bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Hemolysis on Nonesterified Fatty Acid and β-Hydroxybutyrate Concentrations in Bovine Blood
Authors: Stokol Tracy , Nydam Daryl V.
Published: May 2011

Nonesterified fatty acid (NEFA) and b-hydroxybutyrate (BHB) assays are used for evaluating dairy herds for negative energy balance and subclinical ketosis, respectively. Hemolysis is a common artifact in samples submitted to diagnostic laboratories. The effect of hemolysis on NEFA and BHB in bovine serum was determined. Hemolysis was introduced into 26 serum samples by adding serial dilutions of a red cell hemolysate, prepared by repeated freeze-thawing of EDTA-anticoagulated bovine blood. NEFA, BHB, and degree of hemolysis (hemolytic index) were measured by an automated chemistry analyzer. Two endpoint assays that differed by inclusion of a sample blank were used for NEFA measurement. A kinetic enzymatic assay with 2 reagent sources was used for BHB measurement. The assessed methods yielded similar NEFA or BHB results in baseline, nonhemolyzed samples (median NEFA: 0.25 mEq/L, median BHB: 3 mg/dL, median hemolytic index: 8 units). NEFA results were adversely affected by hemolysis, with values increasing significantly with higher degrees of hemolysis. Median values increased above a critical medical decision limit (0.40 mEq/L) at a hemolytic index of 506 units (marked hemolysis). This increase was prevented by inclusion of a sample blank. Result interpretation was affected in individual animals when samples were moderately hemolyzed (median hemolytic index: 258 units). In contrast, BHB results were unaffected by hemolysis with either reagent source. Thus, assays for measuring NEFAs should include a sample blank and NEFA results should not be interpreted in moderately to markedly hemolyzed bovine samples, because result accuracy cannot be assured.
Comparison of Cytogenetics and Polymerase Chain Reaction Based Detection of the Amelogenin Gene Polymorphism for the Diagnosis of Freemartinism in Cattle
Authors: McNiel Elizabeth A. , Madrill Nicole J. , Treeful Amy E. , Buoen Lance C. , Weber Alvin F.
Published: May 2011

A polymerase chain reaction (PCR) assay which detects a sex-based polymorphism in the bovine amelogenin locus was modified and compared to conventional cytogenetic analysis for diagnosis of freemartinism (XX/XY chimerism) in cattle. The PCR assay is more sensitive than cytogenetic analysis for detection of XY cells, with the limit of detection of the assay falling between 0.2% and 1% XY cells. Seventy-three heifer blood samples submitted for evaluation of freemartinism to the University of Minnesota Diagnostic Laboratory were tested using both cytogenetic and PCR techniques. Poor-quality samples precluded successful lymphocyte culture and recovery of mitotic nuclei for cytogenetic evaluation in 17 cases (23%). Two of these samples (2.7%) also failed to amplify with PCR. There was 100% agreement in the results from the 56 samples that were suitable for testing using both techniques. This PCR-based assay provides an alternative to the more laborious cytogenetic evaluation for diagnosis of freemartinism.
Isolation of Equine Herpesvirus-5 from Blood Mononuclear Cells of a Gelding
Authors: Bell Stephanie A. , Balasuriya Udeni B. R. , Nordhausen Robert W. , MacLachlan N. James
Published: May 2011

Horses are commonly infected by herpesviruses, but isolation of equine herpesvirus-5 (EHV-5) has only infrequently been reported. We describe the isolation and characterization of a strain of EHV-5 from the blood mononuclear cells of a healthy adult horse in California. The virus was initially identified by EHV-5 specific polymerase chain reaction (PCR), and it caused lytic infection of cultured rabbit kidney cells only after repeated serial passage. Virions with characteristic herpesvirus morphology were readily demonstrated in cell culture lysate by transmission electron microscopy. A portion of the glycoprotein B gene of this strain of EHV-5 had 99% identity to the published EHV-5 sequence, and it was clearly distinguishable from other EHV (1–4) by virus-specific PCR assays. Prevalence of EHV-5 infection in a group of young racehorses was estimated at 64% using the EHV-5 specific PCR on nasopharyngeal secretions.
Goat Paratuberculosis in Chile: First Isolation and Confirmation of Mycobacterium Avium Subspecies Paratuberculosis Infection in a Dairy Goat
Authors: Kruze Juan , Salgado Miguel , Paredes Enrique , Mella Armin , Collins Michael T.
Published: May 2011

In October 2004, 41 goats >2 years old from a Saanen dairy goat herd located in Purranque County, 10th Region, Chile, were sampled and tested for paratuberculosis. While collecting samples it was observed that several goats were thin and emaciated. One goat was sufficiently debilitated to warrant humane euthanasia. This animal was brought to the Veterinary School at the Universidad Austral de Chile for necropsy. The goat selected for necropsy was a 12-year-old doe. The animal showed classical clinical signs of caprine paratuberculosis: emaciation despite willingness to eat, dry and rough hair coat, and no evidence of diarrhea. Gross pathology and histopathology of the necropsied goat were consistent with paucibacillary paratuberculosis. Bacteriology, serology, and PCR confirmed the diagnosis. This is the first published report of goat paratuberculosis in Chile confirming a case of caprine paucibacillary paratuberculosis.
Distribution of Equine Infectious Anemia in Horses in the North of Minas Gerais State, Brazil
Authors: Bicout Dominique J. , Carvalho Regina , Chalvet-Monfray Karine , Sabatier Philippe
Published: May 2011

The paper examines the prevalence of equine infectious anemia (EIA) in horse populations in the northern part (comprising 89 cities) of Minas Gerais State, Brazil, from January 2002 to December 2004. Data on 8,981 agar gel immunodiffusion test results from the region were used as input for a statistical and autoregressive analysis model to construct a city-level map of the distribution of EIA prevalence. The following EIA prevalence (P) levels were found: 49 cities with 0 < P ≤ 0.5%, 26 with 0.5% < P ≤ 1.5%, 10 with 1.5% < P ≤ 5%, and 4 with 5% < P ≤ 25%.
Chlorophacinone Exposure Causing an Epizootic of Acute Fatal Hemorrhage in Lambs
Authors: Piero Fabio Del , Poppenga Robert H.
Published: May 2011

This report describes an epizootic of chlorophacinone toxicosis in lambs with severe acute hemorrhages. Eleven lambs, approximately 1–2 months of age, suddenly developed epistaxis, respiratory distress, and facial and cervical swelling. Affected animals died within 1–2 hours from the onset of clinical signs. Two lambs were available for complete postmortem examination. Gross lesions included mucosal and organ pallor, icterus, melena, and lung edema, as well as thymic, cervical muscle, and intra-articular hemorrhage. Histologically hepatocellular centrolobular necrosis was observed. The anticoagulant chlorophacinone was detected in the livers at 0.58 ppm and 0.50 ppm (wet weight), respectively. The source of exposure to chlorophacinone was old bait material placed between the wall studs of the building housing the ewes and lambs. The lambs were able to reach the bait through a hole in the plywood interior wall of the building.
Acute Anhydrous Ammonia Intoxication in Cattle
Authors: Fitzgerald Scott D. , Grooms Daniel L. , Scott M. A. , Clarke K. R. , Rumbeiha Wilson K.
Published: May 2011

Intoxication with anhydrous ammonia (AA) is a common occupation-related health problem affecting farmers and fertilizer applicators, however, very few descriptions of animal exposure to this toxicant exist. Thieves entered a feedlot and damaged a liquid AA storage tank, resulting in the accidental leakage of gas from a valve. Overnight, 12 cattle were found dead, with a total mortality of 64 cattle out of 260 on the premises dying or euthanized in <1 week. Signs of affected cattle included blindness, drooling, inappetence, respiratory distress, recumbency, and death. Two cattle were submitted live for diagnostic evaluation 3 days after initial exposure. Gross lesions included corneal ulcers, fibrinonecrotizing rhinitis, hemorrhages within the nasal sinuses, and anterioventral bronchopneumonia. Microscopic lesions consisted predominantly of degeneration and necrosis of superficial epithelium lining nasal passages, trachea, and pulmonary airways. This case illustrates the acute AA effects on cattle, which is likely to be an increasingly encountered problem because of on-farm storage of AA and its access by illicit drug manufacturers.
Monensin Toxicosis in Water Buffaloes (Bubalus Bubalis)
Authors: Rozza Daniela Bernadete , Vervuert Ingrid , Kamphues Josef , Farias da Cruz Cláudio Estêvão , Driemeier David
Published: May 2011

The consumption of monensin-containing feed resulted in deaths of water buffaloes from a feedlot in which cattle and buffaloes were kept together. The monensin formulation was recommended only for use in cattle. Anorexia, muscular weakness, dyspnea, and recumbency were the major clinical findings. The most significant gross lesions were focal pale areas in semitendinosus and semimembranosus muscles, in which segmental necrosis of myofibers was seen microscopically. To compare susceptibilities of species to monensin, 3 bovine calves and 3 buffalo calves were orally dosed. At 5, 7.5, and 10 mg/kg of monensin, only the buffaloes became ill and died. Clinical signs initiated 18–20 h postdosing and were comparable to those from field cases. Gross changes consisted of ascites, hydrothorax, hydropericardium, hepatomegaly, and focal pale areas in the myocardium and to a lesser degree in semitendinosus and semimembranosus muscles. Histopathological changes also resembled those from the field cases, but were especially pronounced in the myocardial cells. The hypothesis that buffaloes could have a lower tolerance to monensin than cattle has been supported by experimental cases.
Renal Tubular Necrosis and Interstitial Hemorrhage (“Turkey-Egg Kidney”) in a Circovirus-Infected Yorkshire Cross Pig
Authors: Imai Denise M. , Cornish Jennifer , Nordhausen Robert , Ellis John , MacLachlan N. James
Published: May 2011

A juvenile Yorkshire cross pig with rapidly progressive acute renal failure was submitted for necropsy. There was marked edema and disseminated petechiation of both kidneys, producing the “turkey-egg” appearance that is characteristic of exotic diseases such as African and classical swine fever. Microscopic findings included renal tubular epithelial necrosis with extensive interstitial edema and hemorrhage; lymphoplasmacytic, eosinophilic, and histiocytic tubulointerstitial nephritis; and numerous botryoid intracytoplasmic inclusions within the renal tubular epithelium and interstitial macrophages. Porcine circovirus 2 (PCV2) was readily identified within these lesions by both PCV2-specific immunohistochemistical staining and electron microscopy. Tests for African and classical swine fever viruses, as well as bacterial cultures, were negative. The striking renal lesions in this pig were attributed to PCV2 infection and are distinct from those that are typical of other PCV2-associated diseases.
Malignant Nonteratoid Ocular Medulloepithelioma in a Llama (Llama Glama)
Authors: Schoeniger Sandra , Donner Ludvik R. , Van Alstine William G.
Published: May 2011

A 6-year-old female llama presented with buphthalmos of its right eye owing to the presence of an intraocular mass. The affected globe was enucleated and submitted for microscopic examination. The intraocular mass was diagnosed as malignant medulloepithelioma. Within the following months, the llama developed soft tissue masses, which completely filled the right orbital cavity and expanded the cranial portion of the right mandibular bone, and enlarged mandibular lymph nodes. Euthanasia was elected 30 months after the initial diagnosis. The carcass was submitted for postmortem examination, which revealed the presence of medulloepithelioma metastases within the right orbit, mandible, mandibular lymph nodes, lungs, liver, and mesenteric and sublumbar lymph nodes. The primary intraocular tumor and its metastases were composed of neoplastic undifferentiated neuroepithelial cells, which formed tubules, Flexner-Wintersteiner and Homer Wright rosettes, and rare solid sheets. Electron microscopy showed that tumor cells were connected by desmosome-like junctions and contained rare intracytoplasmic basal bodies. Neoplastic cells were positive for vimentin, nestin, microtubule-associated protein 1B, S-100 protein, and glial fibrillary acidic protein (GFAP). To the best of the authors' knowledge, this is the first report of a malignant nonteratoid ocular medullopithelioma with distant metastases in a llama and of the ultrastructural and extended immunohistochemical characterization of a nonteratoid medulloepithelioma in this species.
Mixed Thymoma in a Cow
Authors: Ecco Roselene , Langohr Ingeborg M. , Túry Erno , Santos Helvécio L. , Jacobina Gabriel C.
Published: May 2011

A neoplastic mass measuring 25 × 8 × 3 cm was found in the mediastinum of a 10-year-old Holstein cow. The neoplasm was encapsulated, multilobulated, and cystic. Several firm adhesions were present between the neoplasm and the right cranial lung lobe and the ipsilateral side of the pericardial sac. Under microscopic examination, the neoplasm was characterized by solid sheets of spindle epithelial cells (pancytokeratin and high molecular weight cytokeratin positive) subdivided by fibrous trabeculae. Numerous individualized or small aggregates of mature T-cell (CD3 positive) lymphocytes were scattered throughout the neoplastic cell population. Based on the gross, histologic, and immunohistochemical findings, the neoplasm was diagnosed as a mixed thymoma.
Hepatic Yersiniosis in a Cougar (Felis Concolor)
Authors: Owston Michael A. , Wu Ching Ching , Ramos-Vara José A.
Published: May 2011

A cougar ( Felis concolor) was diagnosed with hepatic yersiniosis by bacterial culture and histopathology. The animal had a 2-week history of anorexia and jaundice before its death. Grossly, the liver exhibited caseo-necrotic foci. Histopathologically, there was necrotizing and suppurative hepatitis, with large numbers of intralesional gram-negative coccobacilli. Additional hepatic lesions included central vein thrombosis, lymphoplasmacytic portal hepatitis, and capsulitis. Yersinia pseudotuberculosis coccobacilli were isolated in pure culture from the hepatic lesions. Because the hepatic lesions in this animal resemble those of other zoonotic diseases, such as plague and tularemia, veterinarians and laboratory personnel who handle samples should take adequate safety precautions. This report is the first to describe the pathology associated with hepatic yersiniosis in a cougar.
Lymphoma and Mycobacterium Avium Infection in a Ferret (Mustela Putorius Furo)
Authors: Saunders Geoffrey K. , Thomsen Bruce V.
Published: May 2011

A 6-year-old, neutered male ferret presented with weight loss. Radiography revealed an enlarged liver and other abdominal masses. The ferret was euthanized, and at necropsy, the stomach wall was thickened, mesenteric lymph nodes were enlarged, and the liver contained multifocal tan nodules. Histopathology confirmed lymphoma and granulomatous inflammation in all affected organs. Acid-fast bacilli were present in the lesions and were confirmed to be Mycobacterium avium by PCR.
Effect of Probe-Site Mismatches on Detection of Virulent Newcastle Disease Viruses Using a Fusion-Gene Real-Time Reverse Transcription Polymerase Chain Reaction Test
Authors: Kim L. Mia , Afonso Claudio L. , Suarez David L.
Published: May 2011

Virulent forms of Newcastle disease virus (NDV) are a major concern for poultry producers around the world and the rapid diagnosis of an outbreak is crucial to any control program. A validated real-time reverse transcription-PCR test (fusion test) directed at the fusion-cleavage site of NDV was developed to differentiate virulent Newcastle disease virus strains from those of low virulence, however one virulent isolate, Dove/Italy/2736/2000, escaped detection during the initial evaluation of the test. The objectives of this study were to determine how this isolate differed from other detectable isolates, to identify other isolates that may fail detection, and to characterize the effect of specific probe-site mutations on the fusion test at a range of annealing temperatures. Using a virulent NDV isolate (Game fowl/US(CA)/2002) as a backbone that has 100% identity to the fusion-test probe, specific changes were made to the fusion-test probe-site to reflect the unique mismatches found in Dove/Italy/2736/2000 and other selected regions of the probe. Mutated clones with mismatches unique to Dove/Italy/2736/2000 at positions 6, 13, and 14 were not detected until annealing temperatures were lowered to 50°C. Those detected at 58°C contained 1–2 mismatches (position 1 and 6, 13 and 14, or 14 only) although increased cycle threshold values compared to the parent clone indicated decreased sensitivity. Data from this study predicts that the fusion test may fail to detect some viruses among lineage 4b and potential solutions to identify this subset of viruses include lowering the annealing temperature or modifying the probe.
A Testing Scheme for the Detection of Mycobacterium Avium subsp. Paratuberculosis in Bovine Feces Utilizing the ESP Para-JEM Liquid Culture System
Authors: Rajeev Sreekumari , Shulaw William , Berghaus Roy , Zhang Yan , Byrum Beverly
Published: May 2011

A testing scheme for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in broth cultures of bovine fecal samples carried out in ESP para-JEM System was evaluated. The scheme included acid-fast staining (on signal-positive and signal-negative samples), and confirmation by PCR for 2 MAP-specific targets and subculture of all acid-fast positive PCR-negative samples. Two hundred and fifty bovine fecal samples were evaluated for the presence of MAP using this scheme. Thirty-seven (15%) of 250 fecal samples had a positive culture result when the proposed testing scheme was used, compared to 14 (6%) positive results when using the standard ESP para-JEM protocol (requiring samples to have a positive signal from the system, a positive acid-fast stain, and a positive IS 900 PCR result), and 20 (8%) positives when conventional culture was performed on Herrold egg yolk (HEY) media. A preliminary comparison of real-time and conventional PCR on DNA extracted from 15 MAP-positive broth cultures by 3 different protocols suggested that conventional PCR may be a better choice for the confirmation of the presence of MAP in the liquid cultures than real-time PCR.
Tissue Distribution of Psittacid Herpesviruses in Latently Infected Parrots, Repeated Sampling of Latently Infected Parrots and Prevalence of Latency in Parrots Submitted for Necropsy
Authors: Tomaszewski Elizabeth K. , Wigle William , Phalen David N.
Published: May 2011

Psittacid herpesvirus-1 (PsHV-1) is the cause of an acute fatal disease in parrots and is implicated as the cause of papillomatous lesions of the digestive tract. Not all infections cause disease and some parrots are infected asymptomatically. Latently infected parrots are potential sources for virus dissemination. Tissues from parrots that died spontaneously with a history of coming from flocks where a PsHV-1 outbreak had occurred were examined for PsHV-1 DNA. Fourteen of 16 parrots examined were infected with at least 1 variant of PsHV-1; of these 13 (93%) had viral DNA in either or both the oral and cloacal mucosa, suggesting that most latently infected parrots could be detected by sampling these sites. Nine of 9 parrots shown to be infected 5 years prior to this study were positive again on repeat sampling and were infected with the same virus genotype. Opportunistic sampling of parrots submitted for diagnostic necropsy indicated that the prevalence of PsHV-1 in parrots in the sampled population was approximately 9.3%. PsHV-1 genotypes 1, 2, and 3 were found in these birds, but genotype 4 was not. Six necropsy specimens were found to be infected with two PsHV-1 genotypes and it was concluded that infection with one serotype did not protect against infection with another. Psittacid herpesvirus 2 (PsHV-2) was identified in 4 African grey parrots and a blue and gold macaw. Prior to this study PsHV-2 had only been found in African grey parrots.
Development and Evaluation of an Indirect Enzyme-Linked Immunosorbent Assay for Detection of Foot-and-Mouth Disease Virus Nonstructural Protein Antibody using a Chemically Synthesized 2B Peptide as Antigen
Authors: Inoue Toru , Parida Satya , Paton David J. , Linchongsubongkoch Wilai , Mackay David , Oh Yooni , Aunpomma Dilok , Gubbins Simon , Saeki Takakiyo
Published: May 2011

Forty peptides were synthesized corresponding to hydrophilic clusters of amino acids within the sequences of foot-and-mouth disease virus (FMDV) nonstructural proteins (NSP). Six peptides were studied in more detail and the most promising, a 2B peptide, was evaluated in enzyme-linked immunosorbent assay (ELISA) using sera from naïve, vaccinated, and vaccinated-and-challenged cattle as well as bovine sera from field outbreaks. The performance of the new NSP peptide ELISA was compared to that of 4 commercial NSP ELISA kits. Antibody to 2B was detectable from the end of the first week to the second week after infection in most of the nonvaccinated animals and by the second to third week in vaccinated-and-challenged animals. The sensitivity of the 2B peptide ELISA was comparable to the 3ABC Ceditest ( Ceditest® FMDV-NS, Cedi Diagnostics B.V.; Chung et al., 2002). With some modification and further validation, this 2B test could be useful as a screening or conformational NSP test in postvaccination surveillance for FMD.
Transmission of Chronic Wasting Disease of Mule Deer to Suffolk Sheep following Intracerebral Inoculation
Authors: Hamir Amir N. , Kunkle Robert A. , Cutlip Randall C. , Miller Janice M. , Williams Elizabeth S. , Richt Juergen A.
Published: May 2011

To determine the transmissibility of chronic wasting disease (CWD) to sheep, 8 Suffolk lambs of various prion protein genotypes (4 ARQ/ARR, 3 ARQ/ARQ, 1 ARQ/VRQ at codons 136, 154, and 171, respectively) were inoculated intracerebrally with brain suspension from mule deer with CWD (CWDmd). Two other lambs were kept as noninoculated controls. Within 36 months postinoculation (MPI), 2 inoculated animals became sick and were euthanized. Only 1 sheep (euthanized at 35 MPI) showed clinical signs that were consistent with those described for scrapie. Microscopic lesions of spongiform encephalopathy (SE) were only seen in this sheep, and its tissues were determined to be positive for the abnormal prion protein (PrPres) by immunohistochemistry and Western blot. Three other inoculated sheep were euthanized (36 to 60 MPI) because of conditions unrelated to TSE. The 3 remaining inoculated sheep and the 2 control sheep did not have clinical signs of disease at the termination of the study (72 MPI) and were euthanized. Of the 3 remaining inoculated sheep, 1 was found to have SE, and its tissues were positive for PrPres. The sheep with clinical prion disease (euthanized at 35 MPI) was of the heterozygous genotype (ARQ/VRQ), and the sheep with subclinical disease (euthanized at 72 MPH) was of the homozygous ARQ/ARQ genotype. These findings demonstrate that transmission of the CWDmd agent to sheep via the intracerebral route is possible. Interestingly, the host genotype may play a notable part in successful transmission and incubation period of CWDmd.
Aplasia Cutis Congenita (Epitheliogenesis Imperfecta) in Swine: Observations from a Large Breeding Herd
Authors: Benoit-Biancamano Marie-Odile , Drolet Richard , D'Allaire Sylvie
Published: May 2011

Epitheliogenesis imperfecta has been reported in several animal species, and its inheritance is suspected to be autosomal recessive. This term has been used to describe two different diseases, namely epidermolysis bullosa and aplasia cutis congenita, which are both grossly characterized by an absence of epidermis or mucosal epithelium and are most frequently reported on the distal limbs and oral cavity. Epitheliogenesis imperfecta has been described in swine, but the literature on the subject is scarce. To better characterize this condition, 70 piglets with congenital skin defects macroscopically compatible with epitheliogenesis imperfecta were examined. In all but 1 case, only 1 piglet per litter was affected. Of the affected piglets, 65 (93%) were male, suggesting a sex-related problem. More than half of the piglets had multiple skin lesions. All defects were located on the caudal half of the body, and none was found in the oral cavity. Most lesions were characterized by an absence of epidermis and part of the dermis and adnexae. Adnexal dysplasia was also observed at several sites, both with and without epitheliogenesis imperfecta, suggesting a developmental problem. Fluid-filled, congenital subcutaneous bullae were noted grossly on 7 piglets; their relationship, if any, with epitheliogenesis imperfecta remains unknown. As the term epitheliogenesis imperfecta has been used in cases of epidermolysis bullosa, the term aplasia cutis congenita seems to be more appropriate to describe these lesions in swine.
Freezing Complete Polymerase Chain Reaction Master Mix Reagents for Routine Molecular Diagnostics
Authors: West Donna Marie , Sawyer Jason
Published: May 2011

The potential of storing complete frozen real-time polymerase chain reactions (PCRs) and real-time reverse transcription PCRs (RT-PCRs), which require only thawing and the addition of template nucleic acid before PCR cycling, was examined. Master mixes containing all necessary reagents at working concentration (except template nucleic acid) were aliquoted into single-reaction volumes and stored at −70°C for periods of up to 8 months. Reactions were removed from storage and nucleic acid template was added and amplified using different real-time PCR instruments. Threshold cycle values were used to monitor changes in assay performance after storage. Results for hybridization probe and TaqMan probe assays showed that freezing complete real-time PCR and RT-PCR reaction mixes was possible without deterioration in assay performance. This approach has advantages for routine molecular diagnostics in areas such as convenience, test consistency, quality control, and ease of use by nonspecialist staff.
Comparison of Direct Fluorescent Antibody Staining and Real-Time Polymerase Chain Reaction for the Detection of Borrelia Burgdorferi in Ixodes Scapularis Ticks
Authors: Gaumond Gwenn , Tyropolis Allison , Grodzicki Sarah , Bushmich Sandra
Published: May 2011

Borrelia burgdorferi, the agent responsible for causing Lyme disease in humans and animals, is transmitted via the bite of infected Ixodes spp. ticks. Ticks removed from humans and animals are routinely tested by diagnostic laboratories to determine if they are infected with these bacteria. The objective of this study was to compare the efficacy of 2 commonly used methods, direct fluorescent antibody staining and real-time polymerase chain reaction (PCR), for the detection of B. burgdorferi in Ixodes scapularis ticks. One hundred and twenty-seven adult I. scapularis ticks collected in Connecticut, a Lyme disease endemic area, were tested, and results were compared. Results showed 24.8% ticks tested positive for Borrelia spp. by fluorescent antibody testing and 32.5% ticks were positive for B. burgdorferi by real-time PCR testing. When ticks were grouped into categories by level of engorgement (unengorged, partially engorged, and fully engorged), 95% of unengorged ticks, 90.5% of partially engorged, and 86.8% of engorged ticks tested were in agreement. Ten of the 127 ticks examined were too dehydrated to be tested by the fluorescent antibody technique; half of these tested positive by PCR. Real-time PCR appears to be the better of these 2 methods for the diagnosis of this bacterial infection in I. scapularis ticks.
Comparison of Three DNA Preparation Methods for Real-Time Polymerase Chain Reaction Confirmation of Mycobacterium Avium subsp. Paratuberculosis Growth in an Automated Broth Culture System
Authors: Sweeney Raymond W. , Whitlock Robert H. , McAdams Susan C.
Published: May 2011

Three methods of harvesting DNA from broth culture tubes for quantitative real-time polymerase chain reaction (qrtPCR) confirmation of Mycobacterium avium subsp. paratuberculosis (MAP) were evaluated. A commercial DNA extraction kit, the boil method (boiling for 5 minutes), or direct addition of broth culture media to the PCR reaction mix were tested. Samples were evaluated at 8 or 11 days of incubation and at the time of instrument-signal culture-positive. In total, when tested at time to instrument signal positive, 10/10 (100%) of samples extracted by the commercial method were positive on qrtPCR, whereas 9/10 (90%) were positive after the boil method, and 6/10 (60%) were positive after the direct method. Increased volumes of egg-yolk emulsion added to the culture tubes prolonged the number of cycles to threshold positive for the samples that were not subjected to commercial extraction or boiling. Samples were not reliably positive when tested at 8 or 11 days of incubation. The boil method appears to represent a reasonable time- and money-saving method to harvest DNA for qrtPCR confirmation of MAP in broth culture at time to instrument signal positive.
Cyclooxygenase-2 Expression in Feline Pancreatic Adenocarcinomas
Authors: Newman Shelley Joy , Mrkonjich Ladonna
Published: May 2011

Cyclooxygenase-2 (COX-II) is an inducible enzyme that is responsible for the production of prostaglandin E2 (PGE2), which is often upregulated in neoplastic conditions. Expression of COX-II is documented in the majority of human pancreatic adenocarcinomas and in many epithelial neoplasms in humans and animals. The purpose of this study was to assess a series of feline pancreatic adenocarcinomas for the expression of COX-II. Eight feline pancreatic adenocarcinomas (5 poorly differentiated ductular variants and 3 well-differentiated acinar variants) were included. Immunohistochemical staining showed that COX-II was expressed in 2 (both poorly differentiated ductular variants) of the 8 neoplasms (25%). Approximately 10% of the epithelial cells from these 2 neoplasms expressed intense cytoplasmic staining. However, because feline pancreatic adenocarcinoma does not appear to consistently express COX-II, it is not a useful prognostic indicator for this group of feline neoplasma. In addition, COX-II inhibitors are not likely to be effective therapeutics for cats with this neoplasm.
Hyperandrogenism from an Ovarian Interstitial-Cell Tumor in an Alpaca
Authors: Gilbert Rosanne , Kutzler Michelle , Valentine Beth A. , Semevolos Stacy
Published: May 2011

An 8-year-old intact female Huacaya alpaca ( Lama pacos) was presented for recent development of male behavior. Serum testosterone concentration was determined to be 969.1 pg/ml by using radioimmunoassay, while the range in 33 healthy female adult intact alpacas was 11.7–62.1 pg/ml. An ovarian mass was suspected, and an exploratory laparotomy was performed. A tan mass was present on the left ovary. Histologically, the mass was composed of closely packed, plump, polygonal cells with central round nuclei with granular chromatin and abundant eosinophilic finely granular to vesiculate cytoplasm. An ovarian benign interstitial (Leydig) cell tumor was diagnosed.
Halicephalobus Gingivalis-Associated Meningoencephalitis in a Thoroughbred Foal
Authors: Bryant Uneeda K. , Lyons Eugene T. , Bain Fairfield T. , Hong Chuen B.
Published: May 2011

A 13-week-old Thoroughbred colt from central Kentucky was euthanized after an acute onset of ataxia, blindness, head tremors, leaning to the right, recumbency, and seizures. Microscopically, there was a verminous meningoencephalitis characterized by an eosinophilic and granulomatous inflammatory reaction primarily affecting the cerebellum. Dispersed within regions of inflammation were numerous cross and longitudinal sections of intact and degenerative small nematodes. The nematodes had dorsoflexed ovaries and ventroflexed vulvas, which are distinguishing features of Halicephalobus gingivalis. Intact nematodes, compatible with H. gingivalis, also were recovered and identified from portions of the brain that had been frozen for 5-week post-necropsy examination via tissue maceration and additional laboratory techniques.
Systemic Blastomycosis in a Horse
Authors: Wilson Julia H. , Olson Erik J. , Haugen Edward W. , Hunt Luanne M. , Johnson Jennifer L. , Hayden David W.
Published: May 2011

Progressive multisystemic disease caused by Blastomyces dermatitidis was diagnosed in a 17-year-old Quarter horse broodmare. The mare had been treated unsuccessfully with antibiotics for mastitis 3 months postpartum. The disease progressed to exudative cutaneous lesions affecting the ventrum, pectoral region, and limbs accompanied by weight loss across several months. Yeast bodies were observed in swabs of the cutaneous exudate, suggesting a clinical diagnosis of blastomycosis. Following referral, pleural effusion, cavitated lung lesions, and hyperproteinemia were identified, and the mare was euthanized because of poor prognosis. Necropsy revealed extensive pyogranulomas in the mammary gland, skin, subcutaneous tissues, and lungs, accompanied by thrombi in major blood vessels of the lungs and hind limbs. Histologically, pyogranulomatous inflammation was evident in many tissues, and fungal organisms were seen in sections of mammary gland, skin, subcutis, pericardium, and lung. Blastomyces dermatitidis was cultured from mammary tissue, lungs, lymph node, and an inguinal abscess. Although blastomycosis is endemic in the area of origin of the mare in northwestern Wisconsin, the disease is extremely rare in horses and hence easily misdiagnosed. Unique features of this case included the extent of mammary gland involvement and the presence of thrombi in multiple sites.
Spontaneous Subcutaneous Myxosarcoma in a Captive European Hedgehog (Erinsceus Europaeus)
Authors: Singh Kuldeep , Blas-Machado Uriel , Cooper Emily J. , Caseltine Shannon L. , Nordhausen Robert
Published: May 2011

A surgically excised biopsy representing a subcutaneous mass on the left side of the neck from a 3-year-old female European hedgehog ( Erinsceus europaeus) was presented. Spontaneous myxosarcoma was diagnosed based on histological, immunohistochemical, and ultrastructural characteristics. The neoplasm grossly consisted of a firm, pale, multilobulated mass with a characteristic clear gelatinous fluid. Histologically, the neoplasm was nonencapsulated and composed of pleomorphic stellate or spindle-shaped vimentin and periodic acid–Schiff-positive cells arranged in loose sheets and occasionally whorls. The neoplastic cells were suspended in Alcian blue–positive stroma and contained infrequent mitotic figures. Evidence of a viral etiology was not detected using electron microscopy and polymerase chain reaction. This is the first case report of a myxosarcoma in a captive European hedgehog.
Augmentation of Secreted and Intracellular Gamma Interferon following Johnin Purified Protein Derivative Sensitization of Cows Naturally Infected with Mycobacterium Avium Subsp. Paratuberculosis
Authors: Stabel Judith R. , Kimura Kayoko , Robbe-Austerman Suelee
Published: May 2011

Measurement of secreted interferon (IFN)-γ has proven to be a valuable tool for the detection of animals infected with mycobacterial pathogens, including Mycobacterium avium subsp. paratuberculosis. Previous reports have suggested that tuberculin skin testing can influence the performance of the IFN-γ assay. In the present study, healthy noninfected cows, and cows subclinically and clinically infected with M. paratuberculosis were administered an intradermal injection of johnin purified protein derivative (JPPD) and effects on secreted and intracellular IFN-γ were observed. Intradermal injection resulted in significant increases in secreted IFN-γ for subclinically infected cows after stimulation of peripheral blood mononuclear cells (PBMC) with concanavalin A or M. paratuberculosis antigen preparations (whole-cell sonicate and JPPD) on days 7 and 10 postinjection. Intracellular IFN-γ was increased after intradermal injection in total PBMC for all treatment groups and was higher within CD4+ and CD8+ subpopulations for infected cows compared to healthy controls throughout the study. When T-cell populations were further defined by CD45RO expression, intracellular IFN-γ was higher within CD8+/CD45RO+ lymphocytes compared to CD4+/CD45RO+ cells for subclinically and clinically infected cows but similar within these subpopulations for healthy controls. These results indicate that intradermal sensitization of cows in the subclinical stage of infection will upregulate expression of IFN-γ, enhancing the sensitivity of this assay. In addition, CD8+ lymphocytes appear to play an important role as a mediator of M. paratuberculosis infection in naturally exposed cattle.
A Prospective, Case Control Study Evaluating the Association between Clostridium Difficile Toxins in the Colon of Neonatal Swine and Gross and Microscopic Lesions
Authors: Yaeger Michael J. , Kinyon Joann M. , Songer J. Glenn
Published: May 2011

Clostridium difficile infection in swine has most often been described in suckling pigs, where it has been associated with mesocolonic edema and typhlocolitis. This prospective study was designed to assess the correlation between the presence of C. difficile toxins (TCd) in the colon contents of neonatal pigs and a number of parameters, including gross evidence of diarrhea, mesocoloninc edema, typhlitis, and colitis. C. difficile was isolated from 51% (66/129) of large intestines and TCd was detected in the colon contents of 50% (65/129) of the piglets. Fifty-eight percent (38/65) of TCd-positive piglets had normal to pelleted colon and rectal contents, whereas 75% (48/64) of TCd-negative pigs had gross evidence of diarrhea. Clostridium difficile toxin-positive animals were significantly more likely to have normal to pelleted feces. Edema of the mesocolon was observed in 38/65 (59%) of TCd-positive piglets. Because a high number of TCd-positive piglets (41%) lacked edema of the mesocolon and a high number of TCd-negative pigs had mesocolonic edema (51%), a statistically significant association between TCd and mesocolonic edema was not identified. Seventy-five percent (49/65) of TCd-positive piglets had colitis and 47/65 (72%) had typhlitis. The association between TCd and both colitis and typhlitis was statistically significant. Apparently healthy piglets were obtained from 5 separate sites. Because TCd was detected in the colon contents of 23/29 (79%) apparently healthy piglets obtained from 5 separate sites, and 70% of TCd-positive control pigs had colitis, C. difficile may represent an important subclinical issue in neonatal swine.
Epidemiological Investigation of the Prevalence and Features of Postweaning Multisystemic Wasting Syndrome in Japan
Authors: Kawashima Kenji , Katsuda Ken , Tsunemitsu Hiroshi
Published: May 2011

To investigate the prevalence and features of postweaning multisystemic wasting syndrome (PMWS) in Japan, an epidemiological study was conducted in 692 weaned pigs with various clinical signs, commonly including wasting or weight loss, collected from 129 swine farms between 2000 and 2003. The presence of PMWS was diagnosed by the detection of characteristic histological lesions and moderate to large amounts of porcine circovirus type 2 (PCV2) antigen within the lesions in multiple lymphoid tissues. Postweaning multisystemic wasting syndrome was positive in 23.4% of pigs (162/692) over the course of the study, and occurred in 50.4% of the farms (65/129). Mortality in 30–120-day-old pigs in the farms positive for PMWS varied from 0.1 to 32.0%. No significant difference in mortality was seen between PMWS-positive and -negative farms ( P = 0.1). However, mortality was significantly higher in the PMWS-positive farms where PMWS was diagnosed in more than 50% of the pigs examined compared to farms negative for PMWS ( P = 0.02). These findings indicate that PMWS has spread widely in Japan. Moreover it may exist in variable forms in swine farms, including an epidemic form or a subtle endemic or sporadic form. A case-control study suggested that risk factors for the occurrence of PMWS include porcine reproductive and respiratory syndrome (PRRS) pneumonias and Mycoplasma hyorhinis infection.
Development of a Real-Time Polymerase Chain Reaction Assay for Rapid Diagnosis of Neuropathogenic Strains of Equine Herpesvirus-1
Authors: Allen George P.
Published: May 2011

This communication reports the development and performance assessment of a rapid diagnostic test for identifying horses actively infected with the neurovirulent pathotype of equine herpesvirus-1 (EHV-1). The test is a real-time polymerase chain reaction (PCR)-based assay that uses EHV-1 pathotype-specific TaqMan® reporter probes for discrimination between neuropathogenic and non-neuropathogenic strains of EHV-1 in equine blood or nasal swabs. The diagnostic performance of the new technique was evaluated by testing specimens collected from 234 horses involved in recent outbreaks of EHV-1 myeloencephalopathy at three separate thoroughbred racetracks and one large riding/boarding stable. Side-by-side comparison of the EHV-1 pathotyping results yielded by the new single-step, PCR-based allelic discrimination technique (24-hour turn-around-time) with those generated by a multi-step, conventional nested PCR followed by nucleotide sequencing of the amplified DNA (4-day turn-around-time) revealed complete agreement between the 2 test methods. The ability to rapidly identify horses infected with neuropathogenic strains of EHV-1 using a single-step, PCR-based method has significant implications for future diagnostic evaluation of suspect animals.
Development of an Assay to Determine Single Nucleotide Polymorphisms in the Prion Gene for the Genetic Diagnosis of Relative Susceptibility to Classical Scrapie in Sheep
Authors: Johnson Mary Lynn , Evoniuk Jessica M. , Stoltenow Charles L. , O'Rourke Katherine I. , Redmer Dale A.
Published: May 2011

The objective of this study was to develop a reliable Taqman® 5′ Nuclease Assay for genotyping sheep for scrapie susceptibility. The sheep prion gene contains 2 single nucleotide olymorphisms (SNPs) that may mediate resistance to classical scrapie, one at codon 136, alanine (A) or valine (V), and another at codon 171, arginine (R) or glutamine (Q). The R allele appears to confer resistance to classical scrapie, with the AA136 RR171 genotype the most resistant to scrapie and QR171 only rarely infected in the US sheep population. The Assays by Design® protocol was used for development of probes and rimers for codon 136 and Primer Express® for codon 171. Commercially available kits were used to isolate genomic DNA from blood or muscle. For validation, 70 SNP determinations for each codon were compared to commercial testing with an error rate of less than 1%. Then, 935 samples from blood ( n = 818) and muscle ( n = 117) were tested for both codons with 928 successful determinations and only 7 samples (<1% of total samples) that needed repeating. Genotypes were AA QQ ( n = 102; 11.0%), AV QQ ( n = 28; 3.0%), AA QR ( n = 396; 42.7%), AV QR (n = 54; 5.8%), and AA RR ( n = 348; 37.5%). Thus, 86% of the sheep tested ( n = 798) contained R at codon 171 and were expected to be scrapie-resistant. This new Taqman® 5′ Nuclease SNP genotyping assay is accurate, easy to perform, and useful in the study of classical scrapie in sheep and its prevention through selective breeding programs to eliminate highly susceptible animals.
Comparative Serological Study of Leptospira Serovar Hardjo Genotypes for Use in the Microscopic Agglutination Test
Authors: Hotka Morgan L. , Wilson Mark A. , Anderson Tammy M. , Tichenor Corrie L. , Miller David A.
Published: May 2011

A comparative serological study was conducted using the Leptospira microscopic agglutination test (MAT). Genotypes hardjoprajitno (HP), hardjo-bovis A (HA), and hardjo-bovis B (HB) were compared to determine which best detects hardjo antibody in cattle serum. A total of 2,431 cattle sera were tested. Sera were collected from 4 geographic regions of the United States. Samples were obtained without knowledge of breed, age, vaccination history, or herd health status. Of the sera collected, 60.7% (1,475) were negative at the 1:100 dilution for all three genotypes. Serological reactivity at the 1:100 dilution was identified in 956 (39.3%) of the sera tested. Considering the 956 positive sera, 941 (98.4%) reacted to HP, whereas the remaining 15 sera (1.6%) reacted to only HA and/or HB. A total of 394/941 (41.9%) HP positive sera failed to react to HA or HB. The results of this study support the conclusion that HP antigen was most sensitive in detection of hardjo antibody.
Diagnosis of Paratuberculosis by Fecal Culture and ELISA on Milk and Serum Samples in Two Types of Chilean Dairy Goat Herds
Authors: Salgado Miguel , Kruze Juan , Collins Michael T.
Published: May 2011

Fecal culture has been the primary method used to diagnose paratuberculosis in goats. It is laborious, slow, and expensive. Validation of enzyme-linked immunosorbent assays (ELISAs) on milk samples could make paratuberculosis testing more widely available for goat farmers. The aim of this study was to determine the accuracy of serum and milk ELISAs for paratuberculosis, relative to fecal culture, in Chilean dairy goats. Eight dairy goat herds were selected. Feces, blood, and milk samples were collected from all female goats >2 years old. Fecal samples were cultured using Herrold egg yolk medium with mycobactin J and antibiotics. Serum and milk samples were tested using a commercial ELISA kit for Mycobacterium avium subsp. paratuberculosis antibody detection. A total of 383 goats were tested by ELISA and fecal culture. The sensitivity of ELISA on serum and milk relative to fecal culture was 74.3% (95% CI: 59.8-88.8) and 60% (95% CI: 43.8-76.2), respectively. The corresponding values for ELISA specificity based on the percentage of non- M. avium subsp. paratuberculosis-infected goats testing ELISA-negative were 98.6% (95% CI: 96.6-100) and 99.3% (95% CI: 97.9-100) on serum and milk, respectively. Proportions of positive results for serum and fecal samples were significantly different, whereas the proportions of positive results for milk and fecal samples were not significantly different. The milk ELISA had a moderate level of agreement with fecal culture results ( Kappa = 0.57). The paratuberculosis ELISA on goat milk samples may be a cost-effective, accurate alternative to fecal culture.
Bovine Cardiac Troponin T is Not Accurately Quantified with a Common Human Clinical Immunoassay
Authors: Willis Monte S. , Snyder Jennifer A. , Poppenga Robert H. , Grenache David G.
Published: May 2011

The detection of myocardial injury in cattle caused by the ingestion of cardiotoxic compounds or cardiac diseases would be facilitated by the availability of a rapid and specific quantitative serum assay for cardiac troponins. Therefore, the accuracy of the only cardiac troponin T (cTnT) immunoassay to receive approval by the US Food and Drug Administration for the measurement of cTnT in human serum was evaluated to quantify the protein in bovine serum. Recovery experiments were performed by the addition of purified bovine cTnT to normal bovine serum. Cardiac troponin T was quantified using an immunoassay commonly used for the measurement of cTnT in human serum. The immunoassay demonstrated a well correlated ( r = 0.99) and linear dose-dependent response to bovine cTnT but with poor accuracy (slope = 0.024; 95% CI = 0.018 to 0.030). The mean recovery of bovine cTnT was 2.4% across a concentration range of 10 ng/ml to 1,000 ng/ml. These studies demonstrate that a commonly used immunoassay for the measurement of cTnT in human serum demonstrates poor accuracy for the quantification of bovine cTnT.