bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of Chlamydophila Abortus DNA Extraction Protocols for Polymerase Chain Reaction Diagnosis in Paraffin-Embedded Tissues
Authors: Ortega Nieves , Navarro José A. , Nicolás Laura , Buendía Antonio J. , Caro María R. , Del Río Laura , Martínez Carlos M. , Cuello Francisco , Salinas Jesús , Gallego María C.
Published: May 2011

Polymerase chain reaction (PCR) has gained increasing importance as a tool for directly demonstrating the presence of Chlamydophila in the placentas of aborted sheep and goats. However, because of the zoonotic potential of the disease, it is advisable to use fixed materials. To evaluate 4 different DNA extraction protocols in paraffin-embedded sections for PCR, previously immunohistochemically diagnosed placental samples from outbreaks of abortions in goats and sheep were used. The samples were also used to evaluate the effect of the duration of fixation in formalin on PCR. A protocol that uses Tris-HCl pH 8.5 with EDTA and subsequent digestion with proteinase K was found to be an easy protocol for obtaining excellent PCR products for Chlamydophila abortus diagnosis from formalin-fixed and paraffin-embedded specimens. It was also found that if samples are fixed in formalin for more than 2 weeks, the PCR technique is affected more adversely than immunohistochemical methods.
Immunohistochemical Diagnosis of Canine Ovarian Epithelial and Granulosa Cell Tumors
Authors: Riccardi Elena , Greco Valeria , Verganti Sara , Finazzi Mario
Published: May 2011

In humans and canines, the morphology of granulosa cell tumors is extremely variable and causes diagnostic difficulties. In human pathology, immunohistochemistry has been widely used for the diagnosis of granulosa cell tumors, whereas, limited studies are present in canine species. The aim of this study was to investigate the expression of cytokeratins, vimentin, and inhibin-α in canine normal ovaries, epithelial ovarian tumors, and granulosa cell tumors to establish an immunohistochemical panel for the differential diagnosis of ovarian tumors. Formalin-fixed, paraffin-embedded tissue sections from 4 normal ovaries, 8 granulosa cell tumors, and 6 epithelial ovarian tumors (2 adenomas and 4 adenocarcinomas) sections were obtained and stained with hematoxylin and eosin and immunohistochemically for cytokeratin AE1/AE3, cytokeratin 7, vimentin, and inhibin-α. In normal ovaries, cytokeratin 7, cytokeratin AE1/AE3, and vimentin were expressed in the surface epithelium. Granulosa cells were negative for cytokeratin 7 and displayed variable expression of vimentin, cytokeratin AE1/AE3, and inhibin-α toward follicular maturation. Granulosa cell tumors were negative for cytokeratin 7 and positive for inhibin-α. Conversely, ovarian epithelial cells tumors were positive for cytokeratin 7 and negative for inhibin-α. Both granulosa and epithelial cell tumors displayed variable expression of vimentin. Cytokeratin AE1/AE3 was expressed by all epithelial-derived tumors and 6 of 8 granulosa cell tumors. The results of this study suggest that useful immunohistochemical markers to distinguish epithelial ovarian tumors from granulosa cell tumors are cytokeratin 7 and inhibin-α.
Cyclooxygenase-2 Immunoreactivity in Equine Ocular Squamous-Cell Carcinoma
Authors: Rassnick Kenneth M. , Njaa Bradley L.
Published: May 2011

Squamous-cell carcinoma (SCC) is the second most common tumor in horses, and 40%-50% may occur in ocular and adnexal structures. Cyclooxygenase (COX)-2 is an inducible enzyme responsible for the production of prostaglandins that control cell growth and the development and progression of cancer. Mechanisms responsible for the initial upregulation of COX-2 in neoplasia are unclear; prolonged sunlight exposure and mutations in the p53 gene may be possibilities. Because the etiopathogenesis of ocular SCC in horses may involve ultraviolet sunlight and p53 mutations, the purpose of this study was to characterize the immunoreactivity of COX-2 in these tumors. Cyclooxygenase-2 expression was found in 6 of 22 (27%) paraffin-embedded equine SCCs. Cyclooxygenase-2 immunoreactivity was associated with the mitotic index ( P < 0.001). Strategies to inhibit COX-2 by the use of topical or systemic COX-2 inhibitors might prove to be a safe and economical treatment in some horses with SCC.
Agreement Among Surgical Pathologists Evaluating Routine Histologic Sections of Digits Amputated from Cats and Dogs
Authors: Wobeser Bruce K. , Kidney Beverly A. , Powers Barbara E. , Withrow Stephen J. , Mayer Monique N. , Spinato Maria T. , Allen Andrew L.
Published: May 2011

Agreement among pathologists interpreting histologic specimens is an area of interest within human pathology, but little work in this area has been reported in the veterinary literature. Agreement among pathologists evaluating routine histologic sections of amputated digits from cats and dogs submitted to multiple diagnostic centers was examined. Histologic sections from surgical specimens were reviewed in a blinded fashion by two pathologists, and a comparison to the original diagnosis, as stated in the diagnostic report, was recorded. A total of 513 cases were reviewed, and complete agreement was reached in 409 (79.7%). Of the 104 instances of disagreement, 77 (74.0%) were considered to be of clinical significance. The diagnosis of keratoacanthoma was disagreed with in 19 of 21 diagnoses (90.4%). No other individual diagnosis was similarly disputed. The overall level of disagreement is large and is similar to that reported in human pathology and suggests that further study of this issue would be useful in veterinary pathology.
Precursor B-1 B Cell Lymphoma in a Newborn Calf
Authors: Yamamoto Shinji , Wada Yoshihiro , Ishikawa Yoshiharu , Kadota Koichi
Published: May 2011

A newborn Holstein female calf had neoplastic lesions in the skin and within the thoracic and abdominal cavities but not in the bone marrow, spleen, thymus, or most lymph nodes. Because the tumor cells were positive for CD79a (B cell marker), CD5 (B-1 cell marker) and terminal deoxynucleotidyl transferase (marker for immature lymphoid precursors), a diagnosis of precursor B-1 B cell lymphoma was made. The diagnosis was strongly supported by the fact that B-1 cells can develop in the fetus, unlike B-2 cells, which are produced after birth. The lymphoma was distinct from the typical calf form of lymphoma of B-2 cell origin, which does not express CD5 and is characterized by generalized lymphadenopathy and involvement of the bone marrow, blood and spleen.
Carcinoma in a Mixed Mammary Tumor in a Llama (Lama Glama)
Authors: Bangari Dinesh S. , Stevenson Gregory W.
Published: May 2011

A 13-year-old female llama was presented to the referring veterinarian for swelling and firmness of the right rear mammary gland, for a duration of 2 months, which had been unresponsive to antibiotics. A formalin-fixed wedge biopsy specimen from the affected quarter was submitted to Purdue University Animal Disease Diagnostic Laboratory for histopathology. Histopathologic examination revealed tubulopapillary acinar or solid nest-like clusters of neoplastic epithelial cells surrounded by whorls and sheets of proliferative myoepithelial cells. Histologic criteria for malignancy observed in neoplastic epithelial cells included marked cellular and nuclear atypia, high mitotic index, and numerous bizarre mitoses. The presence of osseous metaplasia in the proliferative mesenchymal component justified classification as a mixed tumor. Positive immunohistochemical staining of neoplastic epithelial cells with anticytokeratin antibody, and proliferative spindloid cells with antiviemtin and antismooth muscle actin antibodies supported the histopathologic diagnosis. The llama was in good health after about 1 year of initial presentation, and metastasis to regional lymph nodes was not reported. Mammary neoplasia is rare in camelids. To the authors' knowledge, this is the first report of a carcinoma in a mixed mammary tumor in a llama.
Evaluation of an Ovine Testis Cell Line (OA3.Ts) for Propagation of Capripoxvirus Isolates and Development of an Immunostaining Technique for Viral Plaque Visualization
Authors: Babiuk Shawn , Parkyn Geoff , Copps John , Larence June E. , Sabara Marta I. , Bowden Timothy R. , Boyle David B. , Kitching R. Paul
Published: May 2011

An ovine testis cell line (OA3.Ts) was evaluated and compared with primary lamb kidney (LK) cells for its utility in capripoxvirus propagation and titration. A comparison of OA3.Ts cell growth kinetics and morphology at low (<33) and high (34−36) passage levels indicated a difference in both characteristics. However, viral titers determined in low and high passage OA3.Ts cells were comparable with those obtained using LK cells. Capripoxvirus infection of OA3.Ts and LK cells resulted in a similar cytopathic effect, which allowed for the detection of discrete viral plaques following immunostaining with capripoxvirus-specific antiserum.
Evaluation of Surveillance Protocols for Detecting Porcine Reproductive and Respiratory Syndrome Virus Infection in Boar Studs by Simulation Modeling
Authors: Rovira Albert , Reicks Darwin , Muñoz-Zanzi Claudia
Published: May 2011

Because porcine reproductive and respiratory syndrome virus (PRRSV) can be transmitted through semen, PRRSV-free boar studs need to be routinely monitored to rapidly detect any potential PRRSV introduction. However, current protocols for monitoring PRRSV in boar studs are diverse, sometimes very costly, and their effectiveness has not been quantified. The objective of this study was to evaluate the ability of different monitoring protocols to detect PRRSV introduction into a negative boar stud by using a simulation modeling approach. A stochastic transmission model was constructed to simulate the spread of PRRSV in a typical negative boar stud in the USA (herd size of 200 boars, 60% annual replacement) and the performance of monitoring protocols by using different sample sizes (10, 30, and 60 samples), sampling frequency (3 times a week, weekly, and biweekly), and diagnostic procedures (PCR on semen, PCR on serum, ELISA on serum, and both PCR and ELISA on serum). The monitoring protocols were evaluated in terms of the time from PRRSV introduction into the boar stud to PRRSV detection. Protocols that used PCR on serum detected the PRRSV introduction earlier than protocols that used PCR on semen, and these were earlier than those that used ELISA on serum. The most intensive protocol evaluated (testing 60 boars 3 times a week by PCR on serum) would need 13 days to detect 95% of the PRRSV introductions. These results support field observations, suggesting that an intensive monitoring protocol needs to be in place in a boar stud to quickly detect a PRRSV introduction.
Development and Application of an Enzyme-Linked Immunosorbent Assay to Detect Antibodies against Prevalent Salmonella Serovars in Swine in Southern Brazil
Authors: Kich Jalusa Deon , Schwarz Patrícia , Silva Luis Eduardo , Coldebella Arlei , Piffer Itamar Antônio , Vizzoto Remídio , Marisa Ribeiro de Itapema Cardoso
Published: May 2011

The implementation of Salmonella control programs in the pork production chain demands rapid and cost-effective methods to assess the prevalence of infection in pig herds. The objective of the present study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) based on S. Typhimurium lipopolysaccharides (LPS) to measure the prevalence of infection caused by Salmonella in swine herds. Coating antigen was produced by phenol extraction of S. Typhimurium culture. After standardization of ELISA test conditions, the assay was validated by testing serum samples on different animal categories: pigs orally inoculated with S. Typhimurium and sentinel animals in contact with them, naturally infected animals, colostrum-deprived piglets, and bacterin-immunized pigs. Seroconversion was observed in inoculated pigs (7 days postinfection [DPI]) and in the sentinels (21 DPI). Nonspecific reactions were not detected in the sera of colostrum-deprived animals. Serum samples from animals immunized with Salmonella Agona, Salmonella Derby, Salmonella Panama, and Salmonella Bredeney bacterins showed marked cross-reaction with the LPS from the serovar Typhimurium. Moreover, positive results obtained with the in-house ELISA were associated with Salmonella isolation in 75 infected pig herds. Comparisons with 2 commercial kits showed a linear correlation coefficient of 0.847 between the in-house ELISA and kit A and 0.922 with kit B but a low agreement in the qualitative results. In conclusion, the newly developed in-house ELISA based on S. Typhimurium LPS can be a useful tool to determine the intensity of Salmonella sp. infection in swine herds.
Outbreaks of Renal Failure Associated with Melamine and Cyanuric Acid in Dogs and Cats in 2004 and 2007
Authors: Brown Cathy A. , Jeong Kyu-Shik , Poppenga Robert H. , Puschner Birgit , Miller Doris M. , Ellis Angela E. , Kang Kyung-Il , Sum Steffen , Cistola Alexis M. , Brown Scott A.
Published: May 2011

Sixteen animals affected in 2 outbreaks of pet food-associated renal failure (2 dogs in 2004; 10 cats and 4 dogs in 2007) were evaluated for histopathologic, toxicologic, and clinicopathologic changes. All 16 animals had clinical and laboratory evidence of uremia, including anorexia, vomiting, lethargy, polyuria, azotemia, and hyperphosphatemia. Where measured, serum hepatic enzyme concentrations were normal in animals from both outbreaks. All animals died or were euthanized because of severe uremia. Distal tubular lesions were present in all 16 animals, and unique polarizable crystals with striations were present in distal tubules or collecting ducts in all animals. The proximal tubules were largely unaffected. Crystals and histologic appearance were identical in both outbreaks. A chronic pattern of histologic change, characterized by interstitial fibrosis and inflammation, was observed in some affected animals. Melamine and cyanuric acid were present in renal tissue from both outbreaks. These results indicate that the pet food-associated renal failure outbreaks in 2004 and 2007 share identical clinical, histologic, and toxicologic findings, providing compelling evidence that they share the same causation.
Differentiation of Moraxella Bovoculi sp. nov. from other Coccoid Moraxellae by the Use of Polymerase Chain Reaction and Restriction Endonuclease Analysis of Amplified DNA
Authors: Angelos John A. , Ball Louise M.
Published: May 2011

Moraxella oris was historically the only coccoid Moraxella identified in cultures of ocular fluid from cattle with infectious bovine keratoconjunctivitis (IBK) and could be morphologically and biochemically differentiated from Moraxella bovis. Moraxella bovoculi sp. nov. is a recently characterized Moraxella isolated from ulcerated eyes of calves with IBK in northern California in 2002. Like Moraxella ovis, M. bovoculi sp. nov. is a gram-negative coccus/diplococcus. All 18 original isolates of M. bovoculi sp. nov. possessed phenylalanine deaminase (PADase) activity and could therefore be differentiated from M. ovis and M. bovis. During the characterization of 44 additional isolates of hemolytic gram-negative cocci that were cultured from ulcerated eyes of IBK-affected calves, 2 PADase-negative isolates were identified that could not be differentiated biochemically from M. ovis; however, the DNA sequence of the 16S-23S intergenic spacer region (ISR) of the isolates matched the 16S-23S ISR DNA sequence of M. bovoculi sp. nov. To facilitate the identification of PADase-negative moraxellae, a polymerase chain reaction (PCR) coupled with restriction enzyme digestion analysis of amplified DNA was developed. Amplification of the 16S-23S ISR followed by AfaI digestion of amplified DNA could differentiate M. bovoculi sp. nov. from M. ovis and other moraxellae. The DNA sequence analysis of the amplified 16S-23S ISR from the 42 PADase-positive isolates of hemolytic gram-negative cocci indicated that all were M. bovoculi sp. nov. and all possessed an AfaI site. A PCR coupled with restriction analysis of amplified DNA can aid in identifying M. bovoculi sp. nov.
Bovine Serum Panel for Evaluating Foot-and-Mouth Disease Virus Nonstructural Protein Antibody Tests
Authors: Parida Satya , Fleming Lucy , Gibson Debi , Hamblin Pip A. , Grazioli Santina , Brocchi Emiliana , Paton David J.
Published: May 2011

A panel of 36 sera has been assembled from experimental cattle that had been infected by inoculation or contact exposure with 4 serotypes of foot-and-mouth disease virus (FMDV) with or without prior vaccination. Virus replication and persistence had been characterized in all of the animals. The proportion of the sera scored positive by 5 tests for antibodies to the nonstructural proteins of FMDV varied, suggesting that the panel can discriminate between the sensitivity with which such tests are able to identify infected cattle. Use of this panel will help in assessment of new tests and quality control of existing methods.
Effectiveness of Capillary Electrophoresis Fluoroimniunoassay of Blood PrPSc for Evaluation of Scrapie Pathogenesis in Sheep
Authors: Everest David J. , Waterhouse Susan , Kelly Tina , Velo-Rego Elena , Sauer Maurice J.
Published: May 2011

Management of prion diseases in livestock would benefit greatly from availability of a validated blood test. A promising immunocapillary electrophoresis technique (also known as capillary electrophoresis fluoroimmunoassay) to detect abnormal prion protein in blood from live sheep is evaluated here. Capillary electrophoresis fluoroimmunoassay was applied to analysis of extracted blood from scrapie-exposed sheep ( n = 87; 347 samples) at various stages of incubation, and to control sheep ( n = 194; 489 samples). Overall, test values for the control and test populations were not significantly different, and a similar proportion of control (7%) and test (10%) sheep were classified as positive. Over 2−3 month intervals from birth until clinical disease, test specificity and sensitivity ranged from 66.7% to 100% and 0% to 66.7%, respectively, indicating poor diagnostic performance at all stages of pathogenesis. In routine application, in its present form, the capillary electrophoresis fluoroimmunoassay procedure proved to be insufficiently robust for use as a blood test for scrapie diagnosis.
Membranoproliferative Glomerulonephritis Type III in a Simultaneous Infection of Leishmania Infantum and Dirofilaria Immitis in a Dog
Authors: Aresu Luca , Valenza Federico , Ferroglio Ezio , Pregel Paola , Uslenghi Federica , Tarducci Alberto , Zanatta Renato
Published: May 2011

In this report a 9-year-old female German Shepherd dog with a membranoproliferative glomerulonephritis (MPGN) type III associated with concomitant infection of Dirofilaria immitis and Leishmania infantum is presented. Light microscopic evaluation of kidney revealed a diffuse hypercellularity and thickening of glomerular basement membrane. Heavy and coarse granular complement C3 deposition and a weaker positive reaction to immunoglobulin G were present along peripheral glomerular basement membrane and in the mesangium in the immunofluorescent study. Transmission electron microscopy revealed deposits in the mesangium, subendothelium, and subepithelium. These lesions are compatible with membranoproliferative glomerulonephritis type III in humans.
Evaluation of the Toxicity of Adonis Aestivalis in Calves
Authors: Woods Leslie W. , George Lisle W. , Anderson Mark L. , Woods Dale M. , Filigenzi Mike S. , Puschner Birgit
Published: May 2011

Toxicosis of Adonis aestivalis is well documented in horses, but little is known of its toxicity in cattle. A. aestivalis (summer pheasant's eye) was collected over multiple years, under different growing conditions, and at various stages of maturity, dried, and administered to calves to evaluate the toxicity of A. aestivalis in cattle. Four 300-lb Holstein, and 2 90-lb, preruminating Jersey calves were administered 1% body weight of ground A. aestivalis via a stomach tube and monitored for clinical signs for 2 weeks and 1 week, respectively. The Holstein calves were then fed 0.2 to 1% body weight A. aestivalis daily for 4 to 5 weeks. The Holstein calves had transient, mild cardiac abnormalities during the feeding trial. Mild, transient gastrointestinal and cardiac signs were noted in the preruminating calves. No gross or microscopic lesions were seen on necropsies performed at the end of the study. Based on the results of this study, cattle do not appear to be as susceptible to toxicosis from A. aestivalis as other species, such as horses and pigs.
Porcine Circovirus Type 2–Associated Disease: Update on Current Terminology, Clinical Manifestations, Pathogenesis, Diagnosis, and Intervention Strategies
Authors: Opriessnig Tanja , Meng Xiang-Jin , Halbur Patrick G.
Published: May 2011

Porcine circovirus type 2 (PCV2)–associated disease (PCVAD) continues to be an important differential diagnosis on pig farms in the United States and worldwide. Case trend analyses indicate that the incidence of PCVAD is on the rise in the United States. Accurate diagnosis is important in order to implement appropriate intervention strategies. PCVAD can manifest as a systemic disease, as part of the respiratory disease complex, as an enteric disease, as porcine dermatitis and nephropathy syndrome, or as reproductive problems. PCVAD may be only a sporadic individual animal diagnosis; however, PCVAD may also manifest as a severe herd problem accelerated and enhanced by concurrent virus or bacterial infections. This article is intended to discuss the most common disease manifestations, pathogenesis, diagnostic approaches, and intervention strategies associated with PCVAD in North America.
Development and Bench Validation of Real-Time Reverse Transcription Polymerase Chain Reaction Protocols for Rapid Detection of the Subtypes H6, H9, and H11 of Avian Influenza Viruses in Experimental Samples
Authors: Das Amaresh , Suarez David L.
Published: May 2011

Real-time reverse transcription polymerase chain reaction (RRT-PCR) is commonly used for the rapid detection, as well as to determine the subtype, of avian influenza viruses (AIVs). There are 16 known serologically distinct hemagglutinin (HA) subtypes of AIV described. Currently, determination of the subtypes of AIVs by RRT-PCR tests has been limited to the H5 and H7 subtypes. In this study, RRT-PCR assays were developed in simplex formats for rapid detection of AIV subtypes H6, H9, and H11. The primers and probes for RRT-PCR were designed from nucleotide sequences of the HA genes, which were either downloaded from GenBank (for H6 and H9) or sequenced for this study. The specificity and sensitivity of the RRT-PCR assays were determined based on the detection of the virus from a proficiency panel consisting of 15 different HA subtypes of AIVs and from serial dilutions of target viral RNA. The subtype-specific RRT-PCR assays were used to detect the virus in cloacal and oropharyngeal swabs of experimental chickens inoculated with H6, H9, and H11 AIVs, and the test results were compared with validated RRT-PCR assays based on the amplification of AI matrix (MA) gene. A high correlation of the matrix test and the specific H6, H9, and H11 by the RRT-PCR assays was observed; kappa coefficients for the agreement of test results in cloacal and oropharyngeal swabs combined were 0.927, 0.962, and 0.981, respectively.
Development and Validation of a Radioimmunoassay for Thyrotropin in Cattle
Authors: Guyot Hughes , Sulon José , Beckers Jean-François , Closset Jean , Lebreton Pascal , de Oliveira Laurent Alves , Rollin Frédéric
Published: May 2011

In mammals, thyrotropin, or thyroid-stimulating hormone (TSH), assay is used for the diagnosis of primary hypothyroidism. Hypothyroidism is the most common type of thyroid disorder in cattle. The aim of this study was to develop and validate, under physiologic and pathologic conditions, a radioimmunoassay (RIA) for bovine TSH (bTSH). Double RIA was performed with purified bTSH and specific bovine antiserum. Laboratory validation included research of minimal detection limit, accuracy, and reproducibility. The physiologic validation included a thyrotropin-releasing hormone (TRH) challenge performed on euthyroid cows and a follow-up of bTSH concentration over a 24-hour period. Furthermore, bTSH concentration was assayed in a large population of healthy dairy and beef cows to define reference interval. The pathologic validation was made by assaying bTSH and thyroid hormones on healthy and goitrous newborn calves. The minimum detection limit (MDL) for bTSH assay was 1.3 μU/ml. The recovery was 101% to 106%. The intra- and interassay coefficients of variation (CVs) ranged from 5% to 11% and 11% to 15%, respectively. The RIA covered the whole range of physiologic bTSH values, as shown by bTSH values induced by TRH-challenge. A pulsatile secretion of bTSH was observed, accompanied by a diurnal variation with lower night values than day values. Reference intervals of bTSH ranged from 1.3 to 13.0 μU/ml for beef and dairy breeds. Finally, bTSH easily discriminated goitrous newborn calves from healthy ones, leading to the definition of a cutoff value of 35 μU/ml. The bTSH assay positively reacted to physiologic and pathologic conditions. The accuracy and precision of the RIA were satisfying.
Validation of Tissue Microarrays for Immunohistochemical Analyses of Canine Lymphomas
Authors: Keller Stefan M. , Keller Barbara C. , Grest Paula , Börger Claas T. , Guscetti Franco
Published: May 2011

In most validation studies of tissue microarrays (TMAs), a fixed number of cores with a given diameter are analyzed to determine the degree of accuracy by which the TMA represents the whole section. The statistical model described in the present study predicts this property for various combinations of 2 core sizes (0.6 mm and 1.2 mm) and different core numbers. The model was based on artificial TMA core biopsies generated from Ki-67 and active caspase-3 immunostains of 40 canine lymphoma samples. Positivity was scored on a continuous scale, and a large number of cells were analyzed with the help of semiautomated cell counting. Despite considerable differences in range and distribution of Ki-67 and active caspase-3 positivity values, the model predictions showed a high degree of agreement for both markers. Comparison of 0.6 mm and 1.2 mm cores indicated that the use of small cores necessitates inclusion of a larger number of samples but requires counting a markedly smaller number of cells. Suitability of TMAs to determine the immunophenotype of the whole section was assessed using 2 different combinations of core sizes and numbers. Both displayed a high degree of concordance with the whole section (κ0.6 = 0.79; κ1.2 = 0.91). The present study provides a basis for the use of TMAs in future high-throughput immunohistochemical investigations of selected markers in canine lymphomas. The statistical model presented can be used to determine an optimal TMA design depending on a desired accuracy.
Temporal Distribution of Porcine Circovirus 2 Genogroups Recovered from Postweaning Multisystemic Wasting Syndrome-Affected and -Nonaffected Farms in Ireland and Northern Ireland
Authors: Allan Gordon M. , McNeilly Francis , McMenamy Michael , McNair Irene , Krakowka Steven G. , Timmusk Sirje , Walls Dermot , Donnelly Maria , Minahin Donal , Ellis John , Wallgren Per , Fossum Caroline
Published: May 2011

Porcine circovirus type 2 (PCV2) is now recognized as the essential infectious component of porcine postweaning multisystemic wasting syndrome (PMWS). PMWS was first recognized in high-status, specific pathogen-free pigs in Canada in 1991 and is now an economically important disease that affects the swine industry around the world. Recently, reports of genomic studies on PCV2 viruses indicated that 2 distinctive genogroups of PCV2 exist. 4,10 This report involves the results of a study on the distribution of predominant PCV2 genogroups recovered from samples taken from PMWS-affected and PMWS-nonaffected farms on the island of Ireland over a 9-year period and the results of a study on PCV2 genogroup recovery from fecal samples taken from a farm in Northern Ireland from 2003 to 2005 that was first diagnosed as PMWS positive in August 2005. The results indicate that, although at least 2 distinct genogroups of PCV2 have been circulating on pig farms on the island of Ireland, there does not appear to be a direct relationship between infection with these different genogroups of PCV2 and the development of PMWS.
Age-Related Lesions in Laboratory-Confined Raccoons (Procyon Lotor) Inoculated with the Agent of Chronic Wasting Disease of Mule Deer
Authors: Hamir Amir N. , Kunkle Robert A. , Miller Janice M. , Cutlip Randall C. , Richt Juergen A. , Kehrli Marcus E. , Williams Elizabeth S.
Published: May 2011

This communication documents age-associated pathologic changes and final observations on experimental transmission of chronic wasting disease (CWD) by the intracerebral route to raccoons ( Procyon lotor). Four kits were inoculated intracerebrally with a brain suspension from mule deer with CWD. Two uninoculated kits served as controls. One CWD-inoculated raccoon was humanely killed at 38 months after inoculation, and 1 control animal died at 68 months after inoculation. Both animals had lesions that were unrelated to transmissible spongiform encephalopathy. Six years after inoculation, none of the 3 remaining CWD-inoculated raccoons had shown clinical signs of neurologic disorder, and the experiment was terminated. Spongiform encephalopathy was not observed by light microscopy, and the presence of abnormal prion protein (PrPd) was not detected by either immunohistochemistry or Western blot techniques. Age- related lesions observed in these raccoons included islet-cell pancreatic amyloidosis (5/6), cystic endometrial hyperplasia (3/4), cerebrovascular mineralization (5/6), neuroaxonal degeneration (3/6), transitional-cell adenoma of the urinary bladder (1/6), and myocardial inclusions (4/6). The latter 2 pathologic conditions were not previously reported in raccoons.
Toxic Hepatopathy in Sheep Associated with the Ingestion of the Legume Tephrosia Cinerea
Authors: Santos Julio César A. , Riet-Correa Franklin , Dantas Antônio F. M. , Barros Severo S. , Molyneux Russell J. , Medeiros Rosane M. T. , da Silva Durval M. , de Oliveira Odací F.
Published: May 2011

A disease known as water belly (barriga d'água), characterized by chronic progressive ascites, affects sheep in the semiarid region of northeastern Brazil. The objectives of this investigation were to study the disease and to determine its cause. Only sheep grazing for long periods in pastures where Tephrosia cinerea represents 80% to 100% of the available forage are affected. Most animals die after a clinical manifestation period of some weeks or months, but others recover when they are withdrawn from the pastures or after the first rains. At necropsy, large amounts of liquid were found in the abdominal cavity, and the liver was hard, with an irregular surface. On histology examination, the main liver lesion was chronic periportal and subcapsular fibrosis with bridging. The disease was produced experimentally in 1 sheep by the administration of large amounts of T. cinerea for 232 days. Another sheep, recovered from the spontaneous disease, had clinical signs after the ingestion of large amounts of the plant for 40 days. Seeds and leaves of the plant were examined by gas chromatography-mass spectrometry for the presence of pyrrolizidine alkaloids with negative results. It is concluded that the disease is caused by the ingestion of T. cinerea.
Diagnostic Evidence of Staphylococcus Warneri as a Possible Cause of Bovine Abortion
Authors: Barigye Robert , Schaan Lynn , Gibbs Penelope S. , Schamber Ev , Dyer Neil W.
Published: May 2011

Following a routine necropsy of a bovine fetus aborted at 5 months of gestation, placenta, fetal tissue samples, and stomach contents were subjected to a number of laboratory tests. Staphylococcus warneri was isolated in pure culture from the lung, liver, and stomach contents, whereas the placenta yielded S. warneri and a number of contaminants. Gross evaluation of agar plates showed predominant colonies to be morphologically consistent with those of S. warneri and the identity of the agent was further confirmed on a Trek Diagnostic Systems Sensititre, gram-positive identification (GPID) plate. Microscopic evaluation of fetal tissue sections showed extensive necrotizing lesions of the tongue, lung, and placenta in which there were numerous coccoid shaped gram-positive bacteria with morphology consistent with Staphylococcus spp. These results provide strong diagnostic evidence of S. warneri as a possible cause of bovine abortion and suggest there should be further investigations into the abortivirulence of this agent.
Brain Heart Infusion Broth May Not Be a Required Component for the Decontamination Process for the Isolation of Mycobacterium Avium Subspecies Paratuberculosis from Fecal Samples using ESP® Broth Cultures
Authors: Rajeev Sreekumari , Berghaus Roy D. , Johnson Jill , Pence Mel , Byrum Beverly , Farrell Troy , Baldwin Charles
Published: May 2011

Based on the authors' laboratory experience indicating that increased bacterial contamination in Mycobacterium avium ssp. paratuberculosis (MAP) cultures may be because of the addition of brain heart infusion broth (BHI) during the decontamination process, this study was designed to examine whether BHI is a required component for the isolation of MAP from ESP® broth cultures. Twenty-six National Veterinary Services Laboratory (NVSL) proficiency test samples supplied for the year 2005 were used for the comparison. Two paired sets of samples were processed in the experiment. In one set, the hexadecylpyridinium chloride monohydrate (HPC) and antibiotic brew were prepared in half strength BHI and for the other set, HPC and antibiotic brew were prepared in sterile distilled water. Culture of the 26 samples using the BHI/HPC decontamination method identified 13 (50%) positives, whereas culture using the water/HPC decontamination method identified 14 (54%) positives. The proportions of samples with a positive result did not differ significantly between the 2 decontamination methods. Although in most cases it took longer to identify a positive result by the BHI method, the difference between methods with respect to the number of days to a positive culture result was not statistically significant. Retrospective data collected from the Animal Disease Diagnostic Laboratory, Ohio also suggest that inclusion of BHI in the decontamination protocol may not have any effect on MAP recovery or contamination rate. Elimination of BHI from broth cultures may increase the sensitivity of MAP isolation, and reduce the cost of testing.
Pathological Findings in Dogs Naturally Infected with Angiostrongylus Vasorum in Newfoundland and Labrador, Canada
Authors: Bourque Andrea C. , Conboy Gary , Miller Lisa M. , Whitney Hugh
Published: May 2011

Fifty-six dogs from St. John's, Newfoundland, Canada, were evaluated for Angiostrongylus vasorum infection. Small numbers of nematodes were found within pulmonary arteries of 6 dogs. Larvae were identified in fecal samples in 2 of 6 dogs. All 6 dogs had multifocal granulomatous pneumonia and sometimes foci of chronic thrombosis, which varied from very mild to severe. One dog had extensive pulmonary lesions resulting in cor pulmonale. Right heart failure was characterized by right ventricular hypertrophy, hepatic congestion, ascites, and hydrothorax. Microscopically, in most cases, eggs, larvae, and sometimes intravascular adults, were present within lung tissue sections. Small foci of granulomatous inflammation with and without larvae were present in kidney and brain in 4 dogs. An additional dog, diagnosed antemortem with angiostrongylosis via fecal examination, was also examined. Pathological findings consisted of severe pyogranulomatous interstitial pneumonia with myriad eggs, larvae, and numerous intravascular pulmonary adult nematodes with extensive arterial thrombosis. Five hundred and seventy-two adult worms were removed from pulmonary arteries. Foci of granulomatous inflammation, often associated with larvae and/or eggs, were present in tracheobronchial lymph nodes, adrenal gland, brain, and kidneys. Severe seizuring noted antemortem was attributed to several large, discrete areas of acute hemorrhagic infarction within the cerebrum and cerebellum. Natural A. vasorum infection in domestic dogs in eastern Newfoundland causes lung pathology of variable severity, which in some cases, may progress to cor pulmonale and which may be associated with extrapulmonary lesions and clinical signs.
Respiratory Disease Associated with Bovine Coronavirus Infection in Cattle Herds in Southern Italy
Authors: Decaro Nicola , Campolo Marco , Desario Costantina , Cirone Francesco , D'abramo Maria , Lorusso Eleonora , Greco Grazia , Mari Viviana , Colaianni Maria Loredana , Elia Gabriella , Martella Vito , Buonavoglia Canio
Published: May 2011

Four outbreaks of bovine respiratory disease (BRD) associated with bovine Coronavirus (BCoV) infection in Italian cattle herds were reported. In 3 outbreaks, BRD was observed only in 2–3-month-old feedlot calves, whereas in the remaining outbreak, lactating cows, heifers, and calves were simultaneously affected. By using reverse transcription polymerase chain reaction (RT-PCR), BCoV RNA was detected in all outbreaks without evidence of concurrent viral pathogens (i.e., bovine respiratory syncytial virus, bovine herpesvirus type 1, bovine viral diarrhea virus, bovine parainfluenza virus). Common bacteria of cattle were recovered only from 2 outbreaks of BRD: Staphylococcus spp. and Proteus mirabilis (outbreak 1) and Mannheimia haemolytica (outbreak 4). A recently established real-time RT-PCR assay showed that viral RNA loads in nasal secretions ranged between 3.10 × 10 2 and 7.50 × 10 7 RNA copies/μl of template. Bovine Coronavirus was isolated from respiratory specimens from all outbreaks except outbreak 1, in which real-time RT-PCR found very low viral titers in nasal swabs.
Evaluation of the Pathogenic Potential of Cervid Adenovirus in Calves
Authors: Woods Leslie W. , Lehmkuhl Howard D. , Hobbs Lea Ann , Parker Jackie C. , Manzer Mike
Published: May 2011

Four 3-month-old Jersey calves and three 3-month-old Holstein calves were inoculated with cervid adenovirus and monitored for clinical signs until necropsied between 10 and 42 days postinoculation. The neonatal Jersey calves had received colostrum, and the Holstein calves were colostrum deprived. Preinoculation and postinoculation serum samples were tested for antibodies to the cervid adenovirus, bovine adenovirus type 6, bovine adenovirus type 7, and goat adenovirus type 1. Virus isolation was performed on kidney, nasal secretion, and/or lung homogenates in fetal white-tailed deer lung cells. Negatively stained preparations of feces from Jersey calves were examined weekly using an electron microscope, and weekly blood samples were collected for complete blood counts. Full necropsies were performed on all calves. A complete selection of tissues was evaluated for microscopic changes, and immunohistochemistry was performed on all tissues using a polyclonal antibody to deer adenovirus. No clinical signs were observed in the calves during the study period. Following inoculation, colostrum-deprived calves developed low antibody titers to deer adenovirus, while the Jersey calves that received colostrum did not. Calves that received colostrum had high antibody titers to bovine adenovirus type 7 and goat adenovirus type 1. No consistent gross or microscopic lesions were seen. Adenovirus was not observed in negatively stained preparations of feces. Immunohistochemistry results did not demonstrate virus in all tissues examined microscopically, and virus was not isolated from lungs, nasal secretions, and kidneys.
Molecular Detection of Betanodavirus in Wild Marine Fish Populations in Korea
Authors: Gomez Dennis Kaw , Baeck Gun Wook , Kim Ji Hyung , Choresca Casiano Hermopia , Park Se Chang
Published: May 2011

Viral nervous necrosis (VNN) is a worldwide disease affecting several species of cultured marine fish. In Korea, VNN has been identified in several species of cultured marine fish. In this study, the authors present data of the amplified nested polymerase chain reaction product (420 bp) of 21 nodavirus strains from different species of apparently healthy wild marine fish on the southern coast of Korea. Phylogenetic analysis based on the partial nucleotide sequence (177 bases) of the RNA2 coat protein gene of 21 strains was highly homologous (93–100%) and closely related to that of the known betanodavirus, redspotted grouper nervous necrosis virus. These results indicate that betanodaviruses occur in large populations of wild marine fish in the southern part of the Korean peninsula, suggesting the importance of these subclinically infected fish as an inoculum source of betanodavirus that is horizontally transmitted to susceptible cultured fish species.
Comparison of Retropharyngeal Lymph Node and Obex Region of the Brainstem in Detection of Chronic Wasting Disease in White-Tailed Deer (Odocoileus Virginianus)
Authors: Keane Delwyn P. , Barr Daniel J. , Keller Jason E. , Hall S. Mark , Langenberg Julie A. , Bochsler Philip N.
Published: May 2011

Chronic wasting disease (CWD) in Wisconsin was first identified in February 2002. By April 2005. medial retropharyngeal lymph node (RLN) tissues had been examined from over 75,000 white-tailed deer for the presence of CWD by either immunohistochemical (IHC) staining for the prion protein associated with CWD (PrPres) or by using enzyme-linked immunosorbent assays with confirmation of positives by IHC staining and had been detected in 469 animals. Obex tissue was also available from 438 of the CWD-positive animals and was CWD positive by IHC staining in 355 (81%). To verify whether false-negative results were possible examining only RLN, both obex and RLN samples were examined for CWD by IHC staining from 4,430 of the white-tailed deer harvested from an area in Wisconsin where the overall deer CWD prevalence was approximately 6.2%. Two hundred and fourteen of the 269 positive deer (79.6%) had deposits of PrPres in both obex and lymphoid tissues. 55 (20.4%) had deposits only in lymphoid tissue, and there were no deer that had deposits only in obex.
Molecular Screening of Canine GM1 Gangliosidosis Using Blood Smear Specimens after Prolonged Storage: Detection of Carriers among Shiba Dogs in Northern Japan
Authors: Yamato Osamu , Jo Eun-Og , Chang Hye-Sook , Satoh Hiroyuki , Shoda Toru , Sato Reeko , Uechi Masami , Kawasaki Naomi , Naito Yoshihisa , Yamasaki Masahiro , Maede Yoshimitsu , Arai Toshiro
Published: May 2011

Molecular screening of GM1 gangliosidosis in Shiba dogs was carried out in northern Japan using blood smear specimens after prolonged storage. Of 125 specimens obtained from 3 veterinary teaching hospitals for this screening, 68 specimens (54%) were adequate for direct amplification in a polymerase chain reaction (PCR)-based DNA test, and the percentage of adequacy was different at each hospital (34%, 73%. and 100%), suggesting that the amount of blood on the smear and the storage condition of specimens may affect adequacy. Of the 68 dogs examined, 2 dogs (2.9%) were heterozygous carriers for this disease and the other dogs were all genotypically normal. The results suggest blood smear specimens can be useful for PCR testing after prolonged storage provided specimens contain a generous amount of blood and have been adequately stored. The study also suggests that GM1 gangliosidosis may be widely prevalent in the Shiba dog population in northern Japan.
Prevalence and Characterization of Bovine Viral Diarrhea Virus in the White-Tailed Deer Population in Indiana
Authors: Pogranichniy Roman M. , Raizman Eran , Thacker H. Leon , Stevenson Gregory W.
Published: May 2011

Bovine viral diarrhea (BVD) is one of the economically important diseases of cattle. For many years, different types of vaccines have been commercially available, yet this disease is hard to control in high-density population areas. Detection and isolation of bovine viral diarrhea virus (BVDV) from any potential reservoir is vital, especially when considering virus eradication from a herd or locale. One potential source is wild ruminants. Ear notches and lymph nodes were collected from the wild population of white-tailed deer (Odocoileus virginianus) during deer hunting season in Indiana and tested for BVDV with a commercial BVD antigen capture enzyme-linked immunosorbent assay. Two samples out of 745 collected samples were positive, and subsequently cp and ncp BVDV was isolated from 1 ear notch and 1 lymph node. These isolates were genotyped as type 1a and 1b based on sequence analysis of the 5′ untranslated region (UTR). The results of the present study indicate that the prevalence of BVDV in the white-tailed deer population of Indiana is about 0.3%. Wild ruminants infected with BVDV should be taken into consideration during an eradication program of BVDV from the livestock population.
Evaluation of Hunter-Harvested White-Tailed Deer for Evidence of Bovine Viral Diarrhea Virus Infection in Alabama
Authors: Passler Thomas , Walz Paul H. , Ditchkoff Stephen S. , Walz Heather L. , Givens M. Daniel , Brock Kenny V.
Published: May 2011

Bovine viral diarrhea virus (BVDV) is one of the most relevant pathogens affecting today's cattle industries. Although great strides have been made in understanding this virus in cattle, little is known about the role of wildlife in the epidemiology of BVDV. While persistently infected cattle are the most important reservoir, free-ranging ungulates may become infected with BVDV as demonstrated by serosurveys and experimental infections. Therefore, free-ranging wildlife may maintain BVDV as the result of an independent cycle and may serve as a reservoir for the virus. Systematic studies on prevalence of BVDV-specific antibodies or frequency of persistent BVDV infection in North American wildlife are sparse, and no information is available from the southeastern United States. The objective of this study was to evaluate blood and skin samples from hunter-harvested white-tailed deer (Odocoileus virginianus) for evidence of BVDV infection. Virus-neutralizing antibodies were detected in 2 of 165 serum samples. Skin biopsy immunohistochemistry (IHC) was performed on samples from 406 deer using a BVDV-specific monoclonal antibody (MAb) (15c5), and BVDV antigen was detected in one sample. A similar IHC staining pattern was obtained using a second BVDV MAb (3.12F1). Viral antigen distribution in the skin sample of this deer resembled that found in persistently infected cattle and in a previously described persistently infected white-tailed deer; thus, the deer was presumed to be persistently infected. Evidence of BVDV infection in free-ranging white-tailed deer should encourage further systematic investigation of the prevalence of BVDV in wildlife.