bluuebunny/crossref_metadata_2025 huggingface dataset:
Abomasal Coccidiosis Associated with Proliferative Abomasitis in a Sheep
Authors: Maratea Kimberly A. , Miller Margaret A.
Published: May 2011

Abomasal coccidiosis was diagnosed in an 11-month-old female sheep that died after a 2-week period of anorexia and diarrhea. The abomasal mucosa was grossly thickened with a nodular surface and focal areas of hemorrhage. Microscopically, the middle to deep abomasal mucosa contained many intact and ruptured giant protozoal schizonts associated with hyperplasia of mucous neck cells, parietal cell atrophy, moderate lymphocytic-plasmacytic inflammation, and fibrosis centered on mineralized remnants of degenerate schizont walls. Sexual tissue stages and oocysts were not present. Microscopic features of schizonts and ultrastuctural features of merozoites were comparable to previous descriptions of Eimeria (Globidium) gilruthi, a coccidian of uncertain taxonomic status historically associated with incidental infections of the abomasum in sheep and goats. The distinctive lesions suggest that, similar to ostertagiasis, heavy coccidial infection of the abomasum should be considered as a cause of anorexia, diarrhea, and proliferative abomasitis in sheep.
Atypical Coccidiosis in South American Camelids
Authors: Chigerwe Munashe , Middleton John R. , Williams Fred , Tyler Jeff W. , Kreeger John M.
Published: May 2011

Reported clinical signs of coccidiosis in South American camelids include anorexia of a few days duration, sudden death, and diarrhea. Antemortem diagnosis of clinical coccidiosis is usually based on clinical signs and supported by detection of coccidial oocysts in feces. This report describes 2 atypical cases of coccidiosis in South American camelids that had no coccidial oocysts detected on antemortem fecal flotation, prolonged weight loss, and normal fecal consistency.
Identification and Characterization of two Bovine Spongiform Encephalopathy cases Diagnosed in the United States
Authors: Richt Jürgen A. , Kunkle Robert A. , Alt David , Nicholson Eric M. , Hamir Amir N. , Czub Stefanie , Kluge John , Davis Arthur J. , Hall S. Mark
Published: May 2011

Bovine spongiform encephalopathy (BSE) is a transmissible spongiform encephalopathy of cattle, first detected in 1986 in the United Kingdom and subsequently in other countries. It is the most likely cause of variant Creutzfeldt-Jakob disease (vCJD) in humans, but the origin of BSE has not been elucidated so far. This report describes the identification and characterization of two cases of BSE diagnosed in the United States. Case 1 (December 2003) exhibited spongiform changes in the obex area of the brainstem and the presence of the abnormal form of the prion protein, PrPSc, in the same brain area, by immunohistochemistry (IHC) and Western blot analysis. Initial suspect diagnosis of BSE for case 2 (November 2004) was made by a rapid ELISA-based BSE test. Case 2 did not exhibit unambiguous spongiform changes in the obex area, but PrPSc was detected by IHC and enrichment Western blot analysis in the obex. Using Western blot analysis, PrPSc from case 1 showed molecular features similar to typical BSE isolates, whereas PrPSc from case 2 revealed an unusual molecular PrPSc pattern: molecular mass of the unglycosylated and monoglycosylated isoform was higher than that of typical BSE isolates and case 2 was strongly labeled with antibody P4, which is consistent with a higher molecular mass. Sequencing of the prion protein gene of both BSE-positive animals revealed that the sequences of both animals were within the range of the prion protein gene sequence diversity previously reported for cattle.
Detection of Antimicrobial Resistance Genes of Pathogenic Salmonella from Swine with DNA Microarray
Authors: Ma Menggen , Wang Hongning , Yu Yong , Zhang Dong , Liu Shigui
Published: May 2011

A glass-based microarray was developed to detect 11 antimicrobial resistance genes that confer resistance to aminoglycosides, tetracyclines, sulfonamides, and chloramphenicols. The target genes for microarray were generated from Salmonella isolates by PCR and confirmed by sequencing. The specificity of the microarray was tested using 11 positive DNA probes. The sensitivity was tested with tetA gene and Salmonella isolates. Using detection threshold of signal-to-noise ratio (S/N) ≥1.5 or median pixel intensity ≥1000, antimicrobial resistance genes carried by 30 Salmonella isolates were detected. Common genes included sul I (76.7%, 23/30), aph(3′)-IIa (60%, 18/30), tetC (60%, 18/30), cat1 (43.3%, 13/30), tetA (40%, 12/ 30) and aadA1 (36.7%, 11/30), and the results were confirmed to be correct by PCR.
Aflatoxicosis in Nine Dogs after Exposure to Contaminated Commercial Dog Food
Authors: Newman Shelley Joy , Smith Joanne R. , Stenske Kate A. , Newman Leslie B. , Dunlap John R. , Imerman Paula M. , Kirk Claudia A.
Published: May 2011

The purpose of this study was to characterize light and electron microscopic findings from 9 dogs that had consumed aflatoxin-contaminated commercial dog food from recalled batches. Four dogs died and 5 were euthanized after signs of liver failure. Analysis of feed and liver samples confirmed exposure to aflatoxin. Of the 9 dogs, 8 had classic signs of liver failure, and 1 had signs of liver failure. Enlarged, pale yellow livers were seen macroscopically at necropsy in the dogs with subacute hepatopathy, and cirrhosis was noted in the dog with chronic hepatopathy. Histopathologic findings included hepatic lipidosis, portal fibroplasia, and biliary hyperplasia, which supported a diagnosis of subacute toxic hepatopathy in the 8 symptomatic animals. Marked lobular atrophy, bridging portal fibrosis, and regenerative hepatocellular nodules characterized the dog with chronic hepatopathy. Electron microscopy revealed marked hepatocellular lipid vacuolation and early fibroplasia in the dogs with acute hepatopathy and marked fibrosis and regeneration in the dog with chronic hepatopathy. Analysis of feed for aflatoxin consistently revealed high levels of aflatoxin B1 (range of 223–579 ppb), and hepatic tissue contained elevated levels of aflatoxin B1metabolite M1 (0.6–4.4 ppb). Although dogs are not commonly affected by aflatoxicosis, they are highly susceptible and can present with classic signs of acute or chronic hepatopathy. Characteristic gross, histologic, and electron microscopic changes help pathologists determine a presumptive toxic insult. Detecting aflatoxins or their metabolites in feed or liver specimens can help confirm the diagnosis of aflatoxicosis.
Toward a Multiplexed Serotyping Immunoassay for Foot-and-Mouth Disease Virus
Authors: Perkins Julie , Clavijo Alfonso , Ortiz Josue I. , Salo Timothy J. , Holland Hilary J. , Hindson Benjamin J. , McBride Mary T.
Published: May 2011

Initial results demonstrating the feasibility of a multiplexed liquid array immunoassay for foot-and-mouth disease viral antigen detection and simultaneous serotype differentiation are presented. Serotype-specific antibodies from rabbit and guinea pig hyperimmunesera were isolated and prepared for use in a multiplexed, bead-based assay. The performance of all of the available antibodies as both capture and detector reagents was evaluated in the multiplexed system to establish a combination exhibiting the highest homotypic responses and lowest heterotypic reactions. The multiplexed assay was evaluated against inactivated cell culture supernatant samples of the same subtype as the virus used to raise the capture and detector antibodies. Distinct serotype differentiation was observed, except in the case of serotype SAT1. Subsequently, cell culture supernatant samples from a larger pool of viral subtypes were analyzed. Distinct serotype differentiation was obtained when analyzing cell culture supernatant samples from viral serotypes C, Asia, and SAT3, irrespective of the subtype. However, limitations of the current antibody pairs were realized in some inconclusive results obtained when analyzing samples from a broader range of O, A, and SAT2 subtypes. The results obtained in this initial study will be used to further optimize the assay using polyvalent or monoclonal antibodies and move toward the analysis of clinical samples.
Prevalence of Neoplasia in Llamas and Alpacas (Oregon State University, 2001–2006)
Authors: Valentine Beth A. , Martin Jeanne M.
Published: May 2011

Prevalence and type of neoplastic disease were determined in 551 camelid submissions (368 alpacas [Lama pacos], 180 llamas [Lama glama], and 3 cases in which species was not identified) over a 5-year period. Forty neoplasms were identified in 38 animals (6.9%). Prevalence of neoplasia in llamas was higher (11%) than in alpacas (4.9%). Mean age of camelids with neoplasia was 9.42 ± 4.9 years. Mean age of alpacas with neoplasia (5.48 ± 3.7 years) was significantly less than of llamas with neoplasia (12.53 ± 3.2 years; P < 0.001). Cutaneous and mucocutaneous fibroma/fibropapilloma was most common (10 animals), followed by cutaneous and mucocutaneous squamous cell carcinoma (6 animals), disseminated lymphoma (5 animals), and fibrosarcoma (4 animals). Four of 5 animals with lymphoma were alpacas, aged 0.21 to 4 years. Lymphoma occurred in 1 aged llama (15 years). Disseminated carcinoma and adenocarcinoma occurred in 4 llamas and 2 alpacas, and included biliary (2), gastrointestinal (2), mammary gland (1), and unknown (1) origin. Mean age of camelids with any type of carcinoma or adenocarcinoma (12.36 ± 2.8 years) was significantly greater than that of camelids with lymphoma (4.24 ± 6.2 years; P = 0.02). Results indicate that neoplasia is relatively common in camelids and that there are differences between llamas and alpacas as regards prevalence of neoplasia, tumor types, and age at diagnosis.
Vertebral Polyostotic Lymphoma in a Young Dog
Authors: Vascellari Marta , Tasca Silvia , Furlanello Tommaso , Carli Erika , Patron Carlo , Mutinelli Franco , Caldin Marco
Published: May 2011

An unusual clinical presentation of lymphoma with vertebral involvement in a dog is reported. A 20-month-old intact female Golden Retriever presented with progressive paraparesis and anorexia. Complete blood count and serum biochemistry profile demonstrated pancytopenia and hypercalcemia. Ventral fusion of the lumbar vertebrae by new bony tissue deposition was evident on X-ray and CT scan. Fine needle aspiration revealed neoplastic lymphoid cells in lymph nodes and bone marrow. Histologically, vertebral bone and osteophytes, liver, bone marrow, kidney, and lymph nodes were diffusely infiltrated by neoplastic, lymphoid cells, with scant cytoplasm and round hyperchromatic nuclei. Polyostotic and medullary T-cell lymphoma with spondylosis was diagnosed. Lymphoma mainly affecting bone is uncommon in the dog. The present case differs from previously described polyostotic lymphomas in clinical signs of the disease, mainly attributable to spondylarthrosis. In addition, lymphomatous proliferation was associated with osteoproductive lesions of the vertebrae.
Arsenic and Metaldehyde Toxicosis in a Beef Herd
Authors: Valentine Beth A. , Rumbeiha Wilson K. , Hensley Terry S. , Halse Richard R.
Published: May 2011

Over a 12-day period, 13 animals in a herd of 110 beef cattle developed ataxia with profound muscle fasciculations progressing to recumbency. Twelve animals (5 adults and 7 calves from 8–10 months of age) died, and 1 cow was euthanized. Hemorrhagic diarrhea occurred in some, but not all, animals. The onset of clinical signs was at least 12 hours after the cattle had gained access to contents of old buildings used for storage, and the majority of deaths occurred within 24 to 48 hours after the onset of clinical signs. Approximately 9 kg of unidentified pellets were found strewn in the barn area where the cattle had been. Autolysis considered more severe than expected for the postmortem interval, suggestive of high body temperature before death, and congestion of body tissues were the only significant findings detected in the cow that was euthanized and submitted for necropsy examination. The clinical history and lack of postmortem lesions were most consistent with toxicity. A toxic level of arsenic (6.18 ppm) was detected in the kidney, and metaldehyde was detected in the liver. The pellets were analyzed and found to contain both arsenic and metaldehyde, consistent with a discontinued molluscicidal product.
Diagnostic Characterization of a Feral Swine Herd Enzootically Infected with Brucella
Authors: Stoffregen William C. , Olsen Steven C. , Wheeler C. Jack , Bricker Betsy J. , Palmer Mitchell V. , Jensen Allen E. , Halling Shirley M. , Alt David P.
Published: May 2011

Eighty feral swine were trapped from a herd that had been documented to be seropositive for Brucella and which had been used for Brucella abortus RB51 vaccine trials on a 7,100-hectare tract of land in South Carolina. The animals were euthanized and complete necropsies were performed. Samples were taken for histopathology, Brucella culture, and Brucella serology. Brucella was cultured from 62 (77.5%) animals. Brucella suis was isolated from 55 animals (68.8%), and all isolates were biovar 1. Brucella abortus was isolated from 28 animals (35.0%), and isolates included field strain biovar 1 (21 animals; 26.3%), vaccine strain Brucella abortus S19 (8 animals, 10.0%), and vaccine strain Brucella abortus RB51 (6 animals, 7.5%). Males were significantly more likely to be culture positive than females (92.9% vs. 60.6%). Thirty-nine animals (48.8%) were seropositive. Males also had a significantly higher seropositivity rate than females (61.9% vs. 34.2%). The relative sensitivity rates were significantly higher for the standard tube test (44.6%) and fluorescence polarization assay (42.6%) than the card agglutination test (13.1%). Lesions consistent with Brucella infection were commonly found in the animals surveyed and included inflammatory lesions of the lymph nodes, liver, kidney, and male reproductive organs, which ranged from lymphoplasmacytic to pyogranulomatous with necrosis. This is the first report of an apparent enzootic Brucella abortus infection in a feral swine herd suggesting that feral swine may serve as a reservoir of infection for Brucella abortus as well as Brucella suis for domestic livestock.
Detection of Mixed Infections with “Candidatus Mycoplasma Haemominutum” and Mycoplasma Haemofelis Using Real-Time TaqMan Polymerase Chain Reaction
Authors: Sykes Jane E. , Drazenovich Nicole L. , Kyles Andrew E. , Ball Louise M. , Leutenegger Christian M.
Published: May 2011

The objective of this study was to evaluate the use of real-time TaqMan PCR assays for detection of coinfections with “ Candidatus Mycoplasma haemominutum” (Mhm), and Mycoplasma haemofelis (Mhf), in vitro and over time in experimentally infected cats. First, the ability of each real-time PCR assay to detect and quantify mixed infections was determined in vitro by testing mixtures of plasmids containing Mhm and Mhf 16S rDNA with each assay. Subsequently, 4 specific pathogen-free (SPF) cats, 2 of which were splenectomized, were inoculated with blood from a cat infected with both Mhm and Mhf. Sixteen blood samples were then collected from each cat over a 55-day period. Each of the 64 postinoculation samples was tested using both conventional polymerase chain reaction (cPCR) and real-time PCR for the 16S rRNA gene of each organism. When applied to mixtures of plasmid DNA from each species, the results of quantitation with each of the real-time PCR assays approximately reflected the number of plasmid copies present. Forty-nine of 64 post-inoculation samples (77%) were positive using both cPCR and real-time PCR, 4 (6%) were positive using cPCR only, and 3 (5%) were positive using real-time PCR only. Both organisms were detected in 23 samples using real-time PCR. Mixed infections were not detected using cPCR. The size of the corresponding cPCR products suggested infection with Mhm in 4 and Mhf in 18 of these samples. The use of multiple separate real-time PCR assays rather than cPCR alone should thus be considered for epidemiologic studies of hemoplasmosis in cats.
Critical Assessment of the Diagnostic Value of Feline α1-Acid Glycoprotein for Feline Infectious Peritonitis Using the Likelihood Ratios Approach
Authors: Saverio Paltrinieri , Alessia Giordano , Vito Tranquillo , Stefano Guazzetti
Published: May 2011

Alpha-1-acid glycoprotein (AGP) increases in the blood of cats with feline infectious peritonitis (FIP), a lethal disease caused by feline coronavirus (FCoV). However, the diagnostic potential of AGP might be limited because AGP also increases in pathophysiological conditions other than FIP. In this retrospective study, the diagnostic potential of serum AGP concentration was evaluated on the basis of the pretest probability of disease, according to the Bayesian approach. Serum AGP levels from cats with FIP (group 1; n = 58) and without FIP (group 2; n = 104) were evaluated. Non-FIP cats were further subgrouped as follows: 2a) inflammation ( n = 26), 2b) asymptomatic FCoV infection ( n = 49), 2c) injection-site sarcoma ( n = 19), 2d) postvaccination ( n = 7), and 2e) specific pathogen free ( n = 3). Standard descriptive analyses by group and empirical receiver-operating characteristic (ROC) curve estimation were performed. Ordinary logistic regression analysis was performed to derive an estimate of the continuous likelihood ratio to produce the posttest probability of disease for any combination of pretest probability and serum AGP value. The comparison of serum AGP levels in the different groups and the analysis of the ROC curve confirmed that serum AGP is a powerful discriminating marker for FIP. The Bayesian approach demonstrated that when the pretest probability of FIP is high, based on history and clinical signs (groups 1 or 2a), moderate serum AGP levels (1.5–2 mg/ml) can discriminate cats with FIP from others, while only high serum AGP levels (>3 mg/ml) can support a diagnosis of FIP in cats with a low pretest probability of disease (groups 2b to 2e).
Genetic Evidence for Multi-Event Imports of Avian Influenza Virus A (H5N1) into Bavaria, Germany
Authors: Rinder Monika , Lang Victoria , Fuchs Cordula , Hafner-Marx Angela , Bogner Karl-Heinz , Neubauer Antonie , Büttner Mathias , Rinder Heinz
Published: May 2011

The almost simultaneous initial detections of avian influenza A H5N1 viruses in central Europe in February 2006, at a time devoid of migratory bird activity, raised the question of the origin of these viruses. This report presents molecular data from Europe providing evidence for multiple and spatially overlapping H5N1 introductions into Bavaria, Germany.
In Situ Hybridization for Lawsonia Intracellularis-Specific 16S rRNA Sequence in Paraffin-Embedded Tissue Using a Digoxigenin-Labeled Oligonucleotide Probe
Authors: Weissenböck Herbert , Mrakovcic Maria , Ladinig Andrea , Fragner Karin
Published: May 2011

An in situ hybridization (ISH) procedure with a digoxigenin-labeled oligonucleotide probe for detection of Lawsonia intracellularis in paraffin-embedded tissue is described. This technique recognized 71% of PCR-positive cases and was thus superior to Warthin-Starry silver stain, which only detected 41%. The presented ISH is of comparable sensitivity to previously published immunohistochemical assays and is recommended for laboratories wishing to diagnose L. intracellularis infections in tissue sections but without access to antibodies.
Effects of Blood Contamination of Cerebrospinal Fluid on Results of Indirect Fluorescent Antibody Tests for Detection of Antibodies against Sarcocystis Neurona and Neospora Hughesi
Authors: Finno Carrie J. , Packham Andrea E. , Wilson W. David , Gardner Ian A. , Conrad Patricia A. , Pusterla Nicola
Published: May 2011

The purpose of this study was to determine the effect of blood contamination of cerebrospinal fluid (CSF) on the results of indirect fluorescent antibody tests (IFATs) for Sarcocystis neurona and Neospora hughesi. The in vitro study used antibody-negative CSF collected from non-neurologic horses immediately after euthanasia and blood samples from 40 healthy horses that had a range of IFAT antibody titers against S. neurona and N. hughesi. Serial dilutions of whole blood were made in seronegative CSF to generate blood-contaminated CSF with red blood cell (RBC) concentrations ranging from 10 to 100,000 RBCs/μl. The blood-contaminated CSF samples were then tested for antibodies against both pathogens using IFAT. Blood contamination of CSF had no detectable effect on IFAT results for S. neurona or N. hughesi at any serologic titer when the RBC concentration in CSF was <10,000 RBCs/μl. At concentrations of 10,000-100,000 RBCs/μl of CSF, positive CSF results (IFAT titer ≥5) for S. neurona and N. hughesi were detected only when the corresponding serum titers were ≥160 and ≥80, respectively. The IFAT performed on CSF is reliable for testing horses for equine protozoal myeloencephalitis caused by S. neurona or N. hughesi, even when blood contamination causes the RBC concentration in CSF to be up to 10,000 RBCs/μl.
Investigation of the Molecular Detection of Vaccine-Derived Equine Herpesvirus Type 1 in Blood and Nasal Secretions from Horses Following Intramuscular Vaccination
Authors: Pusterla Nicola , Chaney Kristin P. , Maes Roger , Wise Annabel G. , Holland Robert , Schott Hal C.
Published: May 2011

The objective of this study was to investigate whether intramuscular vaccination of healthy adult horses with a killed or a modified live equine herpesvirus type 1 (EHV-1) vaccine could induce transient positive PCR results in either blood or secretions collected on a nasopharyngeal swab. Four horses in each group received either a single killed or a modified-live vaccine intramuscularly. Two local commingled and 2 distant nonvaccinated controls were included for each group. All horses were observed daily for evidence of clinical abnormalities throughout the study periods. Blood and nasopharyngeal swabs were collected twice before vaccination and once weekly for 4 weeks after vaccination and submitted for PCR testing for EHV-1 by 2 independent laboratories using different real-time PCR methodologies. Serum samples collected from all horses on the vaccination day and 21 days later were tested for antibodies against EHV-1 using a serum neutralization test. Whereas the 2 vaccine strains tested positive in both EHV-1 PCR assays, nasopharyngeal swabs and whole blood collected from vaccinated and control horses had negative PCR test results for EHV-1 during the entire study period. Serum neutralization testing revealed a 2- to 4-fold increase in titers for all vaccinated horses, whereas titers in control horses were largely unchanged. The use of seropositive horses before immunization and the sampling frequency of 7 days may have prevented the occasional molecular detection of the vaccine virus in whole blood and nasopharyngeal secretions. However, the study results demonstrate that detection of EHV-1 DNA by PCR in vaccinated and unvaccinated healthy horses is not a common event.
Microalbuminuria and Comparison of Serologic Testing for Exposure to Borrelia Burgdorferi in Nonclinical Labrador and Golden Retrievers
Authors: Goldstein Richard E. , Cordner Amy P. , Sandler Jody L. , Bellohusen Beth A. , Erb Hollis N.
Published: May 2011

Canine Lyme disease is caused by the spirochete Borrelia burgdorferi after transmission by an Ixodes tick, typically resulting in joint pain, fever and lethargy. Lyme nephritis is a poorly characterized syndrome associated with severe glomerular and tubular renal injury and poor clinical outcome in young to middle-aged dogs positive for exposure to B. burgdorferi. The aims of this study were to identify associations between natural exposure to B. burgdorferi and the presence of microalbuminuria in nonclinical young Labrador and Golden Retrievers and to compare two commonly used serologic tests available to document B. burgdorferi exposure: the Western blot and the commercial point-of-care C6 peptide enzyme-linked immunosorbent assay (ELISA) tests. Microalbuminuria was assessed using a commercial point-of-care ELISA specific for canine albumin. Blood and urine samples from 268 asymptomatic Labrador and Golden Retrievers were included. Of these, 18.7% were positive for B. burgdorferi exposure according to the C6 ELISA; 21.2% were positive for natural exposure to B. burgdorferi and 11.5% for vaccinal antibodies according to the Western blot. The agreement rate was 93% between the two tests (kappa = 0.78, P < 0.0001) for natural exposure. Urine from 6.1% of the dogs was positive for microalbuminuria. There was no association between microalbuminuria and exposure to B. burgdorferi based on results of a Western blot ( P = 0.57) or C6 ELISA ( P = 0.53). Microalbuminuria is likely not a consequence of B. burgdorferi exposure in young nonclinical Labrador and Golden Retrievers.
Polypoid Uterine Leiomyosarcomas in a Sheep
Authors: Vemireddi Vimala , Langohr Ingeborg M. , Thacker Herbert L.
Published: May 2011

Leiomyosarcoma was diagnosed in the uterus surgically removed from a 3-year-old pet Suffolk ewe with a history of bleeding from the vulva, spontaneous lactation, and nursing behavior. The uterus contained multiple well-circumscribed, soft, intraluminal polypoid masses of variable sizes (0.5–4 cm). The masses were red, with white, smooth, and glistening cut surfaces. Histologically they comprised variably dense sheets of moderately pleomorphic, plump spindle cells embedded in richly vascularized stroma. The mitotic index was usually low (0-1/high-power field), but in some polyps there were up to 10 mitoses/high-power field. Neoplastic cells stained positive for alpha smooth muscle actin (α-SMA) by immunohistochemistry. Ultrastructural features of neoplastic cells included the presence of basal lamina, scant microfilaments, contracted nuclei with blunt ends, and flat intercellular junctions. Uterine leiomyosarcoma was diagnosed based on cellular morphology and atypia and positive immunohistochemistry for α-SMA.
Diagnosis of Amanita Toxicosis in a Dog with Acute Hepatic Necrosis
Authors: Puschner Birgit , Rose Heidi H. , Filigenzi Michael S.
Published: May 2011

Poisoning with amanitin-containing hepatotoxic mushrooms demands extensive efforts from clinicians, toxicologists, and pathologists. Presumptive diagnoses are established by positive identification of the suspect mushroom along with the occurrence of consistent clinical signs. If the animal dies, hepatic lesions may suggest exposure to amanitin-containing mushrooms; however, lesions are nonspecific. A 15-week-old female Dachshund was presented to the referring veterinarian for acute onset of lethargy that quickly progressed to sternal recumbency. Despite supportive care, the dog remained lethargic and died approximately 12 hours after initial presentation. A pale tan liver was noted at necropsy. Microscopically, the liver showed panlobular coagulative necrosis of hepatocytes. A presumptive diagnosis of amanitin poisoning was based on suspect history of exposure to mushrooms, clinical signs, and pathologic findings. Exposure to amanitin was confirmed through detection of α-amanitin in the liver by liquid chromatography/mass spectrometry. The objective of this case report is to illustrate the essential components to a successful diagnostic work-up of a suspect case of hepatotoxic mushroom poisoning. Although hepatotoxic mushroom poisoning has been documented in dogs before, confirmatory techniques for biologic specimens have not been used previously in diagnostic investigations.
Systemic Nocardiosis in a Reindeer (Rangifer Tarandus Tarandus)
Authors: Vemireddi Vimala , Sharma Alok , Wu Ching Ching , Lin Tsang Long
Published: May 2011

Systemic infection by Nocardia asteroides was diagnosed in a reindeer ( Rangifer tarandus tarandus). At necropsy, the animal had enlarged tracheobronchial lymph nodes, pleural and peritoneal effusions, and numerous, multifocal to coalescing, yellow, firm nodules with inspissated pus in the lung, pleura, omentum, liver, heart, adrenal glands, and left kidney. Microscopically, the nodules consisted of foci of pyogranulomatous inflammation. Microscopic lesions were present in the grossly affected organs as well as spleen and brain. Sections stained with Gram and modified Fite-Faraco histochemical stains had numerous gram-positive, variably acid fast, beaded and branching filamentous organisms in the necrotic centers. N. asteroides was isolated from the lung, bronchial lymph nodes, liver, and left kidney. To the authors' knowledge, this is the first report of systemic N. asteroides infection in a reindeer.
Toxinotypes of Clostridium Perfringens Isolated from Sick and Healthy Avian Species
Authors: Crespo Rocio , Fisher Derek J. , Shivaprasad H. L. , Fernández-Miyakawa Mariano E. , Uzal Francisco A.
Published: May 2011

Currently, the factors/toxins responsible for Clostridium perfringens-associated avian enteritis are not well understood. To assess whether specific C. perfringens' toxinotypes are associated with avian enteritis, the isolates of C. perfringens from 31 cases of avian necrotic or ulcerative enteritis submitted between 1997 and 2005 were selected for retrospective analysis using multiplex PCR. C. perfringens was isolated from chickens, turkeys, quail, and psittacines. The toxinotypes of isolates from diseased birds were compared against the toxinotype of 19 C. perfringens isolates from avian cases with no evidence of clostridial enteritis. All C. perfringens isolates were classified as type A regardless of species or disease history. Although many isolates (from all avian groups) had the gene encoding the C. perfirngens beta2 toxin, only 54% produced the toxin in vitro when measured using Western blot analysis. Surprisingly, a large number of healthy birds (90%) carried CPB2-producing isolates, whereas over half of the cpb2-positive isolates from diseased birds failed to produce CPB2. These data from this investigation do not suggest a causal relationship between beta2 toxin and necrotic enteritis in birds.
Characteristics of Diagnostic Tests Used in the 2002 Low-Pathogenicity Avian Influenza H7N2 Outbreak in Virginia
Authors: Elvinger François , Akey Bruce L. , Senne Dennis A. , Pierson F. William , Porter-Spalding Barbara A. , Spackman Erica , Suarez David L.
Published: May 2011

An outbreak of low-pathogenicity avian influenza (LPAI) H7N2 occurred in 2002 in the Shenandoah Valley, a high-density poultry production region in Virginia. Infected flocks were identified through a combination of observation of clinical signs and laboratory diagnostic tests designed to detect avian influenza (AI) antibodies, virus, or H7-specific RNA. In this report, fitness for purpose of 3 virus/RNA detection assays used during the outbreak was examined: 1) antigen capture enzyme immunoassay (AC-EIA), 2) real-time reverse transcription polymerase chain reaction (RRT-PCR), and 3) virus isolation (VI). Results from testing 762 turkey and 2,216 chicken tracheal swab pooled specimens were analyzed to determine diagnostic sensitivities and specificities of these tests under field conditions using Bayesian techniques for validation of diagnostic tests in the absence of a “gold standard.” Diagnostic sensitivities (with 95% probability intervals) in turkeys of AC-EIA and RRT-PCR, in reference to VI, were 65.9 (50.6; 81.3)% and 85.1 (71.9; 95.7)% and of VI 92.9 (78.0; 98.8)% in reference to AC-EIA or 88.7 (76.0; 97.2)% in reference to RRT-PCR; in chickens, diagnostic sensitivities were 75.1 (45.6; 94.2)%, 86.3 (65.9; 97.1)%, and 86.2 (65.8; 97.1)% or 86.3 (66.4; 97.2)%, respectively. Specificities were 99.1 (97.9; 99.8)%, 98.9 (98.0; 99.5)%, and 98.6 (97.4; 99.4)% or 98.8 (97.8; 99.5)% in turkeys and between 99.25% and 99.27% with probability intervals of approximately ±0.4% for all tests in chickens. Simultaneous use of AC-EIA and RRT-PCR contributed significantly to the rapid control of the outbreak, but the AI RRT-PCR assay with >85% sensitivity and ∼99% specificity, combined with relatively low cost and fast turnaround, could be used as the sole diagnostic test in outbreaks of LPAI.
Detection of Botulinum Type E Toxin in Channel Catfish with Visceral Toxicosis Syndrome Using Catfish Bioassay and Endopep Mass Spectrometry
Authors: Gaunt Patricia S. , Kalb Suzanne R. , Barr John R.
Published: May 2011

Visceral toxicosis of catfish (VTC) is a syndrome characterized by sudden mortality in apparently healthy market- and brooder-sized catfish ( Ictalurus punctatus). This paper reports the design of a catfish neutralization assay to detect botulinum in catfish with VTC and verification by Endopep mass spectrometry (Endopep-MS). Sera from 6 affected catfish were incubated with botulinum antitoxin serotypes A, B, C, D, E, or F. For each serum sample, 3 experimental fingerlings were injected intracoelomically with each serotype-serum mixture and placed separately in an aquarium. Three fish were injected with VTC-affected serum only, and 3 fish were injected with unaffected serum only and also placed in separate aquaria. Signs of morbidity and mortality were seen in fish injected with sera combined with serotype A, B, C, or D, as well as in positive controls. No morbidity or mortality was seen in fish injected with sera combined with antitoxin serotypes E or F or negative control serum. Sera from affected and unaffected catfish were sent to Centers for Disease Control and Prevention for detection and differentiation of botulinum neurotoxin. Aliquots of 0.5 ml of sera were incubated with magnetic beads coated with antibodies to botulinum, and the beads were subjected to the Endopep-MS reaction. Sera from affected catfish tested positive for botulinum E. Sera from 34 unaffected catfish tested negative for botulinum. Although there was not enough botulinum present in affected samples to obtain exact quantification, the estimated quantity of botulinum E in these sera samples was between 0.01 and 0.5 mouse LD50/ml.
Development and Evaluation of an Immunochromatographic Strip for Detection of Streptococcus Suis Type 2 Antibody
Authors: Yang Junxing , Jin Meilin , Chen Jianfeng , Yang Ying , Zheng Pei , Zhang Anding , Song Yunfeng , Zhou Hongbo , Chen Huanchun
Published: May 2011

In this study, an immunochromatographic strip (ICS) was developed for the detection of antibody against Streptococcus suis serotype 2 (SS2). Colloidal gold particles labeled with staphylococcal protein A (SPA), which can bind to the FC fragment of mammalian immunoglobulin, were used as the detector reagent. The capsular polysaccharide (CPS) of SS2 and affinity-purified IgG from a healthy naive pig were immobilized on test and control regions of a nitrocellulose membrane, respectively. The ICS was used to 1) detect anti-CPS antibody in 14 sera taken from 4 SS2-infected pigs, 24 sera from pigs hyperimmunized with SS2, and 68 sera from pigs inoculated or infected with bacteria other than SS2; 2) determine anti-CPS antibody titers of 20 positive sera for comparison with enzyme-linked immunosorbent assay (ELISA); and 3) detect anti-CPS antibody in 226 clinical sera taken from diseased pigs also for comparison with ELISA. An ELISA used as a reference test determined the specificity and sensitivity of the ICS to be 97.1% and 86.3%, respectively. There was excellent agreement between the results obtained by ELISA and the ICS (kappa = 0.843). Additionally, there was strong agreement between the results of bacterial isolation from pig tonsils and ICS test (kappa = 0.658). Because it is rapid and easy to use, the test is suitable for the serological surveillance of SS2 at farms.
Comparative Evaluation of Rose Bengal Plate Agglutination Test, Mallein Test, and Some Conventional Serological Tests for Diagnosis of Equine Glanders
Authors: Naureen Abeera , Saqib Muhammad , Muhammad Ghulan , Hussain Muhammad H. , Asi Muhammad N.
Published: May 2011

The Rose Bengal plate agglutination test (RBT) was evaluated for the diagnosis of equine glanders, and its diagnostic efficiency was compared with that of mallein and other serological tests, including indirect hemagglutination test (IHAT), complement fixation test (CFT), and modified counter immunoelectrophoresis test (mCIET). Sera from 70 naturally infected culture-positive, 96 potentially exposed cohorts, and 110 healthy equines were tested. All tests but mCIET showed 100% specificity when testing the sera from glanders-negative equines. The calculated sensitivities of RBT, IHAT, CFT, mCIET, and mallein test when testing culture-positive equines were 90.0, 97.1, 91.4, 81.4, and 75.7%, respectively. The RBT was significantly ( P < 0.05) more sensitive than the mallein test and mCIET. The positive and negative predictive values of each test (RBT, IHAT, CFT, mallein test, and mCIET) were as follows: 100 and 94, 100 and 98.2, 100 and 96.7, 100 and 86.6, and 90.5 and 88.6, respectively. On comparing glandered and nonglandered animals, the highest agreement (0.987) was found between RBT and CFT followed by RBT and IHAT (0.940), RBT and mallein test (0.871), and RBT and mCIET (0.852). Because the RBT is simpler and rapid to perform, the inclusion of the test as a supplementary test for the diagnosis of glanders in field conditions is recommended.
Association of Myocarditis with High Viral Load of Porcine Circovirus Type 2 in Several Tissues in Cases of Fetal Death and High Mortality in Piglets. A Case Study
Authors: Brunborg Inger M. , Jonassen Christine M. , Moldal Torfinn , Bratberg Bjørn , Lium Bjørn , Koenen Frank , Schönheit Jürgen
Published: May 2011

During a period of 1.5 months, a newly established pig herd experienced a high number of mummifications and stillbirths, a high neonatal mortality rate, and many piglets with congenital tremors or hind leg ataxia. After clinical and histological investigations, the submitted animals were divided into 4 groups: mummified or stillborn (N = 6), live born with myocarditis (N = 5) (average age 22.8 days), live born without myocarditis (N = 14) (average age 20.0 days), and control animals from a different herd (N = 5) (newborn). Statistically significant differences were observed in the mean porcine circovirus 2 (PCV2) load among the 4 groups in the liver ( P < 0.0001). The presence of PCV2 antigen within the myocardial lesions was confirmed by immunohistochemistry. A high load of PCV2 DNA was observed in myocardium, liver, and spleen from mummified or stillborn piglets (>1 × 107 copies per 500 ng DNA), lower in piglets with myocarditis (>1 × 105 copies per 500 ng DNA), and even further lower in pigs without myocarditis (<1 × 105 copies per 500 ng DNA), whereas no PCV2 DNA was detected in the control animals. Myocardium, liver, and spleen were well suited for routine testing of fetuses and young piglets by quantitative real-time polymerase chain reaction. Neither porcine parvovirus nor encepaholomyocarditis virus was detected. These results indicate that the PCV2 infection might have been of etiological importance for the fetal deaths and piglet mortality observed in this herd.
Evaluation of the i-STAT Portable Clinical Analyzer in Chickens (Gallus Gallus)
Authors: Steinmetz Hanspeter W. , Vogt Rainer , Kästner Sabine , Riond Barbara , Hatt Jean-Michel
Published: May 2011

The i-STAT portable clinical analyzer (PCA) was evaluated for performance in avian species. With the EG7+ cartridge, which provided results for hydrogen ion concentration, oxygen tension, carbon dioxide tension, sodium, potassium, ionized calcium, hematocrit, and various calculated parameters, analytical accuracy and precision were tested by comparing obtained values to those of established traditional blood gas and chemistry analyzers. Deming's regression and bias plots were used to compare i-STAT results with those obtained by laboratory professionals using benchtop analyzers. The reliability of the i-STAT PCA with EG7+ cartridges was good, with 0–5.7% system failures in measured values. Regression statistics were good for all blood gas analytes and acceptable for electrolytes and calculated parameters, except for potassium and base excess, for which the regression data or the discrepancy between the methods was too large. The system was reliable and easy to use and had an overall acceptable accuracy in avian species. These features, together with portability and small required blood volumes, make the i-STAT suitable for point-of-care use in critical avian patients, although single values require careful interpretation.
Detection of Newcastle Disease Virus RNA by Reverse Transcription-Polymerase Chain Reaction Using Formalin-Fixed, Paraffin-Embedded Tissue and Comparison with Immunohistochemistry and in situ Hybridization
Authors: Wakamatsu Nobuko , King Daniel J. , Seal Bruce S. , Brown Corrie C.
Published: May 2011

The usefulness of reverse transcription-polymerase chain reaction (RT-PCR) from formalin-fixed, paraffin-embedded (FFPE) tissues was examined and compared to the immunohistochemistry (IHC) and in situ hybridization (ISH) assays for detection of Newcastle disease virus (NDV). Spleen and lung tissues were collected from chickens experimentally infected with either of 2 NDV isolates: a low virulent virus (LaSota) and a virulent virus (from the 2002–2003 California outbreak). The tissues were harvested immediately postmortem and fixed in 10% neutral buffered formalin for approximately 52 hours. Also, just before euthanasia, oral and cloacal swabs were collected for virus isolation. RNA was obtained from the FFPE tissues by digestion with proteinase K and subsequent extraction with phenol, chloroform, and isoamyl alcohol. By seminested RT-PCR with primers for the NDV matrix gene, a 232-base pair (bp) product was generated and visualized by electrophoresis. The results of PCR were compared to those of IHC for viral nucleoprotein and ISH for matrix gene (850 bp) on 3-μm sections and to those of virus isolation from swabs. All samples from infected chickens were positive by RT-PCR, including samples that were negative by both IHC and ISH. The RT-PCR positives included tissue from chickens that were no longer shedding virus detectable by virus isolation. The RT-PCR was an effective and sensitive method to detect NDV in FFPE tissues. To the authorsn' knowledge, this is the first report of NDV detection in FFPE tissues as a diagnostic approach possibly suitable for archival materials.
Real-Time Reverse Transcription-Polymerase Chain Reaction Detection of Newcastle Disease Virus Using Light upon Extension Fluorogenic Primers
Authors: Antal Márta , Farkas Tibor , Germán Péter , Belák Sándor , Kiss István
Published: May 2011

A real-time reverse transcriptase (RT)-PCR assay, applying light upon extension (LUX) fluorogenic primers, was developed for rapid and efficient detection of Newcastle disease virus (NDV). The method, which targets the fusion (F) protein gene of the viral genome, gave positive signal with all NDV isolates tested (32/32), while negative results were obtained with heterologous pathogens (35/35), including 13 avian influenza virus isolates. The detection limit of the assay was approximately 10+1.2 egg infectious dose (EID)50/0.2 ml and 10+2.2 EID50/0.2 ml for virus suspensions and spiked chicken fecal samples, respectively. As expressed in plasmid copy number, the procedure has a sensitivity of approximately 20 copies of the plasmid harboring the target gene. Due to its high specificity, sensitivity, and relative simplicity, the LUX RT-PCR assay provides a novel, rapid, and practical tool for the detection of NDV.
Evaluation of Tongue as a Complementary Sample for the Diagnosis of Parvoviral Infection in Dogs and Cats
Authors: McKnight Christy A. , Maes Roger K. , Wise Annabel G. , Kiupel Matti
Published: May 2011

Diagnosis of canine parvovirus type 2 and feline panleukopenia virus infection in dogs and cats may be hampered by the severity of enteric lesions, secondary bacterial overgrowth, and rapid onset of autolysis. In contrast to small intestine, tongue epithelium is less sensitive to postmortem changes. Sections of tongue and small intestine from 11 dogs and 11 cats with a clinical history and gross and microscopic lesions compatible with canine and feline parvoviral infection were examined for parvoviral infection using real-time polymerase chain reaction (PCR), immunohistochemistry (IHC), and direct fluorescent antibody testing (FA). Parvoviral DNA was detected by PCR in both small intestine and tongue of all but 1 dog. Nineteen of 22 animals (86%) with suspect or positive FA staining in the small intestine also had positive FA and IHC staining in the tongue. Three of 3 dogs (100%) whose carcasses had been frozen and thawed prior to necropsy had more consistently positive staining in tongue than in small intestine by FA and IHC. These data confirm tongue as an excellent complementary sample for parvoviral testing in dogs and cats, especially in cases in which postmortem autolysis has occurred.
Comparison of Surface Plasmon Resonance Imaging and Enzyme-Linked Immunosorbent Assay for the Detection of Antibodies against Iridovirus in Rock Bream (Oplegnathus Fasciatus)
Authors: Cho Ho Seong , Kim Tae Jung
Published: May 2011

A protein chip based on surface plasmon resonance imaging (SPRI) was developed for detecting fish iridovirus antibody using a recombinant 50-kDa fragment of major capsid protein (MCP) as an antigen. The diagnostic potential of SPRI for measuring antibodies to the iridovirus MCP was compared with that of a conventional enzyme-linked immunosorbent assay (ELISA) using 40 juvenile rock bream ( Oplegnathus fasciatus) serum samples in a nursery. There was a strong positive correlation between the SPRI and ELISA ( n = 40, r = 0.939, P < 0.01). Therefore, this recombinant 50-kDa MCP can be used as an antigen for serological studies, and the SPRI, which is a label-free and high-throughput method, is potentially a valuable tool in the serodiagnosis of an iridoviral infection.
Application of Polymerase Chain Reaction Fingerprinting to Differentiate Ornithobacterium Rhinotracheale Isolates
Authors: Thachil Anil J. , Velayudhan Binu T. , Lopes-Berkas Vanessa C. , Halvorson David A. , Nagaraja Kakambi V.
Published: May 2011

Ornithobacterium rhinotracheale (ORT) is an infectious respiratory pathogen of chickens, turkeys, and wild birds. There are 18 serotypes of ORT reported worldwide. In this study, enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction and random amplified polymorphic DNA assay with Universal M13 primer-based fingerprinting techniques were investigated for their ability to differentiate ORT isolates. The authors examined 50 field isolates and 8 reference strains of ORT for their genetic differences. The fingerprint patterns were compared with serotyping results of ORT by the agar gel precipitation test. M13 fingerprinting revealed different patterns for 6 reference serotypes of ORT that were tested, namely, C, D, E, I, J, and K. Ornithobacterium rhinotracheale reference serotypes A and F yielded indistinguishable fingerprints with M13 fingerprinting. The ERIC 1R technique discerned only 5 of the 8 reference serotypes of ORT. Distinct fingerprints were also found within the ORT serotypes with both techniques. From 58 isolates of ORT that were fingerprinted belonging to 8 ORT serotypes, 10 different fingerprints were obtained with M13 fingerprinting and 6 different fingerprints were obtained with ERIC 1R fingerprinting. M13 fingerprinting technique was found to be more discriminative in differentiating ORT isolates than the ERIC 1R fingerprinting technique. These results suggest that fingerprinting techniques may be a more discerning tool for characterizing ORT isolates than the serological test using the agar gel precipitation test. This fingerprinting technique could potentially be a valuable tool in identifying an isolate from a clinical outbreak of ORT infection for development of an autogenous vaccine.