1N1 pandemic strain isolated from pigs also was tested in simplex RT-PCR and realtime RT-PCR assays with the H1 primers and probes. The efficacy of the multiplex RT-PCR and real-time RT-PCR shows the suitability of multiplex RT-PCR and real-time RT-PCR for rapid subtype identification and monitoring in North American pigs of Influenza A virus.
Development of a Peptide-Based Immunochromatographic Strip for Differentiation of Serotype O Foot-and-Mouth Disease Virus—Infected Pigs from Vaccinated Pigs
Authors: Yang Suzhen , Yang Jifei , Zhang Gaiping , Qiao Songlin , Wang Xuannian , Zhao Dong , Li Xuewu , Deng Ruiguang , Zhi Aimin , You Leiming , Chai Sujun , Teng Man
Published: May 2011

An immunochromatographic strip for discriminating Foot-and-mouth disease virus (FMDV) infected from vaccinated pigs was developed based on synthetic peptide. Five peptides designed from the amino acid sequences of nonstructural proteins (NSP) of FMDV were synthesized, and pep5 located in NSP 3B reacted strongly with serum from FMDV-infected pigs but did not react with serum samples from healthy vaccinated pigs. An immunochromatographic strip was developed by using colloidal gold labeled with pep5 as the detector. Staphylococcal protein A and rabbit against peptide-conjugated ovalbumin antibody immunoglobulin G were blotted on the nitrocellulose membrane for the test and control lines. In comparison with 2 commercial NSP enzyme-linked immunosorbent assays, the peptide-based strip showed good specificity and sensitivity. The apparent agreements of this new assay with Ceditest® ELISA and UBI® ELISA were 98.59% and 96.63%, respectively. These results indicate that the strip can be adequately used to discriminate FMDV-infected animals from vaccinated animals.
Isolation and Partial Sequencing of Equid Herpesvirus 5 from a Horse in Iceland
Authors: Thorsteinsdóttir Lilja , Torfason Einar G. , Torsteinsdóttir Sigurbjörg , Svansson Vilhjálmur
Published: May 2011

Horses are hosts to 2 types of gammaherpesviruses, Equid herpesvirus 2 and 5 (EHV-2 and EHV-5, respectively). Both EHV-2 and EHV-5 are common in horses in Iceland. An Icelandic EHV-5 isolate was recovered by sequential culture in primary fetal horse kidney and rabbit kidney cells. Glycoprotein B, glycoprotein H, and DNA terminase genes of the isolate were fully sequenced, and the DNA polymerase gene was partly sequenced. To date, the glycoprotein B gene of EHV-5 was the only gene that has been reported to be completely sequenced in addition to small parts of the glycoprotein H, DNA polymerase, and DNA terminase genes. The present report, therefore, is a significant addition to previously reported EHV-5 sequences.
Improvement and Optimization of a Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay for the Detection and Typing of Vesicular Stomatitis Virus
Authors: Hole Kate , Velazques-Salinas Lauro , Clavijo Alfonso
Published: May 2011

An improvement to a previously reported real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assay for the detection of Vesicular stomatitis virus (VSV) is described. Results indicate that the new assay is capable of detecting a panel of genetically representative strains of VSV present in North, Central, and South America. The assay is specific for VSV and allows for simultaneous differentiation between Vesicular stomatitis Indiana virus and Vesicular stomatitis New Jersey virus. This realtime RT-PCR is able to detect current circulating strains of VSV and can be used for rapid diagnosis of VSV and differentiation of VSV from other vesicular diseases, such as foot-and-mouth disease.
Flow Cytometric Analysis of Peripheral Blood and Tumor-Infiltrating Regulatory T Cells in Dogs with Oral Malignant Melanoma
Authors: Tominaga Makiko , Horiuchi Yutaka , Ichikawa Mika , Yamashita Masao , Okano Kumiko , Jikumaru Yuri , Nariai Yoko , Kadosawa Tsuyoshi
Published: May 2011

It is well known that tumor-infiltrating lymphocytes (TILs) and peripheral blood lymphocytes (PBLs) from patients with advanced-stage cancer have a poor immune response. Regulatory T cells (Tregs), characterized by the expression of a cluster of differentiation 4 and intracellular FoxP3 markers, can inhibit antitumor immunoresponse. In the present study, the prevalence of Tregs in peripheral blood and tumor tissue from dogs with oral malignant melanoma was evaluated by triple-color flow cytometry. The percentage of Tregs in the peripheral blood of the dogs with malignancy was significantly increased compared with healthy control dogs, and the percentage of Tregs within tumors was significantly increased compared with Tregs in peripheral blood of dogs with oral malignant melanoma. This finding suggests that the presence of tumor cells induced either local proliferation or selective migration of Tregs to tumor-infiltrated sites. A better understanding of the underlying mechanisms of Treg regulation in patients with cancer may lead to an effective anticancer immunotherapy against canine malignant melanoma and possibly other tumors.
Cottontail Rabbit Papillomavirus in Langerhans Cells in Sylvilagus spp.
Authors: Weiner Cristina M. , Rosenbaum Matthew D. , Fox Karen , Mason Gary , Bielefeldt-Ohmann Helle , Veeramachaneni D. N. Rao , Vandewoude Sue
Published: May 2011

A wildlife sanctuary presented an adult female cottontail rabbit ( Sylvilagus spp.), age unknown, to the Colorado State University Pathology service for postmortem examination. Gross examination revealed numerous pigmented wartlike lesions arising from the skin of the head surrounding the ears, eyes, nares, mouth, and dorsum. Masses were firm, friable, and easily detached from the underlying skin. Differential diagnoses included Cottontail rabbit papillomavirus, Rabbit fibroma virus, and Myxoma virus. Histological examination revealed multiple papillary masses lined by stratified squamous epithelial cells with central cores of fibrovascular connective tissue and parakeratotic hyperkeratosis. Cells of the Stratum spinosum were frequently swollen with abundant perinuclear, cytoplasmic, clearing, and occasional intranuclear basophilic, glassy, spherical inclusions up to 3 μm in diameter. The lesions were consistent with Cottontail rabbit papillomavirus infection. Papilloma virus antigens were identified by immunohistochemistry. In addition, papillomavirus particles were identified by transmission electron microscopy within Langerhans cells of the epidermis, suggesting a unique mechanism for systemic dissemination of the virus. The present case report highlights the finding of viral particles within the Langerhans cells and suggests a novel mechanism of pathogenesis.
Cystic Echinococcosis Due to Echinococcus Equinus in a Horse from Southern Germany
Authors: Blutke Andreas , Hamel Dietmar , Hüttner Marion , Gehlen Heidrun , Romig Thomas , Pfister Kurt , Hermanns Walter
Published: May 2011

In Europe, cystic echinococcosis is rare in horses and is mostly diagnosed at slaughter or postmortem examination. Equine cystic echinococcosis can be caused by various Echinococcus taxa, but only Echinococcus equinus (the “horse strain”) is known to produce fertile cysts. In Europe, E. equinus appears to be endemic in Great Britain, Ireland, Spain, and Italy and has sporadically been reported in Belgium and Switzerland. The present report describes the first case of a molecularly confirmed E. equinus infection in a horse foaled and raised in Germany. The 19-year-old mare was presented for examination of inappetence, emaciation, and respiratory symptoms. X-ray radiographs of the thorax showed 2 well-circumscribed tumor-like masses, each approximately 10 cm in diameter in the caudal lung field. The horse was euthanized as its condition rapidly deteriorated. Necropsy revealed 2 thick-walled hydatid cysts, each 7–8 cm in diameter in the lung. The tri-layered cyst walls consisted of an outer adventitial layer, a laminated acellular intermediate layer, and an inner germinal membrane. Grossly, the cysts contained a clear, amber liquid with hydatid sand. Light microscopy of the hydatid sand revealed free protoscoleces, intact and ruptured brood capsules, calcareous corpuscles, and debris. Samples of protoscoleces underwent molecular characterization, and the diagnosis of E. equinus was confirmed by restriction fragment length polymorphism—polymerase chain reaction and sequence analysis of the complete mitochondrial nicotinamide adenine dinucleotide dehydrogenase subunit 1 gene.
Gastrointestinal Leiomyosarcoma in an Egyptian Fruit Bat (Rousettus Aegyptiacus)
Authors: Bradford Carol , Jennings Ryan , Ramos-Vara José
Published: May 2011

A 10-year-old female Egyptian fruit bat ( Rousettus aegyptiacus) was evaluated for weakness and lethargy. Despite clinical improvement after supportive therapy, the bat was found dead the following day. Gross necropsy identified a mass associated with the duodenal wall and right renal cortex. Microscopically, the duodenal and gastric tunica muscularis and serosa and the right renal cortex were effaced by interlacing bundles of neoplastic spindle cells that were diffusely and strongly positive for vimentin and alpha-smooth muscle actin. The neoplastic cells also had mild to moderate cytoplasmic labeling for laminin and S100 and were negative for c-kit and desmin. On the basis of cell morphology and the immunophenotype, this tumor was diagnosed as a gastrointestinal leiomyosarcoma.
Malignant B-Cell Lymphoma with Mott Cell Differentiation in a Ferret (Mustela Putorius Furo)
Authors: Gupta Aradhana , Gumber Sanjeev , Schnellbacher Rodney , Bauer Rudy W. , Gaunt Stephen D.
Published: May 2011

A 3.5-year-old, male, neutered ferret ( Mustela putorius furo) was presented with a 3-day history of lethargy and anorexia. Splenic aspirates revealed high numbers of intermediate-sized lymphocytes and Mott cells interpreted as lymphoma with Mott cells. The ferret was euthanized because of a poor clinical prognosis. Postmortem examination revealed markedly enlarged spleen and lymph nodes, with multifocal white nodules in the liver parenchyma. Histologically, the spleen had multifocal large nodules composed of neoplastic lymphocytes with frequent Mott cells. Similar neoplastic cells were present in the sections of liver, lymph nodes, and bone marrow. These cells were cluster of differentiation (CD)3-negative, CD79α-positive, and lambda light-chain—positive. Electron microscopy revealed that the cytoplasm of the neoplastic Mott cells had increased, disorganized, dilated, rough endoplasmic reticulum containing electron-dense immunoglobulin. On the basis of cytologic, histopathologic, immunohistochemical, and electron microscopic findings, a malignant B-cell lymphoma with Mott cell differentiation was diagnosed.
Nonsteroidal Anti-Inflammatory Drug—Associated Renal Papillary Necrosis in a White-Tailed Deer (Odocoileus Virginianus)
Authors: Stern Adam W. , Ritchey Jerry W. , Hall Brittany , Ketz-Riley Cornelia J. , Genova Suzanne G.
Published: May 2011

Renal papillary necrosis was diagnosed during postmortem examination of a juvenile white-tailed deer ( Odocoileus virginianus) from Oklahoma. The deer was surgically treated for a Salter Harris type II fracture of the proximal tibia of the left hind limb. The animal was administered multiple nonsteroidal anti-inflammatory drugs (NSAIDs), including meloxicam, flunixin meglumine, and ketoprofen for pain management. At postmortem examination, gross lesions included a proximal tibial Salter Harris type II fracture with an associated fibrinonecrotizing myositis and bilateral renal papillary necrosis. Histologically, the kidneys exhibited coagulation necrosis of the renal medulla and renal papilla, thrombosis of renal blood vessels, and interstitial medullary edema. The gross and microscopic lesion coupled with the clinical history of multiple NSAID administration suggests NSAID-induced renal papillary necrosis.
Characterization and Phylogenetic Analysis of Bovine Viral Diarrhea Virus in Brain Tissues from Nonambulatory (Downer) Cattle in Korea
Authors: Oem Jae-Ku , Chung Joon-Yee , Roh In-Soon , Kim Hye-Ryoung , Bae You-Chan , Lee Kyung-Hyun , Jin Young-Hwa , Lee O-Soo
Published: May 2011

Between August 2008 and May 2009, 386 brain and serum samples from adult cattle (2–7 years old) showing a variety of clinical signs of downer cow syndrome were received by the National Veterinary Research and Quarantine Service. All brain samples were tested for the presence of Bovine viral diarrhea virus (BVDV) by reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and antigen capture ELISA (Ag-ELISA). The BVDV nucleic acid was detected in 54 of 386 (15.5%) brain samples tested by RT-PCR. Positive results were detected in 14 (3.67%) and 13 (3.4%) of samples tested by IHC and Ag-ELISA, respectively. Both BVDV nucleic acid and antigen were detected in 11 cattle (2.9%) by all 3 diagnostic tests; however, antibodies against BVDV were not detected in these 11 cattle. A molecular classification of the identified viral strains ( n = 40) was also carried out. Neighbor-joining phylogenetic analysis revealed that most of the identified viruses belonged to BVDV genotype 1a ( n = 10), 1b ( n = 16), and 2a ( n = 8). The remaining strains were subtypes 1c ( n = 1), 1n ( n = 4), and 1m ( n = 1). Interestingly, most of the BVDV-1b strains ( n = 9) identified in brain samples were confirmed by all 3 diagnostic tests. Further studies should be performed to determine why the BVDV-1b strain was found in brain samples that were positive using all 3 diagnostic tests.
Expression and Validation of D-Erythrulose 1-Phosphate Dehydrogenase from Brucella abortus: A Diagnostic Reagent for Bovine Brucellosis
Authors: Eoh Hyungjin , Jeon Bo-Young , Kim Zhiyeol , Kim Seung-Cheol , Cho Sang-Nae
Published: May 2011

Brucella abortus is a bacterium of brucellosis causing abortion in cattle. The diagnosis of bovine brucellosis mainly relies on serologic tests using smooth lipopolysaccharide (S-LPS) from B. abortus. However, the usefulness of this method is limited by false-positive reactions due to cross-reaction with other Gram-negative bacteria. In the present study, the eryC gene encoding B. abortus D-erythrulose 1-phosphate dehydrogenase, which is involved in the erythritol metabolism in virulent B. abortus strain but is absent from a B. abortus vaccine strain (S19), was cloned. Recombinant EryC was expressed and purified for the evaluation as a diagnostic reagent for bovine brucellosis. Other B. abortus proteins, Omp16, PP26, and CP39 were also purified and their seroreactivities were compared. Recombinant EryC, Omp16, PP26, and PP39 were all reactive to B. abortus-positive serum. The specificity of recombinant Omp 16, PP26, CP39, and EryC, were shown to be approximately 98%, whereas that of B. abortus whole cell lysates was shown to be 95%. The sensitivity of Omp16, PP26, CP39, and EryC were 10%, 51%, 64%, and 43%, respectively, whereas that of B. abortus whole cell lysates was 53%. These results suggested that B. abortus EryC would be a potential reagent for diagnosis for bovine brucellosis as a single protein antigen.
Use of an Indirect Enzyme-Linked Immunosorbent Assay for Detection of Antibodies in Sheep Naturally Infected with Salmonella Abortusovis
Authors: Wirz-Dittus Sophie , Belloy Luc , Doherr Marcus G. , Hüssy Daniela , Sting Reinhard , Gabioud Patricia , Waldvogel Andreas S.
Published: May 2011

An indirect enzyme-linked immunosorbent assay (ELISA) was modified and validated to detect antibodies against Salmonella Abortusovis in naturally infected sheep. The ELISA was validated with 44 positive and 45 negative control serum samples. Compared with the immunoblot, the sensitivity and specificity of the assay were 98% and 100%, respectively. To follow antibody levels over time, samples from 12 infected ewes were collected at 1, 3, and 10 months after abortion. All animals showed antibody levels above the cutoff value throughout the observation period. One and 3 months after abortion, high antibody levels could be detected in all but one animal, whereas after 10 months, 9 animals had markedly lower but still positive antibody levels. The test characteristics and evidence for the persistence of detectable antibody levels in all infected animals for up to 10 months indicates that the ELISA can be used for herd surveillance testing.
Development of Microarray and Multiplex Polymerase Chain Reaction Assays for Identification of Serovars and Virulence Genes in Salmonella Enterica of Human or Animal Origin
Authors: Peterson Greg , Gerdes Bryan , Berges Jami , Nagaraja Tiruvoor G. , Frye Jonathan G. , Boyle David S. , Narayanan Sanjeev
Published: May 2011

Salmonella enterica is an important enteric pathogen consisting of many serovars that can cause severe clinical diseases in animals and humans. Rapid identification of Salmonella isolates is especially important for epidemiologic monitoring and controlling outbreaks of disease. Although immunologic and DNA-based serovar identification methods are available for rapid identification of isolates, they are time consuming or costly or both. In the current study, 2 molecular methods for identification of Salmonella serovars were developed and validated. A 70-mer oligonucleotide spotted microarray was developed that consisted of probes that detected genes responsible for genetic variation among isolates of Salmonella that can be used for serotyping. A multiplex polymerase chain reaction (PCR) assay was also developed, which is capable of identifying 42 serovars, thus providing a valuable prediction of the pathogenicity of the isolates by detecting the presence of virulence genes sseL, invA, and spvC. The gene spvC was the best predictor of pathogenicity. In a blind study, traditional serologic methods were correlated at 93.3% with the microarray-based method and 100% with the multiplex PCR-based serovar determination.
Experimental Rayless Goldenrod (Isocoma pluriflora) Toxicosis in Goats
Authors: Stegelmeier Bryan L. , Davis T. Zane , Green Benedict T. , Lee Stephen T. , Hall Jeffery O.
Published: May 2011

Rayless goldenrod ( Isocoma pluriflora) sporadically poisons livestock in the southwestern United States. Similarities with white snakeroot ( Ageratina altissima) poisoning and nearly identical chemical analyses led early researchers to conclude that tremetol, a mixture of benzofuran ketones, is the rayless goldenrod toxin. The toxicity of these ketone toxins have not been fully characterized nor are the pathogenesis and sequelae of poisoning completely understood. The objective of the current study was to characterize and describe the clinical and pathologic changes of rayless goldenrod toxicity in goats. Fifteen goats were gavaged with rayless goldenrod to obtain benzofuran ketone doses of 0, 10, 20, 40, and 60 mg/kg/day. After 7 treatment days, the goats were euthanized, necropsied, and tissues were processed for microscopic studies. After 5 or 6 days of treatment, the 40-mg/kg and 60-mg/kg goats were reluctant to move, stood with an erect stance, and became exercise intolerant. They had increased resting heart rate, prolonged recovery following exercise, and increased serum aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and creatinine kinase activities. All treated animals developed skeletal myopathy with dose-related distribution and severity. The goats dosed with 20 mg/kg and higher also developed myocardial degeneration and necrosis. Although skeletal myonecrosis was patchy and widely distributed, the quadriceps femoris was consistently damaged, even in low-dosed animals. Myocardial lesions were most severe in the papillary muscles of 60-mg/kg—dosed animals. This indicates that goats are highly susceptible to rayless goldenrod poisoning, and that the characteristic lesion of poisoning is skeletal and cardiac myonecrosis.
An improved Real-Time Polymerase Chain Reaction for the Simultaneous Detection of All Serotypes of Epizootic Hemorrhagic Disease Virus
Authors: Clavijo Alfonso , Sun Feng , Lester Thomas , Jasperson Dane C. , Wilson William C.
Published: May 2011

Epizootic hemorrhagic disease virus (EHDV) is a significant pathogen of wild and sometimes domestic ungulates worldwide. Rapid and reliable methods for virus detection and identification play an essential part in the control of epizootic hemorrhagic disease (EHD). In the present study, a 1-step real-time polymerase chain reaction (PCR) group-specific assay was developed. The assay detects genome segment 5 (NS1) from all of the 8 serotypes of EHDV. Assay sensitivity was evaluated relative to a conventional gel-based nested PCR using cell culture–derived virus and diagnostic samples from clinically affected white-tailed deer ( Odocoileus virginianus). The assay reliably amplified the NS1 gene from any of the EHDV strains tested, including isolates from each of the 8 EHDV serotypes. No cross-reactions were detected when all 24 serotypes of Bluetongue virus, a closely related member of the genus Orbivirus, were tested. A panel of 76 known EHDV-positive clinical samples was used to compare the performance of the assay relative to a previously reported real-time PCR assay. Results indicated that there was no statistically significant difference between the threshold cycle values obtained with both assays. A collection of 178 diagnostic samples submitted for EHD diagnosis was also used for test evaluation. The assay could be applied for rapid detection of EHDV in clinical samples from susceptible ruminants during an outbreak of the disease. In addition, this PCR assay has the benefits of being reliable and simple and could provide a valuable tool for studying the epidemiology of EHDV infection in susceptible ruminants by facilitating the detection of EHDV, regardless of the serotype.
Detection of Strain Variation in Isolates of Rhodococcus Equi from an Affected Foal Using Repetitive Sequence–Based Polymerase Chain Reaction
Authors: Bolton Tim , Kuskie Kyle , Halbert Natalie , Chaffin Keith , Healy Mimi , Lawhon Sara , Jackson Amanda , Cohen Noah
Published: May 2011

Rhodococcus equi is an important pathogen of foals aged 1–6 months. Evidence exists that foals are exposed to a wide diversity of R. equi strains in their environment. However, limited data are available regarding the extent to which genotypic variation exists among isolates infecting individual foals. Therefore, electrophoresis of repetitive sequence–based polymerase chain reaction (rep-PCR) amplicons in an automated microfluidics chip format was used to genotype 9 virulent R. equi isolates obtained from distinct anatomic locations in a single foal. Four of the isolates were obtained from different regions of the lung, and 5 were from abscessed intra-abdominal lymph nodes (LNs). Six distinct genotypes were identified among the 9 isolates. None of the pulmonary isolates was identical; however, a pulmonary isolate was found to be identical to an isolate recovered from a small intestinal LN, and another pulmonary isolate was identical to an isolate from a mesenteric LN. These results indicate that foals can be infected with multiple strains of virulent R. equi. Furthermore, identical strains can be found in multiple, remote anatomic locations in an infected foal, and this can occur for >1 strain in the same foal. The automated system used in the current study provided a rapid, reproducible, and discriminating method for typing R. equi isolates.
A Duplex Real-Time Polymerase Chain Reaction Assay for Differentiation between Bolbophorus Damnificus and Bolbophorus Type II Species Cercariae
Authors: Griffin Matt J. , Wise David J. , Yost Marlena C. , Doffitt Cynthia M. , Pote Linda M. , Greenway Terrence E. , Khoo Lester H.
Published: May 2011

A duplex quantitative real-time polymerase chain reaction (qPCR) assay was developed to differentiate between Bolbophorus damnificus and Bolbophorus type II species cercariae. Both trematode species are prevalent throughout the commercial catfish industry, as both infect the ram's horn snail, Planorbella trivolvis, which is commonly found in catfish ponds. Identification of cercaria to species is important in catfish disease challenge experiments, as only B. damnificus has been shown to have negative impacts on channel catfish. Oligonucleotide primers and fluorescence resonance energy transfer hydrolysis probes were designed to amplify the 18S small subunit ribosomal DNA gene of each species. The quantification cycle indicative of the number of cercariae in the sample prep was determined, and standard curves correlating to cercaria numbers were established. For both species, the assay was found to be highly repeatable and reproducible, with a linear dynamic range covering 7 orders of magnitude. The sensitivity limit of the assay was ∼1/256th of a cercaria, regardless of species, and there was no remarkable interference between the 2 assays when run simultaneously within the same reaction. In a field study, identification of cercaria by the duplex real-time qPCR assay was in complete agreement with previously established end-point PCR protocols, demonstrating the assay to be a more rapid, quantifiable means of parasite identification.
Secondary Pentobarbital Poisoning in Two Dogs: A Cautionary Tale
Authors: Kaiser Amanda M. , McFarland Warner , Siemion Roger S. , Raisbeck Merl F.
Published: May 2011

Two dogs, a 13-year-old spayed female and a 7-year-old neutered male, were diagnosed with pentobarbital poisoning. Follow-up investigation determined that the source of pentobarbital was the carcass of a horse that had been euthanized more than 2 years previously and that was also apparently responsible for the death of a least 1, and possibly 2, other dogs. The fact that the horse carcass remained lethally toxic more than 2 years after it was euthanized reemphasizes the necessity of proper disposal of euthanized animals.
Arthritis and Osteomyelitis Associated with Enterococcus Cecorum Infection in Broiler and Broiler Breeder Chickens in Ontario, Canada
Authors: Stalker Margaret J. , Brash Marina L. , Weisz Alexandru , Ouckama Rachel M. , Slavic Durda
Published: May 2011

In August 2008, an Ontario broiler chicken flock experienced an outbreak of lameness in 4-week-old birds, with morbidity reaching 7% by day 3. Necropsy examination and histopathology revealed arthritis of the hock, stifle, and coxofemoral joints, and femoral and vertebral osteomyelitis. Enterococcus cecorum was isolated from the lesions and identified by 16S ribosomal RNA sequencing. In October 2008, a second case of E. cecorum osteomyelitis involved a flock of 9-week-old broiler breeder chickens, with 2% of the male birds showing reluctance to walk. Necropsy examination revealed osteomyelitis and abscessation of the body of the caudal thoracic vertebra in affected birds, with impingement on the overlying spinal cord.
Fatal Septicemia Caused by Helicobacter Infection in a Pancake Tortoise (Malacochersus Tornieri)
Authors: Stacy Brian A. , Wellehan James F. X.
Published: May 2011

Bacteria of the genus Helicobacter are associated with disease in humans and animals. Reports of infection in reptiles are very limited. In the present study, pathological findings and molecular characterization are reported for a Helicobacter species associated with septicemia in a pancake tortoise ( Malacochersus tornieri). Disseminated infection resulted in regional cellulitis and edema of the head and neck, and pericarditis. Spiral bacteria were identified in cytological preparations and Warthin-Starry-stained sections of pathological lesions. Amplification of partial sequence of the 16S ribosomal gene using polymerase chain reaction identified the organism as Helicobacter and suggest that it is a novel species.
Severe Zinc Responsive Dermatosis in a Litter of Pharaoh Hounds
Authors: Campbell Gregory A. , Crow Dennis
Published: May 2011

A litter of 3-month-old Pharaoh Hound puppies presented to the referring veterinarian with severe generalized erythematous-crusted papules with pruritus, accompanied by exfoliation and erythema of footpads, inappetence, lethargy, and retarded growth. Three of 5 puppies (2 male and 1 female) were affected. Representative areas were biopsied from 1 affected male puppy and were routinely processed. Histologically, there was marked epidermal hyperplasia with a disorganized appearance of the epidermis and massive parakeratotic hyperkeratosis, compatible with zinc-responsive dermatosis. Low serum zinc concentrations were documented, and the affected animals partially responded to intravenous zinc supplementation but did not respond to oral supplementation. One male puppy died as a result of unrelated causes and was necropsied. The remaining 4 puppies were followed over 2 years. Growth was stunted, and enamel hypoplasia of permanent dentition developed compared with unaffected littermates. Intravenous zinc supplementation at 3-4 week intervals was required to prevent further skin lesion development. One dog died at 3 years of age of renal failure.
Tubular Hypoplasia of the Aorta and Right Atrioventricular Valve Dysplasia in a Bulldog
Authors: Robinson Nicholas A. , Armíen Aníbal G.
Published: May 2011

A Bulldog puppy that died at 1 day of age was presented for postmortem evaluation. Macroscopically, there was marked hypoplasia of the ascending, transverse, and proximal segments of the descending thoracic aorta and almost complete secondary thrombosis of the left ventricle causing a functional stenosis of the left atrioventricular valve. Separately, there was right atrioventricular valve dysplasia with secondary dilation of the right atrium. Microscopically, the left ventricular outflow tract was occluded by chondroid metaplasia, fibrosing recanalization of a left ventricular thrombus, and isolated Purkinje fiber degeneration and necrosis.