bluuebunny/crossref_metadata_2025 huggingface dataset:
Accuracy of Three Diagnostic Tests for Determining Mycobacterium Bovis Infection Status in Live-Sampled Wild Meerkats (Suricata Suricatta)
Authors: Drewe Julian A. , Dean Gillian S. , Michel Anita L. , Pearce Gareth P.
Published: May 2011

Accurate diagnosis of Mycobacterium bovis infection (bovine tuberculosis [bTB]) in live animals is notoriously problematic. The aim of the present study was to evaluate the diagnostic potential of 2 new serologic tests (multiantigen print immunoassay [MAPIA] and lateral flow immunoassay rapid test [RT]) in comparison with mycobacterial culture of tracheal washes for determining M. bovis infection status in a free-ranging population of wild meerkats ( Suricata suricatta). During a longitudinal study lasting 2.5 years, 240 individually identifiable meerkats were each sampled up to 8 times under anesthesia every 3 months. Diagnostic accuracy was determined through Bayesian and maximum likelihood estimations of sensitivity, specificity, and likelihood ratios for each diagnostic test when used independently and in parallel to classify the disease status of individual meerkats in the absence of a gold standard. Culture of tracheal washes was highly specific (0.99; 95% confidence interval [CI] = 0.77, 1.00) but of low sensitivity (0.36; 95% CI = 0.24, 0.50) for diagnosing M. bovis-infected individuals. The longitudinal nature of the study with repeated sampling of the same individual animals served simultaneously to improve chances of detecting infection and increase confidence in a negative result in individual animals repeatedly testing negative. Although MAPIA and RT were individually of limited diagnostic use, interpreting the results of these 2 tests in parallel produced estimates of sensitivity (0.83; 95% CI = 0.67, 0.93) and specificity (0.73; 95% CI = 0.62, 0.82) high enough to usefully inform decision making when determining exposure to bTB in wild meerkats and potentially other species in which bTB poses a diagnostic challenge.
Experimental Oral Transmission of United States Origin Scrapie to Neonatal Sheep
Authors: Hamir Amir N. , Kunkle Robert A. , Greenlee Justin J. , Richt Juergen A.
Published: May 2011

Scrapie, a transmissible spongiform encephalopathy (TSE), is a naturally occurring fatal neurodegenerative disease of sheep and goats. The current study documents incubation periods, pathologic findings, and distribution of abnormal prion proteins (PrPSc) by immunohistochemistry and Western blot in tissues of genetically susceptible and resistant neonatal lambs inoculated with pooled brain homogenates from 13 U.S. origin scrapie-affected ewes. Nine Suffolk lambs with genotypes AA/RR/QQ (n = 5) and AA/RR/QR (n = 4) at codons 136, 154, and 171, respectively) were orally inoculated, within 12 hr of birth, with 1 ml of a 10% (w/v) brain homogenate prepared from scrapie-affected sheep brains. Inoculated animals were euthanized when advanced clinical signs of scrapie were observed. All QQ sheep developed clinical signs of scrapie, with a mean survival time of 24 months. Spongiform lesions in the brains and PrPSc deposits in the central nervous system and lymphoid tissues were present in these sheep. None of the QR sheep succumbed to the disease. A previous study that used a larger volume (30 ml of 10% brain suspension) of the same inoculum in 4-month-old Suffolk lambs of susceptible genotype documented longer survival periods (average 32 months), and only 5 of 9 inoculated sheep developed scrapie. Findings of this study suggest that orally exposed neonatal lambs of a susceptible (QQ) genotype exhibit a higher attack rate and shorter incubation period than older (4-month-old) lambs exposed to a larger dose (30×) of the same inoculum.
Evaluation of a SRS2 Sandwich Commercial Enzyme-Linked Immunosorbent Assay for the Detection of Anti-Neospora Caninum Antibodies in Bovine and Canine Sera
Authors: Ghalmi Farida , China Bernard , Kaidi Rachid , Losson Bertrand
Published: May 2011

Neospora caninum is a parasite responsible for abortion in cows and neuromuscular disease in dogs. Serology is the most widely used technique to evaluate the prevalenceof N. caninum in different host populations. A sandwich enzyme-linked immunosorbent assay (ELISA) based on the use of an anti-SRS2 monoclonal antibody was evaluated against the indirect fluorescent antibody test for 100 canine sera and against a well-characterized ELISA for 102 bovine sera. In cattle sera, the relative sensitivity and relative specificity were 100%. In dog sera, the relative specificity and relative sensitivity were 94% and 86%, respectively. The kappa value was 1 for bovine sera and 0.77 for canine sera. The seroprevalence was 3.9% in bovine sera and 21–23% in canine sera. The SRS2 sandwich ELISA was considered a valuable tool in both species.
Tracheal Hypoplasia with a Discrete Subaortic Septal Ridge in a Rottweiler Puppy
Authors: Reed Scott D. , Evans Dawn E.
Published: May 2011

Concurrent tracheal hypoplasia and discrete subaortic stenosis are described in a 12-week-old Rottweiler puppy that presumably died of pulmonary edema. A brief literature review and comparison to previously published cases of tracheal hypoplasia in other breeds is presented along with a description of a subaortic septal ridge and comparison to the analogous condition in humans.
Multiple Atypical Mucosal Xanthomas in a Dog Similar to Human Verruciform Xanthoma
Authors: Balme Emmanuelle , Thuilliez Céline , Lejeune Typhaine , Chateau-Escoffier Laurence , Bernex Florence
Published: May 2011

A case of atypical mucosal xanthomas in a 14-year-old Shi-Tzu dog is reported. Grossly, 1–6-mm granular plaques or slightly elevated papillary nodules were found in the oral cavity (dorsal and ventral sides of the tongue, inner upper lip) and in the upper digestive tract (esophagus, stomach). Microscopically, subepithelial aggregates of large foamy cells were found in strong association with papillary epithelial hyperplasia. By immunohistochemistry, the majority of these cells tested positive for lysozyme and ACM1. The cells had cytoplasmic lipid content that stained positively with oil red O. These findings confirmed a monocyte/macrophage lineage. On the basis of macroscopic observations, microscopic changes, and the absence of a clinical metabolic defect, the condition in this dog appears similar to that of humans with oral verruciform xanthoma. The pathogenesis of these xanthomas in the dog remains obscure, although this condition appears to affect people and dogs with an Asian origin.
Malignant Acoustic Schwannoma in a Dog
Authors: Ottinger Therese , Lindberg Ronny , Ekman Stina
Published: May 2011

A malignant schwannoma of the right acoustic nerve of a dog is described. The neoplasm was found during necropsy of a 12-year-old, spayed, female Labrador Retriever that experienced a sudden onset of severe pain in the neck and upon opening the mouth. Concurrent mild hind limb ataxia also was present. The tumor had infiltrative growth, and the spindle-shaped neoplastic cells were arranged in sheets and concentric whorls. Immunohistochemical staining of the neoplastic spindle cells was positive for S-100 antigen, neuron-specific enolase, and glial fibrillary acidic protein.
A Simple Ductal Mammary Papilloma in a Male Maned Wolf (Chrysocyon Brachyurus)
Authors: Cassali Geovanni D. , Bertagnolli Angélica C. , Ferreira Enio , Malta Marcelo C. C.
Published: May 2011

A 1-cm-diameter nodule was identified in the left inguinal mammary gland of a 9-year-old male maned wolf ( Chrysocyon brachyurus). The mass was surgically excised and examined histologically. Microscopically, the neoplasm consisted of papillary proliferations of epithelial cells on well-defined fibrovascular stalks. A myoepithelial layer was located between the single layer of epithelial cells and the fibrovascular stalk. This histologic appearance was compatible with a diagnosis of simple ductal mammary papilloma. Immunohistochemical staining was positive for p63, cytokeratins AE1/AE3, and estrogen receptors. The clinical and histologic observations in the present case indicate that male maned wolves may develop mammary tumors that are similar to those observed in domestic dogs and humans.
Spontaneous Cutaneous Mast Cell Tumor with Lymph Node Metastasis in a Richardson's Ground Squirrel (Spermophilus Richardsonii)
Authors: He Xi Jun , Uchida Kazuyuki , Tochitani Tomoaki , Uetsuka Koji , Miwa Yasutsugu , Nakayama Hiroyuki
Published: May 2011

A 4-year-old female Richardson's ground squirrel ( Spermophilus richardsonii) presented with multicentric nodules arising from the skin of the middle of the tail and lumbosacral regions. Histologically, the nodules were composed of a proliferation of spindloid to pleomorphic cells that sometimes formed sheets and fascicular to storiform patterns. Diffuse infiltration of eosinophils was also noted. The results of immunohistochemistry indicated positive labeling for vimentin, mast cell tryptase, c-kit, and Ki-67. Toluidine blue stain revealed fine, metachromatic, cytoplasmic granules. The histologic diagnosis was mast cell tumor. The neoplasm recurred and metastasized to the right lumbar lymph node 1 month later.
Isolation of Helcococcus Ovis from Sheep with Pleuritis and Bronchopneumonia
Authors: Zhang Yan , Cui Jing , Parkinson Anne , Hayes Jeff , Ott Kristy , Byrum Beverly
Published: May 2011

Helcococcus ovis is a newly established species in the genus Helcococcus. The clinical significance of this organism in sheep has not been reported. In the current report, isolation of H. ovis from a 6-month-old mixed-breed ewe lamb that died of respiratory disease is described. Pathologic examination revealed severe, focally extensive, chronic necrotizing pleuritis with intralesional coccobacilli and mild, multifocal, subacute mucopurulent bronchopneumonia, indicating a bacterial etiology. A Gram-positive bacterium was isolated in heavy growth from the lung tissue. DNA sequence analysis on the 16S rDNA gene demonstrated that the isolate was H. ovis. To the authors' knowledge, this is the first report of isolation of H. ovis associated with infection in sheep with pleuritis and bronchopneumonia.
Enzyme-Linked Immunospot: An Alternative Method for the Detection of Interferon Gamma in Johne's Disease
Authors: Begg Douglas J. , de Silva Kumudika , Bosward Katrina , Di Fiore Lyrissa , Taylor Deborah L. , Jungersen Gregers , Whittington Richard J.
Published: May 2011

To date, the sensitivity of the interferon gamma (IFN-γ) enzyme-linked immunosorbent assay (ELISA) to detect Johne's disease (JD) has been poor, especially in the early stages of disease. To improve the sensitivity of IFN-γ detection in the early stages of infection, an alternate assay needs to be developed. The enzyme-linked immunospot (ELISPOT) assay is a highly sensitive technique for the detection of cytokines and has the potential to improve the diagnosis of JD. Of the variables examined, choice of capture antibody and the method by which the peripheral blood mononuclear cells were isolated significantly affected the ability to enumerate IFN-γ–secreting cells. The ELISPOT assay was as sensitive as or better than the IFN-γ ELISA at detecting ovine JD and could also detect disease at early time points postinoculation. The IFN-γ ELISPOT could distinguish infected from unexposed animals; however, neither the IFN-γ ELISA nor the ELISPOT assay could distinguish between sheep experimentally infected with Mycobacterium avium subspecies paratuberculosis and those exposed to the bacterium but diagnosed as uninfected at necropsy.
Comparative Efficacy of Conventional and TaqMan Polymerase Chain Reaction Assays in the Detection of Capripoxviruses from Clinical Samples
Authors: Balamurugan Vinayagamurthy , Jayappa Kallesh Danappa , Hosamani Madhusudhan , Bhanuprakash Veerakyathappa , Venkatesan Gnanavel , Singh Raj Kumar
Published: May 2011

Sheeppox and goatpox are economically important viral diseases of sheep and goats, respectively. Both diseases are reportable to the World Organization for Animal Health. To implement a control and eradication program for these diseases, a rapid and user-friendly diagnostic tool is imperative for screening. Therefore, in the present study, TaqMan quantitative polymerase chain reaction (qPCR) and conventional PCR assays targeting the DNA polymerase ( DNA pol) gene were developed for the detection of Capripoxvirus DNA from clinical specimens of sheep and goats. The 2 assays used different primer sets. Conventional PCR yielded a specific product of 134 bp, whereas qPCR yielded a 180-bp product. The specificity of amplified DNA pol gene products was confirmed by their size and by sequence analysis. The 2 assays were specific for Sheeppox virus and Goatpox virus. However, in comparison to conventional PCR, the qPCR was more rapid, specific, and 100 times more sensitive, with a detection limit as low as 0.042 pg of purified DNA. The qPCR assay was more sensitive (84.05%) than conventional PCR (76.06%) when used on clinical samples ( n = 71) from sheep and goats.
Naturally Occurring Sheep-Associated Malignant Catarrhal Fever in North American Pigs
Authors: Alcaraz Ana , Warren Amy , Jackson Carney , Gold Jennifer , McCoy Morgan , Cheong Soon Hon , Kimball Sam , Sells Steve , Taus Naomi S. , Divers Tom , Li Hong
Published: May 2011

Two cases of sheep-associated malignant catarrhal fever (MCF) in pigs were diagnosed on a small farm in New York, and in Kentucky. In both cases, the initial diagnosis was based on histopathological changes representing typical lymphoproliferative vasculitis in multiple tissues of the affected pigs. Ovine herpesvirus 2 (OvHV-2) DNA was detected by polymerase chain reaction in the tissues of affected pigs in both cases. The amplified sequences were identical between the clinically affected pigs and the OvHV-2-infected sheep. Additional virological and bacteriological examination showed that the affected pigs were negative for agents that cause Aujeszky's disease, classical swine fever, porcine enterovirus, and rabies. An antibody against a conserved epitope among MCF viruses was detected in 1 clinically affected pig and 2 unaffected cohort pigs, as well as in all the associated sheep. To the authors' knowledge, this is the first report of porcine MCF in North America, and suggests that OvHV-2 is associated with clinical MCF in pigs.
Blood Chimerism Confounds Genetic Relative Susceptibility Testing for Classical Scrapie in Sheep
Authors: Schneider David A. , Tibary Ahmed , Raudsepp Terje , Das Pranab J. , O'Rourke Katherine I.
Published: May 2011

Classical scrapie disease is a transmissible spongiform encephalopathy of sheep that is enzootic in the United States. Susceptibility of sheep to classical scrapie is linked to single nucleotide polymorphisms in the prion protein gene ( PRNP), forming the basis for genetic testing strategies used by national efforts to eradicate scrapie. Such efforts are occasionally hampered by inconclusive results stemming from the detection of “complex” genotypes. Naturally occurring cases of ovine chimerism are thought to account for some of these instances. In the current report, 4 naturally occurring ovine chimeras are documented through cytogenetic and molecular analyses. All 4 of these sheep had chimeric cells circulating in their blood. Blood and alternate tissue samples of ear punch and hair bulbs from one of these chimeras was submitted in batch with similar samples from control sheep for routine scrapie genetic relative susceptibility testing. A complex PRNP genotype was detected in the blood of the chimeric female but not in the alternate tissue samples or in the control sheep samples. The results demonstrate that naturally occurring blood chimerism can confound current testing efforts. The potential impacts of undetected chimeras on current scrapie eradication efforts are discussed.
Performance of Real-Time Reverse Transcription Polymerase Chain Reaction for the Detection of Foot-and-Mouth Disease Virus during Field Outbreaks in the United Kingdom in 2007
Authors: Reid Scott M. , Ebert Katja , Bachanek-Bankowska Katarzyna , Batten Carrie , Sanders Anna , Wright Caroline , Shaw Andrew E. , Ryan Eoin D. , Hutchings Geoffrey H. , Ferris Nigel P. , Paton David J. , King Donald P.
Published: May 2011

Rapid and accurate diagnosis is essential for effective control of foot-and-mouth disease (FMD). The present report describes the practical steps undertaken to deploy a real-time reverse transcription polymerase chain reaction (real-time RT-PCR) to process the samples received during the outbreaks of FMD in the United Kingdom in 2007. Two independent real-time RT-PCR assays targeting different regions (5′UTR and 3D) of the FMD virus (FMDV) genome were used to confirm the presence of FMDV in clinical samples collected from the first infected premises. Once the FMDV strain responsible had been sequenced, a single real-time RT-PCR assay (3D) was selected to test a total of 3,216 samples, including material from all 8 infected premises. Using a 96-well automated system to prepare nucleic acid template, up to 84 samples could be processed within 5 hr of submission, and up to 269 samples were tested per working day. A conservative cut-off was used to designate positive samples, giving rise to an assay specificity of 99.9% or 100% for negative control material or samples collected from negative premises, respectively. For the first time, real-time RT-PCR results were used to recognize preclinical FMD in a cattle herd. Furthermore, during the later stages of the outbreaks, the real-time RT-PCR assay supported an active surveillance program within high-risk cattle herds. To the authors' knowledge, this is the first documented use of real-time RT-PCR as a principal laboratory diagnostic tool following introduction of FMD into a country that was FMD-free (without vaccination) and highlights the advantages of this assay to support control decisions during disease outbreaks.
Clinical Findings and Serum Cardiac Troponin I Concentrations in Horses after Intragastric Administration of Sodium Monensin
Authors: Divers Thomas J. , Kraus Marc S. , Jesty Sophy A. , Miller Andrew D. , Mohammed Hussni O. , Gelzer Anna R. M. , Mitchell Lisa M. , Soderholm L. Vincent , Ducharme Normand G.
Published: May 2011

Six adult horses were administered sodium monensin, 1.0–1.5 mg/kg, via gastric gavage. Anorexia and/or diarrhea occurred within 24 hr after monensin administration in all 6 horses. Cardiac disease and dysfunction were evaluated by both elevations in heart rate, echocardiography, and an increase in serum concentrations of cardiac troponin I (cTnI), occurred in 4 horses. The development and severity of cardiac disease was likely affected by the monensin dose, vehicle (water or corn oil) mixed with monensin, and/or whether the monensin was administered to fed or fasted horses. Initial increases in cTnI concentrations occurred between 24 and 72 hr after monensin administration. The 2 horses with the highest cTnI concentrations died or were euthanized within 5 days after monensin administration and had severe cardiac disease. One horse had increased cTnI concentrations from day 2 to day 16, but no apparent change in ventricular contractile function was evident on echocardiography. The fourth diseased horse did not return to cTnI reference intervals until day 27 after monensin administration, and the ventricular function was still abnormal just before euthanasia 9 months later. Cardiac troponin I measurements could be useful in managing farm outbreaks of accidental monensin feeding by the early identification of horses with cardiac disease.
Comparison of Three Rapid Commercial Canine Parvovirus Antigen Detection Tests with Electron Microscopy and Polymerase Chain Reaction
Authors: Schmitz Silke , Coenen Christina , Matthias König , Heinz-Jürgen Thiel , Neiger Reto
Published: May 2011

Different antibody-based tests for rapid detection of Canine parvovirus antigens in feces are commercially available, allowing quick diagnosis in a clinical setting. However, the diagnostic accuracy of these tests compared with standard methods has not been evaluated so far. In the current study, 3 commercial tests were compared with immune-electron microscopy (IEM) and polymerase chain reaction (PCR). Dogs were divided into 3 groups: group A, samples from dogs with acute hemorrhagic diarrhea ( n = 50); group B, dogs with chronic diarrhea ( n = 10); and group C, dogs with no evidence of gastrointestinal disease ( n = 40). Specificity of all 3 commercial tests versus PCR and IEM was good to excellent (92.2–100%). Sensitivity, in contrast, was poor: 15.8–26.3% versus PCR and 50–60% versus IEM. In group A, 10 dogs were positive by IEM and 24 dogs were positive by PCR. Positive PCR results were also obtained from animals in control groups (group B, 1 dog; group C, 5 dogs). No dog in group B or C was positive by IEM. In conclusion, the rapid tests are useful to diagnose canine parvoviral enteritis, but they do not rule out parvovirus infection in an animal with typical clinical signs. In addition, a small percentage of healthy dogs and dogs with chronic diarrhea showed positive PCR results; this may be due to asymptomatic/persistent infection or intestinal passage of virus. The significance of this finding remains unclear.
Reverse Transcription Loop-Mediated Isothermal Amplification for the Detection of Porcine Reproductive and Respiratory Syndrome Virus
Authors: Rovira Albert , Abrahante Juan , Murtaugh Michael , Claudia Muñoz-Zanzi
Published: May 2011

Porcine reproductive and respiratory syndrome virus (PRRSV) is an important pathogen of swine. The objective of the current study is to investigate the feasibility of using reverse transcription loop-mediated isothermal amplification (RT-LAMP) for the detection of PRRSV. The RT-LAMP is a recently described DNA amplification technique reported to be simple, inexpensive, fast, and accurate. The RT-LAMP reaction was set up using 2 sets of primers that were designed to detect North American and European strains of PRRSV and performed successfully in a simple heat block. The specificity of the amplified product was demonstrated by restriction analysis. The RT-LAMP was able to detect 5 different PRRSV isolates. However, the limit of detection ranged between 10 2 and 10 4 50% tissue culture infective dose/ml. The RT-LAMP was further evaluated using serum samples from animals of known infection status. The ability of RT-LAMP to detect PRRSV in serum from acutely infected animals was evaluated with 114 serum samples from 18 experimentally inoculated boars. Forty-nine of these samples tested positive by RT-LAMP, while 94 were positive by reverse transcription polymerase chain reaction (RT-PCR). The diagnostic specificity, evaluated with 100 known negative serum samples, was estimated as 99%. The feasibility of RT-LAMP to detect PRRSV was demonstrated in the current study. The RT-LAMP reaction could be performed in just 1 hr with a simple and inexpensive heat block. However, the sensitivity of this technique was significantly lower than that of RT-PCR.
Actinomyces Hyovaginalis—Associated Lymphadenitis in a Nubian Goat
Authors: Schumacher Vanessa L. , Hinckley Lynn , Gilbert Kelly , Risatti Guillermo R. , Alfredo Sanchez Londoño , Smyth Joan A.
Published: May 2011

A 6-year-old Nubian goat with a history of progressive weight loss and cough was presented for necropsy. The goat tested negative for antibodies to caseous lymphadenitis and caprine arthritis and encephalitis by hemagglutination inhibition assay and enzyme-linked immunosorbent assay, respectively. Postmortem examination revealed marked enlargement and, with histopathology, a fibrinopurulent necrotizing lymphadenitis of a tracheobronchial lymph node, with an appearance similar to that reported in cases of caseous lymphadenitis. An organism characterized by molecular methods as Actinomyces hyovaginalis was isolated together with Staphylococcus spp. and Streptococcus spp. from the lesion. No Corynebacterium pseudotuberculosis was recovered. To the authors' knowledge, this is the first isolation of A. hyovaginalis from a goat. Although the exact contribution of A. hyovaginalis to the lesion remains to be established, this case demonstrates that A. hyovaginalis should be considered in cases of caseous lymphadenitis–type lesions, especially when C. pseudotuberculosis has been excluded.
Generalized Tularemia in a Vervet Monkey (Chlorocebus Aethiops) and a Patas Monkey (Erythrocebus Patas) in a Zoo
Authors: Miklós Gyuranecz , László Fodor , Lászlo Makrai , István Szöke , Katalin Jánosi , Krisztalovics Katalin , Károly Erdélyi
Published: May 2011

Generalized tularemia was diagnosed in a vervet monkey ( Chlorocebus aethiops) and a patas monkey ( Erythrocebus patas), both of which died suddenly in the Szeged Zoo, Szeged, Hungary. Macroscopic lesions in each animal included disseminated, grayish-white foci in the lungs, lymph nodes, spleen, liver, and kidney. All focal lesions were characterized microscopically as purulent to pyogranulomatous to granulomatous inflammation with necrosis. Francisella tularensis subsp. holarctica strains were isolated from tissue samples on modified Francis agar after mouse passage and identified by a commercial carbon-source utilization test and polymerase chain reaction-based amplification and sequencing of a portion of the 16S ribosomal RNA gene.
Histopathologic Criteria to Confirm White-nose Syndrome in Bats
Authors: Meteyer Carol Uphoff , Buckles Elizabeth L. , Blehert David S. , Hicks Alan C. , Green D. Earl , Shearn-Bochsler Valerie , Thomas Nancy J. , Gargas Andrea , Behr Melissa J.
Published: May 2011

White-nose syndrome (WNS) is a cutaneous fungal disease of hibernating bats associated with a novel Geomyces sp. fungus. Currently, confirmation of WNS requires histopathologic examination. Invasion of living tissue distinguishes this fungal infection from those caused by conventional transmissible dermatophytes. Although fungal hyphae penetrate the connective tissue of glabrous skin and muzzle, there is typically no cellular inflammatory response in hibernating bats. Preferred tissue samples to diagnose this fungal infection are rostral muzzle with nose and wing membrane fixed in 10% neutral buffered formalin. To optimize detection, the muzzle is trimmed longitudinally, the wing membrane is rolled, and multiple cross-sections are embedded to increase the surface area examined. Periodic acid–Schiff stain is essential to discriminate the nonpigmented fungal hyphae and conidia. Fungal hyphae form cup-like epidermal erosions and ulcers in the wing membrane and pinna with involvement of underlying connective tissue. In addition, fungal hyphae are present in hair follicles and in sebaceous and apocrine glands of the muzzle with invasion of tissue surrounding adnexa. Fungal hyphae in tissues are branching and septate, but the diameter and shape of the hyphae may vary from parallel walls measuring 2 μm in diameter to irregular walls measuring 3–5 μm in diameter. When present on short aerial hyphae, curved conidia are approximately 2.5 μm wide and 7.5 μm in curved length. Conidia have a more deeply basophilic center, and one or both ends are usually blunt. Although WNS is a disease of hibernating bats, severe wing damage due to fungal hyphae may be seen in bats that have recently emerged from hibernation. These recently emerged bats also have a robust suppurative inflammatory response.
Infectious and Lethal Doses of H5N1 Highly Pathogenic Avian Influenza Virus for House Sparrows (Passer Domesticus) and Rock Pigeons (Columbia Livia)
Authors: Brown Justin D. , Stallknecht David E. , Berghaus Roy D. , Swayne David E.
Published: May 2011

Terrestrial wild birds commonly associated with poultry farms have the potential to contribute to the spread of H5N1 highly pathogenic avian influenza (HPAI) virus within or between poultry facilities or between domesticated and wild bird populations. This potential, however, varies between species and is dependent on several virus and host factors, including habitat utilization, susceptibility, and viral shedding patterns. To provide data on susceptibility and shedding patterns of house sparrows ( Passer domesticus) and rock pigeons ( Columba livia), 20 birds from each species were inoculated with decreasing concentrations of A/whooper swan/Mongolia/244/05 (H5N1) HPAI virus, and the birds were evaluated for morbidity, mortality, viral shedding, and seroconversion over a 14-day trial. The house sparrows were highly susceptible to the H5N1 HPAI virus as evidenced by low infectious and lethal viral doses. In addition, house sparrows excreted virus via the oropharynx and cloaca for several days prior to the onset of clinical signs. Based on these results, house sparrows could play a role in the dissemination of H5N1 HPAI virus in poultry. In contrast, pigeons were resistant to the HPAI virus, requiring a high concentration of virus to produce infection or death. When infection did occur, the duration of viral shedding was brief, and viral titers were low. The data suggests that pigeons would contribute little to the transmission and spread of H5N1 HPAI virus in poultry.
Lung Pathology and Infectious Agents in Fatal Feedlot Pneumonias and Relationship with Mortality, Disease Onset, and Treatments
Authors: Fulton Robert W. , Blood K. Shawn , Panciera Roger J. , Payton Mark E. , Ridpath Julia F. , Confer Anthony W. , Saliki Jeremiah T. , Burge Lurinda T. , Welsh Ronald D. , Johnson Bill J. , Reck Amy
Published: May 2011

This study charted 237 fatal cases of bovine respiratory disease (BRD) observed from May 2002 to May 2003 in a single Oklahoma feed yard. Postmortem lung samples were used for agent identification and histopathology. Late in the study, 94 skin samples (ear notches) were tested for Bovine viral diarrhea virus (BVDV) by immunohistochemistry (IHC). Bovine respiratory disease morbidity was 14.7%, and the mortality rate of all causes was 1.3%, with more than half (53.8%) attributed to BRD (0.7% total of all causes). The agents isolated were the following: Mannheimia haemolytica (25.0%), Pasteurella multocida (24.5%), Histophilus somni (10.0%), Arcanobacterium pyogenes (35.0%), Salmonella spp. (0.5%), and Mycoplasma spp. (71.4%). Viruses recovered by cell culture were BVDV-1a noncytopathic (NCP; 2.7%), BVDV-1a cytopathic (CP) vaccine strain (1.8%), BVDV-1b NCP (2.7%), BVDV-2a NCP (3.2%), BVDV-2b CP (0.5%), and Bovine herpesvirus 1 (2.3%). Gel-based polymerase chain reaction (PCR) assays were 4.6% positive for Bovine respiratory syncytial virus and 10.8% positive for Bovine coronavirus. Bovine viral diarrhea virus IHC testing was positive in 5.3% of the animals. The mean values were determined for the treatment data: fatal disease onset (32.65 days), treatment interval (29.15 days), number of antibiotic treatments (2.65), number of different antibiotics (1.89), and day of death (61.81 days). Lesions included the following: 1) duration: acute (21%), subacute (15%), chronic (40.2%), healing (2.8%), normal (18.1%), and autolyzed (2.8%); 2) type of pneumonia: lobar bronchopneumonia (LBP; 27.1 %), LBP with pleuritis (49.1 %), interstitial pneumonia (5.1 %), bronchointerstitial pneumonia (1.4%), septic (0.9%), embolic foci (0.5%), other (2.8%), normal (10.3%), and autolyzed (2.8%); and 3) bronchiolar lesions: bronchiolitis obliterans (39.7%), bronchiolar necrosis (26.6%), bronchiolitis obliterans/bronchiolar necrosis (1.4%), other bronchiolar lesions (6.5%), and bronchiolar lesion negative (25.7%). Statistically significant relationships were present among the agents, lesions, and the animal treatment, disease onset, and mortality data. Clinical illnesses observed in this study were lengthier than those reported 16–20 years ago, based on fatal disease onset, treatment interval, and day of death.
Rapid Bilateral Intraocular Cocktail Sampling Method for West Nile Virus Detection in Dead Corvids
Authors: Lim Arleen K. , Dunne Gundula , Gurfield Nikos
Published: May 2011

Corvids can be a sensitive indicator for West Nile virus (WNV) prevalence and are a component of many WNV surveillance programs. An improved sampling procedure using a bilateral intraocular cocktail (BIC) was developed for testing corvid carcasses for WNV. This new procedure was substantially faster than harvesting internal organs, required less specialized equipment and training, and yielded excellent diagnostic sensitivity.
Detection of Avian Leukosis Virus Genome by a Nested Polymerase Chain Reaction using DNA and RNA from Dried Feather Shafts
Authors: Hatai Hitoshi , Ochiai Kenji , Umemura Takashi
Published: May 2011

The nested polymerase chain reaction (nPCR) using frozen feather pulp is useful for detecting fowl glioma-inducing virus (FGV), which belongs to the Avian leukosis virus family, and it has recently been suggested that FGV has spread to ornamental chickens kept in Japanese zoological gardens. In the current study, the practicality of using DNA and RNA from dried feather shafts as PCR samples was examined to establish a simple method for tissue preservation. Feather shafts were collected from 7 FGV-positive chickens and stored at room temperature for 30 days. DNA and RNA were extracted from these dried materials. All DNA and complementary DNA (cDNA) prepared from the RNA showed positive results for chicken β-actin and FGV, respectively. Screening for FGV was performed on Japanese fowls kept in zoological garden N. Of the feather shafts collected from 57 birds, 1 sample tested positive for FGV according to PCR of DNA and cDNA samples from the dried feather shafts. This positive bird originated from zoological garden A and had brain lesions suggestive of fowl glioma. The results suggest that DNA and RNA from dried feather shafts can be used in nPCR to detect the FGV genome.
Naturally Occurring Influenza Infection in a Ferret (Mustela Putorius Furo) Colony
Authors: Patterson Abby R. , Cooper Vickie L. , Yoon Kyoung-Jin , Janke Bruce H. , Gauger Phillip C.
Published: May 2011

Tissue samples from 2 juvenile ferrets ( Mustela putorius furo) from a colony that was undergoing an outbreak of respiratory disease were submitted to the Iowa State University Veterinary Diagnostic Laboratory. Microscopic examination of lung samples revealed bronchointerstitial pneumonia with necrotizing bronchiolitis. Influenza A virus was detected in sections of formalin-fixed lung by immunohistochemistry and reverse transcription polymerase chain reaction assay. A field investigation of the premises and analysis of additional samples led to the confirmation and characterization of an influenza virus with high homology to contemporary reassortant H1N1 swine influenza viruses. Although ferrets have been used extensively to research the virulence and transmissibility of avian, human, and swine influenza virus strains, no published information exists on naturally occurring outbreaks of swine influenza in ferrets.
Herpesvirus Infections in Rock Hyraxes (Procavia Capensis)
Authors: Galeota Judith A. , Napier Julia E. , Armstrong Douglas L. , Riethoven Jean-Jack , Rogers Douglas G.
Published: May 2011

Seven juveniles and 3 adults from a closed group of 19 rock hyraxes ( Procavia capensis) housed in a zoo's indoor rock exhibit died or were euthanized after developing blepharoconjunctivitis and orofacial ulcers over a 2-week period. Histopathologic examination of dermal ulcers and ulcerated tongues revealed amphophilic to basophilic intranuclear inclusion bodies in epithelial cells bordering ulcers. Epithelial cells with inclusion bodies were often characterized by cytomegaly and karyomegaly, and many cells had formed syncytia. Examination of inclusion bodies in tongue epithelium by transmission electron microscopy revealed icosahedral nucleocapsids, approximately 80–95 nm in diameter, with morphologic features consistent with herpesvirus. Cytopathic effect (CPE) typical of alphaherpesvirus infection was seen in bovine turbinate, equine dermal, and Vero cell monolayers after inoculation with homogenates of the skin lesions, but CPE was not seen after inoculation onto Madin-Darby canine kidney or swine testicle cell monolayers. Polymerase chain reaction analysis using degenerate primers that targeted a portion of the herpesvirus polymerase gene generated a product of approximately 227 base pairs. The product was cloned, sequenced, and then analyzed using BLAST. At the nucleotide level, there was 86%, 77%, and 76% shared identity with Eidolon herpesvirus 1, Human herpesviruses 1 and 2, and Cercopithecine herpesvirus 2, respectively. Herpesvirus infections in rock hyraxes have not been characterized. The data presented in the current study suggest that a novel alphaherpesvirus caused the lesions seen in these rock hyraxes. The molecular characteristics of this virus would tentatively support its inclusion in the genus Simplexvirus.
Persistent Truncus Arteriosus with Dissecting Aneurysm and Subsequent Cardiac Tamponade in a Lamb
Authors: Haist Verena , von Altrock Alexandra , Beineke Andreas
Published: May 2011

The present case report describes the necropsy and histopathology findings of a 3-week-old lamb with persistent truncus arteriosus (PTA). This rare cardiac malformation was characterized by the presence of a common arterial trunk arising from the right ventricle and overriding a ventricular septal defect. The pulmonary arteries originated from a short common trunk from this PTA, which subsequently continued as the thoracic aorta. The death of the lamb was attributed to a rupture of the PTA with subsequent cardiac tamponade. Histologically, a dissecting aneurysm with elastic fiber fragmentation in the wall of the PTA was identified as the underlying pathologic condition. Altered hemodynamic forces with subsequent secondary vasculopathy, as well as congenital primary vasculopathies, should be considered as an underlying pathogenetic mechanism.
Mycotic Pododermatitis and Mycotic Pneumonia in Commercial Turkey Poults in Northern California
Authors: Stoute Simone T. , Bickford Arthur A. , Walker Richard L. , Charlton Bruce R.
Published: May 2011

Seven 5-week-old broad-breasted white commercial meat turkeys were submitted to the California Animal Health and Food Safety laboratory in Turlock with a history of respiratory illness. The primary diagnostic findings were mycotic pododermatitis and mycotic pneumonia. The unique feature of this case was the colonization of footpad epidermis and subcutis by fungal hyphae in commercial turkey species. No fungal cultures were undertaken at the time of the necropsy; therefore, paraffin-embedded tissue sections of lung and footpads were used to extract, amplify, and sequence mycotic DNA. A mixed population of fungi was identified in both lung and footpads by polymerase chain reaction amplification of part of the large subunit ribosomal RNA gene using broad-range fungal primers and DNA sequencing. In footpads, sequences matching Cryptococcus saitoi and Cladosporium and Cudoniella species were identified. It is believed that these fungi were opportunistic pathogens originating from the litter. The fungi identified from lungs were Aspergillus species, most closely matching Aspergillus flavus and Arxiozyma telluris (most likely a contaminant). Mycotic pododermatitis in avian species is considered a rare pathologic finding, and few documented reports are available. The on-farm prevalence of footpad lesions was estimated at 3%, and there was no associated increase in the incidence of lameness or weight depression in affected birds. Microscopically, a granulomatous inflammatory reaction associated with fungal hyphae was observed in lung parenchyma. Disruption of keratinized epidermis, encrustations, and acute inflammation were also noted in footpads invaded with fungal hyphae.
Nephrotoxicosis in Iberian Piglets Subsequent to Exposure to Melamine and Derivatives in Spain between 2003 and 2006
Authors: González Jorge , Puschner Birgit , Pérez Valentín , Ferreras María C. , Delgado Laetitia , Muñoz María , Pérez Claudia , Reyes Luis E. , Velasco Javier , Fernández Víctor , García-Marín Juan F.
Published: May 2011

Between November 2003 and September 2006, 300 to 400 45–60-day-old Iberian piglets developed anorexia, polydipsia, and lethargy. Piglets were from 5 different farms in the western part of Spain. Morbidity was between 40% and 60%, and mortality ranged from 20% to 40% of the total population of postweaning piglets. In the 9 piglets in which postmortem examinations were conducted, kidneys were enlarged with yellow foci in the cortex and medulla. Microscopically, these foci were accumulations of crystals within the lumina of dilated distal tubules and collecting ducts, causing flattening of the renal tubular epithelial cells. The crystals displayed a multicolored birefringence under cross-polarized light. The multinucleated giant cells surrounding the crystals, interstitial fibrosis, and nonsuppurative infiltrates indicated a chronic inflammatory response. Toxicologic analysis of fixed kidney tissues from 4 piglets demonstrated the presence of melamine, ammeline, ammelide, and cyanuric acid. Ammelide concentrations were highest, ranging from 39,000 to 92,000 mg/kg, followed by ammeline (20,000–34,000 mg/kg), melamine (9,200–29,000 mg/kg), and cyanuric acid (2,200–9,100 mg/kg). The clinical, histologic, and toxicologic findings in affected piglets were similar to those reported in dogs and cats that died of melamine and melamine analogue-associated renal failure in 2004 and 2007. To the authors' knowledge, this is the first documented report of poisoning due to melamine and its analogues in pigs and demonstrates that contamination of pig feed occurred as early as 2003.
Extraintestinal Pathogenic Escherichia Coli-Induced Pneumonia in Three Kittens and Fecal Prevalence in a Clinically Healthy Cohort Population
Authors: Highland Margaret A. , Byrne Barbara A. , DebRoy Chitrita , Samitz Eileen M. , Peterson Tracy S. , Oslund Karen L.
Published: May 2011

Three kittens, ages 5, 9, and 17 weeks, were found dead by separate caregivers and were submitted for necropsy. At gross necropsy, each kitten had hemorrhagic or bloody fibrinoserous thoracic fluid and differing distributions of pulmonary consolidation. On histologic examination, the pulmonary lesion in each kitten was similar and was characterized by acute necrotizing and hemorrhagic pneumonia and pleuritis, with numerous intralesional small Gram-negative rods. A pure culture of a distinct serotype of Escherichia coli was identified in lung tissue from each kitten (O4:H5, O6:H7, O6:H5). Lung isolates, genotyped by polymerase chain reaction, carried genes that are characteristic of extraintestinal pathogenic E. coli (ExPEC), including cnf-1, papG allele I, papA, papC, sfa, fim, hlyD, malX, iroN, fyuA, kpsMII, and ompT. Escherichia coli isolates from the intestines of 2 of the kittens were 100% related to the respective lung isolate, as determined by pulsed-field gel electrophoresis. Cultures of fecal samples collected from a clinically healthy cohort population of kittens revealed 16 of 19 tested kittens (84%) to be shedding hemolytic E. coli. Ten different serotypes were identified from 43 hemolytic E. coli fecal isolates from the cohort population, each of which had a genetic profile consistent with that typical of ExPEC. To the authors’ knowledge, this is the first report to describe a cluster of isolated cases of pneumonia in kittens caused by distinct serotypes of ExPEC and to evaluate the prevalence of hemolytic E. coli carrying ExPEC-associated genes in the feces of a cohort population of kittens.
Optimal Probe Size and Fixation Time for the Detection of Porcine Circovirus–2 DNA by in Situ Hybridization in Formalin-Fixed, Paraffin-Embedded Tissue
Authors: Ha Yooncheol , Chae Chanhee
Published: May 2011

Probe size and fixation time for detecting Porcine circovirus–2 (PCV–2) by in situ hybridization in formalin-fixed, paraffin-embedded lymph nodes from experimentally infected pigs were optimized. In situ hybridization using a 169–base pair (bp) probe detected significantly fewer PCV–2–positive cells than when using 8 other larger probes ( P < 0.05). The difference in hybridization intensity between smaller probes (169 and 225 bp) and larger probes (416, 473, 571, 631, 693, and 753 bp) was statistically significant ( P < 0.05). The PCV–2–positive cells were consistently detected in lymph nodes fixed up to 3 days; thereafter, the number of positive cells declined. The PCV–2–positive cells were detected in lymph nodes fixed for up to 730 days. The difference in hybridization intensity between samples fixed for a short term (1 or 3 days) and a longer term (4–730 days) was statistically significant ( P < 0.05). The data demonstrates that the optimal probe size and fixation time for detecting PCV–2 in formalin-fixed, paraffin-embedded lymph nodes is 473 bp and 1–3 days, respectively.
Standardization and Validation of an Immunoperoxidase Assay for the Detection of African Horse Sickness Virus in Formalin-Fixed, Paraffin-Embedded Tissues
Authors: Clift Sarah J. , Williams Mark C. , Gerdes Truuske , Smit Marie M. E.
Published: May 2011

An immunoperoxidase assay for the detection of African horse sickness virus (AHSV) in formalin-fixed tissues is a valuable tool in the study of the pathogenesis of the disease, as well as a useful addition to existing diagnostic tests when only preserved tissues are available. An assay that uses Hamblin antiserum in a basic avidin-biotin complex detection system was standardized and validated in accordance with the guidelines of the American Association of Veterinary Laboratory Diagnosticians Subcommittee on Standardization of Immunohistochemistry. Using 128 positive cases of African horse sickness confirmed by viral isolation and serotyping and 119 negative cases from countries where the disease has never occurred, diagnostic sensitivity and diagnostic specificity were 100% in the prime target tissues of heart and lung. There was no variation in the ability of the assay to detect all 9 serotypes of AHSV, and there was no cross-reactivity with other orbiviruses in formalin-fixed tissues. The only cross-reactivity observed was in the lungs of 2 negative cases infected with Rhodococcus equi. The assay gave good results on tissues that had been fixed in formalin for up to 365 days. Nonspecific staining was minimal provided that the standard procedures for processing and staining tissues were followed. Good immunohistochemical results were also obtained on samples fixed as long as 24 hr after death. The assay, therefore, provides a robust diagnostic tool for detection of AHSV in formalin-fixed tissues, provided the analysis is done by an experienced pathologist.
Development of Quantitative Real-Time Polymerase Chain Reaction for Detection of and Discrimination between Erysipelothrix Rhusiopathiae and Other Erysipelothrix Species
Authors: To Ho , Koyama Tomohiro , Nagai Shinya , Tuchiya Kotaro , Nunoya Tetsuo
Published: May 2011

Quantitative real-time polymerase chain reaction (qPCR) assays were developed and validated in combination with enrichment culture for the detection and discrimination of Erysipelothrix rhusiopathiae and other Erysipelothrix species from tissue samples. The targets for SYBR green qPCR assays were the 16S ribosomal RNA gene for Erysipelothrix species and a gene involved in capsular formation for E. rhusiopathiae. The specificity of the assays was assessed with Erysipelothrix species and other related bacterial species. The limit of detection was found to be 5 colony-forming units per reaction. Amplification of DNA extracted from spleen and joint samples spiked with increasing quantities of Erysipelothrix cells was shown to be equally sensitive to DNA extracted from a pure bacterial culture. The assays were evaluated with 88 tissue samples from 3 experimentally infected pigs and 50 mice and with 36 tissue samples from 3 naturally infected pigs and 11 noninfected pigs. Results were compared with those of direct qPCR and conventional culture. The qPCR after enrichment increased the diagnostic sensitivity over that of culture and qPCR, thereby significantly reducing the total time taken for the detection of E. rhusiopathiae and other Erysipelothrix species. Therefore, this technique could be used for practical applications.