bluuebunny/crossref_metadata_2025 huggingface dataset:
Protein Profiling of Urine from Dogs with Renal Disease Using ProteinChip Analysis
Authors: Forterre Simone , Raila Jens , Schweigert Florian J.
Published: May 2011

Measurement of total urinary proteins in individuals that tested positive by urinary dipstick is a typical method for assessing the presence of potentially serious renal disorders. In the absence of such overt proteinuria, however, measurement of specific urinary proteins may be useful in the diagnosis of nephropathies and may provide greater insight into the pathogenesis. The urine of 28 dogs (16 with renal disease and 12 healthy) was evaluated to determine whether specific low—molecular-weight proteins or the pattern of protein excretion could also be used as a marker of tubular dysfunction in dogs. Specific proteins were assessed by immunological methods, whereas protein profiles were determined by surface-enhanced laser desorption/ionization time–of-flight mass spectrometry (MS). In particular, changes in the excretion of retinol-binding protein (RBP) and Tamm-Horsfall protein (THP) appear to be of clinical relevance in the diagnosis of canine kidney diseases. The pattern of urinary protein and peptides revealed specific changes in abundance in dogs with renal disease at molecular masses (kD) of 11.58, 12.41, 12.60, 14.58, 20.95 (RBP), 27.85, and 65.69 (albumin). In conclusion, comparable proteins as in humans might be used as urinary markers for proximal (RBP) and distal (THP) tubular dysfunction in dogs. Surface-enhanced laser desorption/ionization time-of-flight MS is a promising tool for the study of kidney physiology and pathophysiology and might aid in the discovery of new biomarkers of renal disease.
Comparison of Polymerase Chain Reaction–Restriction Fragment Length Polymorphism Assay and Enzyme Assay for Diagnosis of GM1-Gangliosidosis in Shiba Dogs
Authors: Yamato Osamu , Kobayashi Asogi , Satoh Hiroyuki , Endoh Daiji , Shoda Toru , Masuoka Yukiko , Hatakeyama Ayano , Jo Eun-Og , Asano Tomoya , Yonemura Madoka , Yamasaki Masahiro , Maede Yoshimitsu
Published: May 2011

In the present study, diagnostic methods for canine GM1-gangliosidosis were examined by comparing a DNA mutation assay with an enzyme assay. Sixty-two Shiba dogs of a pedigree with GM1-gangliosidosis were differentiated into 3 genotypes, i.e., normal, heterozygous, and homozygous affected dogs, using a DNA mutation assay, which consists of polymerase chain reaction amplification and the determination of restriction fragment length polymorphisms. The β-galactosidase activity in leukocytes, umbilical cords, and plasma was measured using 4-methylumbelliferyl β-D-galactoside and p-nitrophenyl β-D-galactoside as artificial substrates and compared among the 3 genotypes. The results showed that it was possible to identify homozygous dogs with the enzyme assay using leukocytes and umbilical cords. When using leukocytes, heterozygous carriers could be differentiated from normal dogs in many cases. However, the use of the DNA mutation assay is essential for a complete determination of heterozygous carriers because of the overlap in the distribution of enzyme activity between these 2 groups. When umbilical cords were used, heterozygous carriers could not be differentiated from normal dogs because of no significant difference in enzyme activity between these 2 groups. The β-galactosidase activity in plasma was not applicable to the diagnosis and genotyping of GM1-gangliosidosis in Shiba dogs.
Pathological and Immunohistochemical Findings in American Crows (Corvus Brachyrhynchos) Naturally Infected with West Nile Virus
Authors: Arno Wünschmann , Shivers Jan , Carroll Larry , Bender Jeff
Published: May 2011

Twenty-one American crows were identified as being West Nile virus (WNV) infected by WNV-specific reverse transcriptase-polymerase chain reaction (RT-PCR) performed on fresh brain tissue (cerebrum and cerebellum of 16 crows) or by WNV-specific immunohistochemistry of various organs (21 crows). Consistent gross lesions attributable to WNV infection were not detected. Common histological lesions included necrosis of spleen and bone marrow. West Nile virus antigen was consistently detected in heart and kidney (100%). In addition, bone marrow (92%), duodenum (89%), proventriculus (87%), liver (86%), lung (85%), spleen (80%), pancreas (61%), and brain (45%) contained WNV antigen-positive cells. Infected cells included cardiomyocytes; neurons; endothelial cells and vascular smooth muscle cells; hematopoietic cells of bone marrow; and macrophages of spleen, liver (Kupffer cells), and lungs. Epithelial cells of renal tubules, duodenum, pancreas, and proventriculus were also infected. The diagnostic histopathologist should consider WNV infection in crows in the absence of any inflammatory lesions. Immunohistochemistry of heart and kidney is as reliable in detecting WNV infection in American crows as RT-PCR of fresh brain tissue.
Lolitrem B Residue in Fat Tissues of Cattle Consuming Endophyte-Infected Perennial Ryegrass Straw
Authors: Miyazaki Shigeru , Ishizaki Ikumi , Ishizaka Masumi , Kanbara Tadashi , Ishiguro-Takeda Yuriko
Published: May 2011

Lolitrems are neurotoxins found in endophyte-infected perennial ryegrass. Lolitrems, primarily lolitrem B, are the causative agents of ryegrass staggers in livestock. To guarantee the safety of meat produced from cattle consuming endophyte-infected perennial ryegrass, lolitrem B concentrations in tissues of Japanese Black cattle were determined using high-performance liquid chromatography. Lolitrem B was not detected in muscle, liver, kidney, or cerebrum of a Japanese Black cow with signs of ryegrass staggers. In contrast, perirenal fat contained 210 ppb lolitrem B. Three cows that received half as much perennial ryegrass straw as the cow with ryegrass staggers showed no clinical signs of ryegrass staggers. However, low concentrations of lolitrem B (less than 150 ppb) were detected in their fat tissue. These observations indicate that human exposure to the neurotoxic effect of lolitrem B through beef is unlikely. The amount of lolitrem B consumed by cattle can be estimated by the determination of lolitrem B in fat tissue.
Abdominal Fat Necrosis in a Pygmy Goat Associated with Fescue Toxicosis
Authors: Smith Geoffrey W. , Rotstein David S. , Brownie Cecil F.
Published: May 2011

Abdominal fat necrosis was diagnosed in an 11-year-old female pygmy goat with a 10-day history of lethargy, anorexia, and progressive abdominal distension. Gross necropsy findings revealed multiple firm, dark yellow, nodular masses of fat throughout the abdominal cavity, which compressed several abdominal organs including the rumen, small intestine, spiral colon, and gall bladder. Histologically, multiple to coalescing adipocyte necrosis, saponification, and infiltration with variable numbers of macrophages, lymphocytes, and plasma cells was observed. Fat necrosis in this case was attributed to tall fescue toxicity based on the presence of high levels of endophyte ( Neotyphodium coenophialum)–infected fescue identified in the goat's pasture. This is the first known report of abdominal fat necrosis in a goat and demonstrates the fat necrosis syndrome of fescue toxicosis in ruminants.
Influence of Age and Production Type on Liver Copper Concentrations in Calves
Authors: Puschner Birgit , Thurmond Mark C. , Choi Young-Ku
Published: May 2011

A retrospective study of necropsy reports was undertaken to assess the relation between liver copper concentration and age, sex, and production class for calves that did not have a history compatible with copper toxicosis or with copper deficiency. Results of a generalized least squares regression analysis of 601 records found that copper concentration was as much as 60 mg/kg wet weight higher for dairy calves than for beef calves ( P < 0.00001, R2 = 0.172). For both beef and dairy calves, copper concentration also was significantly associated with linear ( P = 0.001) and nonlinear (age2, P = 0.002; age3, P = 0.008) functions of age, where the concentration increased for the first 2 months, then declined until 9 months of age, after which it began to increase. Sex was not associated with copper concentration ( P = 0.22). The 95% confidence intervals of the mean concentration for a group of calves and the 95% prediction intervals of a single concentration value for an individual animal are presented. As a consequence of the significant influence of both production class (dairy or beef) and age on liver copper concentration of calves with no evidence of copper toxicosis or copper deficiency, diagnosis of copper imbalances based on liver copper concentration in calves should consider the diagnostic covariates of age and production class.
The Pathology of Peracute Experimental Clostridium Perfringens Type D Enterotoxemia in Sheep
Authors: Uzal F. A. , Kelly W. R. , Morris W. E. , Bermudez J. , Baisón M.
Published: May 2011

The pathological findings in sheep with peracute experimental Clostridium perfringens type D enterotoxemia are described. Of 16 animals inoculated intraduodenally with a whole culture of this microorganism and a starch solution in the abomasum, 12 developed clinical signs including increased respiratory efforts, recumbency, paddling, bleating, convulsions, blindness, and opisthotonus. Diarrhea was not observed in any of the animals. The time lapse between the beginning of intraduodenal infusion and onset of clinical signs varied between 30 minutes and 26 hours, and the clinical course varied between 1 and 9 hours. Gross postmortem changes were observed in these 12 animals and included pulmonary edema; excess pericardial, peritoneal, or pleural fluid with or without strands of fibrin; liquid small intestinal contents; leptomeningeal edema; cerebellar coning; and subcapsular petechiae on kidneys. Histological changes consisted of severe edema of pleura and interlobular septa and around blood vessels and airways and acidophilic, homogeneous, proteinaceous perivascular edema in the brain. Five of 12 animals (42%) with clinical signs consistent with enterotoxemia lacked specific histological lesions in the brain. None of the intoxicated or control animals developed nephrosis. Glucose was detected in the urine of 3 of 6 animals that were tested for this analyte. These results stress the importance of the use of histological examination of the brain, coupled with epsilon toxin detection, for a definitive diagnosis of C. perfringens type D enterotoxemia in sheep.
Efficacy of Immunologic Assays for the Detection of Johne's Disease in Dairy Cows Fed Additional Energy during the Periparturient Period
Authors: Stabel Judith R. , Goff Jesse P.
Published: May 2011

This study was designed to evaluate if the immunosuppression typically observed during the immediate periparturient period (3 weeks before and after calving) in dairy cows influences the effectiveness of diagnostic tests for the detection of Johne's disease; and, if providing additional energy to the cows during this period would minimize any immunosuppressive effects. Twelve dairy cows naturally infected with Mycobacterium paratuberculosis were fitted with rumen cannulas in late gestation and assigned to treatment groups: control, n = 6; or stuffed, n = 6. Cows in the control group were allowed to consume feed ad libitum. Cows assigned to the stuffed treatment group were also fed ad libitum but received additional total mixed ration by manually stuffing their rumens with refused feed to maintain a dry matter intake of 2% body weight/day before calving and 2.5% body weight/day after calving. Parturition had a significant impact on immune function with significant reductions in M. paratuberculosis-specific antibodies detected in the serum and milk regardless of treatment group. Similarly, in vitro immunoglobulin production was decreased at calving for both treatment groups. In addition, stuffing cows modulated cell-mediated immune function by reducing antigen-specific lymphocyte proliferation and interferon- γ production after calving. Shedding of M. paratuberculosis in the milk was apparent in 58% (7/12) of cows after parturition with no difference noted between control and stuffed animals. Parturition had no major effect on fecal shedding of cows regardless of treatment. These data suggest that parturition had a significant effect on immune function parameters including diagnostic tests for paratuberculosis. Furthermore, providing additional energy to cows with Johne's disease did not preclude immunosuppressive effects during the periparturient period.
Fibrosarcoma with Lung and Lymph Node Metastases in an Asian Elephant (Elephas Maximus)
Authors: Liu Chen-Hsuan , Chang Chih-Hua , Chin Shih-Chien , Chang Pen-Heng , Zhuo Yi-Xing , Lee Chin-Cheng
Published: May 2011

A case of fibrosarcoma with lung and lymph node metastases in a 54-year-old female Asian elephant ( Elephas maximus) is described. After pododermatitis of 2 years duration in the right forefoot, a mass developed in the lateral toenail. At postmortem, metastasis to the right axillary lymph node and both lungs was noted. Microscopic examination of primary and metastatic sites revealed infiltrating bundles of spindle cells, with fairly distinct cell borders, variable amounts of eosinophilic cytoplasm, and elongate or oval nuclei. Tumor cells were often arranged in interwoven bundles and herringbone patterns. Mitotic figures were numerous and frequently bizarre. The diagnosis of fibrosarcoma with lung and lymph node metastases was made on the basis of histologic features and positive immunohistochemical staining for vimentin.
Atypical Hypoplastic Left Ventricular Syndrome in a Calf
Authors: Lemberger Karin Y. , Mohr Kayla R. , Andrews John J.
Published: May 2011

An 8-day-old male Angus calf was presented to the University of Illinois, Veterinary Teaching Hospital, Urbana, IL, for lethargy, weakness, and poor suckle reflex. Clinical evaluation revealed a strong left-sided heart murmur and a split S2 sound. The calf died within 48 hours. Necropsy revealed a combination of the following cardiac defects: left ventricular hypoplasia, high ventricular septal defect, left auricular atresia with mitral valve aplasia, patent foramen ovale, patent ductus arteriosus, and pulmonary trunk atresia. Mild suppurative pneumonia with pulmonary edema and congestion were also present. This combination of defects appears to be similar to the hypoplastic left heart syndrome in humans and is reported here for the first time in cattle.
Isolation and Antimicrobial Susceptibilities of Bacterial Pathogens from Bovine Pneumonia: 1994–2002
Authors: Welsh Ronald D. , Dye Laura B. , Payton Mark E. , Confer Anthony W.
Published: May 2011

Between 1994 and 2002, a total of 390 (46.3%) Mannheimia haemolytica, 292 (34.7%) Pasteurella multocida, and 160 (19.0%) Histophilus somni were isolated at the Oklahoma Animal Disease Diagnostic Laboratory from lungs from 6–18-month-old beef cattle with pneumonia. The ratio of M. haemolytica isolations to P. multocida isolations decreased from 3.1 in 1994 to 0.8 in 2000 while increasing to 1.5 in 2002. Mannheimia haemolytica isolations significantly ( P < 0.05) decreased from 62.5% in 1994 to between 30.6% and 50.4% in 1998–2002. Pasteurella multocida isolations significantly ( P < 0.05) increased from 20.0% in 1994 to between 28.6% and 47.4% in 1998–2002. Histophilus somni isolations were <19% except in 1998 (40.8%) and 1999 (23%). Antimicrobial susceptibilities for M. haemolytica significantly declined for erythromycin ( P = 0.0001), florfenicol ( P = 0.0004), spectinomycin ( P = 0.0001), and tilmicosin ( P = 0.03). For P. multocida, antimicrobial susceptibilities significantly declined for erythromycin ( P = 0.0001), florfenicol ( P = 0.004), spectinomycin ( P = 0.03), sulfachloropyridizine ( P = 0.028), tetracycline ( P = 0.017), tilmicosin ( P = 0.0001), and trimethoprim/sulfamethoxazole ( P = 0.0003). Antimicrobial susceptibilities for H. somni were variable for spectinomycin and sulfachloropyridizine, whereas susceptibilities to other antibiotics remained consistently high.
Bovine Arsenic Toxicosis
Authors: Neiger Regg , Nelson Nicole , Miskimins Dale , Caster Jim , Caster Larry
Published: May 2011

A ranch in central South Dakota had a number of dead calves because of arsenic poisoning. The clinical picture included diarrhea, central nervous system signs, and death. Gross necropsy findings included adequate body fat, stomachs full of normal-appearing ingesta, and large amounts of greenish brown watery fluid in the intestine and colon. Microscopically there was severe lymphoid tissue necrosis in the mesenteric lymph nodes and gut-associated lymphoid tissue. Chemical analysis of kidneys showed no significant amounts of lead; however, kidney arsenic concentrations were 25 to 44 ppm. The source was a small pile of Paris Green (common name for cupric acetoarsenite) found in an old dump site in the pasture.
Molecular Diagnosis of Paramyxovirus Infection in Snakes Using Reverse Transcriptase–Polymerase Chain Reaction and Complementary Deoxyribonucleic Acid:Ribonucleic Acid in Situ Hybridization
Authors: Sand Matthew A. , Latimer Kenneth S. , Gregory Christopher R. , Rakich Pauline M. , Jacobson Elliott , Pennick Kate E.
Published: May 2011

Identification of ophidian paramyxovirus (OPMV) nucleic acid was accomplished in 11 of 14 snakes by a reverse transcriptase–polymerase chain reaction (RT-PCR) assay that detected a 153-bp region of the OPMV genome in total RNA extracted from paraffin-embedded tissues and cell culture. The RT-PCR protocol amplified a portion of the OPMV RNA genome, producing a 153-bp complementary DNA (cDNA) product from both fresh and paraffin-embedded tissue samples. In addition, cDNA:RNA in situ hybridization localized OPMV in formalin-fixed, paraffin-embedded tissue specimens to specific tissues and cells. This latter technique increased the degree of specificity with which a diagnosis of OPMV could be made.
Suspected Metastatic Coccygeal Chordoma in a Ferret (Mustela Putorius Furo)
Authors: Munday John S. , Brown Cathy A. , Richey Lauren J.
Published: May 2011

A chordoma was removed from the tail base of a 6.5-year-old ferret ( Mustela putorius furo). A nodule was observed in the area of tumor development when the ferret was purchased at 3 months of age. Although the nodule did not enlarge for 2 years, slow, steady growth of the tumor was observed for 4 years before surgical removal. Eight months after removal of the chordoma, the ferret developed 2 cutaneous masses. One was adjacent to the vulva, close to where the chordoma had been removed from, whereas the other was in the nasofacial region. After 4 months of slow growth, both masses were removed and both were histologically and immunohistochemically consistent with chordoma. Over the next 8 weeks, additional masses developed in the facial, maxillary gingival, and scapular regions. Enlargement of the gingival mass caused dysphagia, and the ferret was euthanized. Although a necropsy was not performed, these additional masses had a clinical appearance and texture that was similar to the 2 previously removed cutaneous chordomas. To the authors' knowledge, this is the first report of a ferret coccygeal chordoma that developed close to the base of the tail. Ferret chordomas have been reported previously to metastasize to the subcutis overlying the tumor. However, this is the first report of a ferret chordoma that metastasized to a location distant to the primary site of neoplasm development. Cell proliferation indices did not predict this metastatic behavior. It is hypothesized that the long clinical period before removal may have predisposed this neoplasm to metastasis. Observations from this case suggest that chordomas in ferrets may have metastatic potential and so should be removed promptly.
Molecular Epidemiology of Two Fowl Cholera Outbreaks on a Free-Range Chicken Layer Farm
Authors: Zhang P. , Fegan N. , Fraser I. , Duffy P. , Bowles R. E. , Gordon A. , Ketterer P. J. , Shinwari W. , Blackall P. J.
Published: May 2011

Two outbreaks of fowl cholera on a multiage free-range egg farm were investigated. The outbreaks occurred in 1994 and 2002. A total of 22 strains of Pasteurella multocida were available for study, 11 from the 1994 outbreak and 11 from the 2002 outbreak. Lesions typical of acute fowl cholera were seen in the 1994 outbreak, whereas both acute and chronic fowl cholera occurred in the 2002 outbreak. The isolates were examined in an extended phenotypic typing methodology, by a P. multocida–specific polymerase chain reaction (PCR), by the Heddleston somatic serotyping scheme, and by restriction endonuclease analysis (REA) typing using the enzyme HpaII. All 22 strains had the same phenotypic properties, all were confirmed as P. multocida by PCR, all were Heddleston serovar 4, and all had the same REA pattern. The results indicate that these 2 outbreaks were caused by the same clone of P. multocida–-despite the 8-year time period between the outbreaks.
Congenital Polycystic Kidney in a White-Tailed Deer (Odocoileus Virginianus)
Authors: Palmer Mitchell V. , Carpenter Jeremy G.
Published: May 2011

Polycystic kidney and liver disease was seen in a stillborn white-tailed deer ( Odocoileus virginianus) fawn. Bilaterally enlarged kidneys were characterized by severe dilatation of all renal tubules. Glomeruli were sparse, small, and located within a dilated Bowman's capsule. The liver was characterized by marked periportal fibrosis, biliary hyperplasia, and bile duct ectasia with dilated ducts containing inspissated bile. The presentation and morphology of this case are most similar to autosomal recessive polycystic disease in humans.
A Multilaboratory Evaluation of a Commercial Enzyme-Linked Immunosorbent Assay Test for the Detection of Antibodies against Mycobacterium Avium Subsp. Paratuberculosis in Cattle
Authors: Dargatz David A. , Byrum Beverly A. , Collins Michael T. , Goyal Sagar M. , Hietala Sharon K. , Jacobson Richard H. , Kopral Christine A. , Martin Barbara M. , McCluskey Brian J. , Tewari Deepanker
Published: May 2011

Five laboratories participated in a study to evaluate sources of variation in results from an enzyme-linked immunosorbent assay (ELISA) for antibodies against Mycobacterium avium subsp. paratuberculosis. Each laboratory repeatedly tested duplicates of a negative, positive (P), and high-positive (HP) serum sample, which were supplied by the United States Department of Agriculture: Animal and Plant Health Inspection Service: Veterinary Services, National Veterinary Services Laboratories, Ames, IA, on all 96-well microtiter plates when routinely testing other samples for M. avium subsp. paratuberculosis antibodies. These 3 sera were aliquoted and sent to the 5 participating laboratories. This study focused on variation in test results because of assay reagents and laboratory techniques and did not account for biologic variability associated with the time course of infection in cattle. Overall, results from 868 microtiter plates were used in the study. For each sample a sample-to-positive (S/P) ratio was calculated according to the manufacturer's directions. The S/P ratio for the P sample ranged from 0.06 to 1.039 (mean = 0.466 and 0.484 for wells 1 and 2, respectively) and those for the HP sample ranged from 2.446 to 8.727 (mean = 4.027 and 3.980 for wells 1 and 2, respectively). The majority of the variation in S/P ratio for the P sample was attributed to kit lot (37.5%), followed by random (unexplained) error (27.0%), laboratory (18.3%), and kit lot by laboratory (11.9%). By eliminating plates in which the separation between negative and positive control ODs was less than 0.4, the proportion of variation attributed to laboratory was reduced markedly. These results confirm that there is variability in M. avium subsp. paratuberculosis ELISA results and that several sources contribute to the observed variability. The study gives a relative estimate of the contribution of various sources to the overall variability observed in the M. avium subsp. paratuberculosis ELISA results with kit lot being a primary contributor. Similar data for other ELISA tests for antibodies to M. avium subsp. paratuberculosis or other antigens also should be developed.
Morphological Variation of “Complex Vertebral Malformation” in Holstein Calves
Authors: Agerholm Jorgen S. , Bendixen Christian , Arnbjerg Jens , Andersen Ole
Published: May 2011

A study was performed to investigate the morphological expression of the inherited syndrome “complex vertebral malformation” (CVM) in Holstein calves. A total of 107 late-term aborted, premature, or neonatal calves suspected of having CVM were necropsied and retrospectively analyzed for the causal mutation in the gene SLC35A3. Sixty-two calves were homozygous affected, 16 were heterozygous, and 29 were homozygous normal. Calves affected by CVM were growth retarded. Vertebral lesions identified by radiography were present in 61 cases, of which 58 also had costal malformation. Malformation of the head, primarily in the form of dysplasia or palatoschisis, was present in 15 cases. Bilateral symmetric flexion of the carpal and metacarpophalangeal joints was present in all cases, whereas posterior arthrogryposis was found in 54 cases. Interventricular septal defects occurred in 33 calves, often in combination with other cardiac malformations. A wide spectrum of additional malformations was found. Other congenital syndromes were in most cases distinguishable from CVM on a morphological basis. However, a calf with a prenatal infection with bovine virus diarrhea virus constituted a phenocopy. The study demonstrated that the morphological expression of CVM is wide, but certain aspects, i.e., growth retardation, vertebral malformation, and symmetric arthrogryposis, are almost constant findings. However, cases without vertebral defects and phenocopies constitute a diagnostic problem. A presumptive diagnosis of CVM can in most cases be based on necropsy findings combined with information on descent and paternal CVM genotype, whereas a definitive diagnosis requires genotyping.
Recognition of the Nor98 Variant of Scrapie in the Swedish Sheep Population
Authors: Gavier-Widén D. , Nöremark M. , Benestad S. , Simmons M. , Renström L. , Bratberg B. , Elvander M. , Segerstad C.Hård af
Published: May 2011

Within the framework of the active surveillance for transmissible spongiform encephalopathies in sheep in Sweden, 4 cases of the atypical form of scrapie, Nor98, were identified during 2003. Nor98 is a recently recognized and poorly understood variant of scrapie, first described in Norway. The cases were positive by the rapid test (enzyme-linked immunosorbent assay). Immunohistochemical staining showed diffuse thin-granular staining of the cerebellar cortex. Western immunoblotting analysis of specimens of brain stem and cerebellum showed a light band of approximately 12 kDa. Typical scrapie was ruled out based on the confirmatory testing. The affected ewes were from 4 different flocks. They were between 7 and 9 years old. Two were of the ARQ/ARQ genotype, 1 ARR/ARQ, and 1 ARR/AHQ. Two ewes had shown ataxia, and the other 2 had no clinical signs. Whole-flock slaughter was applied, and testing of the flock mates did not reveal additional cases. Nor98 differs from typical scrapie in its epidemiology, frequency of genotypes of sheep affected, clinical signs, microscopic lesions, distribution of scrapie prion protein in the brain, and characteristics of the immunostaining and immunoblotting profiles.
Granulomatous Pneumonia Caused by Pythium Insidiosum in a Central American Jaguar, Panthera Onca
Authors: Camus Alvin C. , Grooters Amy M. , Aquilar Roberto F.
Published: May 2011

A 7-month-old, male jaguar presented with dyspnea and leukocytosis unresponsive to antibiotic therapy. Radiographs revealed unilateral pulmonary consolidation. An exploratory thoracotomy was performed, and the left lung, which contained a large multilobular mass with extensive fibrosis and numerous caseonecrotic foci, was removed. Microscopically, eosinophilic granulomatous inflammation surrounded broad (4.4–8.3 μm) rarely septate hyphae. A diagnosis of Pythium insidiosum infection was confirmed by immunohistochemistry, immunoblot serology, culture, and polymerase chain reaction. Dyspnea recurred despite treatment, and the animal succumbed 3 weeks after surgery. Necropsy findings indicated that death resulted from occlusion of the right main stem bronchus by a fungal granuloma. The oomycete P. insidiosum typically causes granulomatous disease of the skin or gastrointestinal tract in animals and arteritis, keratitis, or cellulitis in humans. Infection is uncommon in felines, and pulmonary involvement is rare. This report details the first case of P. insidiosum infection in an exotic felid and provides the first description of primary pulmonary pythiosis in any species.
Diffuse Cystic Endometrial Hyperplasia and Metastatic Endometrial Adenocarcinoma in a Vietnamese Pot-Bellied Pig (Sus Scrofa)
Authors: Harmon B. G. , Munday J. S. , Crane M. M.
Published: May 2011

A 16-year-old female Vietnamese pot-bellied pig was euthanized after a period of inappetence and weight loss. Diffuse cystic endometrial hyperplasia and endometrial adenocarcinoma with metastasis to lymph nodes, liver, and lung were diagnosed. This report follows the recent description of cystic endometrial hyperplasia and uterine leiomyomas in 3 aged female Vietnamese pot-bellied pigs. The findings in this report and previous reports suggest that pigs may develop some similar age-related uterine lesions as do women.
Evaluation of Five Different Antigens in Enzyme-Linked Immunosorbent Assay for the Detection of Avian Pneumovirus Antibodies
Authors: Maherchandani Sunil , Patnayak Devi P. , Muñoz-Zanzi Claudia A. , Lauer Dale , Goyal Sagar M.
Published: May 2011

Five different antigens were evaluated in enzyme-linked immunosorbent assay (ELISA) tests for the detection of avian pneumovirus (APV) antibodies. Two of the 5 antigens were prepared from recent APV isolates from Minnesota. The 2 older isolates were passage 63 of a strain currently used as a live, attenuated vaccine and a Colorado strain isolated for the first time in the United States and currently used in an ELISA test. The fifth antigen is based on an APV recombinant N-protein. Basic parameters and positive-negative threshold of the assays were established for all 5 antigens on the basis of data obtained by testing 46 known negative and 46 known positive serum samples. Subsequently, 449 field samples were tested by all 5 ELISAs. The optical density difference (ODD) was calculated by subtracting optical density of the sample in the negative antigen well from that in the positive antigen well. In the current ELISA test based on the Colorado strain, an ODD of 0.2 is considered to be the cutoff value to classify samples as negative or positive. In this study, however, use of different cutoffs, based on ODD of negative control plus 3 SD or values estimated from Receiver operating characteristic analysis, was considered to be more appropriate for the various antigens used. Overall person-to-person and day-to-day variability was found to be large for all tests using either ODD or sample to positive ratio to report results. In addition, results suggest that antigenicity of the APV isolates in the United States has not changed between 1997 and 2000.
Comparison of Two Swine Mycoplasma Hyopneumoniae Enzyme-Linked Immunosorbent Assays for Detection of Antibodies from Vaccinated Pigs and Field Serum Samples
Authors: Ameri-Mahabadi Mehrdad , Zhou En-Min , Hsu Walter H.
Published: May 2011

Mycoplasma hyopneumoniae (Mhyo) causes mycoplasmal pneumonia, an economically important disease of swine. Serodiagnosis of Mhyo is based on the current available commercial enzyme immunoassays for detection of swine antibodies against Mhyo, which are the indirect enzyme-linked immunosorbent assay (ELISA) and the blocking ELISA (B-ELISA). Because of the limited information available for these ELISAs, these 2 assays were compared by testing 347 serum samples collected from vaccinated pigs at 0, 13, 28, 43, and 62 days postimmunization (DPI), 50 samples from nonvaccinated pigs, and 1,013 field serum samples. The results of comparison study showed that the specificity for both ELISAs was 99.2% generated from 139 non-vaccinated negative samples. The sensitivities for indirect ELISA generated from samples collected from animals that received the vaccine at DPI 13, 28, 43, and 62 were 0%, 95.7%, 88.4%, and 92.6%, respectively, whereas the sensitivities for B-ELISA were 0%, 98%, 100%, and 97%, respectively. The overall agreement of 96.7% and 80.3% was generated between 2 ELISAs from negative and vaccinated pigs and from field samples, respectively.
Molecular Detection of Leptospira Kirschneri in Tissues of a Prematurely Born Foal
Authors: Vemulapalli Ramesh , Langohr Ingeborg M. , Sanchez Alfredo , Kiupel Matti , Bolin Carol A. , Wu Ching-Ching , Lin Tsang-Long
Published: May 2011

Leptospirosis was identified to be the possible cause of premature birth in a foal on a farm with a history of repeated abortions. Using an appropriate polymerase chain reaction (PCR) assay, the presence of Leptospira kirschneri was detected in the tissues of the prematurely born foal. Further confirmation of L. kirschneri was obtained by nucleotide sequence analysis of the PCR-amplified DNA fragment and the partial 16S ribosomal RNA gene sequence. This report further supports mounting evidence that a PCR assay capable of detecting L. kirschneri should be included in routine diagnostic investigations in which Leptospira spp. infection is suspected.
Use of a Novel Virus Inactivation Method for a Multicenter Avian Influenza Real-Time Reverse Transcriptase—Polymerase Chain Reaction Proficiency Study
Authors: Spackman Erica , Suarez David L.
Published: May 2011

Proficiency assessments are important elements in quality control for diagnostic laboratories. Traditionally, proficiency testing for polymerase chain reaction (PCR)–based assays has involved the use of clinical samples, samples “spiked” with live agents or DNA plasmids. Because of government regulations and biosecurity concerns, distribution of live high-consequence pathogens of livestock and poultry, such as avian influenza, is not possible, and DNA plasmids are not technically suitable for evaluating RNA virus detection. Therefore, a proficiency testing panel using whole avian influenza in a diluent containing a phenolic disinfectant that inactivates the virus while preserving the RNA for at least 8 weeks at −70 C was developed and used in a multicenter proficiency assessment for a type A influenza real-time reverse transcriptase (RT)–PCR test. The test, which was highly standardized, except for variation in the real-time RT-PCR equipment used, was shown to be highly reproducible by proficiency testing in 12 laboratories in the United States, Canada, and Hong Kong. Variation in cycle threshold values among 35 data sets and 490 samples was minimal (CV = 5.19%), and sample identifications were highly accurate (96.7% correct identifications) regardless of real-time PCR instrumentation.
Culture Methods Differ on the Isolation of Salmonella Enterica Serotypes from Naturally Contaminated Swine Fecal Samples
Authors: Rostagno Marcos H. , Gailey Jared K. , Hurd H. Scott , Mckean James D. , Leite Romulo C.
Published: May 2011

Four culture methods (A, B, C, and D) were comparatively evaluated for their ability to isolate Salmonella enterica from pooled swine fecal samples ( n = 100). None of the methods was able to isolate Salmonella from all positive samples. The relative sensitivity of the culture methods evaluated was 82%, 94%, 95%, and 78% for methods A, B, C, and D, respectively. The comparison of sensitivities showed that methods B and C performed significantly better ( P < 0.05) than methods A and D. Although relative sensitivities of methods B and C were equal, from the 89 positive samples concomitantly detected by both, 35 (39.3%) had different serotypes (no match) isolated by each method. On the basis of the results of this study, it was concluded that culture methods differ on the isolation of S. enterica serotypes from naturally contaminated swine fecal samples. Depending on the objective(s) of investigations on the ecology and epidemiology of S. enterica in swine populations, a method or a combination of methods should be considered for more reliable results.
A Comparison of Rapid Bovine Spongiform Encephalopathy Testing Methods on Autolyzed Bovine Brain Tissue
Authors: Wear Angus , Henderson Kirstine , Webster Kath , Patel Indu
Published: May 2011

In 1999, the European Union (EU) approved 3 rapid methods for the testing of bovine brain samples for the presence of bovine spongiform encephalopathy (BSE). The evaluation that led to the approval did not include an analysis of autolyzed material. Member states of the EU have active surveillance programs for BSE, which target fallen stock as well as other categories of cattle. Autolysis is a common feature of fallen stock samples because there can be a considerable delay between death and collection of samples. Therefore, it is important to know whether these tests perform optimally on autolyzed samples. The Veterinary Laboratories Agency (VLA) selected 250 positive fallen stock samples. These had been detected during routine testing using the Prionics®-Check Western blot and confirmed as BSE cases by immunohistochemistry or electron microscopy. Samples were graded according to the degree of autolysis and then tested by the 3 methods: Prionics®-Check Western blot, Platelia test, and Enfer test. All 3 methods correctly classified the samples as positive BSE cases, therefore alleviating doubt about their ability to do so. Subsequent EU validation exercises, such as those conducted in 2002–2003, have included the testing of autolyzed material. It is important that all new methods be evaluated on autolyzed tissue before approval for official use.
Application of Nox-Restriction Fragment Length Polymorphism for the Differentiation of Brachyspira Intestinal Spirochetes Isolated from Pigs and Poultry in Australia
Authors: Townsend Kirsty M. , Giang Vo Ngan , Stephens Carol , Scott Paul T. , Trott Darren J.
Published: May 2011

Sixty-nine intestinal spirochetes isolated from pigs and poultry in eastern Australia were selected to evaluate the effectiveness of a species-specific PCR-based restriction fragment length polymorphism (RFLP) analysis of the Brachyspira nox gene. For comparative purposes, all isolates were subjected to species-specific PCRs for the pathogenic species Brachyspira hyodysenteriae and Brachyspira pilosicoli, and selected isolates were examined further by sequence analysis of the nox and 16S ribosomal RNA genes. Modifications to the original nox-RFLP method included direct inoculation of bacterial cells into the amplification mixture and purification of the PCR product, which further optimized the nox-RFLP for use in a veterinary diagnostic laboratory, producing sufficient product for both species identification and future comparisons. Although some novel profiles that prevented definitive identification were observed, the nox-RFLP method successfully classified 45 of 51 (88%) porcine and 15 of 18 (83%) avian isolates into 5 of the 6 recognized species of Brachyspira. This protocol represents a significant improvement over conventional methods currently used in veterinary diagnostic laboratories for rapid specific identification of Brachyspira spp. isolated from both pigs and poultry.
High-Throughput Real-Time RT-PCR Assay to Detect the Exotic Newcastle Disease Virus during the California 2002–2003 Outbreak
Authors: Crossley Beate M. , Hietala Sharon K. , Shih Liu-Mei , Lee Lou , Skowronski Evan W. , Ardans Alex A.
Published: May 2011

During the 2002–2003 Exotic Newcastle Disease (END) outbreak in Southern California, a high-throughput real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) system was developed to respond to the large diagnostic and surveillance sample workload. A 96-well RNA extraction method, using magnetic bead technology, combined with a 96-well RRT-PCR assay, allowed 1 technician to process and test more than 400 samples per day. A 3-technician team could complete testing on approximately 1,900 samples per day. The diagnostic sensitivity of the high-throughput RRT-PCR assay was 0.9967 (95% CI 0.9937–0.9997) based on 926 virus isolation confirmed positive samples. Diagnostic specificity using an initial 434 virus isolation confirmed negative samples was 100%. A diagnostic specificity of 0.9999 (95% CI 0.9999, >0.9999) was subsequently calculated on the basis of 2 false-positive results among 65,343 surveillance samples collected after the final END-positive case was confirmed in May 2003. Assay performance over 500 replicates, including reproducibility of the combined extraction and RRT-PCR amplification steps yielded a standard deviation of 0.70 RRT-PCR cycle thresholds (Ct) and a standard deviation of 0.59 Ct for the RRT-PCR steps alone. The high-throughput RRT-PCR developed for END contributed significantly to the 2002–2003 END control effort, reducing the predicted timeline for eradication from 3 years to just 11 months, primarily because of the large number of samples that could be rapidly tested. The 96-well approach described for high-throughput END RRT-PCR could be adapted to other rapid, high-volume testing needs, as required for potential foreign animal disease responses or intensive surveillance efforts.
C-Reactive Protein Measurement in Canine Saliva
Authors: Parra María Dolores , Tecles Fernando , Subiela Silvia Martínez , Cerón José Joaquín
Published: May 2011

An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 μg/ml and 0.0091 μg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7–9.9% and 8.5–16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental ( R2 = 0.76) and clinical studies ( R2 = 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments.
Immunohistochemical and Ultrastructural Detection of Intestinal Spirochetes in Thoroughbred Horses
Authors: Shibahara Tomoyuki , Kuwano Atsutoshi , Ueno Takanori , Katayama Yoshinari , Ohya Tatsuo , Taharaguchi Sadao , Yamamoto Shinji , Umemura Takashi , Ishikawa Yoshiharu , Kadota Koichi
Published: May 2011

Studies of equine intestinal spirochetes have long focused on intestinal contents alone, but intestinal spirochetosis has been reported recently in a 21-month-old Thoroughbred colt in Japan. To define the clinical and pathological significances of intestinal spirochetosis in several horses, an epizootiologic survey with histologic, immunohistochemical, and ultrastructural methods was conducted for Brachyspira antigen–containing intestinal spirochetes in 12 diseased or injured Thoroughbred horses, aged from 35 days to 17 years. Brachyspira antigen–containing spirochetes were found in 7 of 12 horses (58.3%) and were more frequent in the cecum than in other parts of the bowel. It was not clear whether the infection was clinically related to diarrhea or dysentery, but histopathology revealed a close association between the bacterial infection and epithelial hyperplasia. Crypt epithelium consisted mainly of goblet cells and showed frequent mitosis throughout its length. Inflammatory cells and congestion were also present. There were numerous spirochetes in the crypts, and some invaded the cecal and colonic epithelia and underlying lamina propria. Ultrastructurally, the spirochetes were divided into 4 types. Three types were identified in degenerative epithelial cells or intracellularly. Brachyspira antigen–containing intestinal spirochetes invading the mucosa were capable of causing epithelial hyperplasia in the cecum and colon in the horses. The findings in this study will increase awareness of the importance of intestinal spirochetosis and may also be helpful for diagnosis and treatment of this condition.
Malignant Catarrhal Fever-like Disease in Sheep after Intranasal Inoculation with Ovine Herpesvirus-2
Authors: Li Hong , O'Toole Donal , Kim Okjin , Oaks J. Lindsay , Crawford Timothy B.
Published: May 2011

A malignant catarrhal fever (MCF)–like disease was induced experimentally in 3 sheep after aerosol inoculation with ovine herpesvirus-2 (OvHV-2). Each of 3 OvHV-2–negative sheep was nebulized with 2 ml of nasal secretions containing approximately 3.07 − 109 OvHV-2 DNA copies from a sheep experiencing an intensive viral-shedding episode. Ovine herpesvirus-2 DNA became detectable by polymerase chain reaction in the peripheral blood leukocytes of all 3 sheep within 3 days, and all 3 seroconverted between 6 and 8 days postinfection (PI). The sheep developed clinical signs, with copious mucopurulent nasal discharge and fever around 14 days PI. One of the 3 clinically affected sheep was euthanized at 18 days PI. Major lesions at necropsy were multifocal linear erosions and ulcers in mucosa of the cheeks, tongue, pharynx, and proximal esophagus and mild disseminated pneumonia. Microscopically, there was extensive moderate superficial histiocytic–lymphocytic rhinitis with epithelial dissociation and degeneration. Moderate multifocal histiocytic bronchointerstitial pneumonia was associated with loss of terminal bronchiolar epithelium. Lymphocytic vasculitis was present only in the lung. The remaining 2 sheep recovered clinically, approximately 25 days PI. The study revealed that clinical signs and lesions resembling MCF can develop when uninfected sheep are exposed to a high dose of aerosolized OvHV-2.
Detection of Echinococcus Multilocularis Infection in a Colony of Cynomolgus Monkeys (Macaca Fascicularis) using Serology and Ultrasonography
Authors: Rehmann Patrick , Gröne Andrea , Gottstein Bruno , Völlm Jörg , Sager Heinz , Janovsky Martin , Bacciarini Luca N.
Published: May 2011

Five animals in a colony of cynomolgus monkeys ( Macaca fascicularis) died or were euthanatized because of alveolar echinococcosis, during a period of 5 years. The remainder of the colony was screened for possible infection with Echinococcus multilocularis, using serology and ultrasonography. A total of 46 animals out of a group of 55 were examined. The presence of anti-Em2 antibodies analyzed with enzyme-linked immunosorbent assay was demonstrated in 3 monkeys. In 2 of these 3 monkeys, multilocular structures compatible with metacestodal cysts in the liver were identified, using ultrasonography. The presence of alveolar echinococcosis was subsequently confirmed at postmortem examination in 1 animal. The other animals are still alive. Two other monkeys were negative in the serological examination but had cystic structures in the liver, which were identified as bile duct cysts at postmortem examination in 1 animal. The other monkey is still alive. These findings suggest that serology for antibodies against the Em2 antigen may represent a useful method in identifying animals that might be infected with E. multilocularis and are therefore at risk of developing fatal alveolar echinococcosis.