bluuebunny/crossref_metadata_2025 huggingface dataset:
Papillary Tumor of the Pineal Region of a Dog
Authors: Williams Jonathan M. , Michal Ulrike , Botteron Catherine , Skerritt Geoffrey , Hetzel Udo
Published: May 2011

A 3-year-old, intact, male Beagle dog developed clinical signs of pleurothotonus and altered head position to the right, neck pain, nystagmus, hyperreflexia of the left forelimb, and hyperextension of both forelimbs. Magnetic resonance imaging enabled a tentative diagnosis of thalamic neoplasia with incidental hydromyelia at the level of the second cervical vertebra. The animal was euthanatized due to the poor prognosis, and a necropsy was performed. A large, well-demarcated, nonencapsulated, and focally infiltrative mass was present in the approximate location of, and effacing, the pineal gland. The mass was composed of densely packed polyhedral neoplastic cells that exhibited epithelial characteristics, such as intercellular junctions, and contained carbohydrate granules and occasionally melanin granules. Immunohistology confirmed that neoplastic cells expressed neuron-specific enolase and, in a small proportion, cytokeratin. These combined findings led to the diagnosis of a papillary tumor of the pineal region, a tumor not previously described in dogs.
Detection of Anti-Influenza A Nucleoprotein Antibodies in Pigs Using a Commercial Influenza Epitope-Blocking Enzyme-Linked Immunosorbent Assay Developed for Avian Species
Authors: Ciacci-Zanella Janice R. , Vincent Amy L. , Prickett John R. , Zimmerman Silvia M. , Zimmerman Jeffrey J.
Published: May 2011

Influenza virus causes acute respiratory disease in pigs and is of concern for its potential public health significance. Many subtypes of influenza virus have been isolated from pigs, and the virus continues to evolve in swine populations. Current antibody assays have limited antigenic recognition, and accurate, broad-spectrum, high through-put screening tests are needed to detect infections in swine herds and to aid in the implementation of control measures. In the current study, a commercial blocking enzyme-linked immunosorbent assay (ELISA) developed for the detection of Influenza A virus nucleoprotein antibodies in avian species was evaluated for the detection of anti-influenza serum antibodies in swine. Serum samples used to evaluate the test were archived samples from influenza research conducted at the U.S. Department of Agriculture–Agricultural Research Service–National Animal Disease Center and included samples from influenza-inoculated pigs (H1N1, H1N2, H2N3, and H3N2), contact-infected pigs, vaccinated pigs, and negative controls. Based on samples of known status ( n = 453), a receiver operating characteristic (ROC) curve analysis of the ELISA results estimated the optimized diagnostic sensitivity and specificity at 96.6% (95% confidence interval [CI]: 92.3, 98.9) and 99.3% (95% CI: 97.6, 99.9), respectively. By using the cutoff established in the ROC analysis, the assay was evaluated in pigs infected with 2 isolates of the 2009 pandemic H1N1 virus. Overall, the assay showed excellent diagnostic performance against the range of influenza subtypes investigated and could serve as a useful screening assay for swine.
Development and Application of an Indirect Immunoperoxidase Assay for the Detection of Duck Swollen Head Hemorrhagic Disease Virus Antigen in Pekin Ducks (Anas Platyrhynchos)
Authors: Li Chuanfeng , Shen Chanjuan , Cheng Anchun , Wang Mingshu , Zhang Na , Zhou Yi , Zhu Dekang , Jia Renyong , Luo Qihui , Chen Xiaoyue
Published: May 2011

An improved indirect immunoperoxidase assay (IPA) was developed to detect antigens of Duck swollen head hemorrhagic disease virus (DSHDV) in paraformaldehyde-fixed, paraffin-embedded tissues of Pekin ducks ( Anas platyrhynchos). This technique used an indirect streptavidin–alkaline phosphatase labeling system with polyclonal antiserum developed against purified DSHDV antigens. Specimens from the experimentally inoculated Pekin ducks with DSHDV and archived paraffin-embedded tissues from natural cases of Duck viral swollen head hemorrhagic disease (DVSHD) were examined by clinical and histological criteria. Positive staining was most widely observed in the cytoplasm of the following organs: immune, digestive, and urinary organs, heart, lung, and trachea, which corresponded to the intracellular distribution of reovirus. The DSHDV antigens were first detected at 4 hr postinoculation in the bursa of Fabricius of infected ducks. Therefore, this method was suitable for the early diagnosis of DVSHD. Immunoperoxidase staining was not present in tissues and organs of sham-inoculated ducks (negative control). The IPA developed in the current study is a convenient, sensitive, and specific means of detecting DSHDV and is applicable to routine diagnosis, retrospective studies, and prospective studies of DSHDV infection in ducks.
Orthoreovirus Infection and Concurrent Cryptosporidiosis in Rough Green Snakes (Opheodrys Aestivus): Pathology and Identification of a Novel Orthoreovirus Strain via Polymerase Chain Reaction and Sequencing
Authors: Landolfi Jennifer A. , Terio Karen A. , Kinsel Michael J. , Langan Jennifer , Zachariah Trevor T. , Childress April L. , Wellehan James F. X.
Published: May 2011

Reoviruses are nonenveloped, segmented, double-stranded RNA viruses capable of infecting a wide range of invertebrate, vertebrate, fungus, and plant hosts. Though sporadic infection has been reported in a variety of reptilian species, infection of rough green snakes ( Opheodrys aestivus) has not been previously described. Five wild-caught, adult rough green snakes were obtained by a zoological institution. Clinical deterioration was first noted in all snakes after 3 weeks in quarantine. Despite treatment, clinical decline progressed, and all 5 snakes died or were euthanized by 48 days post-arrival. Moderate, multifocal, acute, necrotizing hepatitis with hepatocellular syncytia was diagnosed in 1 snake. Two additional snakes had severe, diffuse, subacute to chronic pancreatitis. All 5 snakes had gastroenteric cryptosporidiosis. Electron microscopic examination of liver from the snake with hepatic lesions revealed scattered hepatocytes containing 1 or more intranuclear clusters of approximately 90 nm in diameter viral particles arranged in loose arrays. Polymerase chain reaction (PCR) amplification of a segment of the reovirus RNA-dependent RNA polymerase gene was performed on RNA extracted from tissues of all 5 snakes. PCR amplification of samples extracted from the snake with hepatic lesions resulted in a 109–base pair (bp) product. Phylogenetic analyses indicated the virus was a novel strain distinct from other reoviruses at a level consistent with species difference. The source of infection was unknown. PCR amplification of samples extracted from the other 4 snakes was negative.
Prevalence and Antimicrobial Susceptibility of Salmonellae Isolates from Reptiles in Taiwan
Authors: Chen Chun-Yu , Chen Wan-Ching , Chin Shih-Chien , Lai Yen-Hsueh , Tung Kwong-Chung , Chiou Chien-Shun , Hsu Yuan-Man , Chang Chao-Chin
Published: May 2011

Pets, including reptiles, have been shown to be a source of Salmonella infection in humans. Due to increasing popularity and variety of exotic reptiles as pets in recent years, more human clinical cases of reptile-associated Salmonella infection have been identified. However, limited information is available with regard to serotypes in different reptiles (turtles, snakes, and lizards) and antimicrobial resistance of Salmonella in pet reptiles. The current study was thus conducted to determine the prevalence of Salmonella colonization in pet reptiles. Salmonella organisms were isolated from 30.9% of 476 reptiles investigated. The isolation prevalences were 69.7% (23/33), 62.8% (27/43), and 24.3% (97/400) in snakes, lizards, and turtles, respectively. A total of 44 different Salmonella serovars were identified. Compared with S. Heron, Bredeney, Treforest, and 4,[5],12:i:-, S. Typhimurium isolates were resistant to many antimicrobials tested, and notably 61.1% of the isolates were resistant to cephalothin. The results indicated that raising reptiles as pets could be a possible source of Salmonella infection in humans, particularly zoonotic Salmonella serovars such as S. Typhimurium that may be resistant to antimicrobials.
Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes
Authors: Postel Alexander , Letzel Tobias , Frischmann Sieghard , Grund Christian , Beer Martin , Harder Timm
Published: May 2011

Real-time reverse transcription loop–mediated isothermal amplification (real-time RT-LAMP) holds substantial potential as a highly sensitive, specific, and easy-to-perform molecular technique for pathogen detection in clinical samples. In the current study, the analytical and diagnostic performance of 2 commercial realtime RT-LAMP kits, Avian Flu H5 and Avian Flu H7, in detecting Avian influenza virus (AIV) infections were evaluated and compared with validated real-time reverse transcription polymerase chain reaction (RT-PCR) assays using RNA from reference virus isolates of subtypes H5 ( n = 24) and H7 ( n = 25) and of phylogenetically related subtypes ( n = 20). When real-time RT-LAMP was carried out according to the recommendations of the manufacturer, 3 out of 24 H5 isolates and 8 out of 25 H7 reference strains were not detected. Prolonging the amplification phase resulted in detection of all H5 isolates but also in false positive detection of 2 non-H5 isolates. Real-time RT-LAMP specific to H7 failed to detect 2 H7 isolates after prolonged amplification. According to the examination of RNA log dilutions, the sensitivity of the real-time RT-LAMP assays, for a number of historic but also recent strains, was considerably lower compared with subtype-specific real-time RT-PCR assays. Application of the real-time RT-LAMP assays for analysis of diagnostic samples from wild birds confirmed their lower sensitivity. Commercial real-time RT-LAMP as tested in this study with a broad range of AIV H5 and H7 strains of phylogenetically diverse yet recent origin, holds some promise for routine veterinary diagnostic purposes, although real-time RT-LAMP was markedly more vulnerable to a reduction of detection limits because of strain-specific sequence variation than subtype-specific real-time RT-PCR.
Comparison of Three Methods for Extraction of Mycobacterium Avium Subspecies Paratuberculosis DNA for Polymerase Chain Reaction from Broth-Based Culture Systems
Authors: Okwumabua Ogi , Shull Eileen , O′Connor Mike , Moua Tou Vue , Danz Tonya , Strelow Kathy
Published: May 2011

Conventional and real-time polymerase chain reaction (PCR) assays were used to measure the recovery of DNA from Mycobacterium avium subspecies paratuberculosis (MAP) extracted with 3 different methods (MagMAX™, DNeasy®, and phenol-chloroform) after growth in a broth-based culture system. Of the 304 samples tested, bacterial DNA was detected in 197 (65%) of samples after MagMAX, 156 (51%) after phenol-chloroform, and 123 (40%) after DNeasy extractions. By acid-fast stain, 177 (58%) of the samples yielded acid-fast–positive bacilli, of which 4 were PCR negative by the 3 extraction methods. The results demonstrated that the amplifiable MAP DNA, as evidenced by the number of PCR-positive cultures and amplicon intensity on ethidium bromide–stained agarose gel, was best for MagMAX, intermediate for phenol-chloroform, and least for DNeasy. When subjected to real-time polymerase chain reaction, the MagMAX extracts produced the best results, thereby making it an excellent kit for the efficient extraction of MAP DNA from the broth-based culture system.
Development of One-Step Real-Time Reverse Transcription Polymerase Chain Reaction Assays for Rapid Detection of Porcine Group C Rotaviruses
Authors: Chun Young-Hyun , Jeong Young-Ju , Park Sang-Ik , Hosmillo Myra , Shin Dong-Jun , Kwon Hyung-Jun , Kang Shien-Young , Woo Sang-Kyu , Kang Mun-Il , Park Su-Jin , Cho Kyoung-Oh
Published: May 2011

Although the widespread occurrence of porcine group C rotaviruses (GCRV) is assumed, precise prevalence remains largely unknown because of the absence of reliable, specific, and rapid diagnostic methods. To detect and quantify porcine GCRV, the authors evaluated and optimized SYBR Green and TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR) assays and applied them to 108 piglet fecal samples. Using serially diluted standard RNA transcripts of porcine GCRV VP6 gene, both SYBR Green and TaqMan real-time RT-PCR assays detected as few as 1 × 101 genome copies/μl (correlation coefficiency >0.99), whereas conventional RT-PCR detected 1.0 × 103 copies/μl. In addition, the conventional assay detected porcine GCRV in 24% (26/108) of fecal samples, whereas the detection rates of both SYBR Green and TaqMan assays were 72% (78 of 108) and 64% (70 of 108), respectively. The current study indicated that both real-time RT-PCR assays were reliable, specific, and rapid methods for the detection of porcine GCRV in porcine fecal samples.
Development and Validation of an Immunohistochemical Method for Rapid Diagnosis of Swine Erysipelas in Formalin-Fixed, Paraffin-Embedded Tissue Samples
Authors: Opriessnig Tanja , Bender Joseph S. , Halbur Patrick G.
Published: May 2011

The objective of the study was to develop an immunohistochemical (IHC) assay for rapid detection of Erysipelothrix rhusiopathiae. Serotypes 1a, 1b, and 2 are most frequently associated with clinical disease in pigs. Antiserum against serotypes 1a, 1b, and 2 was produced in rabbits, pooled, and applied to formalin-fixed, paraffin-embedded tissue sections of pigs (lungs, heart, spleen, and skin). The results obtained with the IHC assay were compared with direct culture on tissue samples from experimentally inoculated pigs either treated ( n = 6) with antibiotics or untreated ( n = 8) as well as on samples from field cases ( n = 170) submitted to the Veterinary Diagnostic Laboratory at Iowa State University. The agreement between direct culture and IHC staining was found to be substantial. The results of the present study indicate that the IHC assay is highly sensitive and specific in detecting E. rhusiopathiae antigen in formalin-fixed, paraffin-embedded tissues. Results indicated that the IHC is particularly useful in cases in which pigs had been treated with antibiotics prior to submission and in which direct cultures of organs were negative. In addition, the IHC was found to be useful for detection of E. rhusiopathiae antigen in skin lesions, which are often culture negative.
In Situ Hybridization for the Detection of Mycoplasma Bovis in Paraffin-Embedded Lung Tissue from Experimentally Infected Calves
Authors: Björn Jacobsen , Hermeyer Kathrin , Jechlinger Wolfgang , Zimmermann Martina , Spergser Joachim , Rosengarten Renate , Marion Hewicker-Trautwein
Published: May 2011

Mycoplasma bovis DNA was detected in lung tissue of experimentally infected calves by in situ hybridization (ISH) with a nonradioactive, digoxigenin-labeled DNA probe. The 171–base pair DNA probe targeting part of the gene of the major immunodominant variable surface protein A, which is conserved among all vsp genes, was generated by polymerase chain reaction. Four calves between 57 and 63 days old were inoculated intratracheally with 30 ml of a suspension of M. bovis strain 1067 containing 7 × 104 colony forming units per milliliter. Two calves inoculated with 30 ml of sterile medium served as control animals. The calves were euthanized and then examined 21 days after inoculation. The ISH method developed in the current study was suitable for the detection of M. bovis DNA in formalin-fixed, paraffin-embedded lung tissue and may be a valuable tool for diagnostic purposes and for further investigating the pathogenesis of M. bovis infection.
Identification of a Novel Coronavirus Possibly Associated with Acute Respiratory Syndrome in Alpacas (Vicugna Pacos) in California, 2007
Authors: Crossley Beate M. , Barr Bradd C. , Magdesian K. Gary , Ing Michelle , Mora Daniel , Jensen David , Loretti Alexandre P. , McConnell Ty , Mock Richard
Published: May 2011

Alpaca respiratory syndrome (ARS) was first recognized in California in October 2007. This syndrome is characterized by acute respiratory signs, high fever, and occasional sudden death, and has mostly been observed in pregnant alpacas ( Vicugna pacos), although all signalments have been affected. A similarity in clinical signs to cases located on the East Coast of the United States was observed; however, a causative agent had not been identified. Preliminary diagnostic submissions to the California Animal Health and Food Safety Laboratory System (CAHFS) were negative for known bacterial, parasitic, fungal, and viral pathogens, as well as for toxins, making the etiology of this disease unknown. However, based on pathologic findings, a viral or toxic etiology was strongly considered. A novel coronavirus was recovered from lung tissue of a clinical case submitted to CAHFS. The coronavirus identity was confirmed in tissue culture by transmission electron microscopy and by sequence analysis of a conserved region within the viral genome. Statistical analysis calculating a serologic association between the serum virus neutralization antibody titer and coronavirus, the presence of exposure history on 40 animals with a history of ARS, and 167 controls provided an odds ratio of 121 (95% confidence interval: 36.54 and 402.84; P < 0.0001). The findings indicate that the ARS-associated coronavirus described is distinct from the previously reported gastrointestinal-associated coronavirus identified in alpaca herds.
The Same Papillomavirus is Present in Feline Sarcoids from North America and New Zealand but Not in Any Non-Sarcoid Feline Samples
Authors: Munday John S. , Knight Cameron G. , Howe Laryssa
Published: May 2011

Feline sarcoids are uncommon dermal neoplasms that are associated with papillomavirus (PV) infection. A single PV type, designated feline sarcoid–associated PV (FeSarPV), was detected in 9 feline sarcoids from North America. As FeSarPV has only been detected within feline sarcoids, the epidemiology of the infection remains unknown. The present study used polymerase chain reaction (PCR) to investigate whether this PV is also present within sarcoids from New Zealand cats. Additionally, as PVs are often host-specific, it was hypothesized that FeSarPV may often asymptomatically infect cats but rarely cause disease. To test this hypothesis, specific PCR primers were designed to investigate the presence of FeSarPV DNA within 120 samples from the skin and mouth of cats without sarcoids. Feline sarcoids from both New Zealand and North America contained FeSarPV DNA sequences. However, FeSarPV DNA was not detected within any non-sarcoid feline sample. To the authors' knowledge, this is the first time that FeSarPV has been reported in a country outside North America. As FeSarPV does not asymptomatically infect cats, feline sarcoids are likely due to cross-species infection. Although the reservoir host of FeSarPV is unknown, the host is present and has contact with cats, in both New Zealand and North America.
Vascular Endothelial Growth Factor Expression and Microvascular Density in Soft Tissue Sarcomas in Dogs
Authors: de Queiroz Genilson Fernandes , Dagli Maria Lucia Zaidan , Fukumasu Heidge , Zavala Arturo Alejandro Zavala , Matera Julia Maria
Published: May 2011

The aim of the current study was to evaluate the expression of vascular endothelial growth factor (VEGF) and the microvascular density in canine soft-tissue sarcomas. Immunohistochemistry for VEGF expression was performed on 20 canine neoplasms by the streptavidin–biotin–peroxidase method using an anti-VEGF mouse monoclonal antibody (ab-119). The volume fraction of microvessels in the sarcomas was quantified in hematoxylin and eosin–stained tissue sections. At least 10 fields of view (40x magnification) per neoplasm were analyzed by positioning a grid with 100 points and counting the microvessels that fell into the intersection points. This percentage was considered the volume fraction of these microvessels in the tumor section. VEGF expression was detected in 65% of the neoplasms. In 92.3% of the neoplasms, the expression occurred in the peritumor region; in 46.15%, in the intratumor region; and in 38.46%, the expression was present in both regions. The cells responsible for VEGF expression were fibroblasts and macrophages in the peritumor region or in the pseudocapsule and neoplastic cells in the intratumor region. Greater intratumoral VEGF was expressed in hemangiopericytomas ( P = 0.04). No difference was present in the volume fraction of tumor microvessels between VEGF-positive and VEGF-negative neoplasms ( P = 0.3416) or for the different types of neoplasms ( P = 0.5). The results of this study suggest that VEGF participates in the angiogenesis of soft-tissue sarcoma in dogs. Additional research will be necessary to elucidate the contribution of VEGF to the progression of malignancy.
An Outbreak of Late-Term Abortions, Premature Births, and Congenital Deformities Associated with a Bovine Viral Diarrhea Virus 1 Subtype b that Induces Thrombocytopenia
Authors: Blanchard Patricia C. , Ridpath Julia F. , Walker Jennifer B. , Hietala Sharon K.
Published: May 2011

Bovine viral diarrhea virus 1 (BVDV-1) subtype b was isolated from premature Holstein calves from a dairy herd that experienced an outbreak of premature births, late-term abortions, brachygnathism, growth retardation, malformations of the brain and cranium, and rare extracranial skeletal malformations in calves born to first-calf heifers. Experimental inoculation of 3 colostrum-deprived calves aged 2–4 months old with this BVDV isolate resulted in thrombocytopenia, lymphopenia, and leukopenia. Outbreaks of brachygnathism are rarely associated with BVDV, and thrombocytopenia is rarely associated with BVDV-1 strains.
Intracellular Yeasts in Endothelial Cells of a Great Blue Heron (Ardea Herodias)
Authors: Millins Caroline , Hill Janet E. , Wobeser Gary
Published: May 2011

Intracellular organisms in the endothelial cells of several organs of an adult great blue heron ( Ardea herodias) were identified as a yeast in the family Saccharomycetales based on ultrastructural morphology and sequence data from the ribosomal RNA operon. Morphologically similar organisms of unknown identity have been described previously in Muscovy ( Cairina moschata) and domestic ( Anas platyrhynchos domestica) ducks.
Atypical Dermatophilus Congolensis Infection in a Three-Year-Old Pony
Authors: Byrne Barbara A. , Rand Courtney L. , McElliott Valerie R. , Samitz Eileen M. , Brault Stephanie A.
Published: May 2011

Dermatophilosis caused by Dermatophilus congolensis causes exudative dermatitis in a variety of species. The infection is generally limited to the cutaneous tissues, where infection is acquired from the environment and bacteria penetrate into keratinized epithelium through epithelial disruption. A 3-year-old pony filly was examined for enlarging mandibular lymph nodes during the preceding 10 months. Biopsy of the node revealed mixed and granulomatous inflammation and thick, filamentous Gram-positive bacteria. Dermatophilus congolensis, confirmed by biochemical testing and sequencing of the ribosomal RNA gene, was cultured from a sample aspirated from the infected site. Dermatophilus congolensis should be considered as a possible etiologic agent associated with lymphadenopathy and granulomatous inflammation in the horse.
An Outbreak of Orthopoxvirus-Associated Disease in Horses in Southern Brazil
Authors: Mário C. S. Brum , Bruno Leite dos Anjos , Carlos E. W. Nogueira , Amaral Lorena A. , Weiblen Rudi , Flores Eduardo F.
Published: May 2011

An outbreak of severe cutaneous disease associated with an orthopoxvirus in horses in southern Brazil is described. Fourteen Crioulo mares and foals from a husbandry farm developed papules, and vesicles progressing to proliferative and exudative lesions on the muzzle, external nares, and external and internal lips. The vesicles eroded, and the proliferative lesions eventually bled and progressed to moist crusts and scars. The clinical signs lasted approximately 6–12 days, after which the animals progressively recovered. Direct electron microscopy of skin biopsies revealed brick-shaped, 250–300-nm virus particles with orthopoxvirus morphology. Histological examination of the lesions revealed vacuolar degeneration of the cells of the stratum spinosum and the presence of large intracytoplasmic, eosinophilic inclusion bodies. Polymerase chain reaction amplification of poxvirus A-type inclusion body gene confirmed the presence of orthopoxvirus DNA in horse tissues. Inoculation of tissue homogenates into the chorioallantoic membrane of chicken embryonated eggs and intraperitoneally in mice resulted in pock-like lesions with the microscopic appearance of poxvirus-induced histopathology. Taken together, these results demonstrate the association of an orthopoxvirus with the outbreak of cutaneous disease in horses. The origin of the agent causing the outbreak is uncertain because no similar condition has been reported in Brazil.
Gingival Osteogenic Melanoma in Two Dogs
Authors: Ellis Angela E. , Harmon Barry G. , Miller Debra L. , Northrup Nicole C. , Latimer Kenneth S. , Uhl Elizabeth W.
Published: May 2011

Osteogenic melanoma is a rare variant of metaplastic malignant melanoma in human medicine and appears to be a similarly rare variant in dogs. Two dogs with oral malignant melanoma with neoplastic bone formation are reported in this study. Both tumors were characterized by malignant melanocytes that transitioned into neoplastic bone at the deep margins of the neoplasm. Immunohistochemical analysis revealed S100- and Melan-A–positive neoplastic cells adjacent to, and occasionally embedded within, an osteoid and chondroblastic matrix. Scattered clusters of neoplastic cells were also positive for osteocalcin. The findings indicate that in dogs, as in humans, neoplastic melanocytes have metaplastic potential and can be osteogenic.
Concurrent Nasal Adenocarcinoma and Rhinosporidiosis in a Cat
Authors: Brenseke Bonnie M. , Saunders Geoffrey K.
Published: May 2011

A 10-year-old, neutered, male Domestic Shorthair cat was presented to the teaching hospital for labored breathing, anorexia, and weight loss of several months duration. External examination revealed distortion of the bridge of the nose and pink fleshy polyps protruding from each nostril. The cat was euthanized and submitted for postmortem examination. In addition to the external findings, the nasal cavity had extensive bone and cartilage loss and contained a tan firm mass in the caudal region of the nasal cavity near the cribriform plate. On histologic examination, the mass was a nasal adenocarcinoma, and the polyps were composed of hyperplastic nasal epithelium and submucosal stroma that contained sporangia consistent with Rhinosporidium seeberi.
Type A Botulism in Horses in the United States: A Review of the past Ten Years (1998–2008)
Authors: Johnson Amy L. , McAdams Susan C. , Whitlock Robert H.
Published: May 2011

The objective of the current retrospective study was to describe naturally occurring type A botulism in horses in the United States. In the past 10 years, the Botulism Laboratory at the University of Pennsylvania's School of Veterinary Medicine has identified 3 isolated cases and 8 outbreaks of type A botulism in horses via samples positive for Clostridium botulinum type A toxin or spores using the mouse bioassay test. Additional information was obtained by review of submission forms and by telephone or email interviews. Almost all type A cases and outbreaks occurred in the western United States, with Oregon and Idaho overrepresented. Type A toxin was identified in only 1 outbreak; all other identified cases and outbreaks were positive for spores but not preformed toxin. Reported clinical signs included progressive muscle weakness, recumbency, decreased tail and/or tongue tone, dysphagia, respiratory distress, and death. Isolated cases involved foals ≤1 month of age; outbreaks involved horses ≥11 months. One hundred and nineteen horses were potentially exposed to the toxin source; 54 out of 119 showed signs of botulism, and 49 out of 54 affected horses were confirmed dead. The number of horses affected per outbreak ranged from 2 to 24. The source of infection was confirmed to be hay or silage in 6 out of 8 outbreaks and was unknown in 2 out of 8 outbreaks. The present report is the first description of outbreaks of type A botulism in horses and has important implications for prevention and treatment. Based on these findings, type A botulism should be considered in suspect cases of equine botulism in the western United States.
Equine Primary Liver Tumors: A Case Series and Review of the Literature
Authors: Beeler-Marfisi Janet , Arroyo Luis , Caswell Jeff L. , DeLay Josepha , Bienzle Dorothee
Published: May 2011

Hepatoblastoma (HB) is an uncommon pediatric liver tumor in humans and horses. In humans, HB is most frequently diagnosed in fetuses, neonates, and young children, whereas hepatocellular carcinoma (HCC) affects juvenile and adult humans. Hepatoblastoma in the horse is rare, with only 9 reported cases. Affected horses ranged in age from late-term aborted fetuses to 3 years. The current study describes 3 new cases of primary liver tumors in horses and reviews findings in relation to other reports on this condition. Tumors classified as HB were identified in a male Standardbred aborted fetus and in a 4-year-old Thoroughbred filly. Hepatocellular carcinoma was diagnosed in a 15-month-old Paint filly. In the Standardbred fetus, the tumor was only present in the liver. In the Thoroughbred and Paint fillies, primary tumors were in the right liver lobe and at the hilus, respectively, and there were metastases to other lobes (HB) and mesenteric lymph nodes (HCC). Tumors were sharply demarcated from adjacent tissue, nonencapsulated, compressive, and invasive. Consisting of cords and nests, or disorganized sheets of epithelial cells, tumors had variable stromal and vascular components. The fetal tumor contained areas of smaller, less differentiated cells with a pronounced mesenchymal component interpreted to be embryonal hepatic tissue. Diagnoses were based on tumor histomorphologic features, resemblance to hepatocyte developmental stages, age of the animal, and patterns of metastasis. Tumors classified as HB were α-fetoprotein immunoreactive. Primary hepatic tumors in the horse are diverse in morphology and include subtypes compatible with classification criteria applied to human tumors.
Measurement of Urinary F2-Isoprostanes as Markers of in Vivo Lipid Peroxidation: A Comparison of Enzyme Immunoassays with Gas Chromatography–Mass Spectrometry in Domestic Animal Species
Authors: Soffler Carl , Campbell Vicki L. , Hassel Diana M.
Published: May 2011

F2-isoprostanes are useful markers for assessing oxidant injury; however, the validity of measuring urinary 15-F2t-isoprostane concentration by enzyme-linked immunosorbent assay (ELISA) has not been evaluated in veterinary species. The current study assesses the agreement between 2 commercially available urinary isoprostane kits and gas chromatography and negative ion chemical ionization–mass spectrometry (GC/NICI-MS). The results indicate that only feline urinary isoprostane measurement by glucuronidase (GL)-ELISA has acceptable agreement with GC/NICI-MS. Urinary isoprostane concentration was highly variable in critically ill animals, but there were too many variations between healthy and critically ill animals to draw meaningful conclusions. Currently, GC/NICI-MS is the only method that can be recommended for the assessment of urinary isoprostanes in dogs, cattle, and horses. Feline urinary isoprostanes can be assessed by GL-ELISA, but caution is still warranted when comparing data from manuscripts using different methods given the relatively low Spearman rank correlation coefficient. Future studies may require large sample sizes or focused inclusion criteria to account for variability in isoprostane concentration.
Optimization of a Whole Blood Gamma Interferon Assay for the Detection of Sheep Infected with Mycobacterium Avium Subspecies Paratuberculosis
Authors: Bosward Katrina L. , Dhand Navneet K. , Begg Douglas J. , Thomson Peter C. , Emery David L. , Whittington Richard J.
Published: May 2011

The capacity of a commercially available gamma interferon (IFNγ) assay to detect infected sheep early in the pathogenesis of Johne's disease enables the removal of such animals from the flock before bacterial shedding and pasture contamination. However, nonspecific IFNγ responses in the assay have meant that to achieve high-test specificity, there has been a reduction in sensitivity. Although the optimal conditions for the use of the assay in cattle have been well documented, there have been few studies optimizing the assay for use in sheep. The current study details the effect of anticoagulant, duration of incubation, cell concentration, blood storage temperature, time of stimulation of cells with antigen relative to time of sample collection, and temperatures during transit on IFNγ synthesis. Maximal IFNγ synthesis occurred with incubation periods of 48 hr in blood collected into heparinized tubes. Decreasing the leukocyte population by diluting the total peripheral blood leukocyte concentration was associated with a decreasing IFNγ response. Conversely, concentrating the peripheral blood concentration 2-fold resulted in an increase in the IFNγ production. In field studies, immediate incubation of blood samples with antigen at 37°C resulted in larger IFNγ responses; however, significantly lower IFNγ values were obtained if the samples were transported at ambient temperature. The results of this study indicate that optimization of the IFNγ assay may enable increased synthesis of IFNγ during the stimulation phase of the assay and that future work may determine whether this translates to increased sensitivity of the assay in detecting early infections in sheep.
Characterization of Fowl Adenovirus Serotype 4 Isolated from Chickens with Hydropericardium Syndrome Based on Analysis of the Short Fiber Protein Gene
Authors: Mase Masaji , Nakamura Kikuyasu , Imada Tadao
Published: May 2011

The sequences of short fiber genes of the Fowl adenovirus serotype 4 (FAdV-4), including isolates from chickens with hydropericardium syndrome (HPS), in Japan, India, and Pakistan were compared. By phylogenetic analysis based on complete nucleotide sequences of this gene, FAdV-4 strains from HPS (HPS–FAdV-4) in Japan, India, and Pakistan fell into a different cluster from FAdV-4 strains not derived from HPS. Hydropericardium syndrome–FAdV-4 isolates were differentiated from other FAdV-4 strains by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis using the enzyme the Alu I. The use of PCR-RFLP analysis of short fiber genes may be useful to distinguish among FAdV-4 strains.
Development of in Situ Hybridization Assay that Differentiates between Two Genotypes of Porcine Circovirus-2 in Formalin-Fixed, Paraffin-Embedded Tissues
Authors: Kim Duyeol , Ha Yooncheol , Oh Yeonsu , Han Kiwon , Noh Seung Ho , Kim Cheolwoo , Kim Sung-Hoon , Chae Chanhee
Published: May 2011

The aim of the current study was to develop a nonradioactive in situ hybridization assay that can differentiate between genotypes 2a and 2b of Porcine circovirus-2 (PCV-2) in formalin-fixed, paraffin-embedded lymph node tissues from pigs with postweaning multisystemic wasting syndrome. Two different digoxigenin-labeled oligonucleotide probes were designed from the PCV-2 open reading frame 2 sequences. The PCV-2a–specific probe did not hybridize with formalin-fixed, paraffin-embedded lymph nodes from naturally PCV-2b–infected pigs and vice versa. Both PCV-2a–specific and PCV-2b–specific probes gave consistent negative signals in lymph nodes from naturally PCV-1–infected pigs. The in situ hybridization assay described in the present study represents a diagnostic tool that can differentiate between the 2 genotypes of PCV-2.
Rapid and Reliable Genotyping Technique for GM1 Gangliosidosis in Shiba Dogs by Real-Time Polymerase Chain Reaction with TaqMan Minor Groove Binder Probes
Authors: Chang Hye-Sook , Arai Toshiro , Yabuki Akira , Hossain Mohammad A. , Rahman Mohammad M. , Mizukami Keijiro , Yamato Osamu
Published: May 2011

Real-time polymerase chain reaction (PCR) with TaqMan minor groove binder (MGB) probes was examined to establish a rapid and reliable genotyping technique for GM1 gangliosidosis in Shiba dogs. This technique was applied to DNA samples extracted from the blood, umbilical cord, or postmortem liver tissue specimens, and to DNA-containing solutions prepared from blood and saliva that had been applied to Flinders Technology Associates filter papers (FTA cards). The amplification of the targeted sequence in all the samples was sufficient to determine the genotypes of GM1 gangliosidosis. Forty-seven DNA samples that had previously been obtained from blood or tissue specimens of Shiba dogs were examined using this real-time PCR technique, and the findings were consistent with the data obtained by the earlier PCR–restriction fragment length polymorphism (RFLP) assay. In addition, the use of this new technique in combination with FTA cards for sampling could markedly shorten the time required for genotyping, as well as simplify the procedure. Furthermore, in the present study, the results of a previous epidemiological screening of 96 Shiba dogs in the Czech Republic were rechecked by this real-time PCR technique using stored crude buccal cell DNA-containing solutions directly as DNA templates. The results provided clear-cut genotyping in all the samples although the earlier PCR-RFLP assay could not determine the genotype in all cases. In conclusion, this new real-time PCR technique is a simple, rapid, and reliable choice for large-scale screening to detect an abnormal allele indicating GM1 gangliosidosis in Shiba dogs.
A Rapid Assay for Detecting Antibody against Bluetongue Virus with a Latex Agglutination Test Using Recombinant VP7 Antigen
Authors: Yang Junxing , Hua Qunyi , Chen Huanchun , Lv Jianqiang , Chen Bing , Ruan Zhouxi
Published: May 2011

A latex agglutination test (LAT) for detecting antibody against Bluetongue virus (BTV) in ruminants was developed using latex beads coupled with recombinant VP7 protein. Compared with competitive enzyme-linked immunosorbent assay (ELISA), the specificity and sensitivity of the LAT were 99.0% and 93.0%, respectively. There was excellent agreement between the results obtained by competitive ELISA and the LAT (kappa = 0.930). Because it is rapid and easy to use, the LAT could be used for BTV antibody detection, especially for screening many serum samples.
Experimental Validation of a Nested Polymerase Chain Reaction Targeting the Genetic Element ISMAP02 for Detection of Mycobacterium Avium Subspecies Paratuberculosis in Bovine Colostrum
Authors: Pithua Patrick , Wells Scott J. , Godden Sandra M. , Sreevatsan Srinand , Stabel Judith R.
Published: May 2011

Colostrum samples experimentally inoculated with Mycobacterium avium subsp. paratuberculosis (MAP; strain K-10) at increasing concentrations between 1 × 101 and 1 × 109 cells/ml were tested for recovery of MAP DNA using a nested ISMAP02 target polymerase chain reaction initially developed for detecting MAP DNA in fecal samples. The following detection rates were achieved for sample replicates inoculated with unsonicated MAP pure stock: 100% between 1 × 107 and 1 × 109 cells/ml, 75% between 1 × 103 and 1 × 106 cells/ml, and 50% between 1 × 101 and 1 × 102 cells/ml replicates. Detection rates achieved for the colostrum sample replicates inoculated with sonicated MAP cell suspension were 75% for 1 × 109 cells/ml, 100% between 1 × 107 and 1 × 108 cells/ml, 75% for 1 × 106 cells/ml, 0 for 1 × 104 cells/ml, and 25% between 1 × 101 and 1 × 103 cells/ml. When negative control colostrum samples were tested, 16 of 18 (89%) samples were correctly detected as negative for MAP DNA using the current assay. In conclusion, the MAP DNA detection rates of the present assay improved with increasing concentrations of MAP in the colostrum sample replicates, although MAP DNA was also detected in 2 of 18 (11%) negative control samples, suggesting an undefined technical problem with the assay or, perhaps, sample contamination during preparation. Overall, the present findings suggest a potential role of the proposed polymerase chain reaction assay to detect MAP in colostrum. However, adoption of this test for use in routine screening of field colostrum for MAP awaits findings from an ongoing field validation study.
Quantitative Detection of Porcine Torque Teno Virus in Porcine Circovirus-2–Negative and Porcine Circovirus–Associated Disease-Affected Pigs
Authors: Lee Sung-Seok , Sunyoung Sunwoo , Jung Hokyung , Shin Jungyeon , Lyoo Young S.
Published: May 2011

Torque teno virus (TTV) is a recently identified virus that has a wide range of host tropisms from humans to shrews. Human TTV and Torque teno mini virus are distributed worldwide, and their high prevalence in human populations has been reported. Pigs have their own species-specific TTV, and like human TTV, a high prevalence of porcine TTV also has been reported. Despite its high prevalence, the role of TTV-related disease or syndrome in humans and pigs has not been determined. In the swine industry, TTV is thought to be one of the agents that aggravate clinical manifestation of Porcine circovirus–associated disease (PCVAD), a newly emerging, economically devastating disease. The purpose of the current study was to quantify TTV viral load in serum obtained from Porcine circovirus-2–negative pigs and PCVAD-affected pigs with real-time quantitative polymerase chain reaction assays and to compare TTV viral load between these groups. Results of this study indicate that there are no statically remarkable differences in TTV viral load between the 2 groups, which indicates that TTV might not be an agent of aggravation in PCVAD.
A Quantitative Polymerase Chain Reaction Assay for Detection and Quantification of Lawsonia Intracellularis
Authors: Drozd Mary , Kassem Issmat I. , Gebreyes Wondwossen , Rajashekara Gireesh
Published: May 2011

Lawsonia intracellularis, a causative agent of porcine proliferative enteropathy, is an obligate intracellular bacterium that is difficult to culture, propagate, and quantify. The intestinal epithelial cell line (IEC-18, derived from rat small intestine crypt cells) has been used to isolate and propagate this pathogen. However, the lack of rapid and simple quantification methods has led to mixed results when using the rat cell line, complicating Lawsonia studies. To overcome these limitations, a SYBR green quantitative polymerase chain reaction (qPCR) assay targeting a unique hypothetical protein was developed for detecting and quantifying L. intracellularis in IEC-18 rat epithelial cells, porcine fecal samples collected from different farms, and experimentally infected pigs. The method was optimized to detect as few as 3 copies per qPCR reaction of the bacterium growing in IEC-18 cells, providing a new and necessary approach to assess the growth of L. intracellularis in these cell lines. Furthermore, the qPCR assay was successful in detecting L. intracellularis in fecal samples collected from pigs with and without a history of Lawsonia infections, as well as in experimentally infected pigs, which further confirmed the suitability of the qPCR assay for studying the epidemiology of this pathogen.
The Interlobular Distribution of Copper in the Liver of Beef Calves on a High-Copper Diet
Authors: Miranda Marta , Benedito José Luis , Gutiérrez Betiana , García-Vaquero Marco , Blanco-Penedo Isabel , Löpez-Alonso Marta
Published: May 2011

The aims of the present study were 1) to evaluate the interlobular distribution of copper (Cu) in the liver of beef calves on a high-Cu diet, 2) to determine whether this distribution differs between Galician Blonds and Holstein Friesians, and 3) to determine whether in vivo needle biopsy provides an appropriate measure of overall hepatic Cu status. Liver biopsies were performed before slaughter on twenty-nine 10-month-old beef calves fed growing and finishing diets supplemented with 35 mg/kg of Cu sulfate (10 Galician Blonds, 9 Holstein Friesians, and 10 Galician Blond × Holstein Friesian crosses). At slaughter, samples taken from 6 regions of the liver (the internal and external faces of the right lobe; the left, caudate, and quadrate lobes; and the processus papillaris) were acid digested, and their Cu contents were determined by inductively coupled plasma atomic emission spectrometry. The highest Cu concentrations were found in the left lobe, followed by the processus papillaris, and the lowest Cu concentrations were found in the caudate and quadrate lobes. Different breeds differ in absolute hepatic Cu levels, but interlobular Cu distribution does not appear to depend on breed, at least when Galician Blonds are compared with Holstein Friesians. In vivo needle biopsy afforded accurate estimates of overall hepatic Cu status.
Natural Fatal Sarcocystis Falcatula Infections in Free-Ranging Eagles in North America
Authors: Wünschmann Arno , Rejmanek Daniel , Conrad Patricia A. , Hall Natalie , Cruz-Martinez Luis , Vaughn Samuel B. , Barr Bradd C.
Published: May 2011

Three bald eagles ( Haliaeetus leucocephalus) and 1 golden eagle ( Aquila chrysaetos) were admitted to rehabilitation facilities with emaciation, lethargy, and an inability to fly. Intravascular schizonts and merozoites were present in 2 bald eagles, mainly in the lung tissue, whereas the third bald eagle and the golden eagle had lymphohistiocytic encephalitis with intralesional schizonts and merozoites. In all eagles, protozoal tissue cysts were present in skeletal musculature or heart. The protozoal organisms were morphologically compatible with a Sarcocystis sp. By immunohistochemistry, the protozoal merozoites were positive for Sarcocystis falcatula antigen in all cases when using polyclonal antisera. Furthermore, the protozoa were confirmed to be most similar to S. falcatula by polymerase chain reaction in 3 of the 4 cases. To the authors' knowledge, this report presents the first cases of natural infection in eagles with S. falcatula as a cause of mortality.
Non–West Nile virus-Associated Mortality in a Population of American Crows (Corvus Brachyrhynchos): A Gross and Histopathologic Study
Authors: Miller Andrew D. , Townsend Andrea K. , McGowan Kevin J. , Clark Anne B. , Glaser Amy L. , Patrican Lisa A. , Dobson Elizabeth , Buckles Elizabeth L.
Published: May 2011

The American crow ( Corvus brachyrhynchos) is a common urban and rural inhabitant of the Northeast and Midwest United States that is commonly infected with West Nile virus (WNV). The current study was initiated to determine non–WNV-associated causes of mortality in the American crow. All animals (40/40) tested negative for WNV infection via polymerase chain reaction and had no evidence of infection based on immunohistochemistry. Common gross necropsy findings included external trauma (6/40), hepatosplenomegaly (6/40), poxviral dermatitis (5/40), and pneumonia (3/40). Common histologic findings included endoparasitism (32/40), multifocal hepatic and splenic necrosis (7/40), pigment accumulation in the spleen (5/40), and disseminated bacterial infection (3/40). The most significant and debilitating diseases included fungal pneumonia and poxvirus-associated lesions. The present report increases the knowledge of diseases present in the American crow population.