bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation of Cokeromyces Recurvatus, Initially Misidentified as Coccidioides Immitis, from Peritoneal Fluid in a Cat with Jejunal Perforation
Authors: Nielsen Cheri , Sutton Deanna A. , Matise Ilze , Kirchhof Nicole , Libal Melissa C.
Published: May 2011

Cokeromyces recurvatus, a zygomycete, was isolated by fungal culture from the peritoneal fluid of a cat with jejunal perforation secondary to intestinal lymphosarcoma. This organism has not been recovered previously from a veterinary patient. The tissue form of C. recurvatus is morphologically similar to those of Coccidioides immitis and Paracoccidioides brasiliensis and may be misdiagnosed as 1 of these organisms on the basis of cytologic or histopathologic specimens, particularly in geographic regions where these organisms are not endemic.
Pyogranuloma Caused by Mycobacterium Asiaticum Infection in a Steer
Authors: Oakey Jane , Forbes-Faulkner Judy , Gilpin Christopher , Murdoch Bronwyn , Johnson Steven
Published: May 2011

In 2003, a steer carcass was condemned at a Central Queensland abattoir because of metastatic tumors. In addition, a granulomatous lesion was found in the mediastinal lymph node. Histological examination showed this to be a pyogranuloma, typically associated with Rhodococcus or the Nocardia/ Streptomyces group. However, in this case, the only etiological agent was an acid-fast bacillus, which would normally be associated with a more fibrous lesion. A number of nucleic acid–based techniques were used, and the isolate was identified as Mycobacterium asiaticum. This organism is a rarely encountered opportunistic pathogen of humans, associated with subtropical climates. This is the first report of this organism causing infection in cattle. The similarities between this case and cases of human disease are discussed.
Natural Infection of a Horse with Fascioloides Magna
Authors: McClanahan S. L. , Stromberg B. E. , Hayden D. W. , Averbeck G. A. , Wilson J. H.
Published: May 2011

A 25-year-old Quarterhorse mare was euthanized for a variety of medical reasons. At necropsy, 7 liver flukes, identified as Fascioloides magna, were recovered from the liver. This is the first report of F. magna in a horse.
A Study of Mutations in the c-kit Gene of 32 Dogs with Mastocytoma
Authors: Riva Federica , Brizzola Stefano , Stefanello Damiano , Crema Simone , Turin Lauretta
Published: May 2011

Mutations in the intracellular juxtamembrane domain of the c-kit gene in 32 dogs with different grades of histologically confirmed mastocytoma were studied. Transcript RNAs extracted from neoplastic tissue surgically collected from dogs of different breeds and from a negative control were reverse transcribed into complementary DNA and amplified by polymerase chain reaction. The region corresponding to the c-kit juxtamembrane domain was sequenced and compared with GenBank sequences. Two different types of mutations were identified within exon 11: a previously underscribed single-nucleotide substitution and a 6-bp deletion. The c-kit juxtamembrane domain sequences of all dogs were grouped in 3 clusters. No mutations were detected in tissues constitutively expressing c-kit (cerebellum and spleen), obtained from dogs not affected by mastocytoma (controls). All the substitutions were found in dogs bearing grade I or II mast cell tumors; the deletion was detected in 1 dog with grade II mastocytoma.
Endometritis and Cystic Endometrial Hyperplasia in a Goat
Authors: Radi Zaher A.
Published: May 2011

Histologic examination was performed on uterine biopsy samples of irregular cystic masses noted during caesarean section of a 2-year-old female Boer goat. Histopathologic examination revealed multifocal erosions of the superficial epithelium and multifocal infiltration of the endometrium by widely scattered viable and degenerate neutrophils, lymphocytes, and plasma cells admixed with mild amounts of cellular debris and hemorrhage. The endometrium was markedly expanded by many irregular cystic and hyperplastic glands. This is the first case report of endometritis and cystic endometrial hyperplasia in a goat in North America.
Canine Cutaneous Clear Cell Adnexal Carcinoma: Histopathology, Immunohistochemistry, and Biologic Behavior of 26 Cases
Authors: Schulman F. Y. , Lipscomb T. P. , Atkin T. J.
Published: May 2011

Thirty tumors including 27 distinctive cutaneous neoplasms and 3 metastatic tumors from 26 dogs were collected from diagnostic submissions to 3 laboratories. Characteristic histopathologic features included location in the subcutis or dermis (or both); lobular, nodular, and nest-like architecture; and a component of epithelioid cells with clear cytoplasm. Additional features present in most cases included follicular dermal papilla-like structures, low mitotic index, nuclear pleomorphism, necrosis, and mineralization. Cytoplasmic periodic acid Schiff—positivity, which was abolished by pretreatment with diastase, indicated the presence of glycogen in all cases. The oil red O stain did not demonstrate cytoplasmic lipid. Melanin granules, accentuated by the Fontana-Masson method, were observed infrequently. A sparsely cellular mucinous stroma and stromal cartilaginous differentiation were uncommon. By immunohistochemistry, neoplastic cells stained positively for cytokeratin (29 of 29), vimentin (28 of 28), S-100 protein (24 of 29), and melan A (8 of 12); results were negative for smooth muscle actin and calponin in all cases. Clinical follow-up information was obtained on all 26 dogs. One tumor recurred, 1 metastasized to a regional lymph node, and 1 metastasized to regional lymph nodes twice. In another case, possible pulmonary metastasis was noted radiographically. The findings are consistent with a poorly differentiated, low-grade, adnexal carcinoma of the skin. Similar canine cutaneous neoplasms have been reported as “clear-cell hidradenocarcinoma” and “follicular stem cell carcinoma.” The authors propose the designation “cutaneous clear cell adnexal carcinoma.”
Genetic Analysis of Feline Caliciviruses Associated with a Hemorrhagic-Like Disease
Authors: Abd-Eldaim Mohamed , Potgieter Leon , Kennedy Melissa
Published: May 2011

Feline calicivirus (FCV) is 1 of the most common causes of upper respiratory tract disease in cats. Other disease syndromes associated with FCV infection have been reported. Recently, calicivirus infection associated with a hemorrhagic-like disease leading to significant mortality in cats has been reported. The clinical signs are similar to those observed with the calicivirus of rabbit hemorrhagic disease. This study characterized 2 FCV isolates associated with hemorrhagic-like disease. Nucleotide sequencing of the complete genome has been done for these 2 isolates as well as for 4 additional isolates representing other disease syndromes. Previously reported sequence data for the entire genome of classical FCV (6 isolates) and a portion of the capsid gene for hemorrhagic-like FCV (3 isolates), isolated in different regions of United States were used in the genetic analysis. Sequence data were used to determine relationships among the isolates and any correlation with phenotype. Nucleotide sequence comparisons of the entire genome and individual open reading frames revealed high homology among all isolates. Data suggest that the virulence may have genetic determinants on the basis of phylogenetic clustering of the isolates associated with hemorrhagic-like disease.
In Vitro Antimicrobial Inhibition Profiles of Mycoplasma Bovis Isolates Recovered from Various Regions of the United States from 2002 to 2003
Authors: Rosenbusch Ricardo F. , Kinyon Joann M. , Apley Michael , Funk Nathan D. , Smith Sean , Hoffman Lorraine J.
Published: May 2011

Antimicrobial therapy continues to be important in reducing losses due to pneumonic forms of Mycoplasma bovis disease in beef and dairy calves. Although M. bovis diseases have been documented as frequent and economically important in the United States, there are no published reports on the antimicrobial activity of approved compounds against US strains. In this study, the authors report on the activity of 9 different antimicrobials against 223 recently recovered isolates of M. bovis. These isolates represent accessions from 5 geographic regions of the United States and were grouped by 4 tissues of origin (milk, respiratory, joint, or ear and eye). A broth microdilution test was used to determine minimum inhibitory concentration (MIC) values by reading redox changes detected in broth with alamarBlue (resazurin) indicator. For each antimicrobial, the median, MIC50, MIC90, mode, and range were calculated, and the values used for comparisons. In the absence of accepted breakpoint values, published MIC cutoff values for animal mycoplasmas as well as Clinical Laboratory Standards Institute interpretive criteria were used as a reference to define in vitro activity. The MIC values from active antimicrobials were found to distribute independently of region of origin of the isolates or of tissue of origin. Enrofloxacin, florfenicol, and spectinomycin were found to be active compounds in vitro. Oxytetracycline and chlortetracycline were active against more than half of the isolates. Very few isolates were inhibited by tilmicosin and none by erythromycin, ampicillin, or ceftiofur. The antimicrobial profiles determined for these US strains were remarkably similar to those reported for European isolates. However, unlike in Europe, there appears to be no diversity of profiles when US isolates are grouped by region or tissue of origin.
Persistence of Neospora Caninum–Specific Immunoglobulin G Antibodies in Bovine Blood and Lung Tissue Stored at Room Temperature
Authors: Jakubek Eva-Britt , Uggla Arvid
Published: May 2011

Blood and tissue samples obtained at necropsy from carcasses showing varying degrees of postmortem decomposition are sometimes used as sources of antibodies for serological tests, primarily in studies involving wild animals or in examination of aborted fetuses. To test the persistence of antibodies in blood and lung tissue kept at room temperature (approximately 20°C) for up to 48 and 118 days, respectively, serum and lung fluid from Neospora caninum–infected and uninfected cattle were tested using an indirect N caninum immunostimulating complex enzyme-linked immunosorbent assay (ELISA). Testing of selected samples was also performed using a commercial Neospora competitive ELISA. Despite severe changes in the gross appearance of the samples, the blood collected through venipuncture from live animals remained essentially sterile, whereas the pieces of lung tissue showed a variety of bacterial and fungal growth. Nevertheless, there was no obvious reduction in antibody levels in either the blood or the tissue samples during the 2 observation periods. This study indicates that specific immunoglobulin G antibodies may be remarkably stable in sterile blood and decomposed lung tissue.
Comparison of Immunohistochemistry and Two Rapid Tests for Detection of Abnormal Prion Protein in Different Brain Regions of Sheep with Typical Scrapie
Authors: Bolea Rosa , Monleón Eva , Schiller Irene , Raeber Alexander J. , Acín Cristina , Monzón Marta , Martín-Burriel Inmaculada , Struckmeyer Thomas , Oesch Bruno , Badiola Juan José
Published: May 2011

One of the “gold standard” techniques for postmortem confirmation of scrapie diagnosis in sheep and goats is immunohistochemical examination of brain tissue. Active surveillance for scrapie is mainly performed by rapid diagnostic tests on the basis of postmortem immunochemical detection of prion protein (PrP) in the obex tissue. The aim of this study was to determine the performance of 2 rapid tests, Prionics®-Check LIA (a chemiluminescence sandwich enzyme-linked immunosorbent assay) and Prionics®-Check Western blot for scrapie diagnosis when applied to brain areas other than the obex, in comparison with the recognized immunohistochemistry. Prion protein was detected in the obex, cervical spinal cord, and thalamus from all the scrapie-positive sheep by the 3 tests. Western blot and LIA were negative in other areas of the brain, although weak immunohistochemical staining was detected. The results show that the 2 rapid tests studied may detect PrP in brain areas other than the obex, although with a lower sensitivity than immunohistochemistry when there is minimal PrP deposition.
Serotyping of US Isolates of Chlamydophila Psittaci from Domestic and Wild Birds
Authors: Andersen Arthur A.
Published: May 2011

The identities of chlamydial strains, which can infect a given host, are important to know for disease prognosis, disease control, and epidemiology. The microimmunofluorescence test (MIFT) was used with a panel of 14 serovar-specific monoclonal antibodies (MAbs) to serotype 150 chlamydial isolates from domestic and wild birds. The isolates were obtained from birds submitted to diagnostic laboratories or during investigation of outbreaks. The 150 US isolates included 96 from the order Psittaciformes, 14 isolates from the order Columbiformes, 2 from the order Passeriformes, 16 from the order Galliformes, 12 from the order Struthioniformes, and 3 from the order Falconiformes. A total of 93, or 97%, of the Psittaciformes isolates were of serovar A; 11, or 79%, of the Columbiformes isolates were of serovar B; 64% of the Galliformes isolates were of serovar D, and all the Struthioniformes isolates were of serovar E. The 3 Falconiformes isolates did not react with any of the MAbs to the avian and mammalian isolates and are presumed to represent a new strain. The results show that specific chlamydial strains are usually associated with certain types of birds and that some serovars may be unusually virulent for certain species of birds. The MIFT using serovar-specific MAbs provides a rapid method to serotype new isolates, making it a useful system for epidemiological studies.
Diagnosis of Transmissible Spongiform Encephalopathies in Animals: A Review
Authors: Gavier-Widén Dolores , Stack Michael J. , Baron Thierry , Balachandran Aru , Simmons Marion
Published: May 2011

Transmissible spongiform encephalopathies (TSEs) in animals include, among others, bovine spongiform encephalopathy (BSE), scrapie, chronic wasting disease, and atypical forms of prion diseases. Diagnosis of TSEs is based on identification of characteristic lesions or on detection of the abnormal prion proteins in tissues, often by use of their partial proteinase K resistance property. Correctly sampling of target tissues is of utmost importance as this has a considerable effect on test sensitivity. Most of the rapid or screening tests are based on ELISA or Western immunoblot (WB) analysis, and many are officially approved. Confirmatory testing is normally performed by use of histologic examination, immunohistochemical analysis, certain WB protocols, or detection of prion fibrils by use of electron microscopy (scrapie-associated fibril). The discriminatory methods for diagnostic use are mostly based on WB technology and provide initial identification of the prion strain, particularly for differentiation of BSE from scrapie in small ruminants. Definitive prion strain characterization is performed by use of bioassays, usually in mice. A burgeoning number of transgenic mice have been developed for TSE studies. Development of new tests with higher sensitivity and of more reliable diagnostic applications for live animals tested for food safety reasons is a rapidly developing field. Ultimately, the choice of a test for TSE diagnosis depends on the rationale for the testing.
Clostridial Enteric Infections in Pigs
Authors: Songer J. Glenn , Uzal Francisco A.
Published: May 2011

Clostridium perfringens types A and C and Clostridium difficile are the principal enteric clostridial pathogens of swine. History, clinical signs of disease, and gross and microscopic findings form the basis for a presumptive diagnosis of C. perfringens type-C enteritis. Confirmation is based on isolation of large numbers of type-C C. perfringens and/or detection of beta toxin in intestinal contents. Diagnosis of C. perfringens type-A infection, however, remains controversial, mostly because the condition has not been well defined and because type-A organisms and their most important major (alpha) toxin can be found in intestinal contents of healthy and diseased pigs. Isolation of large numbers of C. perfringens type A from intestinal contents, in the absence of other enteric pathogens, is the most reliable criterion on which to base a diagnosis. Recently, beta2 (CPB2) toxin-producing C. perfringens type A has been linked to disease in piglets and other animals. However, implication of CPB2 in pathogenesis of porcine infections is based principally on isolation of C. perfringens carrying cpb2, the gene encoding CPB2, and the specific role of CPB2 in enteric disease of pigs remains to be fully defined. Clostridium difficile can also be a normal inhabitant of the intestine of healthy pigs, and diagnosis of enteric infection with this microorganism is based on detection of its toxins in feces or intestinal contents.
Development of a Real-Time PCR for Detection of Mycoplasma Bovis in Bovine Milk and Lung Samples
Authors: Cai Hugh Y. , Bell-Rogers Patricia , Parker Lois , Prescott John F.
Published: May 2011

A real-time polymerase chain reaction (PCR) assay using hybridization probes on a LightCycler platform was developed for detection of Mycoplasma bovis from individual bovine mastitis milk and pneumonic lung tissues. The detection limit was 550 colony forming units (cfu)/ml of milk and 650 cfu/25 mg of lung tissue. A panel of bovine Mycoplasma and of other bovine-origin bacteria were tested; only M. bovis strains were positive, with a melting peak of 66.6°C. Mycoplasma agalactiae PG2 was also positive and could be distinguished because it had a melting peak of 63.1°C. In validation testing of clinical samples, the relative sensitivity and specificity were 100% and 99.3% for individual milks and 96.6% and 100% for the lung tissue. Using M. bovis real-time PCR, the M. bovis culture-positive milk samples were estimated to contain between 5 × 104 and 7.7 × 108 cfu/ml and the M. bovis culture-positive lungs between 1 × 103 and 1 × 109 cfu/25 mg. Isolation, confirmed with the real-time PCR and colony fluorescent antibody test, showed that at the herd level, the proportion of samples positive for M. bovis isolation in mastitis milk samples submitted to the Mastitis Laboratory, Animal Health Laboratory, University of Guelph, Ontario, Canada, was 2.4% (5/201). We conclude that this probe-based real-time PCR assay is a sensitive, specific, and rapid method to identify M. bovis infection in bovine milk and pneumonic lungs.
Development of a SYBR Green Real-Time Polymerase Chain Reaction Assay for Quantitative Detection of Babesia Gibsoni (Asian Genotype) DNA
Authors: Matsuu Aya , Ono Satomi , Ikadai Hiromi , Uchide Tsuyoshi , Imamura Saiki , Onuma Misao , Okano Shozo , Higuchi Seiichi
Published: May 2011

A real-time fluorogenic polymerase chain reaction (PCR) assay based on SYBR green that allows for sensitive, reproducible, and accurate quantification of Babesia gibsoni (Asian genotype). DNA from peripheral blood of infected dogs was developed. Standard curves were created by plotting the input amount of a standard template, constructed with plasmid DNA containing 182 base pairs (bp) of the p18 gene, against threshold cycle numbers. The curves showed a wide dynamic range (1,000,000-fold input) and high correlation values (>0.99). The PCR amplification efficacy of the standard template was similar to that of intact genomic DNA obtained from peripheral blood with B. gibsoni infection. The detection limit of the assay was 9 parasites/μl of blood with B gibsoni infection. The intra-assay and interassay coefficients of variation of the threshold cycles ranged from 0.70% to 1.89% and from 1.18% to 1.92%, respectively. This assay system was found to be reproducible and accurate for the quantification of parasite DNA in experimentally infected dogs and far more sensitive than traditional microscopic examination.
Comparison of Six RNA Extraction Methods for the Detection of Classical Swine Fever Virus by Real-Time and Conventional Reverse Transcription–PCR
Authors: Deng Ming Y. , Wang He , Ward Gordon B. , Beckham Tammy R. , McKenna Thomas S.
Published: May 2011

Six RNA extraction methods, i.e., RNAqueous kit, Micro-to-midi total RNA purification system, NucleoSpin RNA II, GenElute mammalian total RNA kit, RNeasy mini kit, and TRIzol LS reagent, were evaluated on blood and 7 tissues from pig infected with classical swine fever virus (CSFV). Each of the 6 extraction methods yielded sufficient RNA for positive results in a real-time reverse transcription–PCR (RT-PCR) for CSFV, and all RNA, except the one extracted from blood by TRIzol LS reagent, yielded positive results in both a conventional RT-PCR for CSFV and a conventional RT-PCR for an endogenous gene encoding β-actin. The RNA extracted from blood by TRIzol LS reagent became positive in both conventional RT-PCR assays when it was diluted to 1:2, 1:4, or up to 1:64 in nuclease-free water. It is concluded that all 6 methods are more or less useful for the detection of CSFV by real-time and conventional RT-PCR in swine blood and tissues. However, some of the 6 reagents offer certain advantages not common to all 6 extraction procedures. For example, RNA extracted by the TRIzol LS reagent constantly had the highest yield; that by the RNAqueous kit had the highest A260/A280 ratio for almost all samples; and that by the NucleoSpin RNA II and the GenElute mammalian total RNA kit was most likely to be free of contaminations with genomic DNA.
Equine Herpesvirus-4 Kinetics in Peripheral Blood Leukocytes and Nasopharyngeal Secretions in Foals Using Quantitative Real-Time TaqMan PCR
Authors: Pusterla Nicola , Leutenegger Christian M. , Wilson W. David , Watson Johanna L. , Ferraro Gregory L. , Madigan John E.
Published: May 2011

Based on the hypothesis that the viral load of cells infected with EHV-4 will likely change during the course of disease, TaqMan PCR was used to investigate and characterize the kinetics of EHV-4 viral DNA load (glycoprotein B gene) and transcriptional activity (glycoprotein B and latency-associated transcripts) in peripheral blood leukocytes (PBLs) and nasopharyngeal secretions (NSs) collected from 11 foals during a field outbreak of respiratory disease. The EHV-4 DNA load in PBLs was low and of short duration after onset of clinical signs. In contrast, the EHV-4 load in NSs remained high for the majority of the foals over a period of 4 weeks. Viral replication determined by detection of mRNA expression of the structural glycoprotein B was detected only in NSs during the first 7 days after onset of clinical signs for most foals. The majority of foals expressed latency-associated transcripts in NS sonly during the first 7 days after onset of clinical signs. Persistence of the expression of latency-associated transcripts in NS, as a reflection of a latent viral state, was not documented during the 28-day study period. Based on these results, it was concluded that lytic infection with EHV-4 can be diagnosed either by high EHV-4 DNA load of glycoprotein B gene or by detection of transcriptional activity of glycoprotein B.
Bovine Viral Diarrhea Virus in Alpaca: Abortion and Persistent Infection
Authors: Carman Susy , Carr Nancy , DeLay Josepha , Baxi Mohit , Deregt Dirk , Hazlett Murray
Published: May 2011

An alpaca herd in eastern Ontario experienced vague signs of illness, including anorexia and lethargy in 9 animals, 2.5 months after the addition of a chronically ill cria and his dam to the farm. Subsequently 2 alpaca had early pregnancy loss; one aborted at 5.5 months gestation and the other at 7 months gestation. Seventeen were found to have serum antibody to bovine viral diarrhea virus (BVDV), with highest titers to BVDV type 1. The fetus that was aborted at 5.5 months gestation, 3 months after the clinical outbreak, was found to be positive for BVDV on immunohistochemical staining, and noncytopathic BVDV type 1b was isolated. Of the 13 cria born alive that season, a single male underweight alpaca cria, born 9 months after the clinical illnesses, was infected with BVDV type 1b. The cria was positive for BVDV at birth, at 3 and 26 days of age and continued to be positive for noncytopathic BVDV using virus isolation, nested reverse transcription PCR, antigen detection ELISA, and immunohistochemical staining until euthanasia at 46 days of age. The cria remained serum antibody negative to both BVDV type 1 and type 2. A diagnosis of persistent infection was made. This is the first report describing persistent infection with BVDV in an alpaca cria.
Persistent Viral Shedding during Asymptomatic Aleutian Mink Disease Parvoviral Infection in a Ferret
Authors: Pennick Kate E. , Stevenson Mary Ann M. , Latimer Kenneth S. , Ritchie Branson W. , Gregory Christopher R.
Published: May 2011

A 2-year-old domestic ferret that appeared clinically healthy was repeatedly seropositive for Aleutian mink disease parvovirus (ADV) over a 2-year observation period. Antibody titers, determined by counter-immunoelectrophoresis, ranged from 1024 to 4096. Viral DNA also was identified in serum, urine, feces, and blood cell fractions by polymerase chain reaction analysis. Ultimately, DNA in situ hybridization revealed ADV DNA in histologic sections of various tissues and organs. These data indicate that this asymptomatic ferret was persistently infected with ADV.
Systemic Cryptococcus Albidus Infection in a Doberman Pinscher
Authors: Labrecque Olivia , Sylvestre Doris , Messier Serge
Published: May 2011

Cryptococcus albidus is a saprophytic, encapsulated yeast usually found in air, both outdoor and indoor, and sometimes on human skin. It is not usually considered to be a primary pathogen. Most cryptococcal infections of humans and animals are caused by Cryptococcus neoformans. Several cases of C. albidus infection have been reported in humans over the past 20 years. In the veterinary literature, 2 equine cases have been described: genital infection and mycotic keratitis. The present report is the first documented case of C. albidus systemic infection in a dog. Veterinarians and diagnosticians should be aware that C. albidus may be a potential canine pathogen.
Eastern Equine Encephalitis in Dogs
Authors: Farrar Michele D. , Miller Debra L. , Baldwin Charles A. , Stiver Shane L. , Hall Clint L.
Published: May 2011

Eastern equine encephalitis virus (EEEV) is an Alphavirus that is endemic in the Southeastern United States. From 1993 to January 2005, the Veterinary Diagnostic and Investigational Laboratory in Tifton, Georgia, performed postmortem examinations on over 101 domestic canines exhibiting clinical neurological disturbances. In 12 of these dogs, brains were histologically suggestive of infection with EEEV. All dogs were less than 6 months of age, with no breed predilection. Clinical signs included pyrexia, depression, nystagmus, and lateral recumbency. Microscopically, brains from all 12 puppies contained infiltrates of lymphocytes, plasma cells, and histiocytes, with occasional neutrophils and random foci of astrocytosis and gliosis. There were mild to moderate perivascular infiltrates of neutrophils along with scattered lymphocytes, plasma cells, and macrophages in the meninges. Viruses isolated from brain homogenates of all 12 puppies were confirmed by indirect fluorescent antibody testing to be EEEV. Additionally, RNA extracted from the brains and viral cultures of 2 dogs were determined by a specific reverse-transcriptase polymerase chain reaction (RT-PCR) to contain EEEV. The single available serum sample exhibited a 1:8 serum neutralization titer to EEEV.
Fatal Necrotizing Fasciitis and Myositis in a Captive Common Bottlenose Dolphin (Tursiops Truncatus) Associated with Streptococcus Agalactiae
Authors: Zappulli Valentina , Mazzariol Sandro , Cavicchioli Laura , Petterino Claudio , Bargelloni Luca , Castagnaro Massimo
Published: May 2011

A common bottlenose dolphin ( Tursiops truncatus) was presented for necropsy after acute onset of gastrointestinal signs and cutaneous lesions that rapidly progressed to death. Gross and microscopic findings were characterized by locally extensive severe necrohemorrhagic fasciitis and cellulitis, and severe necrotizing myositis in the head and dorsocranial thorax, with numerous disseminated gram-positive cocci. Streptococcus agalactiae was isolated from the lesions and from visceral organs (liver and lung), and it was identified by standard microbiology techniques. This communication is the first report of necrotizing fasciitis in a marine mammal associated with S. agalactiae.
Longitudinal Study of ELISA Seroreactivity to Mycobacterium Avium subsp. Paratuberculosis in Infected Cattle and Culture-Negative Herd Mates
Authors: Sweeney Raymond W. , Whitlock Robert H. , McAdams Susan , Fyock Terry
Published: May 2011

Two thousand nine hundred fifty-two serum samples, collected once or twice annually from 545 cows of known fecal culture status were tested for antibodies to Mycobacterium avium subsp. paratuberculosis using a commercially available enzyme-linked immunosorbent assay (ELISA) test. Overall, 13.5% of the samples from 282 infected cows had positive ELISA results, but when tested multiple times, 38.3% of the cows had at least 1 serum sample with positive results. Among 263 fecal culture–negative cows, 98.1% of the serum samples had negative ELISA results, but when tested multiple times, 7.8% of the cows had at least 1 positive ELISA sample. Fecal culture was positive on a test before the first positive ELISA in 50 cows, ELISA was positive before fecal culture in 12 cows, and in 38 cows, both tests became positive at the same testing time. An additional 174 cows were positive on fecal culture and always negative on ELISA until culled. For cows that had ELISA sample:positive (S/P) ratios below the cutoff point, the change in S/P between sequential tests was evaluated to determine whether a rise in S/P could predict infection status. In this study, change in S/P was not a useful predictor of infection status in seronegative cows.
High Throughput Detection of Bluetongue Virus by a New Real-Time Fluorogenic Reverse Transcription—Polymerase Chain Reaction: Application on Clinical Samples from Current Mediterranean Outbreaks
Authors: Jiménez-Clavero Miguel Angel , Agüero Montserrat , Miguel Elena San , Mayoral Tomás , López Maria Cruz , Ruano María José , Romero Esther , Monaco Federica , Polci Andrea , Savini Giovanni , Gómez-Tejedor Concepción
Published: May 2011

A real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for the detection of bluetongue virus (BTV) in blood samples. A combination of primers specific for a highly conserved region in RNA segment 5 (based on Mediterranean BTV sequences) and a DNA probe bound to 5′-Taq nuclease-3′ minor groove binder (TaqMan© MGB) was used to detect a range of isolates. This real-time RT-PCR assay could detect 5.4 × 10−3 tissue culture infectious doses (TCID50) of virus per milliliter of sample, which was comparable to our current BTV diagnostic nested RT-PCR assay. The assay detected all recent Mediterranean isolates (including serotypes 2, 4, and 16), BTV vaccine strains for serotypes 2 and 4, and 15 out of the 24 BTV reference strains available (all serotypes), but did not detect the related orbiviruses epizootic hemorrhagic disease and African horse sickness viruses. Following assay evaluation, the ability of this assay to identify BTV in recent isolates (2003, 2004) from ovine and bovine samples from an epizootic outbreak in Spain was also tested. Minor nucleotide changes (detected by sequencing viral genomes) within the probe-binding region were found to have a profound effect on virus detection. This assay has the benefits of being fast and simple, and the 96-well format enables large-scale epidemiological screening for BTV, especially when combined with a high-throughput nucleic acid extraction method.
Diseases and Pathogens Associated with Mortality in Ontario Beef Feedlots
Authors: Gagea Mihai I. , Bateman Kenneth G. , van Dreumel Tony , McEwen Beverly J. , Carman Susy , Archambault Marie , Shanahan Rachel A. , Caswell Jeff L.
Published: May 2011

This study determined the prevalence of diseases and pathogens associated with mortality or severe morbidity in 72 Ontario beef feedlots in calves that died or were euthanized within 60 days after arrival. Routine pathologic and microbiologic investigations, as well as immunohistochemical staining for detection of bovine viral diarrhea virus (BVDV) antigen, were performed on 99 calves that died or were euthanized within 60 days after arrival. Major disease conditions identified included fibrinosuppurative bronchopneumonia (49%), caseonecrotic bronchopneumonia or arthritis (or both) caused by Mycoplasma bovis (36%), viral respiratory disease (19%), BVDV-related diseases (21%), Histophilus somni myocarditis (8%), ruminal bloat (2%), and miscellaneous diseases (8%). Viral infections identified were BVDV (35%), bovine respiratory syncytial virus (9%), bovine herpesvirus-1 (6%), parainfluenza-3 virus (3%), and bovine coronavirus (2%). Bacteria isolated from the lungs included M. bovis (82%), Mycoplasma arginini (72%), Ureaplasma diversum (25%), Mannheimia haemolytica (27%), Pasteurella multocida (19%), H. somni (14%), and Arcanobacterium pyogenes (19%). Pneumonia was the most frequent cause of mortality of beef calves during the first 2 months after arrival in feedlots, representing 69% of total deaths. The prevalence of caseonecrotic bronchopneumonia caused by M. bovis was similar to that of fibrinosuppurative bronchopneumonia, and together, these diseases were the most common causes of pneumonia and death. M. bovis pneumonia and polyarthritis has emerged as an important cause of mortality in Ontario beef feedlots.
Naturally Occurring Mycoplasma Bovis—Associated Pneumonia and Polyarthritis in Feedlot Beef Calves
Authors: Gagea Mihai I. , Bateman Kenneth G. , Shanahan Rachel A. , van Dreumel Tony , McEwen Beverly J. , Carman Susy , Archambault Marie , Caswell Jeff L.
Published: May 2011

Mycoplasma bovis is perceived as an emerging cause of mortality in feedlot beef cattle. This study examined the lesions and infectious agents in naturally occurring M. bovis–associated bronchopneumonia and arthritis and the relationship of this condition with bovine viral diarrhea virus (BVDV) infection. Standardized pathologic, immunohistochemical, and microbiologic investigations were conducted on 99 calves that died or were euthanized within 60 days after arrival in 72 feedlots. Cranioventral bronchopneumonia with multiple foci of caseous necrosis was identified in 54 of 99 calves, including 30 with concurrent fibrinosuppurative bronchopneumonia typical of pneumonic pasteurellosis. Mycoplasma bovis was consistently identified in these lesions by culture and immunohistochemistry, but also commonly in healthy lungs and those with pneumonia of other causes. Focal lesions of coagulation necrosis, typical of pneumonic pasteurellosis, were often infected with both Mannheimia haemolytica and M. bovis. Arthritis was present in 25 of 54 (46%) calves with M. bovis pneumonia, and all calves with arthritis had pneumonia. BVDV infection was more common in calves with lesions of bacterial pneumonia than in those dying of other causes, but BVDV infection was not more common in calves with caseonecrotic bronchopneumonia than those with fibrinosuppurative bronchopneumonia. Retrospective analysis identified cases of M. bovis pneumonia in the early 1980s that had milder lesions than the current cases. The findings suggest that, in at least some calves, M. bovis induces caseonecrotic bronchopneumonia within the lesions of pneumonic pasteurellosis.
Experimentally Induced Infection of Reindeer (Rangifer Tarandus) with Mycobacterium Bovis
Authors: Palmer Mitchell V. , Waters W. Ray , Thacker Tyler C. , Stoffregen William C. , Thomsen Bruce V.
Published: May 2011

In the USA, all species of Cervidae are included in the USDA's uniform methods and rules for the eradication of bovine tuberculosis and, therefore, are subject to regulations regarding intradermal tuberculin testing. In reindeer ( Rangifer tarandus), infection with Mycobacterium bovis is exceedingly rare and the response of reindeer to infection with M. bovis in pathologic and immunologic terms is unknown. The objectives of the study reported here were to describe the pathologic changes associated with M. bovis infection in reindeer and evaluate the effectiveness of intradermal tuberculin testing as a means of diagnosis of tuberculosis in reindeer. Thirteen reindeer were inoculated intratonsilarly with 105 colony-forming units (CFU) of M. bovis, and 4 noninoculated reindeer served as negative controls. The comparative cervical test (CCT) was done on all reindeer 90 and 240 days after inoculation. Thirteen months after inoculation, all reindeer were euthanized and examined. All experimentally inoculated reindeer developed lesions in the medial retropharyngeal lymph nodes. The CCT accurately identified all M. bovis-inoculated reindeer, but false-positive results were common among negative-control reindeer. Modifications of the method for interpretation of the CCT decreased false-positive results without increasing false-negative results. Reindeer are susceptible to infection with M. bovis; however, lesions are fewer in number, less severe in nature, and less widely disseminated than are those seen in white-tailed deer ( Odocoileus virginianus). Comparative cervical skin testing of reindeer can be highly sensitive, but has low specificity. Specificity can be improved by modification of criteria for interpretation of the CCT.
Comparative Toxicosis of Sodium Selenite and Selenomethionine in Lambs
Authors: Tiwary Asheesh K. , Stegelmeier Bryan L. , Panter Kip E. , James Lynn F. , Hall Jeffery O.
Published: May 2011

Excess consumption of selenium (Se) accumulator plants can result in selenium intoxication. The objective of the study reported here was to compare the acute toxicosis caused by organic selenium (selenomethionine) found in plants with that caused by the supplemental, inorganic form of selenium (sodium selenite). Lambs were orally administered a single dose of selenium as either sodium selenite or selenomethionine and were monitored for 7 days, after which they were euthanized and necropsied. Twelve randomly assigned treatment groups consisted of animals given 0, 1, 2, 3, or 4 mg of Se/kg of body weight as sodium selenite, or 0, 1, 2, 3, 4, 6, or 8 mg of Se/kg as selenomethionine. Sodium selenite at dosages of 2, 3, and 4 mg/kg, as well as selenomethionine at dosages of 4, 6, and 8 mg/kg resulted in tachypnea and/or respiratory distress following minimal exercise. Severity and time to recovery varied, and were dose dependent. Major histopathologic findings in animals of the high-dose groups included multifocal myocardial necrosis and pulmonary alveolar vasculitis with pulmonary edema and hemorrhage. Analysis of liver, kidney cortex, heart, blood, and serum revealed linear, dose-dependent increases in selenium concentration. However, tissue selenium concentration in selenomethionine-treated lambs were significantly greater than that in lambs treated with equivalent doses of sodium selenite. To estimate the oxidative effects of these selenium compounds in vivo, liver vitamin E concentration also was measured. Sodium selenite, but not selenomethionine administration resulted in decreased liver vitamin E concentration. Results of this study indicate that the chemical form of the ingested Se must be known to adequately interpret tissue, blood, and serum Se concentrations.
Reverse Transcription-Polymerase Chain Reaction on Pooled Samples to Detect Bovine Viral Diarrhea Virus by using Fresh Ear-Notch—Sample Supernatants
Authors: Kennedy James A. , Mortimer Robert G. , Powers Barbara
Published: May 2011

Ear-notch samples from 3,599 yearling heifers were collected to detect persistently infected (PI) animals with suspect bovine viral diarrhea virus (BVDV). Individual immunohistochemistry (IHC), individual antigen-capture enzyme-linked immmunosorbent assay (AC-ELISA), and reverse transcription-polymerase chain reaction (RT-PCR) tests with pooled ear-notch supernatants were compared with samples from 3,016 heifers, whereas RT-PCR ear-notch pools and individual AC-ELISA tests were compared with samples from all 3,599 heifers. Four heifers were identified positive by both IHC and AC-ELISA, whereas the remaining heifers were identified negative by both tests. When supernatant from ear notches from 100 animals was randomly pooled and RT-PCR was accomplished on each pool, RT-PCR identified 2 pools that contained 1 positive AC-ELISA sample and 1 pool that contained 2 positive AC-ELISA samples. Further evaluation of the pooled RT-PCR ear-notch supernatant detected 100% ( n = 36) samples spiked with supernatant from a single randomly selected positive AC-ELISA ear notch. Although follow-up confirmatory tests were not completed, all 3 methods correlated 100% in detecting suspect PI animals, with a kappa value of 1. The use of RT-PCR on pooled ear-notch supernatant could provide an initial, rapid, cost-effective method of screening cattle herds for BVDV PI animals. Subsequent serial testing with an AC-ELISA to evaluate individual samples included in the positive pool could minimize the length of time other animals are exposed to the virus.
Comparison of Three Monoclonal Antibodies for use in Immunohistochemical Detection of Bovine Spongiform Encephalopathy Protease-Resistant Prion Protein
Authors: Salguero Francisco J. , Segundo Fayna Díaz-San , Brun Alejandro , Cano María J. , Torres Juan M.
Published: May 2011

Confirmatory diagnosis of prion diseases in humans and animals relies on the histopathological examination and immunodetection of the protease-resistant isoform of prion protein (PrPres). The generation of novel PrP-specific monoclonal antibodies (MAbs) has greatly improved diagnostic methodology and basic research on prion diseases as well. In this study, the performance of 3 different PrP-specific MAbs in recognizing brain PrPres deposits from cows affected with bovine spongiform encephalopathy (BSE) was compared by using a standard immunohistochemical technique under different pretreatment conditions. All antibodies showed similar reactivity after denaturing treatment. However, greater differences were found among them after proteinase K treatment, even in the absence of a denaturing step. In fact, 1 MAb (2A11) was able to react with PrPres deposits in the absence of a denaturing step, yielding the strongest signal and confirming the usefulness of MAb 2A11 in immunohistochemistry for the diagnosis of BSE.
Compressive Myelopathy due to Intervertebral Disk Extrusion in a Llama (Lama Glama)
Authors: Valentine Beth A. , Saulez Montague N. , Cebra Christopher K. , Fischer Kay A.
Published: May 2011

A 12-year-old intact female llama was euthanized following acute onset of spastic tetraparesis and recumbency with inability to rise. Postmortem examination revealed caudal cervical spinal cord compression due to a mass within the ventral spinal canal arising from the C6–C7 intervertebral disk space and attached to an irregularly thickened annulus fibrosis. On histopathologic examination, the mass was composed of amorphous acellular basophilic to amphophilic material admixed with irregularly arranged collagen bundles. The amorphous material was metachromatic and contained multiple small foci of markedly vacuolated round cells, characteristic of origin from the nucleus pulposus. Severe necrosis of all white matter tracts with astrocytic reaction was present in the overlying spinal cord segment. Ascending and descending Wallerian degeneration and dissecting interstitial astrogliosis were present within white matter tracts above and below the lesion, respectively. The diagnosis was compressive myelopathy due to chronic extrusion of the nucleus pulposus of the C6–C7 inter-vertebral disk. To the authors' knowledge, this is the first report of intervertebral disk disease in a camelid.
Detection and Serotype-Specific Differentiation of Vesicular Stomatitis Virus Using a Multiplex, Real-Time, Reverse Transcription-Polymerase Chain Reaction Assay
Authors: Hole Kate , Clavijo Alfonso , Pineda Luis A.
Published: May 2011

A multiplex, real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was developed that allowed simultaneous detection and rapid differentiation of vesicular stomatitis virus strains—New Jersey (VSV-NJ) and Indiana 1, 2, and 3 (VSV-IN1–3). This assay involves use of a set of VSV universal primers located in the L gene that amplify VSV-IN1–3 and VSV-NJ using probes that allow differentiation of the major serotypes Indiana and New Jersey. The assay was evaluated using reference VSV, foot-and-mouth disease virus, swine vesicular disease virus, and vesicular exanthema of swine virus. To estimate diagnostic sensitivity, 159 epithelial samples collected between 1996 and 2002 from naturally infected cattle in Colombia were used. The assay cut off was calculated by testing RNA extracted from 150 virus-negative bovine tissues consisting of tongue, soft palate, muzzle, coronary band, and lymph node. All infected cattle were test positive for VS by results of real-time RT-PCR analysis; results for 156 of 159 (98.1%) agreed with the serotype determination from the complement-fixation test. Amplification did not occur in any of the negative bovine epithelial samples, allowing the cut-off values for the assay to be set. The real-time RT-PCR assay was documented to be sensitive and specific for the detection of VSV-NJ and VSV-IN (1–3) strains from field samples in a single reaction, thereby supporting use of this assay in the differential diagnosis of vesicular virus diseases in cattle.
Comparison of Two Automated Immunohistochemical Procedures for the Diagnosis of Scrapie in Domestic Sheep and Chronic Wasting Disease in North American White-Tailed Deer (Odocoileus Virginianus) and Mule Deer (Odocoileus Hemionus)
Authors: Baszler T. V. , Kiupel M. , Williams E. S. , Thomsen B. V. , Gidlewski T. , Montgomery D. L. , O'Rourke K. I. , Hall S. M.
Published: May 2011

Two commercially available automated immunohistochemistry platforms, Ventana NexES and DakoCytomation Autostainer Universal Staining System, were compared for diagnosing sheep scrapie and cervid chronic wasting disease. Both automated platforms used the same antiprion protein monoclonal primary antibodies, but different platform-specific linker and amplification reagents and procedures. Duplicate sections of brainstem (at the level of the obex) and lymphoid tissue (retropharyngeal lymph node or tonsil) from the same tissue block were immunostained for the comparison. Examination of 1,020 tissues from 796 sheep revealed 100% concordance of results between the Ventana NexES and DakoCytomation platforms for diagnosing sheep scrapie from lymphoid tissue (103/103 positive; 405/405 negative) and brainstem (120/120 positive; 392/392 negative). Similarly, examination of 1,008 tissues from 504 white-tailed deer revealed 100% concordance between the Ventana NexES and DakoCytomation platforms for diagnosing chronic wasting disease from lymphoid tissue (104/104 positive; 400/400 negative) and brainstem (104/104 positive; 400/400 negative). Examination of 1,152 tissues from 482 mule deer revealed a concordance of 98.6% in lymphoid tissue and 99.9% in brainstem between the Ventana NexES and DakoCytomation platforms for diagnosing chronic wasting disease. The results indicate equivalence or near equivalence between the DakoCytomation and Ventana NexES autostainer platforms for identification of the disease-associated prion protein (PrPd)-positive and PrPd-negative brain and lymphoid tissues in sheep, white-tailed deer, and mule deer.