bluuebunny/crossref_metadata_2025 huggingface dataset:
Quality Control Validation, Application of Sigma Metrics, and Performance Comparison between Two Biochemistry Analyzers in a Commercial Veterinary Laboratory
Authors: Farr Alison J. , Freeman Kathleen P.
Published: May 2011

A review of the literature pertinent to interpretation of biochemistry data and quality control (QC) and proficiency testing data from 2 biochemistry analyzers was used to determine clinical quality requirements for biochemistry assays, characterize the performance of and calculate sigma metrics for the analytes run on the 2 analyzers, and perform QC validation in order to determine the needs for statistical QC for each analyzer. Quality requirements suitable for the analytes based on the needs of the authors' laboratory are presented. These requirements may or may not be appropriate for other laboratories, depending on the needs of the clients, species, and equipment performance capability. The majority of the analytes were easily controlled using the 13s control rule, with a sigma metric approaching or exceeding 6 and with a high probability of error detection and a low probability of false rejection. Some analytes could not be controlled using the 13s rule, and additional control rules with a greater number of control data points were required. There were differences between performances of the 2 analyzers. The findings in the present study emphasize the need for QC specific for the analyte and the clinical decision level and the need for separate QC validation on every instrument.
Pathologic Findings in Equine Muscle (Excluding Polysaccharide Storage): A Necropsy Study
Authors: Valentine Beth A.
Published: May 2011

Gross and histopathologic evaluation of skeletal muscle was performed in 229 equids (217 horses, 8 ponies, 3 donkeys, and 1 mule) 1 year of age or older undergoing postmortem examination at Oregon State University in a 2.5-year period. Animals were evaluated for grossly evident muscle lesions, and muscle samples were fixed in formalin, processed routinely, and stained with hematoxylin and eosin (HE) and periodic acid-Schiff (PAS) for glycogen. Muscle lesions were detected in 149 animals (65%). Chronic myopathic change (excessive fiber size variation and internal nuclei) was evaluated in horses without polysaccharide storage myopathy and was the most common finding (36 animals; 15.7%). Chronic myopathic change was more common in older animals. Generalized muscle atrophy was present in 30 animals (13.1%). Myonecrosis was attributed to endotoxic injury (11 animals; 4.8%), bone fracture (8 animals; 3.5%), bacterial infection (5 animals; 2.2%), muscle rupture (3 animals; 1.3%), selenium deficiency (2 animals; 0.9%), and exertional rhabdomyolysis (1 horse; 0.4%); cause was not determined in 9 animals (3.9%). Intramyofiber protozoa were detected in 19 horses and ponies (8.3%). Denervation atrophy was detected in 14 animals (6.1%). Neoplasia involving muscle occurred in 3 animals (1.3%), injection site reactions were detected in 4 animals (1.7%), and focal lymphocytic infiltrates were found in 6 animals (2.6%). Other findings were ring fibers (2 horses; 0.9%), fiber splitting (2 horses; 0.9%), and fat infiltration (1 horse; 0.4%). Skeletal muscle lesions are common in equids examined at postmortem. Transverse sections stained with HE and PAS are invaluable when evaluating equine muscle.
Effects of T Lymphocytes, Interleukin-1, and Interleukin-6 on Renal Fibrosis in Canine End-Stage Renal Disease
Authors: Yhee Ji-Young , Yu Chi-Ho , Kim Jong-Hyuk , Sur Jung-Hyang
Published: May 2011

Canine end-stage renal disease (ESRD) is defined as the almost complete failure of renal function or irreversible destruction and is characterized by extensive glomerular sclerosis, tubular atrophy, interstitial inflammation, and fibrosis. Renal fibrosis is a common pathway leading to kidney failure. Infiltrating immunocytes in the end-stage kidney and several related factors are involved in renal fibrogenesis. A total of 18 renal tissue samples were obtained from canine patients with ESRD using biopsy and necropsy procedures. The extent of renal fibrosis was histopathologically examined by Masson trichrome staining. T-cell and B-cell localization and macrophage lineages were determined by immunohistochemical staining. Additionally, interleukin-1 (IL-1), IL-2, and IL-6 levels in the canine ESRD kidney were immunohistochemically evaluated and compared with expression patterns in the normal kidney. Significant fibrosis and infiltrating immunocytes consistent with lymphocytes were observed. Although the B-cell count was increased in the end-stage kidney, immunostaining patterns disclosed a marked increase in the number of CD3+ cells. Furthermore, the remarkable increase in IL-1 and IL-6 levels suggests that T cells in the kidneys of dogs with ESRD spontaneously express these cytokines. In this study, the correlation between the degree of renal fibrosis and cytokines in canine ESRD was examined. The present study shows that T lymphocytes and IL-6 play important roles in renal fibrosis.
Analytical Validation of a Real-Time Reverse Transcription Polymerase Chain Reaction Test for Pan-American Lineage H7 Subtype Avian Influenza Viruses
Authors: Spackman Erica , Ip Hon S. , Suarez David L. , Slemons Richard D. , Stallknecht David E.
Published: May 2011

A real-time reverse transcription polymerase chain reaction test for the identification of the H7 subtype in North American Avian influenza viruses (AIVs) was first reported in 2002; however, recent AIV surveillance efforts in wild birds and H7 outbreaks in poultry demonstrated that the 2002 test did not detect all H7 AIVs present in North and South America. Therefore, a new test, the 2008 Pan-American H7 test, was developed by using recently available H7 nucleotide sequences. The analytical specificity of the new assay was characterized with an RNA panel composed of 19 H7 viruses from around the world and RNA from all hemagglutinin subtypes except H16. Specificity for North and South American lineage H7 viruses was observed. Assay limits of detection were determined to be between 103 and 104 gene copies per reaction with in vitro transcribed RNA, and 100.0 and 100.8 50% egg infectious doses per reaction. The 2008 Pan-American H7 test also was shown to perform similarly to the 2002 test with specimens from chickens experimentally exposed to A/Chicken/BritishColumbia/314514-2/04 H7N3 highly pathogenic AIV. Furthermore, the 2008 test was able to detect 100% ( n = 27) of the H7 AIV isolates recovered from North American wild birds in a 2006–2007 sample set (none of which were detected by the 2002 H7 test).
Comparison of Five Culture Methods for Salmonella Isolation from Swine Fecal Samples of Known Infection Status
Authors: Love Brenda C. , Rostagno Marcos H.
Published: May 2011

The current study was conducted to evaluate 5 bacteriologic culture methods (methods 1, 2, 3, 4, and 5) for recovery of Salmonella enterica from swine feces, both for sensitivity of detection (ability to recover Salmonella from a positive sample) and for specificity (not to inadvertently identify an organism as a Salmonella species in a negative sample). Fifty-six negative samples and 46 positive samples were processed using each of the 5 methods, which differed primarily in the combinations of enrichment media used. All negative samples were negative for Salmonella when cultured by all 5 methods (100% specificity). Two of the methods (methods 1 and 4) resulted in the recovery of significantly less ( P < 0.05) Salmonella when compared with the remaining 3 methods (methods 2, 3, and 5). No one method was successful in recovering Salmonella from all positive samples, although recovery with method 2 was statistically similar to the total number of positive samples analyzed (42 vs. 46 Salmonella-positive samples, P > 0.05). This study shows that culture methods differ significantly in their performance regarding the isolation of Salmonella from swine fecal samples.
Persistent Bovine Viral Diarrhea virus Infection in Wild Cervids of Colorado
Authors: Duncan Colleen , Van Campen Hana , Soto Sonia , LeVan Ivy K. , Baeten Laurie A. , Miller Michael W.
Published: May 2011

Bovine viral diarrhea virus (BVDV) is a significant viral pathogen of domestic cattle. Worldwide, there is evidence of BVDV exposure and infection in wild ungulates; however, the frequency and significance of such events are unknown. To determine the prevalence and distribution of Colorado deer, elk, and moose persistently infected (PI) with BVDV, a cross-sectional study was conducted using full-thickness ear tissue samples collected from animals presented to the Colorado Division of Wildlife for chronic wasting disease surveillance in the 2005–2006 hunting season. Tissue from 5,597 harvested animals (2,934 mule deer, 2,516 elk, 141 white-tailed deer, and 6 moose) was paraffin-embedded and stained for BVDV using immunohistochemistry. A single adult male mule deer had BVDV antigen in the skin; staining distribution was consistent with that seen in PI cattle. Skin and lymph node were also positive for viral RNA by polymerase chain reaction, and the virus was determined to be a type 1. The prevalence of BVDV PI cervids in Colorado is very low. However, the identification of a naturally infected adult PI animal in the wild suggests that the virus infects free-ranging populations. The source of the BVDV is unknown and is assumed to be spillover from cattle or maintenance within wildlife populations. Consideration of a potential wild animal reservoir is important in the design and implementation of BVDV management practices in cattle.
Bronchopneumonia Associated with Extraintestinal Pathogenic Escherichia Coli in a Horse
Authors: DebRoy Chitrita , Roberts Elisabeth , Jayarao Bhushan M. , Brooks Jason W.
Published: May 2011

Extraintestinal pathogenic Escherichia coli (ExPEC) strains carrying distinct virulence attributes are known to cause diseases in humans and animals and infect organs other than the gastrointestinal tract. A fatal case of bronchopneumonia in a 12-year-old female Quarterhorse was investigated. Following postmortem examination, E. coli, Enterococcus sp., and Klebsiella pneumonia were isolated from the lungs, which contained multifocal intra-alveolar accumulations of neutrophils and macrophages with edema, hemorrhage, and fibrin. The strain of E. coli belonged to O2:H21 and carried virulence genes cnfl, sfa, foc, fimA, and papG allele I that are known to be associated with ExPEC strains. The strain was resistant to several antimicrobials including clindamycin, erythromycin, oxacillin, penicillin, and rifampin. This is the first report, to the authors' knowledge, in which ExPEC O2:H21 has been associated with fatal bronchopneumonia in a horse.
Bilateral Perirenal Hemorrhage in two Stone's Sheep (Ovis Dalli Stonei): A Possible Manifestation of Malignant Catarrhal Fever
Authors: Himsworth Chelsea G. , Harms Jane N. , Wobeser Gary , Hill Janet
Published: May 2011

Malignant catarrhal fever (MCF)-like disease was diagnosed at postmortem in 2 Stone's sheep ( Ovis dalli stonei). On gross examination, the predominant abnormality in both sheep was severe perirenal hemorrhage and multiple renal infarcts. Microscopically, there was severe, multisystemic lymphocytic arteritis. Both sheep were positive for Ovine herpesvirus 2 (OvHV-2) on polymerase chain reaction, and partial sequencing of the viral DNA polymerase genes from the 2 sheep revealed >99% homology, with 96% similarity to the reference GenBank OvHV-2 viral sequence. Based on the histological lesions, polymerase chain reaction results, and viral DNA polymerase gene sequencing, a diagnosis of OvHV-2-mediated MCF was made. Massive perirenal hemorrhage has not been described previously as a manifestation of MCF.
Chronic Wasting Disease in a Wisconsin White-Tailed Deer Farm
Authors: Keane Delwyn P. , Barr Daniel J. , Bochsler Philip N. , Hall S. Mark , Gidlewski Thomas , O'Rourke Katherine I. , Spraker Terry R. , Samuel Michael D.
Published: May 2011

In September 2002, chronic wasting disease (CWD), a prion disorder of captive and wild cervids, was diagnosed in a white-tailed deer ( Odocoileus virginianus) from a captive farm in Wisconsin. The facility was subsequently quarantined, and in January 2006 the remaining 76 deer were depopulated. Sixty animals (79%) were found to be positive by immunohistochemical staining for the abnormal prion protein (PrPCWD)in at least one tissue; the prevalence of positive staining was high even in young deer. Although none of the deer displayed clinical signs suggestive of CWD at depopulation, 49 deer had considerable accumulation of the abnormal prion in the medulla at the level of the obex. Extraneural accumulation of the abnormal protein was observed in 59 deer, with accumulation in the retropharyngeal lymph node in 58of 59 (98%), in the tonsil in 56 of 59 (95%), and in the rectal mucosal lymphoid tissue in 48 of 58 (83%). The retina was positive in 4 deer, all with marked accumulation of prion in the obex. One deer was considered positive for PrPCWD in the brain but not in the extraneural tissue, a novel observation in white-tailed deer. The infection rate in captive deer was 20-fold higher than in wild deer. Although weakly related to infection rates in extraneural tissues, prion genotype was strongly linked to progression of prion accumulation in the obex. Antemortem testing by biopsy of rectoanal mucosal-associated lymphoid tissue (or other peripheral lymphoid tissue) may be a useful adjunct to tonsil biopsy for surveillance in captive herds at risk for CWD infection.
Genotyping North American Animal Mycobacterium Bovis Isolates Using Multilocus Variable Number Tandem Repeat Analysis
Authors: Martinez Lorene R. , Harris Beth , Black William C. , Meyer Robert M. , Brennan Patrick J. , Vissa Varalakshmi D. , Jones Robert L.
Published: May 2011

Multilocus variable number tandem repeat analysis (MLVA) has recently emerged as a genotyping method that is both robust and highly discriminatory for the differentiation of Mycobacterium tuberculosis complex (MTBC) strains, including Mycobacterium bovis. However, MLVA assessment of M. bovis isolates recovered from animals in North America has been limited. Using an epidemiologically diverse set of 41 North American M. bovis animal isolates, MLVA, based on 27 published variable number tandem repeat (VNTR) loci, was evaluated. Nineteen loci displayed polymorphism, which resulted in differentiation of 21 unique MLVA genotypes. A subset of 6 loci differentiated the isolates into 14 genetically related groups that displayed remarkable concordance with the epidemiological data gathered via traditional trace-back methods. In most cases, MLVA exhibited greater resolution than spoligotyping, which differentiated the isolates into 11 groups. MLVA genotyping of M. bovis shows great potential as a molecular typing tool for characterizing the epidemiology of M. bovis animal infections in North America. However, the greatest resolution was achieved by using a combination of both MLVA and spoligotyping.
Comparison of Three Enzyme-Linked Immunosorbent Assays to Detect Porcine Circovirus-2 (PCV-2)—Specific Antibodies after Vaccination or Inoculation of Pigs with Distinct PCV-1 or PCV-2 Isolates
Authors: Patterson Abby R. , Johnson John , Ramamoorthy Sheela , Meng Xiang-Jin , Halbur Patrick G. , Opriessnig Tanja
Published: May 2011

Porcine circovirus-2 (PCV-2) serology is frequently used to determine PCV-2 status and optimal timing of PCV-2 vaccination in the field. The objectives of the current study are to determine the diagnostic accuracy of 3 currently available commercial anti-immunoglobulin G (IgG) PCV-2 enzyme-linked immunosorbent assays (ELISAs) and to compare the ability of the 3 assays to detect and differentiate between anti—PCV-2a and anti—PCV-2b antibodies, as well as anti—PCV-2 and anti—PCV-1 antibodies. Fifty-five 3-week-old, conventional pigs were randomly allocated to 7 groups: 1) negative controls ( n = 7), 2) PCV-2a ( n = 8; inoculated with PCV-2 ISU-40895), 3) PCV-2b ( n = 8; inoculated with PCV-2 NC-16845), 4) PCV-1 ( n = 8), 5) vaccine A ( n = 8; Ingelvac® CircoFLEX™), 6) vaccine B ( n = 8; Circumvent® PCV2), and 7) vaccine C ( n = 8; Suvaxyn® PCV2 One Dose). Blood samples were collected weekly, and all sera were tested by 3 different anti-PCV-2 IgG ELISAs. The results indicated that all ELISAs had area under the receiver operating curve values greater than 0.94, detected both anti-PCV-2a and −2b antibodies with no differentiation, and did not detect anti-PCV-1 antibodies in infected animals. One of the ELISAs was able to distinguish pigs vaccinated with vaccine B from pigs inoculated with either PCV-2a or PCV-2b.
Serum Total Sialic Acid in Pigs: New Possibilities for an Old Inflammatory Biomarker
Authors: Soler Laura , Silvia Martínez-Subiela , Tecles Fernando , Gutierrez Ana , Fuentes Pablo , Joaquín Jose Cerón
Published: May 2011

The potential use of total sialic acid (TSA) as an acute-phase biomarker in pigs was investigated through clinical and analytic validation of an automated commercial enzymatic-colorimetric assay. Intra- and interassay precision, accuracy, and detection limit were determined for the analytical validation of the method. The intra- and interassay coefficients of variation (%CV) were 2.27% and 3.99%, respectively, whereas the linearity under dilution calculation resulted in a coefficients of correlation of r = 0.995 and r = 0.999 for TSA high- and low-content serum samples, respectively, which indicated that the automated method was reliable to measure TSA in porcine serum samples. Average serum TSA concentrations in diseased pigs (median: 1779.16 mg/l; 5th percentile: 556.62 mg/l; 95th percentile: 963.06 mg/l) were 2.5-fold higher compared with healthy animals (median: 692.78 mg/l; 5th percentile: 1278.74 mg/l; 95th percentile: 2536.12 mg/l), which indicated that TSA can be considered as a moderate acute phase marker in swine. Therefore, TSA may be useful for detecting inflammatory conditions and could be included in biochemical profiles used to monitor the general status of pigs as a moderate acute phase marker.
Presumed Primary Muscular Lymphoma in a Dog
Authors: Thuilliez Céline , Watrelot-Virieux Dorothée , Chanut Franck , Fournel-Fleury Corinne , Ponce Frédérique , Marchal Thierry
Published: May 2011

A case of presumed primary muscular lymphoma in an 8-year-old, intact, male Newfoundland dog is reported. The dog was presented for evaluation of an infiltrating ventral cervical mass, respiratory distress, and anorexia of 1-month duration. Fine-needle aspiration of the mass revealed anaplastic large cell lymphoma. Despite chemotherapy, health status declined and the animal was euthanized a few weeks later. At necropsy, the mass infiltrated the cervical muscles and extended ventrally to the left forelimb and cranially to the tongue and laryngeal musculature. Other muscles were infiltrated by the same neoplasm (diaphragm and intercostal, abdominal, and gluteal muscles) indicating a probable multicentric origin. Histological examination confirmed the diagnosis of anaplastic large cell lymphoma, which showed a strong muscular tropism. Immunohistochemical staining revealed neoplastic cell reactivity for cluster of differentiation 3 (CD3) and Ki-67 antigens (70% and 90%, respectively). The neoplastic cells were negative for CD79a. The presumed histological diagnosis in this dog was primary muscular anaplastic large T-cell lymphoma.
Abortion in a Horse Following Neorickettsia Risticii Infection
Authors: Coffman Elizabeth A. , Abd-Eldaim Mohamed , Craig Linden E.
Published: May 2011

A pregnant 18-year-old Quarterhorse mare presented with fever, anorexia, tachycardia, tachypnea, and gastrointestinal hypermotility at day 68 of gestation. Potomac horse fever was diagnosed based on polymerase chain reaction (PCR) analysis of whole blood and a high antibody titer to Neorickettsia risticii. The mare made a rapid clinical recovery following antibiotic therapy, but aborted 98 days later. Necropsy on the aborted fetus revealed lymphohistiocytic colitis, lymphadenitis, myocarditis, and hepatitis. The placenta was grossly and histologically normal. Formalin-fixed lymph node, thymus, liver, and colon taken from the aborted fetus were positive by PCR for N. risticii DNA. Potomac horse fever is a common disease in horses that may result in delayed abortion. The microscopic lesions in the fetus are characteristic, and the diagnosis can be confirmed by PCR on formalin-fixed tissues.
Immunohistochemical Features of a Feline Spinal Cord Gemistocytic Astrocytoma
Authors: Aloisio Fabio , Levine Jonathan M. , Edwards John F.
Published: May 2011

A gemistocytic astrocytoma is described in the spinal cord of a 3-year-old, spayed, female, Domestic Shorthair cat. The neoplasm was tested for glial fibrillar acidic protein (GFAP), epidermal growth factor receptor (EGFR), and p53 protein expression. On histologic exanimation, the neoplastic cells were pleomorphic, with distinct cell borders and abundant cytoplasm that frequently extended into variably sized fibrillar processes. Neoplastic cells were strongly positive for GFAP and negative for EGFR. Eight percent (mean percentage) of neoplastic cells were p53 positive. Only 1 p53-positive cell was found in the normal spinal cord. The increased expression of p53 in the neoplasm was suggestive of an abnormal biological behavior of the p53 protein and its possible involvement in the neoplastic process of feline astrocytoma.
Mandibular and Maxillary Osteomyelitis and Myositis in a Captive Herd of Red Kangaroos (Macropus Rufus)
Authors: Brookins Milagros D. , Rajeev Sreekumari , Thornhill Timothy D. , Kreinheder Kurt , Miller Debra L.
Published: May 2011

Jaw infections in macropods are common and will result in mortality if not promptly diagnosed and aggressively treated. They have most often been reported in wallabies; however, in the current case, the gross and histopathologic findings, microbial culture, and management of jaw infections in a population of red kangaroos ( Macropus rufus) housed in a zoological park are described. Three red kangaroos, among a group of 23, were submitted for necropsy after death after progressively invasive and nonresolving jaw infections. Extensive bone and soft-tissue inflammation and necrosis were observed in all animals. A mixed population of aerobic and anaerobic bacteria was cultured; however, Pseudomonas spp. was consistently isolated from the sites of infection in all animals. Parental administration of gentamicin and penicillin, along with daily oral flushing of the wounds with Betadine, removal of all rough forage, and hand feeding soft-pelleted feed, was effective in controlling the progression of disease in 1 affected animal. This case documents an important disease in an additional macropod species and identifies predisposing factors, possible etiologies, and treatment and/or management options.
Association between Mycobacterium Avium Subsp. Paratuberculosis Infection among Offspring and their Dams in Nondomestic Ruminant Species Housed in a Zoo
Authors: Witte Carmel L. , Hungerford Laura L. , Rideout Bruce A.
Published: May 2011

The objective of the present study was to determine whether offspring of dams infected with Mycobacterium avium subsp. paratuberculosis (Map) have an increased risk for Map infection. Antemortem and postmortem disease surveillance data were used to identify positive and test-negative ruminants born at the Zoological Society of San Diego (ZSSD) between 1991 and 2007 and to estimate cumulative lifetime incidence. A matched case-control study, nested within the population, was conducted and conditional logistic regression analyses were used to quantify the association between infection status of offspring and their dams. Cases (infected ruminants, n = 47) were matched to controls (test-negative ruminants, n = 152) by species, birth date, birth enclosure, and follow-up time to control for confounding factors. The overall cumulative lifetime incidence of infection was estimated at 2.2%, but it decreased over time and varied by species. There was a significant association between infection status of offspring and their dams (odds ratio [OR] = 6.8, P < 0.01), which is consistent with studies in domestic livestock species. The association was stronger for animals whose dam was diagnosed within 2 years of their birth (OR = 9.0, P < 0.01) than for animals whose dam was diagnosed more than 2 years after their birth (OR = 6.0, P < 0.01) compared to animals with test-negative dams. For positive animals born to a positive dam, 85.3% of the Map infections were attributable to having a positive dam. For the entire population of ZSSD ruminants, 36.8% of the cases were attributable to having a positive dam. Findings will help guide future management of Map infection in zoo ruminant populations.
Cerebral Injury from Intracarotid Injection in an Alpaca (Vicugna Pacos)
Authors: Valentine Beth A. , Riebold Thomas W. , Wolff Peregrine L. , Sanchez Andrea L.
Published: May 2011

A mixture of ketamine, xylazine, and butorphanol was inadvertently injected into the right carotid artery of a 1-year-old alpaca. Injection was followed by a brief period of recumbency and seizure activity. The alpaca recovered, but was euthanatized 72 hr later because of development of progressive neurologic deficits. Pathologic findings were confined to the right cerebrum, meninges, thalamus, and hippocampus. Cerebrocortical edema with astrocytic reaction, perivascular hemorrhage and neutrophilic infiltration, and fibrinoid necrosis of vasculature within the meninges and thalamus were the most prominent lesions. Neuronal necrosis was mild. Astrocytic reaction within the right cerebral cortex was confirmed with immunohistochemistry for glial fibrillary acidic protein.
Serum Acute Phase Protein Concentrations in Female Dogs with Mammary Tumors
Authors: Tecles Fernando , Caldín Marco , Zanella Anna , Membiela Francisco , Tvarijonaviciute Asta , Subiela Silvia Martínez , Cerón José Joaquín
Published: May 2011

Acute phase proteins (APPs) are proteins whose concentrations in serum change after any inflammatory stimulus or tissue damage. The aim of the current study was to evaluate 3 positive APPs (C-reactive protein, serum amyloid A, and haptoglobin) and 1 negative APP (albumin) in female dogs with mammary neoplasia. Acute phase proteins were studied in 70 female dogs aged 8–12 years in the following groups: healthy ( n = 10); mammary tumors in stages I ( n = 19), II ( n = 5), III ( n = 6), IV ( n = 5), and V ( n = 7); and with mammary neoplasia plus a concomitant disease ( n = 18). In animals with mammary neoplasia, significant increases of positive APPs were only detected in those that had metastasis or a neoplasm with a diameter greater than 5 cm and ulceration. Dogs with mammary neoplasia and a concomitant disease also had high C-reactive protein concentrations. Albumin concentration was decreased in animals with metastasis and with a concomitant disease. The results of the present study indicate that the acute phase response could be stimulated in female dogs with mammary gland tumors because of different factors, such as metastasis, large size of the primary mass, and ulceration or secondary inflammation of the neoplasm.
Using Roquefortine C as a Biomarker for Penitrem A Intoxication
Authors: Tiwary Asheesh K. , Puschner Birgit , Poppenga Robert H.
Published: May 2011

Penitrem A is a well-recognized tremorgenic mycotoxin produced by several Penicillium spp. However, most natural cases of penitrem A intoxication have been associated with Penicillium crustosum. Another Penicillium sp., Penicillium roqueforti, is used for the production of blue cheese and is found in silage and feeds. Penicillium roqueforti produces a mycotoxin, roquefortine C, which is also produced by P. crustosum. In contrast to a tremorgenic syndrome produced by penitrem A, roquefortine C toxicosis is characterized by a paralytic syndrome. Two cases of penitrem A intoxication in dogs are presented to investigate the use of roquefortine C as a biomarker for penitrem A exposure. The vomitus, serum, and urine were analyzed for roquefortine C and penitrem A. Results suggest that roquefortine C can be a sensitive biomarker for penitrem A intoxication. However, the detection of roquefortine C in the absence of penitrem A could merely suggest ingestion of blue cheese or spoilt silage or feed. A review of the literature did not identify any case positive for penitrem A but negative for roquefortine C. In cases in which both mycotoxins were detected, roquefortine C concentration was always higher than penitrem A concentration. In contrast, several cases have been described where the clinical history suggested penitrem A intoxication, but only roquefortine C was detected. In conclusion, roquefortine C can serve as a sensitive biomarker for penitrem A intoxication, but the clinical presentation needs to be considered for proper interpretation of its detection in the absence of penitrem A.
Spinal Lymphoma and Pulmonary Filariasis in a Pet Domestic Rabbit (Oryctolagus Cuniculus Domesticus)
Authors: Reed Scott D. , Shaw Shannon , Evans Dawn E.
Published: May 2011

Spinal lymphoma and concurrent pulmonary filariasis are reported in a pet rabbit. The rabbit presented for pelvic limb paralysis resulting from extradural spinal lymphoma, presumably rising from the body of the sixth lumbar vertebra. The neoplasm was subsequently immunophenotyped as a B-cell lymphoma. Pulmonary filariasis was an incidental finding at necropsy.
Evidence for Clostridium Septicum as a Primary Cause of Cellulitis in Commercial Turkeys
Authors: Tellez Guillermo , Pumford Neil R. , Morgan Marion J. , Wolfenden Amanda D. , Hargis Billy M.
Published: May 2011

Etiology and methods of immunoprophylaxis against common field cellulitis in commercial turkeys were evaluated. It was determined that intravenous administration (∼10 8 cells/ml) of 1 of 4 isolates of Clostridium septicum from cellulitis lesions rapidly caused the classic lesions of cellulitis followed by death within 36 hr at high doses. When the supernatant alone was injected into turkey poults, signs of depression and ataxia were temporarily observed for up to 20 hr after injection, but no cellulitis lesions were detected. An enzyme-linked immunosorbent assay was developed to measure antibody levels against the isolated C. septicum. This assay was used to predict susceptibility to infection. An experimental formalin-killed bacterin/toxoid was produced from the challenge strain of C. septicum with inactivation timed to allow toxin accumulation and ∼108 cells/ml. This bacterin/toxoid given at day of hatch generated a rapid and persistent antibody response against the homologous C. septicum in the vaccinated birds at 9 weeks ( P < 0.001). The ability of this experimental vaccine to protect birds in the field as well as its ability to evaluate unvaccinated flocks to establish the time of seroconversion and the relationship to clinical disease are currently under evaluation.
Metastatic Angioinvasive Lymphoma (Lymphomatoid Granulomatosis) in a Cat
Authors: Rogers Douglas G. , Aliano Valerie A.
Published: May 2011

A subcutaneous mass removed from the right rear leg of a 17-year-old, spayed, female Domestic Shorthair cat was characterized histopathologically by granulomatous inflammation, sheets of large atypical lymphoid cells, and necrosis. The walls of the small and medium caliber blood vessels were invaded transmurally by atypical lymphoid cells. A diagnosis of angioinvasive lymphoma (AIL), or lymphomatoid granulomatosis, was made based on histopathologic findings. The cat was euthanized 2 months later because of recurrence of the mass and elevated serum alanine aminotransferase activity, and a necropsy was performed. The histopathologic lesion of AIL was seen in the skin and subcutis of the right rear leg, and neoplastic cell infiltrates were seen in adjacent skeletal muscle, right superficial inguinal lymph node, liver, and spleen. By immunohistochemistry, variable numbers of neoplastic cells expressed B-lymphocyte antigen 36 (BLA36) or cluster of differentiation (CD)3 markers, indicative of B- and T-cell lineages, respectively. Neoplastic cells were uniformly positive for vimentin and uniformly negative for cytokeratins and myeloid/histiocytic antigen. Although the cat had received a Rabies virus vaccine subcutaneously in the right rear leg 6 months earlier, the AIL lesion was not typical of vaccine-induced sarcomas. The AIL in this cat was unusual because the extrapulmonary metastases involved the skin and subcutis.
Systemic Adenovirus Infection in Sulawesi Tortoises (Indotestudo Forsteni) Caused by a Novel Siadenovirus
Authors: Rivera Sam , Wellehan James F. X , McManamon Rita , Innis Charles J. , Garner Michael M. , Raphael Bonnie L. , Gregory Christopher R. , Latimer Kenneth S. , Rodriguez Carlos E. , Diaz-Figueroa Orlando , Marlar Annajane B. , Nyaoke Akinyi , Gates Amy E. , Gilbert Kelly , Childress April L. , Risatti Guillermo R. , Frasca Salvatore
Published: May 2011

A novel siadenovirus was identified in the Sulawesi tortoise ( Indotestudo forsteni). A group of 105 Sulawesi tortoises was obtained by the Turtle Survival Alliance. Many of the tortoises were in poor health. Clinical signs included anorexia, lethargy, mucosal ulcerations and palatine erosions of the oral cavity, nasal and ocular discharge, and diarrhea. Initial diagnostic tests included fecal testing for parasites, complete blood count and plasma biochemical analysis, mycoplasma serology, and polymerase chain reaction (PCR) testing for intranuclear coccidia and chelonian herpesvirus. Treatment included administration of antibiotics, antiparasitic medications, parenteral fluids, and nutritional support. Tissue samples from animals that died were submitted for histopathologic evaluation. Histopathologic examination revealed systemic inflammation and necrosis associated with intranuclear inclusions consistent with a systemic viral infection in 35 tortoises out of 50 examined. Fecal testing results and histopathologic findings revealed intestinal and hepatic amoebiasis and nematodiasis in 31 animals. Two of 5 tortoises tested by PCR were positive for Chlamydophila sp. Aeromonas hydrophila and Escherichia coli were cultured from multiple organs of 2 animals. The mycoplasma serology and PCR results for intranuclear coccidia and chelonian herpesvirus were negative. Polymerase chain reaction testing of tissues, plasma, and choanal/cloacal samples from 41 out of 42 tortoises tested were positive for an adenovirus, which was characterized by sequence analysis and molecular phylogenetic inference as a novel adenovirus of the genus Siadenovirus. The present report details the clinical and anatomic pathologic findings associated with systemic infection of Sulawesi tortoises by this novel Siadenovirus, which extends the known reptilian adenoviruses to the chelonians and extends the known genera of reptilian Adenoviridae beyond Atadenovirus to include the genus Siadenovirus.
The Use of Geographic Information System (GIS) and Non-GIS Methods to Assess the External Validity of Samples Postcollection
Authors: Richardson Esther , Good Margaret , McGrath Guy , More Simon J.
Published: May 2011

External validity is fundamental to veterinary diagnostic investigation, reflecting the accuracy with which sample results can be extrapolated to a broader population of interest. Probability sampling methods are routinely used during the collection of samples from populations, specifically to maximize external validity. Nonprobability sampling (e.g., of blood samples collected as part of routine surveillance programs or laboratory submissions) may provide useful data for further posthoc epidemiological analysis, adding value to the collection and submission of samples. As the sample has already been submitted, the analyst or investigator does not have any control over the sampling methodology, and hence external validity as routine probability sampling methods may not have been employed. The current study describes several Geographic Information System (GIS) and non-GIS methods, applied posthoc, to assess the external validity of samples collected using both probability and nonprobability sampling methods. These methods could equally be employed for inspecting other datasets. Mapping was conducted using ArcView 9.1. Based on this posthoc assessment, results from the random field sample could provide an externally valid, albeit relatively imprecise, estimate of national disease prevalence, of disease prevalence in 3 of the 4 provinces (all but Ulster, in the north and northwest, where sample size was small), and in beef and dairy herds. This study provides practical methods for examining the external validity of samples postcollection.
Evaluation of a 384–Well Format for High-Throughput Real-Time Reverse Transcription Polymerase Chain Reaction for Avian Influenza Testing
Authors: Ferro Pamela J. , Osterstock Jason , Norby Bo , Fosgate Geoffrey T. , Lupiani Blanca
Published: May 2011

As concerns over the global spread of highly pathogenic avian influenza H5N1 have heightened, more countries are faced with increased surveillance efforts and incident response planning for handling a potential outbreak. The incorporation of molecular techniques in most diagnostic laboratories has enabled fast and efficient testing of many agents of concern, including avian influenza. However, the need for high-throughput testing remains. In this study, the use of a 384–well format for high-throughput real-time reverse transcription polymerase chain reaction (real-time RT-PCR) testing for avian influenza is described. The analytical sensitivity of a real-time RT-PCR assay for avian influenza virus matrix gene with the use of both 96– and 384–well assay formats and serial dilutions of transcribed control RNA were comparable, resulting in similar limits of detection. Of 28 hunter-collected cloacal swabs that were positive by virus isolation, 26 (92.9%) and 27 (96.4%) were positive in the 96– and 384–well assays, respectively; of the 340 hunter-collected swabs that were negative by virus isolation, 45 (13.2%) and 23 (6.8%) were positive in the 96– and 384–well assays, respectively. The data presented herein supports the utility of the 384–well format in the event of an avian influenza outbreak for high-throughput real-time RT-PCR testing.
Evaluation of Clinical Laboratory Standards Institute Interpretive Criteria for Methicillin-Resistant Staphylococcus Pseudintermedius Isolated from Dogs
Authors: Schissler Jennifer R. , Hillier Andrew , Daniels Joshua B. , Cole Lynette K. , Gebreyes Wondwossen A.
Published: May 2011

The Clinical and Laboratory Standards Institute published in 2008 new interpretive criteria for the identification of methicillin resistance in staphylococci isolated from animals. The sensitivity of the 2008 interpretive criteria for mecA gene-positive Staphylococcus pseudintermedius, compared with the previous criteria of 2004, was investigated. Thirty clinical isolates of methicillin-resistant S. pseudintermedius from dogs were used. The presence of the mecA gene was determined by polymerase chain reaction. The minimum inhibitory concentration for oxacillin was determined by broth microdilution. The 2008 breakpoint of 4 μg/ml for methicillin resistance resulted in a diagnostic sensitivity of 73.3% (22/30). The 2004 breakpoint guideline of 0.5 μg/ml resulted in a diagnostic sensitivity of 97% (29/30). For oxacillin disk diffusion, the 2008 interpretive criterion of 10 mm for methicillin resistance resulted in a sensitivity of 70% (21/30). If intermediate isolates (11 or 12 mm) were considered resistant, the sensitivity was 93% (28/30). Application of the 2004 interpretive criterion of 17 mm resulted in a diagnostic sensitivity of 100% (30/30). For cefoxitin disk diffusion, the interpretive criterion of 21 mm for methicillin resistance (as used for Staphylococcus aureus) resulted in a diagnostic sensitivity of 6.7% (2/30). The interpretive criterion of 24 mm (as used for coagulase-negative staphylococci) resulted in a diagnostic sensitivity of 43.3% (13/30). With the use of 2008 interpretive criteria, all 3 tests produced what we consider to be an unacceptable level of false negative results. Our findings also suggest that cefoxitin disk diffusion is an inappropriate screening test for methicillin resistance of canine S. pseudintermedius.
Gastric B-Cell Lymphoma with Mott Cell Differentiation in a Dog
Authors: De Zan Gabrita , Zappulli Valentina , Cavicchioli Laura , Martino Linda Di , Ros Eriberta , Conforto Giorgia , Castagnaro Massimo
Published: May 2011

A gastric lymphoid tumor with involvement of regional lymph nodes and spleen was diagnosed in an 8-year-old crossbreed male dog with a 6-month history of gastrointestinal disease. Despite surgical excision and palliative therapy (prednisolone and cimetidine), the dog was euthanized due to worsening of clinical signs. At necropsy, multiple white, solid, nodular, infiltrative masses were observed in the stomach, duodenum, spleen, liver, and lungs in association with generalized lymph node enlargement. Cytology, histology, histochemistry, immunohistochemistry, and electron microscopy revealed that the neoplastic cell population was composed of B lymphocytes that contained variable amounts of round periodic acid-Schiff-positive cytoplasmic globules consistent with Russell bodies. The tumor most likely represented a variant of B-cell neoplasia with extensive Mott cell differentiation.
Gastric Cryptosporidiosis in Freshwater Angelfish (Pterophyllum Scalare)
Authors: Murphy Brian G. , Bradway Daniel , Walsh Timothy , Sanders George E. , Snekvik Kevin
Published: May 2011

A freshwater angelfish ( Pterophyllum scalare) hatchery experienced variable levels of emaciation, poor growth rates, swollen coelomic cavities, anorexia, listlessness, and increased mortality within their fish. Multiple chemotherapeutic trials had been attempted without success. In affected fish, large numbers of protozoa were identified both histologically and ultrastructurally associated with the gastric mucosa. The youngest cohort of parasitized fish was the most severely affected and demonstrated the greatest morbidity and mortality. The protozoa were morphologically most consistent with Cryptosporidium. All of the protozoan life stages were identified ultrastructurally and protozoal genomic DNA was isolated from parasitized tissue viscera and sequenced. Histological, ultrastructural, genetic, and phylogenetic analyses confirmed this protozoal organism to be a novel species of Cryptosporidium.
Evaluation of Three Rapid Diagnostic Tests Used in Bovine Spongiform Encephalopathy Monitoring in Italy
Authors: Carra Elena , Taddei Roberta , Barbieri Ilaria , Botti Giuliana , Tranquillo Vito , Iori Alessandra , Gibelli Lucia , Cerioli Monica , Cavadini Patrizia , Gelmetti Daniela , Pongolini Stefano , Capucci Lorenzo
Published: May 2011

In 2001, a compulsory active surveillance system was started in the European Union to assess the prevalence of bovine spongiform encephalopathy (BSE) in the cattle population. The aim of the current study was to report on the field performances of 3 rapid tests: a Western blot (WB), a chemiluminescence enzymelinked immunosorbent assay (ELISA), and an immunochromatographic assay, routinely used at 3 laboratories of the Istituto Zooprofilattico Sperimentale of Lombardia and Emilia Romagna, over 8 years of BSE monitoring activity. A total of 2,802,866 samples from slaughtered animals and 202,453 samples from fallen stock were tested by 1 of 3 tests. Positive results of the rapid tests were confirmed by histopathological examination, immunohistochemistry, and confirmatory WB. The field performances (i.e., initial reactive and false-positive rates) and practical aspects regarding resources and applicability of the tests to high-throughput routine testing laboratories were evaluated. The 3 tests proved to be reliable tools when applied to slaughtered samples, showing no or very low false-positive rates (<1 per 100,000 negative samples tested) and low retesting frequencies (0.02-0.26%). When samples from fallen stock were analyzed, performances of the immunochromatographic assay, and especially the chemiluminescence ELISA, were negatively affected, resulting in higher false-positive and retesting rates. On the other hand, both tests are less expensive, much easier to use, provide more rapid results, and adapt well to application in routine laboratories as compared with WB. In the authors' experience, the immunochromatographic assay was a good compromise between performance and convenience.
Comparison of Four Methods to Quantify Equid Herpesvirus 1 Load by Real-Time Polymerase Chain Reaction in Nasal Secretions of Experimentally and Naturally Infected Horses
Authors: Pusterla Nicola , Hussey Stephen B. , Mapes Samantha , Leutenegger Christian M. , Madigan John E. , Ferraro Gregory L. , Wilson W. David , Lunn D. Paul
Published: May 2011

The objective of the current study was to compare the performance of 4 methods to quantify Equid herpesvirus 1 (EHV-1) by real-time polymerase chain reaction (PCR) in nasal secretions from experimentally and naturally infected horses. Nasal secretions were collected on the challenge day and daily thereafter for 13 days from 4 experimentally infected horses. Additional nasal swabs were collected from 30 horses with clinical signs consistent with natural EHV-1 infection. Absolute quantitation of EHV-1 target molecules was performed using standard curves for EHV-1 and equine glyceraldehyde-3-phosphate dehydrogenase, and DNA yield, and was expressed as EHV-1 glycoprotein B (gB) gene copies per million nucleated nasal cells, EHV-1 gB gene copies per entire swab, EHV-1 gB gene copies per 1 μl of purified DNA, and EHV-1 gB gene copies per 1 ng of template DNA. The study results showed that all 4 calculation methods yielded comparable results between experimentally and naturally infected horses, and that the different methods were significantly correlated with each other. Reporting of quantitative results for EHV-1 viral load in nasal swabs collected from infected horses constitutes an important advance in both the research and diagnostic fields, allowing one to determine the infectious risk of affected horses, disease stage, or response to antiviral therapy. However, protocols that normalize the PCR results against a preselected volume of DNA or nasal secretions are likely to be more prone to variations than protocols that calculate the load for the entire swab, incorporate a housekeeping gene, or use a constant amount of extracted DNA.
Broadly Reactive Pan-Paramyxovirus Reverse Transcription Polymerase Chain Reaction and Sequence Analysis for the Detection of Canine Distemper Virus in a Case of Canine Meningoencephalitis of Unknown Etiology
Authors: Schatzberg Scott J. , Li Qiang , Porter Brian F. , Barber Renee M. , Claiborne Mary Kate , Levine Jonathan M. , Levine Gwendolyn J. , Israel Sarah K. , Young Benjamin D. , Kiupel Matti , Greene Craig , Ruone Susan , Anderson Larry , Tong Suxiang
Published: May 2011

Despite the immunologic protection associated with routine vaccination protocols, Canine distemper virus (CDV) remains an important pathogen of dogs. Antemortem diagnosis of systemic CDV infection may be made by reverse transcription polymerase chain reaction (RT-PCR) and/or immunohistochemical testing for CDV antigen; central nervous system infection often requires postmortem confirmation via histopathology and immunohistochemistry. An 8-month-old intact male French Bulldog previously vaccinated for CDV presented with multifocal neurologic signs. Based on clinical and postmortem findings, the dog's disease was categorized as a meningoencephalitis of unknown etiology. Broadly reactive, pan-paramyxovirus RT-PCR using consensus-degenerate hybrid oligonucleotide primers, combined with sequence analysis, identified CDV amplicons in the dog's brain. Immunohistochemistry confirmed the presence of CDV antigens, and a specific CDV RT-PCR based on the phosphoprotein gene identified a wild-type versus vaccinal virus strain. This case illustrates the utility of broadly reactive PCR and sequence analysis for the identification of pathogens in diseases with unknown etiology.