bluuebunny/crossref_metadata_2025 huggingface dataset:
Correlation between Herrold Egg Yolk Medium Culture and Real-Time Quantitative Polymerase Chain Reaction Results for Mycobacterium Avium Subspecies Paratuberculosis in Pooled Fecal and Environmental Samples
Authors: Aly Sharif S. , Mangold Beverly L. , Whitlock Robert H. , Sweeney Raymond W. , Anderson Randall J. , Jiang Jiming , Schukken Ynte H. , Hovingh Ernest , Wolfgang David , Van Kessel Jo Ann S. , Karns Jeffrey S. , Lombard Jason E. , Smith Julia M. , Gardner Ian A.
Published: May 2011

Real-time quantitative polymerase chain reaction (qPCR) testing for Mycobacterium avium subspecies paratuberculosis (MAP) in fecal samples is a rapid alternative to culture on Herrold egg yolk medium (HEYM), the traditional antemortem reference test for MAP. Although the sensitivity and specificity of these 2 tests have been estimated based on dichotomized test results, the correlation between real-time qPCR threshold cycle (Ct) values and colony-forming units (CFU) on HEYM for fresh and thawed samples has not been evaluated. The objectives of the present study were to estimate the correlation and association between Ct and CFU in fresh and thawed pooled fecal and environmental samples. Results of HEYM culture of 1,997 pooled fecal samples from cows in 14 herds, and 802 environmental samples from 109 dairies nationwide were negatively (inversely) correlated with their respective real-time qPCR results. The Spearman's rank correlation between Ct and CFU was good (–0.66) in fresh and thawed pooled fecal samples, and excellent (–0.76) and good (–0.61) in fresh and thawed environmental samples, respectively. The correlation varied from good (–0.53) to excellent (–0.90) depending on the number of samples in a fecal pool. Truncated regression models indicated a significant negative association between Ct and CFU in fecal pools and environmental samples. The use of real-time qPCR instead of HEYM can yield rapid, quantitative estimates of MAP load and allow for incorporation of real-time qPCR results of pooled and environmental samples in testing strategies to identify dairy cow groups with the highest MAP shedding.
Canine Reference Intervals for Coagulation Markers Using the STA Satellite® and the STA-R Evolution® Analyzers
Authors: Geffré Anne , Grollier Sophie , Hanot Camille , Vergez François , Trumel Catherine , Braun Jean-Pierre
Published: May 2011

The aim of the current study was to determine canine reference intervals for prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, and antithrombin (AT) according to international recommendations. The STA Satellite® coefficients of variation of within-laboratory imprecision were 3.9%, 1.3%, 6.9%, and 5.1% for PT, APTT, fibrinogen, and AT, respectively. At 4°C, citrated specimens were stable up to 8 hr for whole blood and 36 hr for plasma, except for APTT, which increased slightly (<1 sec). Nonparametric reference intervals determined in citrated plasma from 139 healthy fasting purebred dogs were 6.9–8.8 sec, 13.1–17.2 sec, 1.24–4.30 g/l, and 104–188% for PT, APTT, fibrinogen, and AT, respectively. Based on Passing–Bablok comparison between STA Satellite and STA-R Evolution® using 60 frozen specimens from a canine plasma bank, the corresponding reference intervals were transferred to the STA-R Evolution: 7.1–9.2 sec, 12.9–17.3 sec, 1.20–4.43 g/l, and 94–159% for PT, APTT, fibrinogen, and AT, respectively.
Effect of Pancreatic Tissue Sampling on Serum Pancreatic Enzyme Levels in Clinically Healthy Dogs
Authors: Cordner Amy P. , Armstrong P. Jane , Newman Shelley J. , Novo Roberto , Sharkey Leslie C. , Jessen Carl
Published: May 2011

Little is known about the potential consequences of pancreatic tissue sampling in dogs. The goal of the present study was to evaluate changes in serum trypsin–like immunoreactivity and canine-specific pancreatic lipase after pancreatic fine-needle aspiration and surgical biopsy in 27 clinically healthy dogs. Presurgical, ultrasound-guided aspiration of the pancreas was performed with the dogs under sedation. Subsequently, all the dogs underwent intraoperative pancreatic fine-needle aspiration and clamshell biopsy. After euthanasia, pancreata were sectioned for histopathologic evaluation. Serum pancreatic enzyme levels were measured at 3 time points: baseline, after ultrasound-guided aspiration, and after intraoperative aspiration and biopsy. No significant differences were detected among mean serum pancreatic lipase values at any point ( P > 0.05). Serum trypsin–like immunoreactivity did not change from baseline (18.2 ± 2.1 μg/dl; mean ± standard error) after ultrasound-guided aspiration (13.6 ± 2.2 μg/dl) but increased significantly after intraoperative sampling (44.8 ± 1.9 μg/dl; P < 0.0005). After surgical biopsy, the 20 dogs that had both ultrasound-guided and intraoperative sampling had a higher mean (SE) serum trypsin–like immunoreactivity (44.8 ± 1.9 μg/dl) than the 7 dogs that had only intraoperative samples taken (36.4 ± 4.1 μg/dl; P < 0.05). All 27 pancreata were grossly normal before intraoperative sampling. Pancreatic sampling was associated with increased serum trypsin–like immunoreactivity and mild, peracute necrosis, inflammation, hemorrhage, and fibrin deposition. Tissue damage from sampling was not sufficient to cause an elevation in canine-specific pancreatic lipase in the time frame evaluated. Further studies are needed to determine longer-term effects of pancreatic sampling on enzyme levels and clinical outcome.
A Novel Rapid Genotyping Technique for Collie Eye Anomaly: SYBR Green–Based Real-Time Polymerase Chain Reaction Method Applicable to Blood and Saliva Specimens on Flinders Technology Associates Filter Paper
Authors: Chang Hye-Sook , Mizukami Keijiro , Yabuki Akira , Hossain Mohammad A. , Rahman Mohammad M. , Uddin Mohammad M. , Arai Toshiro , Yamato Osamu
Published: May 2011

Collie eye anomaly (CEA) is a canine inherited ocular disease that shows a wide variety of manifestations and severity of clinical lesions. Recently, a CEA-associated mutation was reported, and a DNA test that uses conventional polymerase chain reaction (PCR) has now become available. The objective of the current study was to develop a novel rapid genotyping technique by using SYBR Green–based real-time PCR for future large-scale surveys as a key part in the strategy to eradicate CEA by selective breeding. First, a SYBR Green–based real-time PCR assay for genotyping of CEA was developed and evaluated by using purified DNA samples from normal, carrier, and affected Border Collies in which genotypes had previously been determined by conventional PCR. This real-time PCR assay demonstrated appropriate amplifications in all genotypes, and the results were consistent with those of conventional PCR. Second, the availability of Flinders Technology Associates filter paper (FTA card) as DNA templates for the real-time PCR assay was evaluated by using blood and saliva specimens to determine suitability for CEA screening. DNA-containing solution prepared from a disc of blood- or saliva-spotted FTA cards was available directly as templates for the real-time PCR assay when the volume of solution was 2.5% of the PCR mixture. In conclusion, SYBR Green–based real-time PCR combined with FTA cards is a rapid genotyping technique for CEA that can markedly shorten the overall time required for genotyping as well as simplify the sample preparation. Therefore, this newly developed technique suits large-scale screening in breeding populations of Collie-related breeds.
The Microanatomy of Healthy Skin from Alpacas (Vicugna Pacos)
Authors: Fleis Rebekah I. , Scott Danny W.
Published: May 2011

The microanatomy of healthy skin from 12 different body sites was investigated in 14 alpacas ( Vicugna pacos). The microanatomy of alpaca skin is typical of domestic animal skin in general, and closely resembles that from llamas.
Characterization of Canine Parvovirus 2 Variants Circulating in Greece
Authors: Ntafis Vasileios , Xylouri Eftychia , Kalli Iris , Desario Costantina , Mari Viviana , Decaro Nicola , Buonavoglia Canio
Published: May 2011

The aim of the present study was to characterize Canine parvovirus 2 (CPV-2) variants currently circulating in Greece. Between March 2008 and March 2009, 167 fecal samples were collected from diarrheic dogs from different regions of Greece. Canine parvovirus 2 was detected by standard polymerase chain reaction, whereas minor groove binder probe assays were used to distinguish genetic variants and discriminate between vaccine and field strains. Of 84 CPV-2–positive samples, 81 CPV-2a, 1 CPV-2b, and 2 CPV-2c were detected. Vaccine strains were not detected in any sample. Sequence analysis of the VP2 gene of the 2 CPV-2c viruses revealed up to 100% amino acid identity with the CPV-2c strains previously detected in Europe. The results indicated that, unlike other European countries, CPV-2a remains the most common variant in Greece, and that the CPV-2c variant found in Europe is also present in Greece.
Heteroduplex Polymerase Chain Reaction is Essential for Canine Receptor Rearrangement Analysis
Authors: Takanosu Masamine , Tadika Tomohiko , Kobayashi Tetsuya
Published: May 2011

Polymerase chain reaction (PCR) for antigen receptor rearrangement is a sensitive technique for detecting lymphocyte-proliferative disorders, but it tends to produce false-positive results, a phenomenon termed pseudoclonality. Heteroduplex analysis, which is useful to distinguish clonal reactions from pseudoclonal ones in dogs, can be applied to avoid misdiagnosis and determine the reliability of results. In the current study, PCR for antigen receptor rearrangement was used to identify clonal proliferation of lymphocytes in duodenal and lymphoid tissue from dogs presenting with chronic vomiting and enlarged peripheral lymph nodes typical of multicentric lymphoma, and the test results were verified with heteroduplex analysis. In the case of almost all of the duodenal samples, even without a histologic diagnosis of lymphoma, a distinct band similar to that observed in the case of lymphoma was obtained for both B- and T-cell clonality. All of the bands obtained from the nonneoplastic duodenum disappeared following heteroduplex analysis of the PCR product, whereas the distinct bands from the lymphoma remained. In the lymph node samples, the pseudoclonal bands that disappeared in the heteroduplex analysis were detected mainly in B cells. In conclusion, heteroduplex analysis with PCR for antigen receptor rearrangement is a suitable tool for diagnosing canine lymphoma and decreasing the possibility of misdiagnosis of pseudoclonality.
A Survey of Porcine Picornaviruses and Adenoviruses in Fecal Samples in Spain
Authors: Buitrago Dolores , Cano-Gómez Cristina , Agüero Montserrat , Fernandez-Pacheco Paloma , Gómez-Tejedor Concepción , Jiménez-Clavero Miguel Ángel
Published: May 2011

In the course of an epidemiologic surveillance program for swine diseases carried out in Spain, 206 cytopathic viruses were isolated from 600 porcine fecal samples between 2004 and 2005. The virus isolates were examined using reverse transcription polymerase chain reaction (RT-PCR) methods specific for different types of porcine picornaviruses, including members of the Teschovirus, Enterovirus, and Sapelovirus genera, and PCR for porcine adenoviruses. Of the 206 isolates, 97 (47%) were identified as teschoviruses, 18 (9%) as sapeloviruses, and 7 (3%) as porcine adenoviruses. Neither Porcine enterovirus B nor Swine vesicular disease virus was found among the isolates. The present study confirms that teschoviruses are highly prevalent in porcine fecal samples, at least in Spain. It also reveals that these viruses commonly circulate among apparently healthy pigs.
Prevalence of Mycoplasma Haemolamae Infection in Peruvian and Chilean Llamas and Alpacas
Authors: Tornquist Susan J. , Boeder Lisa , Rios-Phillips Carolina , Alarcon Virgilio
Published: May 2011

Mycoplasma haemolamae is a hemotropic mycoplasma that affects red blood cells of llamas ( Lama glama) and alpacas (Lama pacos). It is variably associated with anemia, and most infections are subclinical. Development of a polymerase chain reaction assay has facilitated detection of this infection in llamas and alpacas in the United States and other countries. Whether the infection occurs in camelids in South America has previously been unknown. The current study documents a 15.8% infection rate among 76 Peruvian llamas, a 19.3% infection rate among Peruvian alpacas at one site, and a 9.26% infection rate in 108 Chilean alpacas from selected herds. All of the camelids tested appeared to be clinically healthy. No gender or species predilection was found. Only 1 positive camelid younger than 18 months was found. Infection is not associated with anemia, and the mean packed cell volume (PCV) in positive Peruvian camelids was slightly higher than the mean PCV in negative Peruvian camelids. In the Chilean alpacas, the positive alpacas had a slightly lower PCV than the negative alpacas, although the mean PCV was not in the anemic range in any of the groups.
Stenotrophomonas Maltophilia as a Causal Agent of Pyogranulomatous Hepatitis in a Buffalo (Bubalus Bubalis)
Authors: Petridou Evanthia , Filioussis George , Karavanis Emmanouel , Kritas Spyridon K.
Published: May 2011

A 7-year-old female buffalo ( Bubalus bubalis) from a local herd in Serres, northern Greece, was presented to a private veterinary clinic with a chronic loss of appetite for 15 days. The clinical examination revealed high fever (41.5°C), lethargy, yellow discoloration of skin and mucous membranes, an abdomen that appeared to be empty, hyperactive rumen motility, and tachypnea. A biochemical profile revealed an elevated total bilirubin concentration and hepatic enzyme activities, whereas globulin, creatinine, and glucose concentrations were within the reference intervals. The animal received a 12-day course of treatment with intramuscular administration of ampicillin and corticosteroids. However, no significant clinical improvement was achieved, and the buffalo was euthanized. Gross necropsy lesions included serous atrophy of adipose tissue and hepatomegaly. Microscopic lesions included necrotizing pyogranulomatous hepatitis with thrombosis, hemorrhages, edema, and fibrosis. Small, nonpigmented, bacterial colonies were harvested in pure culture from the liver and were confirmed as Stenotrophomonas maltophilia by polymerase chain reaction. The bacterium was sensitive to ciprofloxacin, enrofloxacin, colistin, polymyxin, trimethoprim/sulfamethaxazole, and chloramphenicol. In contrast, resistance to ticarcillin, piperacillin, imipenem, ceftazidime, amikacin, gentamicin, tobramycin, and tetracycline was displayed. The bacterial strain carried the L1 metallo-β-lactamase (L1) and tet35 genes, which contribute to high-level resistance to β-lactams and tetracycline, respectively. Although S. maltophilia is widely believed to be a contaminant, the present report suggests that the isolation, identification, and susceptibility testing of this multidrug-resistant bacterium may be of clinical importance in diagnostic samples.
Paraneoplastic Hypercalcemia in a Dog with Benign Renal Angiomyxoma
Authors: Gajanayake Isuru , Priestnall Simon L. , Benigni Livia , English Kate , Summers Brian A. , Garden Oliver A.
Published: May 2011

An 11-year-old, male, neutered crossbred Collie dog was presented for a history of polydipsia and polyuria. Diagnostic investigations revealed total and ionized hypercalcemia and an increased concentration of parathyroid hormone–related peptide. Abdominal ultrasonography and contrast-enhanced computed tomography of the abdomen revealed a right-sided, cystic-appearing renal mass. Cytological examination of ultrasound-guided aspirates of the mass revealed high numbers of spindle cells. The mass was removed en bloc via an ureteronephrectomy. Histopathological examination of the mass revealed neoplastic spindle cells in loosely packed and interlacing streams within a myxomatous stroma. Immunohistochemical examination with vimentin, von Willebrand Factor, and α-smooth muscle actin confirmed the mass to be a renal angiomyxoma. A minority of the neoplastic spindle cells showed positive cytoplasmic parathyroid hormone–related peptide immunostaining. The hypercalcemia resolved following surgery, and the parathyroid hormone–related peptide concentration returned to within the reference interval. The dog was no longer polydipsic or polyuric 1 year following surgery. The present report describes a previously unreported renal neoplasm causing paraneoplastic hypercalcemia and highlights the possibility of paraneoplastic hypercalcemia being caused by a benign neoplasm.
Natural Cases of 2009 Pandemic H1N1 Influenza A Virus in Pet Ferrets
Authors: Swenson Sabrina L. , Koster Leo G. , Jenkins-Moore Melinda , Killian Mary L. , DeBess Emilio E. , Baker Rocky J. , Mulrooney Donna , Weiss Robin , Galeota Judith , Bredthauer Annette
Published: May 2011

Respiratory swab samples were collected from 5 pet ferrets ( Mustela putorius furo) exhibiting influenza-like illness. The ferrets represented 3 households in 2 states. In each case, the owners reported influenza-like illness in themselves or family members prior to the onset of a similar illness in the ferrets. Realtime reverse transcription polymerase chain reaction assays designed for the detection of the 2009 H1N1 Influenza A virus were conducted in the state animal health laboratories. The assays included detection of the matrix gene of Influenza A virus and neuraminidase gene specific for 2009 H1N1 virus. Samples were positive for both screening assays. The samples were confirmed positive by the National Veterinary Services Laboratories. The history of illness in family members prior to illness in the ferrets suggests that Influenza A virus was transmitted from humans to the ferrets.
Focal Symmetrical Encephalomalacia in a Goat
Authors: Oliveira Diego M. , Pimentel Luciano A. , Pessoa André F. , Dantas Antônio F. M. , Uzal Francisco , Riet-Correa Franklin
Published: May 2011

Focal symmetrical encephalomalacia (FSE) is the most prominent lesion seen in the chronic form of enterotoxemia caused by Clostridium perfringens type D in sheep. However, this lesion has not been reported in goats. The current paper reports a case of FSE in a goat from the state of Paraíba in the Brazilian semiarid region. As reported by the farmer, 30, 4–48-month-old animals from a flock of 150 goats died after showing nervous signs, including blindness and recumbence, for periods varying between 1 and 14 days. The flock was grazing native pasture supplemented with wheat and corn bran. Additionally, lactating goats were supplemented with soybeans. A 4-month-old goat with nervous signs was examined clinically and then necropsied 3 days after the onset of clinical signs. Bilateral, focal, and symmetrical areas of brown discoloration were observed in the internal capsule and thalamus. Histologic lesions in these areas consisted of multifocal, bilateral malacia with a few neutrophils; endothelial cell swelling; perivascular edema; and hemorrhages. The etiology of these lesions was not determined. However, FSE is considered pathognomonic for C. perfringens type D enterotoxemia in sheep, and it is speculated that this microorganism was the etiologic agent in the present case. The flock had been vaccinated against type D enterotoxemia only once, approximately 3 months before the beginning of the outbreak. Insufficient immunity due to the incorrect vaccination protocol, low efficacy of the vaccine used, and a diet including large amounts of highly fermentable carbohydrates were suspected to be predisposing factors for this outbreak.
Meningoencephalitis Caused by Mycoplasma Edwardii in a Dog
Authors: Ilha Marcia R. S. , Rajeev Sreekumari , Watson Cindy , Woldemeskel Moges
Published: May 2011

A 6-week-old, female, mixed-breed dog with a clinical history of sudden onset of neurologic signs was presented for necropsy. The dog was diagnosed with suppurative and histiocytic meningoencephalitis based on necropsy findings and histopathology. Mycoplasma sp. was isolated in pure culture from the brain and meninges and was identified as Mycoplasma edwardii using DNA sequencing.
Juxtaductal Coarctation of Both Pulmonary Arteries in a Cat
Authors: Szatmári Viktor , Freund Matthias W. , Kroeze Edwin J. B. Veldhuis , Strengers Jan
Published: May 2011

A 4-year-old, male, neutered cat was referred because of recurrent episodes of dyspnea. Physical examination revealed a harsh systolic murmur, with the point of maximal intensity in the left heart base, with an intensity of 4 out of 6. Echocardiographic diagnosis was severe supravalvular pulmonary artery stenosis. A selective right ventricular angiocardiogram showed an absence of arterial blood flow to the left lung lobes. A balloon dilatation of the localized stenosis of the right pulmonary artery was attempted with cardiac catheterization. However, when the catheter was passed through the stenosis, the blood flow to the lungs temporarily completely ceased, which led to death. Postmortem examination revealed a circumscribed stenosis of both pulmonary arteries at the site of the bifurcation, where the ligamentum arteriosum was attached. Histopathology showed that the localized ridge-like stenosis at the pulmonary artery bifurcation was caused by connective tissue. The suspected cause of this congenital anomaly is the presence of ectopic ductal tissue in the wall of the pulmonary artery. When the ductus arteriosus closes at birth, pulmonary artery stenosis developed because of constriction of the ectopic ductal tissue.
Atypical Transmissible Spongiform Encephalopathies in Ruminants: A Challenge for Disease Surveillance and Control
Authors: Seuberlich Torsten , Heim Dagmar , Zurbriggen Andreas
Published: May 2011

Since 1987, when bovine spongiform encephalopathy (BSE) emerged as a novel disease in cattle, enormous efforts were undertaken to monitor and control the disease in ruminants worldwide. The driving force was its high economic impact, which resulted from trade restrictions and the loss of consumer confidence in beef products, the latter because BSE turned out to be a fatal zoonosis, causing variant Creutzfeldt—Jakob disease in human beings. The ban on meat and bone meal in livestock feed and the removal of specified risk materials from the food chain were the main measures to successfully prevent infection in cattle and to protect human beings from BSE exposure. However, although BSE is now under control, previously unknown, so-called atypical transmissible spongiform encephalopathies (TSEs) in cattle and small ruminants have been identified by enhanced disease surveillance. This report briefly reviews and summarizes the current level of knowledge on the spectrum of TSEs in cattle and small ruminants and addresses the question of the extent to which such atypical TSEs have an effect on disease surveillance and control strategies.
Diagnostic Assays Used to Control Small Ruminant Lentiviruses
Authors: Herrmann-Hoesing Lynn M.
Published: May 2011

The serologic diagnostic tests, such as the agar gel immunodiffusion assay and various types of enzyme-linked immunosorbent assays (ELISAs), have contributed to the reduction of small ruminant lentivirus (SRLV) infections worldwide. Because there are no treatments or efficacious vaccines, the serologic diagnostic tests have supported most of the eradication efforts by testing and removal or separation of adult animals that generate antibodies to SRLVs. With the advent of molecular diagnostics, standard and quantitative polymerase chain reaction (PCR)—based assays for the detection of provirus in peripheral blood cells are becoming more common and aid in the detection of infected goats and sheep before antibody detection by ELISA in some animals. Performance of the serologic and molecular diagnostic tests is dependent upon a number of factors, including the format of the assay, the percentage of identity between the viral nucleotide sequences in a flock or herd of a certain geographic region and the sequences used to generate SRLV test reagents, and the intrinsic pathogenesis or amount of provirus and SRLV antibody generated in a species or individual small ruminant. In addition, small ruminant genomics may help with establishing genetic markers of SRLV infection and disease, which could also aid eradication or reduction of SRLVs from herds and flocks throughout the world.
Development, Validation, and Utilization of a Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Antibodies against Brucella Species in Marine Mammals
Authors: Meegan Jenny , Field Cara , Sidor Inga , Romano Tracy , Casinghino Sandra , Smith Cynthia R. , Kashinsky Lizabeth , Fair Patricia A. , Bossart Gregory , Wells Randall , Dunn J. Lawrence
Published: May 2011

A competitive enzyme-linked immunosorbent assay (cELISA) was developed by using a whole-cell antigen from a marine Brucella sp. isolated from a harbor seal ( Phoca vitulina). The assay was designed to screen sera from multiple marine mammal species for the presence of antibodies against marine-origin Brucella. Based on comparisons with culture-confirmed cases, specificity and sensitivity for cetacean samples tested were 73% and 100%, respectively. For pinniped samples, specificity and sensitivity values were 77% and 67%, respectively. Hawaiian monk seal ( Monachus schauinslandi; n = 28) and bottlenose dolphin ( Tursiops truncatus; n = 48) serum samples were tested, and the results were compared with several other assays designed to detect Brucella abortus antibodies. The comparison testing revealed the marine-origin cELISA to be more sensitive than the B. abortus tests by the detection of additional positive serum samples. The newly developed cELISA is an effective serologic method for detection of the presence of antibodies against marine-origin Brucella sp. in marine mammals.
Inflammatory Myopathy with Severe Tongue Atrophy in Pembroke Welsh Corgi Dogs
Authors: Toyoda Kaoru , Uchida Kazuyuki , Matsuki Naoaki , Sakai Hideo , Kitagawa Masato , Saito Miyoko , Sasaki Jun , Nakayama Hiroyuki
Published: May 2011

A disease characterized by tongue and facial muscle atrophy has been recognized sporadically among Pembroke Welsh Corgi (PWC) dogs in Japan. The present study describes the pathologic findings of this canine syndrome. Histopathologic examinations were performed in 2 dogs, including a case of muscular biopsy. Identification and characterization of autoantibodies were attempted by fluorescent antibody test (FAT) and Western blot (WB) by using sera from 7 PWC dogs with typical clinical features, 6 PWC dogs with other clinical signs, and 2 from other breeds with polymyositis. Clinically, the 7 affected PWC dogs exhibited dysphagia with severe tongue atrophy, facial muscular atrophy, and occasional walking difficulty. Histopathologic examinations of the 2 dogs with clinical symptoms revealed moderate to severe inflammatory lesions characterized by lymphohistiocytic infiltration and muscular atrophy in the tongue and/or femoral muscles. The tongue lesions were very severe and accompanied by diffuse fatty infiltration. There were no major lesions in the nervous tissues examined. By FAT, an autoantibody against the cross striation of skeletal muscle was detected in sera from 5 affected PWC dogs. By using WB analysis, the autoantibodies recognized a 42-kDa molecule in striated muscle but not in the nervous tissues. All of the findings indicated that the unique disease of PWC dogs might be generalized inflammatory myopathy, whereas the detailed etiology concerning the dominant involvement of tongue muscles and the role of the autoantibody in the canine disease remain to be clarified.
A Diagnostic Algorithm for Detection of Antibodies to Influenza A Viruses in Dogs in Italy (2006–2008)
Authors: De Benedictis Paola , Anderson Tara C. , Perez Andrés , Viale Elisabetta , Veggiato Crispina , Caenazzo Silvia Tiozzo , Crawford P. Cynda , Capua Ilaria
Published: May 2011

Since 2004, there have been several reports of Influenza A virus (FLUAV) infection in dogs. Dogs have been infected with equine influenza H3N8, avian influenza H3N2 and H5N1, and the pandemic H1N1 virus. Because ofrecent avian and equine influenza outbreaksin Italy, the objectives of the present study were to estimate the level of exposure of Italian dogs to influenza A viruses and to assess a diagnostic algorithm for detection of FLUAV exposure in dogs. Sera collected from 6,858 dogs from 2006 to 2008 were screened in a competitive enzyme-linked immunosorbent assay (cELISA) for antibodies to the highly conserved influenza A nucleoprotein. Samples positive in the cELISA were confirmed by testing in hemagglutination inhibition (HI) and fluorescent antibody test (FAT). Two seropositive dogs had antibodies to H3 hemagglutinin proteins, consistent with exposure torecent canine and equine subtype H3N8 viruses. Using a Bayesian model, the sensitivity and specificity of the cELISA were estimated as 93.98% (probability intervals [PI]: 81.67–99.08%) and 98.71% (PI: 98.43–98.96%), respectively. After accounting for the imperfect sensitivity and specificity ofthe cELISA, the Bayesian posterior prevalence of FLUAV exposure among tested Italian dogs was 0.5% (PI: 0.1–1.4%). The study results indicate that screening with a cELISA for influenza A nucleoprotein antibody, followed by confirmatory testing with HI and/or FAT, is a highly sensitive and highly specific approach for diagnosing FLUAV exposure in dogs.
A Multiphasic Typing Approach to Subtype Streptococcus Equi Subspecies Equi
Authors: Lanka Saraswathi , Borst Luke B. , Patterson Sheila K. , Maddox Carol W.
Published: May 2011

The objective of the present investigation was to differentiate between strains of Streptococcus equi subspecies equi implicated in abscess formation in vaccinated horses. Streptococcus equi isolates recovered from clinical specimens associated with equine strangles cases submitted to the University of Illinois Veterinary Diagnostic Laboratory were compared with S. equi isolates representing at least 12 lots of a commercial modified live vaccine (MLV) to determine whether the isolates obtained from the abscesses were vaccine or wild type. Genotyping techniques evaluated included enterobacterial repetitive intergenic consensus polymerase chain reaction (PCR), repetitive extragenic palindrome PCR, BOX element PCR, ribotyping, and pulsed-field gel electrophoresis (PFGE). Phenotypic evaluations were performed using the Biolog GP2 Microplate (hereafter, Biolog). In cases where Biolog and PFGE results did not coincide, a single nucleotide polymorphism located in the upstream regulatory region of szp gene was used to identify the S. equi strains. PFGE and Biolog successfully differentiated wild-type S. equi strains isolated from clinical submissions from isolates of the MLV. PFGE genotyping enabled further subtyping of the wild-type strains, whereas Biolog combined with szp sequencing was useful in differentiating the MLV strain from its wild-type progenitor. Deletion of a single guanine residue located in the upstream regulatory region of the szp gene appears to be conserved among vaccine isolates, and shows a 98.5% correlation to Biolog identification. This multiphasic approach can be used to answer specific diagnostic questions pertaining to the source of infection and/or outbreak, or to address quarantine concerns.
Comparison of the Fibers of Fowl Adenovirus A Serotype 1 Isolates from Chickens with Gizzard Erosions in Europe and Apathogenic Reference Strains
Authors: Marek Ana , Schulz Evelyn , Hess Claudia , Hess Michael
Published: May 2011

A total of 18 samples from 4 outbreaks of gizzard erosions in broiler chickens in Europe were used in the current study. Fowl adenoviruses were found in samples from all 4 outbreaks, and isolates were identified as Fowl adenovirus A (FAdV-A) serotype 1. As described earlier, polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the long fiber gene was conducted. However, all 18 samples showed the same pattern as apathogenic FAdV-1 strains: Ote and chicken embryo lethal orphan (CELO) viruses. Nucleotide and amino acid sequences of the long and short fiber of several isolates from broiler chickens with gizzard erosions were analyzed, and 100% identity between the field isolates on the protein level was revealed. Only 1 nonsynonymous mutation (T→A) was present in the long fiber of studied isolates compared to the CELO strain. The same mutation was also present in the Ote strain. Four nonsynonymous mutations were present in the long fiber of studied isolates compared to Ote strain. In the short fiber, 6 nonsynonymous mutations were found in the studied isolates compared to the CELO strain. However, the short fiber of pathogenic isolates was 100% identical to apathogenic Ote strain. In conclusion, the usefulness of PCR-RFLP analysis of the long fiber gene of FAdV-1 isolates in distinguishing between those that induce gizzard erosions and those that do not remains questionable for the isolates obtained from European poultry flocks. The role of certain FAdV-1 strains with their long and short fiber in pathogenicity regarding gizzard erosions is still not clear.
Clinical Course of a Malignant Peripheral Nerve Sheath Tumor in a Siberian Tiger (Panthera Tigris Altaica)
Authors: Steinmetz Hanspeter W. , Rütten Maja , Ruess-Melzer Katja , Ohlerth Stefanie , Lischer Christoph , Oevermann Anna , Bode-Lesniewska Beata , Hatt Jean-Michel
Published: May 2011

A 14-year-old male Siberian tiger ( Panthera tigris altaica) was admitted with an ulcerating mass on the right thoracic wall. Radiographic and computed tomographic evaluation indicated 2 isolated cutaneous masses without any signs of metastasis. Histology of a Tru-Cut biopsy revealed an anaplastic sarcoma with giant cells. Both tumors were resected with appropriate normal tissue margins. The size of the defect did not allow primary closure of the wound; therefore, a mesh expansion technique was attempted. Three months later, the tiger had to be euthanized due to extensive metastasis to the lungs. Histomorphological features and immunohistochemical results confirmed the diagnosis of malignant peripheral nerve sheath tumor. In contrast to domestic animal experience, the tumor had spread extensively to the lungs without local reccurrence in a short period of time. Correct diagnosis requires various immunohistochemical evaluations of the tumor tissue.
Necrotizing Sialometaplasia of the Parotid Gland in a Dog
Authors: Kim Ha-Young , Woo Gye-Hyeong , Bae You-Chan , Park Yeon-Hee , Joo Yi-Seok
Published: May 2011

Necrotizing sialometaplasia (NS) is a self-limiting, benign, ischemic, inflammatory disease that is most often described in the submandibular glands of dogs, with clinical and histologic features that resemble malignancy. Unilateral swelling of the parotid salivary gland in a 7-year-old Cocker Spaniel dog was diagnosed as NS. The dog also had otitis externa on the same side as the parotid gland lesions. The main histologic features were included lobular necrosis of salivary tissue; fibrinoid necrosis of some arteries; marked squamous metaplasia of duct and/or acinar epithelium, with intercellular bridge formation; preservation of salivary lobular morphology; and variable inflammation and fibrosis. Etiologic factors for NS in both humans and animals remain obscure.
Chromophobe Renal Cell Carcinoma with Sarcomatoid Transformation in a Dog
Authors: Kobayashi Naohito , Suzuki Kazuhiko , Murakami Hironobu , Kagawa Eriko , Aoki Itirou , Nagashima Yoji
Published: May 2011

A 12-year-old spayed female Siberian husky dog presented with hematuria and weight loss. An abdominal ultrasonographic examination revealed a left renal tumor measuring 8 cm in diameter, and a nephrectomy was performed. The resected kidney contained a cavitated tumor with a white solid region. Histologically, this tumor was composed of large polygonal cells with abundant and cloudy cytoplasm and focal sarcomatoid change. The neoplastic epithelial cells were reactive with colloidal iron staining; Dolichos biflorus agglutinin, peanut agglutinin, and Ulex europaeus agglutinin I lectins; and cluster of differentiation 10 and c-KIT antigens but not for periodic acid-Schiff or vimentin stain. Neoplastic sarcomatoid cells stained positive for vimentin. Because these histopathologic features are identical to those of human chromophobe renal cell carcinoma, the present case was diagnosed as canine chromophobe renal cell carcinoma.
Spontaneous Pneumothorax in a Dog Secondary to Dirofilaria Immitis Infection
Authors: Oliveira Cintia , Rademacher Nathalie , David Andrew , Vasanjee Sunil , Gaschen Lorrie
Published: May 2011

A 5-year-old female spayed Labrador Retriever dog was referred to the Louisiana State University Veterinary Teaching Hospital for treatment of pneumothorax. Thoracic radiographs and computed tomography showed spontaneous pneumothorax, thoracic lymphadenopathy, pulmonary hypertension, and multifocal pulmonary bulla lesions. At surgery, numerous adult nematodes protruded from the parenchyma of the left caudal and accessory lung lobes and pulmonary arteries. On histopathology, multiple adult filarid nematodes were observed within the pulmonary blood vessels. Broad foci of necrosis of the pulmonary parenchyma were present. The tunica intima of the pulmonary arteries was markedly thickened by intimal fibrosis and medial hypertrophy. The final diagnosis was severe Dirofilaria immitis infection that resulted in pulmonary vascular lesions and focally extensive infarcts of the pulmonary parenchyma with bulla formation and rupture causing spontaneous pneumothorax. The dog received antibiotic and steroid therapy, as well as adulticide treatment, and recovered.
Bovine Subclinical Mastitis Caused by Mannheimia Granulomatis
Authors: Blum Shlomo , Freed Mor , Zukin Nona , Shwimmer Adin , Weissblit Limor , Khatib Nehaya , Bojesen Anders Miki , Larsen Jesper , Friedman Shmuel , Eladé Daniel
Published: May 2011

Mannheimia granulomatis was isolated for 10 months from the milk of a cow with elevated somatic cell counts. The infection was self-limiting. Phenotypic and molecular characteristics of the isolate were determined.
Spontaneous Odontoameloblastoma in a Female Sprague Dawley Rat
Authors: Burrough Eric R. , Myers Ronald K. , Whitley Elizabeth M.
Published: May 2011

The present report describes a spontaneously developing, highly infiltrative, and expansile neoplasm in the mandible of a 9-month-old female Sprague Dawley rat. The lesion consisted of abundant odontogenic epithelium with induction of osteodentin in the adjacent mesenchyme and was classified therefore as an odontoameloblastoma. Odontoameloblastomas are reported uncommonly in domestic animals, and the histologic and histochemical features of this neoplasm are reported herein along with a review of the veterinary literature.
Lycoperdonosis in Two Dogs
Authors: Alenghat Theresa , Pillitteri Cara A. , Bemis David A. , Kellett-Gregory Lindsay , Jackson Karen V. , Kania Stephen A. , Donnell Robert L. , Van Winkle Thomas
Published: May 2011

Lycoperdonosis is a rare respiratory disease that results from the inhalation of spores released from the Lycoperdon (puffball) mushroom. In the present study, 2 cases of confirmed canine lycoperdonosis are described. The first case presented to the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania, and the second case was submitted for postmortem examination to the University of Tennessee Veterinary Teaching Hospital. Both dogs presented in respiratory distress, and owners reported that the dogs had been playing or digging in areas with puffball mushrooms prior to the onset of clinical signs. In the initial case, thoracic radiographs revealed a diffuse interstitial and multifocal alveolar pulmonary pattern. Despite aggressive medical treatment and mechanical ventilation, the dog continued to worsen and was euthanized. Postmortem examination revealed firm lung lobes and enlarged tracheobronchial lymph nodes. Histologically, there was a severe diffuse histiocytic and pyogranulomatous bronchointerstitial pneumonia. Throughout the lung and lymph nodes, most commonly within macrophages, were round, 3–5μm in diameter, Gomori methenamine silver—positive structures, consistent with Lycoperdon spores. An approximately 750–base pair DNA fragment was amplified from lung of both cases by polymerase chain reaction using primers specific to yeast ribosomal DNA, and the sequence of the fragment was determined to be most closely related to Lycoperdon pyriforme. Importantly, reexamination of an endotracheal wash from the initial case revealed intrahistiocytic spores, suggesti
Endocardial Fibrosarcoma in a Reticulated Python (Python Reticularis)
Authors: Gumber Sanjeev , Nevarez Javier G. , Cho Doo-Youn
Published: May 2011

A female, reticulated python ( Python reticularis) of unknown age was presented with a history of lethargy, weakness, and distended coelom. Physical examination revealed severe dystocia and stomatitis. The reticulated python was euthanized due to a poor clinical prognosis. Postmortem examination revealed marked distention of the reproductive tract with 26 eggs (10–12 cm in diameter), pericardial effusion, and a slightly firm, pale tan mass (3–4 cm in diameter) adhered to the endocardium at the base of aorta. Based on histopathologic and transmission electron microscopic findings, the diagnosis of endocardial fibrosarcoma was made.
Selection of a Cutoff Value for Real-Time Polymerase Chain Reaction Results to Fit a Diagnostic Purpose: Analytical and Epidemiologic Approaches
Authors: Caraguel Charles G. B. , Stryhn Henrik , Gagné Nellie , Dohoo Ian R. , Hammell K. Larry
Published: May 2011

Diagnostic laboratories frequently select a subjective cutoff value for real-time amplification assays, above which a threshold cycle (Ct) value is deemed false. Commonly, higher Ct values are interpreted as amplification or fluorescence artifacts, or cross contaminations. Although the implementation of Ct cutoff might be reasonable, its justification and selection should be based on evidence. The current article reviewed evidence-based strategies to select Ct cutoffs grouped in analytical and epidemiologic approaches. Analytical strategies use criteria gathered during the assay development and include fluorescence threshold, reaction end-cycle, limit of detection, and artifact investigation. Variability in amplification efficacy across test runs may induce some instability in an intended Ct cutoff and requires some standardization or normalization procedures. Epidemiologic strategies use criteria based on either the probability or the cost of a false test result associated with a specified cutoff. Cutoffs, depending on the intended purpose of the test, can be selected graphically to minimize the probability of either false-positive or false-negative results by using two-graph receiver operating characteristics curves. The assay's diagnostic sensitivity and specificity may vary with the tested population, thus, the estimated two-graph receiver operating characteristics curve is population dependent and should be established for the targeted population. Although the selection of a cutoff based on misclassification cost depends on infection prevalence, the selection based on predictive values does not. To optimize the test average diagnostic performance, the Ct cutoff should be selected when diagnostic odds ratio is maximal. Epidemiologic approaches were illustrated by selecting Ct cutoffs for a real-time assay for Infectious salmon anemia virus.
Combination of Reverse Transcription Real-Time Polymerase Chain Reaction and Antigen Capture Enzyme-Linked Immunosorbent Assay for the Detection of Animals Persistently Infected with Bovine Viral Diarrhea Virus
Authors: Yan Lifang , Zhang Shuping , Pace Lanny , Wilson Floyd , Wan Henry , Zhang Michael
Published: May 2011

Bovine viral diarrhea virus (BVDV) is an economically important pathogen of cattle. A successful control program requires early detection and removal of persistently infected (PI) animals. The objective of the current study was to develop, validate, and apply a cost-effective testing scheme for the detection of BVDV PI animals in exposed herds. Pooled samples were screened by using a real-time reverse transcription polymerase chain reaction (real-time RT-PCR), and individual positives were identified with an antigen capture enzyme-linked immunosorbent assay (ACE). The detection limits of the optimized realtime RT-PCR were 10 and 100 RNA copies per reaction for BVDV-1 and BVDV-2, respectively. The semiquantitative results of real-time RT-PCR and ACE or real-time RT-PCR and immunohistochemistry were moderately correlated. The threshold cycle of real-time RT-PCR performed on pooled samples was significantly correlated with the pool size ( R2 = 0.993). The least-cost pool sizes were 50 at a prevalence of 0.25–0.5% and 25 at a prevalence of 0.75–2.0%. By using the combined real-time RT-PCR and ACE procedure, 111 of 27,932 samples (0.4%) tested positive for BVDV. At this prevalence, cost reduction associated with the application of real-time RT-PCR and ACE ranged from 61% to 94%, compared with testing individual samples by ACE, immunohistochemistry, or real-time RT-PCR. Real-time RT-PCR screening also indicated that 92.94% of PI animals were infected with BVDV-1, 3.53% with BVDV-2, and 3.53% with both BVDV-1 and BVDV-2. Analysis of the 5′-untranslated region of 22 isolates revealed the predominance of BVDV-1b followed by BVDV-2a.
An Efficient DNA Extraction Method for Polymerase Chain Reaction–Based Detection of Mycobacterium Avium Subspecies Paratuberculosis in Bovine Fecal Samples
Authors: Zhang Michael Z. , Zhang Shuping
Published: May 2011

Due to the lipid rich cell wall of Mycobacterium avium subspecies paratuberculosis (MAP), the complex nature of bovine feces, and intermittent organism shedding by infected cattle, it is difficult to recover a sufficient amount of high-quality MAP DNA from fecal samples, directly affecting the sensitivity of downstream polymerase chain reaction (PCR) tests. In the current study, a DNA extraction method, designated the Mississippi Veterinary Research and Diagnostic Laboratory (MVRDL) method, was developed for PCR-based detection of MAP in bovine fecal samples. The MVRDL method combined multiple procedures, including chemical pretreatment, 1-tube cell lysis and extraction, chelex matrix absorption, and mini-column purification. The DNA yield and purity, as measured by spectrophotometry, was 3.36 fg per colony forming unit (CFU) MAP and A260/280 absorbance ratio of 2, respectively. This method was further evaluated by real-time PCR. A linear correlation was found between cycle-threshold (Ct) and log input CFU (ranging from 7.2 to 7.2 × 107 CFU per ml or CFU per g). The detection limit of the real-time PCR assay was 3 CFU per ml of MAP culture or per g of MAP-spiked feces. In addition, the MVRDL method was validated by performing 7 Johne's direct fecal PCR proficiency tests administered by the National Veterinary Service Laboratories. Based on culture results as the “gold standard,” the specificity of MVRDL PCR was 100%, and the sensitivity was 98.46% for samples containing more than 1.5 CFU per tube of fecal cultures. To the authors' knowledge, this is the most efficient MAP DNA extraction method in comparison with all previously published protocols.