bluuebunny/crossref_metadata_2025 huggingface dataset:
Pathogenesis of Liver Lesions Caused by Experimental Infection with Piscirickettsia Salmonis in Juvenile Atlantic Salmon, Salmo Salar L
Authors: Almendras Felipe E. , Fuentealba I. Carmen , Markham R F. Frederick , Speare David J.
Published: May 2011

Piscirickettsia salmonis, the etiologic agent of salmonid rickettsial septicemia (SRS), or piscirickettsiosis, causes substantial economic losses to the salmon industry. The pathogenesis of the disease has not been fully characterized. The aim of this study is to describe the hepatic lesions associated with experimental P. salmonis infection in Atlantic salmon juveniles. Fish were maintained in fresh water and inoculated intraperitoneally (IP), orally, or on the gill surface with P. salmonis. A group of uninfected fish was kept as control. Liver samples from 5 fish in each inoculated group and 3 controls were collected weekly and processed for histological and immunohistochemical examination. Thickening of the liver capsule by inflammatory cells was a characteristic histologic feature of IP inoculated fish. Three weeks post-IP inoculation, 8 fish had died and 2 fish were sampled. Histological changes at this time consisted of vasculitis, presence of fibrin thrombi, vacuolated hepatocytes and focal areas of necrosis. Leukocytes containing intracytoplasmic basophilic microorganisms were seen within hepatic sinusoids. Vasculitis and intracytoplasmic vacuoles were prominent features in fish inoculated orally and on the gill surface. The presence of P. salmonis within hepatocellular vacuoles, endothelial cells, and leucocytes was confirmed by immunohistochemistry. The intracellular location of P. salmonis and the vascular damage seen in infected fish are characteristic of rickettsial infections. Histological lesions induced by experimental infection with P. salmonis using the oral and gill surface routes were similar to those observed in natural outbreaks of piscirickettsiosis. The tropism of P. salmonis for endothelial cells explains the vascular lesions observed in SRS, whereas hepatic lesions are due to ischemic necrosis and direct injury by intracytoplasmic organisms.
Bovine Viral Diarrhea Virus in New World Camelids
Authors: Belknap Ellen B. , Collins James K. , Larsen R. Scott , Conrad Kevin P.
Published: May 2011

A virus known to cause multiple problems in cattle, bovine viral diarrhea virus, was isolated from 3 different cases in New World camelids. Virus isolation, immunoperoxidase staining, and fluourescent antibody staining were used to detect the virus. The herds involved were screened for antibody titers to bovine viral diarrhea and virus isolation from the buffy coat. Bovine viral diarrhea virus should be considered as a cause of death in young and old New World camelids.
Immunohistochemical Detection of Avian Pneumovirus in Formalin-Fixed Tissues
Authors: Jirjis Faris F. , Noll Sally L. , Halvorson David A. , Nagaraja Kakambi V. , Shaw Daniel P.
Published: May 2011

An immunohistochemical staining technique (IHC) was developed to detect avian pneumovirus (APV) antigen in formalin-fixed, paraffin-embedded tissue sections using streptavidin-biotin immunoperoxidase staining. Samples of nasal turbinates and infraorbital sinuses were collected from 4-week-old poults experimentally inoculated with APV and from older turkeys infected during naturally occurring outbreaks of avian pneumovirus. Tissue was fixed in 10% buffered neutral formalin, embedded in paraffin, sectioned and stained. Inflammatory changes were observed microscopically in the mucosa and submucosa of the nasal turbinates and infraorbital sinuses of both experimentally inoculated poults and naturally infected birds. Viral antigen was detected by IHC in the ciliated epithelial cells of nasal turbinates and infraorbital sinuses.
Prevalence of the Enteroaggregative Escherichia Coli Heat-Stable Enterotoxin 1 Gene and Its Relationship with Fimbrial and Enterotoxin Genes in E. Coli Isolated from Diarrheic Piglets
Authors: Choi Changsun , Kwon Donghyok , Chae Chanhee
Published: May 2011

A total of 720 Escherichia coli strains isolated from diarrheic piglets on 756 swine farms were screened for the presence of the enteroaggregative E. coli heat-stable enterotoxin 1 (EAST1) gene by polymerase chain reaction (PCR). Escherichia coli strains that carried EAST1 genes were also tested by PCR for the presence of 4 fimbriae (F4, F5, F6, F41), 2 heat-stable enterotoxins (STa and STb), and 1 heat-labile enterotoxin (LT) gene. One hundred sixty-four (22.7%) of the 720 E. coli isolates carried genes for EAST1. Of these 164 isolates, 62 (37.8%) carried EAST1 genes only, 11 (6.7%) carried genes for at least 1 of the fimbrial adhesins, 51 (31.1%) carried genes for at least 1 of the enterotoxins, and 40 (23.8%) carried genes for at least 1 of the fimbrial adhesins and enterotoxins. Forty-six percnt of strains that carried EAST1 genes carried STa genes, and 16% of strains that carried EAST1 genes carried F4. The isolation rate of enterotoxigenic E. coli strains carrying genes for EAST1 gene was 63%. The 6 major genotypes observed in this study (in decreasing order) were EAST1+, EAST1+STa+, EAST1+STa+STb+, EAST1+STa+F5+, EAST1+STa+F4+, and EAST1+STb+F4+. EAST1 is widely prevalent among diarrheagenic strains of E. coli and may represent an important virulence determinant in the pathogenesis of enteric colibacillosis of preweaned pigs.
A Comparison of Diagnostic Assays for the Detection of Type a Swine Influenza Virus from Nasal Swabs and Lungs
Authors: Swenson Sabrina L. , Vincent Lani L. , Lute Brandi M. , Janke Bruce H. , Lechtenberg Kelly F. , Landgraf John G. , Schmitt Beverly J. , Kinker David R. , McMillen Janis K.
Published: May 2011

Nasal swabs and lung samples from pigs experimentally infected with H1N1 swine influenza virus (SIV) were examined for the presence of SIV by the indirect fluorescent antibody assay, immunohistochemistry, cell culture virus isolation, egg inoculation, and 2 human enzyme immunoassays (membrane enzyme immu-noassay, microwell enzyme immunoassay). Egg inoculation was considered to be the gold standard for assay evaluation. The 2 human enzyme immunoassays (EIA) and egg inoculation agreed 100% for the prechallenge nasal swabs. Agreement on SIV identification in nasal swabs with egg inoculation following challenge was considered to be good to excellent for membrane EIA (kappa = 0.85) and microwell EIA (kappa = 0.86). Agreement on SIV identification in lung tissue with egg inoculation following challenge was good to excellent for membrane EIA (kappa = 0.75), fair for microwell EIA, fluorescent antibody, and cell culture virus isolation (kappa = 0.48, 0.64, 0.62, respectively), and poor for immunohistochemistry (kappa = 0.36). No assay was 100% accurate, including the “gold standard,” egg inoculation. In light of this information, it is important to consider clinical signs of disease and a thorough herd history in conjunction with diagnostic results to make a diagnosis of SIV infection.
Maximal Predicted Duration of Viremia in Bluetongue Virus—Infected Cattle
Authors: Singer Randall S. , MacLachlan N. James , Carpenter Tim E.
Published: May 2011

Central to the development of rational trade policies pertaining to bluetongue virus (BTV) infection is determination of the risk posed by ruminants previously exposed to the virus. Precise determination of the maximal duration of infectious viremia is essential to the development of an appropriate quarantine period prior to movement of animals from BTV-endemic to BTV-free regions. The objective of this study was to predict the duration of detectable viremia in BTV-infected cattle using a probabilistic modeling analysis of existing data. Data on the duration of detectable viremia in cattle were obtained from previously published studies. Data sets were created from a large field study of naturally infected cattle in Australia and from experimental infections of cattle with Australian and US serotypes of BTV. Probability distributions were fitted to the pooled empirical data, and the 3 probability distributions that provided the best fit to the data were the gamma, Weibull, and lognormal probability distributions. These asymmetric probability distributions are often well suited for decay processes, such as the time to termination of detectable viremia. The analyses indicated a > 99% probability of detectable BTV viremia ceasing after #x2264; 9 weeks of infection in adult cattle and after a slightly longer interval in BTV-infected, colostrum-deprived newborn calves.
Tissue Distribution and Genetic Typing of Porcine Circoviruses in Pigs with Naturally Occurring Congenital Tremors
Authors: Stevenson Gregory W. , Kiupel Matti , Mittal Suresh K. , Choi Jiwon , Latimer Kenneth S. , Kanitz Charles L.
Published: May 2011

Congenital tremors (CT) type A2 is associated with porcine circovirus (PCV) and deficient and abnormal myelin. The aim of this study was to determine the tissue distribution and genetic type of PCV in 1–2-day-old pigs with naturally occurring CT type A2 using in situ hybridization, polymerase chain reaction (PCR), and indirect fluorescent antibody tests on frozen tissue sections. CT-affected and clinically normal pigs were selected from 4 farms in the midwestern USA that were undergoing outbreaks of CT type A2. All CT and most normal pigs were infected with PCV. PCV was widely distributed in tissues of infected pigs and was most common in tissues of the central nervous system and liver. In all infected pigs, there were more PCV-infected cells in brain and spinal cord than in nonneural tissues. CT pigs had many more PCV-infected cells in the brain and spinal cord than did clinically normal pigs because of a more diffuse distribution and a larger proportion of infected cells. The cells most commonly infected with PCV in brain and spinal cord were large neurons. In nonneural tissues, macrophages were the most frequent cell type infected. PCR analysis demonstrated only PCV type 2 and not PCV type 1 in all PCV-infected pigs on all 4 farms.
Incidence of Polysaccharide Storage Myopathy in Draft Horse-Related Breeds: A Necropsy Study of 37 Horses and a Mule
Authors: Valentine Beth A. , Habecker Perry L. , Patterson Jon S. , Njaa Bradley L. , Shapiro Janet , Holshuh H.J. , Bildfell Robert J. , Bird Karyn E.
Published: May 2011

Skeletal muscle samples from 38 draft horse–related animals 1–23 years of age were evaluated for evidence of aggregates of glycogen and complex polysaccharide characteristic of equine polysaccharide storage myopathy (EPSSM). Cardiac muscle from 12 of these horses was also examined. Antemortem serum levels of creatine kinase (CK) and aspartate aminotransferase (AST) from 9 horses with EPSSM and 5 horses without EPSSM were compared. Skeletal muscle from 17 horses contained inclusions of periodic acid–Schiff (PAS)-positive, amylase-resistant complex polysaccharide. Similar inclusions were also present in the cardiac muscle of 1 horse. A vacuolar myopathy with aggregates of PAS-positive, amylase-sensitive glycogen was seen in 8 other horses, and these findings are also considered diagnostic for EPSSM. Antemortem serum activities of CK and AST were often higher in EPSSM horses than in horses without EPSSM. Using the presence of amylase-resistant complex polysaccharide as the criterion for diagnosis of EPSSM, the incidence in this population was 45%. Inclusion of horses with aggregates of glycogen but no amylase-resistant complex polysaccharide as representative of the range of pathologic findings in horses with EPSSM resulted in a 66% incidence in this population.
Double in Situ Hybridization for Simultaneous Detection of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus (PCV)
Authors: Sirinarumitr Theerapol , Sorden Steven D. , Morozov Igor , Paul Prem S.
Published: May 2011

A double in situ hybridization method for the simultaneous detection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus (PCV) genomes in the same tissue section was applied to lung tissues from 9 pigs in which PRRSV and PCV coinfection had been previously demonstrated. Paraffin-embedded tissue sections were simultaneously hybridized with a digoxigenin-labeled antisense RNA probe for PRRSV and a fluorescein-labeled antisense RNA probe for PCV, and hybridization was detected with anti-digoxigenin alkaline phosphatase/fast red and anti-fluorescein peroxidase/diaminobenzidine, respectively. PRRSV and PCV genomes were identified in the same pulmonary cell types as reported previously in all 9 pigs. In all pigs, PCV-positive cells outnumbered PRRSV-positive cells. A small proportion of alveolar macrophages contained both PRRSV and PCV genomes.
Usefulness of a Commercially Available Enzyme Immunoassay for Shiga-Like Toxins I and II as a Presumptive Test for the Detection of Escherichia Coli O157:H7 in Cattle Feces
Authors: Hyatt Doreene R. , Galland John C. , Gillespie Jerry R.
Published: May 2011

The performance of a commercially available enzyme immunoassay (EIA) for determining the presence of Shiga toxin I and II in human diarrheal stool samples was evaluated for use as a presumptive test for the presence of Escherichia coli O157:H7 in nondiarrheal bovine fecal samples collected from 10 Kansas cow–calf ranches. The prevalence of E. coli O157:H7 in 2,297 samples, as determined by selective bacterial culture, was 1.6%. The sample prevalence of non- E. coli O157:H7 Shiga toxin–producing bacteria, as detected by the Shiga toxin EIA, was 5.8%. Only 2 of 136 samples that tested positive with the Shiga toxin EIA were positive for E. coli O157:H7 by culture. Compared with bacterial culture, the sensitivity of the Shiga toxin EIA was 5.5% and the specificity was 94.1%. Agreement between the 2 tests, as measured by the kappa statistic, was poor (κ = −;0.002). Although the Shiga toxin EIA was not a good presumptive test for the determination of E. coli O157:H7 in bovine fecal samples because of its low sensitivity (5.5%), it might be a useful test for the detection of Shiga toxin producing non- E. coli O157:H7 organisms in bovine feces.
Sensitivity and Specificity of the Complement Fixation Test for Detection of Cattle Persistently Infected with Anaplasma Marginale
Authors: Bradway Daniel S. , de Echaide Susana Torioni , Knowles Donald P. , Hennager Steven G. , McElwain Terry F.
Published: May 2011

The complement fixation (CF) test commonly is used to identify cattle infected with Anaplasma marginale prior to interstate or international movement. Estimates of the accuracy of the CF test in detecting animals persistently infected with A. marginale vary widely. In this study, the sensitivity and specificity of the CF test for detection of carrier animals was determined using serum from 232 cattle previously defined as A. marginale positive or negative by nested polymerase chain reaction methods and hybridization. Considering results from 2 independent laboratories and interpreting a 1:5 suspect reaction as positive, the best estimate of CF test sensitivity was 20%, with a specificity of 98%. Using a 1:10 cutoff, sensitivity decreased to 14% and specificity increased to 99%. Results of this study indicate that the CF test is ineffective for identifying cattle persistently infected with A. marginale and thus is inadequate for anaplasmosis regulatory and surveillance programs.
An Enzyme-Linked Immunosorbent Assay for the Convenient Serodiagnosis of Contagious Equine Metritis in Mares
Authors: Katz Jonathan , Geer Patricia
Published: May 2011

An enzyme-linked immunosorbent assay (ELISA) was developed for the serodiagnosis of contagious equine metritis (CEM), a sexually transmitted disease caused by Taylorella equigenitalis. Antigen preparation was simple, and antigens derived from both classical and atypical forms of T. equigenitalis enabled detection of antibody responses elicted in horses experimentally exposed to either form of the bacterium. Sera serially obtained from these horses from 0 to 63 days postexposure were tested by the traditional complement fixation test (CFT) for CEM and with the ELISA, using both antigens separately. There was close agreement between CFT and ELISA methodologies during the postexposure time period used to detect CEM serodi-agnostically in regulatory animal health testing programs. Unlike the CFT, which requires an overnight incubation step, the ELISAs are more convenient and can be completed in 3 hours.
Immunohistochemical Detection of Scrapie Prion Proteins in Clinically Normal Sheep in Pennsylvania
Authors: Kim Hyun , O'Rourke Katherine I. , Walter Mark , Purchase H. Graham , Enck John , Shin Tae Kyun
Published: May 2011

Following diagnosis of scrapie in a clinically suspect Suffolk sheep, 7 clinically normal flockmates were purchased by the Pennsylvania Department of Agriculture to determine their scrapie status using an immu-nohistochemical procedure. Two of the 7 euthanized healthy sheep had positive immunohistochemical staining of the prion protein of scrapie (PrP-Sc) in their brains, nictitating membranes, and tonsils. The PrP-Sc was localized in the areas of the brain where, histopathologically, there was neurodegeneration and astrocytosis. The PrP-Sc occurred within germinal centers of the affected nictitating membranes and tonsils and was located in the cytoplasm of the dendrite-like cells, lymphoid cells, and macrophages. These results confirm that immunohistochemical examination of the nictitating membrane can be used as a screen for the presence of scrapie infection in clinically normal sheep at a capable veterinary diagnostic laboratory. In sheep with a PrP-Sc-positive nictitating membrane, the diagnosis of scrapie should be confirmed by histopathology and immunohistochemical examination of the brain following necropsy. Following full validation, immunohistochemistry assays for detection of PrP–Sc in nictitating membrane lymphoid tissues can improve the effectiveness of the scrapie control and eradication program by allowing diagnosis of the disease in sheep before the appearance of clinical signs.
Immunohistochemistry of Ultimobranchial Thyroid Carcinomas in Seven Slaughtered Cows and One Bull
Authors: Harmon Barry G. , Kelley Lynda C.
Published: May 2011

Eight thyroid gland epithelial tumors were found in 7 cows and 1 bull in a retrospective study of thyroid gland lesions in slaughtered cattle. All tumors were classified as ultimobranchial thyroid carcinomas based on morphology and immunohistochemistry. All tumors consisted of solid sheets and nests of polygonal to oval epithelial cells, with more sparsely dispersed colloid-filled follicles. Connective tissue separating nests of epithelial cells varied from delicate fibrovascular stroma to dense collagenous stroma. Fusiform epithelial cells with rare neural fibers and ganglion cells were present in 1 tumor. Cells within solid areas of these tumors were immunoreactive for calcitonin, calcitonin gene-related peptide, neuron-specific enolase, and synaptophysin. Colloid and follicle cells were immunoreactive for thyroglobulin. Few follicle cells also were reactive for calcitonin gene–related peptide. Neoplastic cells invaded the fibrous capsules in all 8 cattle. These tumors represented proliferation of a mixed population of undifferentiated cells, C cells, and thyroid follicular epithelial cells, presumably derived from the thyroid ultimobranchial bodies. These ultimobranchial carcinomas in slaughtered cattle are comparable to ultimobranchial tumors described in dairy bulls and the intermediate type of thyroid gland carcinomas (mixed thyroid medullary carcinomas) described in human beings.
Evaluation of Canine Serum Thyrotropin (TSH) Concentration: Comparison of Three Analytical Procedures
Authors: Marca M. C. , Loste A. , Orden I. , González J. M. , Marsellá J. A.
Published: May 2011

A comparative study of 3 analytical methods (immunoradiometric assay, enzyme immunometric assay, and chemiluminescent immunometric assay) for canine serum thyrotropin (TSH) was performed. Ninety-six dogs were included in the study. The within- and between-run precision was evaluated for each method, and correlations for the results obtained with each method were examined. The best within- and between-run precision was obtained with the chemiluminescent immunometric assay. Satisfactory correlations for the 3 analytical procedures were obtained but varied in relation to serum TSH concentration.
A One-Tube Nested Polymerase Chain Reaction for the Detection of Mycobacterium Bovis in Spiked Milk Samples: An Evaluation of Concentration and Lytic Techniques
Authors: Antognoli M. C. , Salman M. D. , Triantis J. , Hernández J. , Keefe T.
Published: May 2011

The objective of this study was to evaluate the use of a one-tube nested polymerase chain reaction (OTN PCR) with 5 concentration and lytic treatments for the detection of Mycobacterium bovis in experimentally inoculated milk samples (spiked samples). OTN PCR and the following treatments were tested in inoculated samples: 1) centrifugation; 2) C18-carboxypropylbetaine + capture resin 1 + Proteinase K (CB18-CH-PK); 3) centrifugation + capture resin 1 + Proteinase K; 4) centrifugation + capture resin 2 + Proteinase K; and 5) centrifugation + immunomagnetic separation (IMS). The OTN PCR and the 5 treatments were evaluated in 2 different sets of spiked milk samples. One set consisted of 10-fold serial dilutions of a phenol-killed M. bovis in milk to final concentrations ranging from 5 to 50,000 cells/ml of milk. The other set of samples consisted of 2.5 serial dilutions of milk spiked with M. bovis to final concentrations ranging from 20.5 to 5,000 cells/ml of milk. Each treatment was repeated 5 times at each cell concentration. CB18-CH-PK and IMS were significantly more sensitive than other treatments. The lowest detection limit for these techniques was 20–50 cells/ml of spiked milk. The specificity of OTN PCR in this study was high as demonstrated by the lack of DNA amplification products when M. bovis cells were not present in the samples. [The OTN PCR used in conjunction with CB18-CH-PK or IMS could be effectively used as a diagnostic and/or screening test for the detection of M. bovis in milk from herds with bovine tuberculosis.]
Prevalence of Ovine and Bovine Respiratory Syncytial Virus Infections in Cattle Determined with a Synthetic Peptide-Based Immunoassay
Authors: Grubbs Steven T. , Kania Stephen A. , Potgieter L. N. D.
Published: May 2011

Subgroup-specific peptide-based enzyme-linked immunosorbent assays from the G-protein of the ovine and bovine respiratory syncytial virus (RSV), respectively, were used to determine the prevalence of the ovine and bovine subgroup strains of RSV infections in cattle. A total of 1,102 bovine serum samples were obtained from 6 diagnostic laboratories located in the northwestern and the southeastern USA and were tested for antibody to either the bovine or ovine subgroups of RSV. Antibody to viruses from each subgroup was present in samples from each region and all states tested. The Southeast had a higher prevalence of the bovine subgroup strains (69.5%). Then did the Northwest (40.9%). The prevalence of the ovine strain was similar for the two regions (16.7% in the southeast, 14.9% in the northwest). The overall prevalence was 56.6% for the bovine strain and 15.9% for the ovine strain. These results suggest members of the ovine subgroup of RSV circulate in the cattle population but with less frequency than those viruses of the bovine subgroup.
Isolation of Genomic DNA from Feathers
Authors: Bello Natalia , Francino Olga , Sánchez Armand
Published: May 2011

The use of feathers in veterinary clinical practice simplifies the sampling of avian genomic DNA, especially when blood extraction is difficult because of the age or the size of the bird. A rapid and accurate protocol was used to isolate high-quality genomic DNA from feathers. The technique includes a lysis step of the feather quill, which differs in temperature and time of incubation depending on the feather size. Purification of genomic DNA is performed with phenol: chloroform: isoamyl alcohol extraction and ethanol precipitation. This protocol consistently provided significant amounts of high-quality genomic DNA from more than 800 birds belonging to 120 different species. Genomic DNA isolated with this method was used for Southern blotting and also in several polymerase chain reaction systems devoted to sex determination and paternity testing.
Comparison of Two Enzyme-Linked Immunosorbent Assays for Serologic Diagnosis of Paratuberculosis (Johne's Disease) in Cattle Using Different Subspecies Strains of Mycobacterium Avium
Authors: Nielsen Søren Saxmose , Houe Hans , Thamsborg Stig Milan , Bitsch Viggo
Published: May 2011

Serologic diagnosis of bovine paratuberculosis (Johne's disease) with currently available tests may give false-positive results due to cross-reactions with avian and bovine tuberculosis viruses and other infectious agents. Indirect enzyme-linked immunosorbent assays (ELISA) for detection of antibodies against paratuberculosis based on antigens from Mycobacterium avium subsp. avium (A-ELISA) and M. avium subsp. paratuberculosis (P-ELISA) were compared. Despite an expected higher specificity for M. a. paratuberculosis in the P-ELISA, the 2 antigens were equally suitable for demonstration of antibody to M. a. paratuberculosis in cattle. Receiver operating characteristic (ROC) curves was used to demonstrate the possible antigenic relationship. The area under the curve (AUC) was calculated for each of the 2 ROC curves. The AUC for the P-ELISA ROC curve was 0.9197, and the AUC for the A-ELISA ROC curve was 0.9149, demonstrating a negligible difference in efficiency of the 2 tests ( z = 0.182).
Experimentally Induced Cholangiohepatopathy by Dosing Sheep with Fractionated Extracts from Brachiaria Decumbens
Authors: Cruz Cláudio , Driemeier David , Pires Viviane Silva , Schenkel Eloir Paulo
Published: May 2011

Cholangiohepatopathy was induced in 5 lambs by oral administration of extracts from signal grass ( Brachiaria decumbens) in Brazil. Grossly there were pale foci multifocally distributed throughout the hepatic parenchyma in 4 lambs. The microscopic changes, which were similar to those produced by other steroidal sapogenins–containing plants such as Tribulus terrestris and Panicum spp., included multifocal chol-angitis, bile duct proliferation, and the presence of crystals in the biliary system.
Genotypic Prevalence of apxIV in Actinobacillus Pleuropneumoniae Field Isolates
Authors: Cho Wan-Seob , Chae Chanhee
Published: May 2011

A total of 90 Actinobacillus pleuropneumoniae field strains from pigs were serotyped by slide agglutination and analyzed for the presence of the apxIV gene by polymerase chain reaction. Of the 90 isolates serotyped, serotypes 2 (47 isolates) and 5 (25 isolates) were the most common, followed by serotype 6 (10 isolates). Three isolates belonged to serotype 7, and 5 isolates could not be typed. All 90 A. pleuropneumoniae field isolates tested carried the apxIV gene. This gene is species specific rather than serotype specific. Therefore, the ApxIV toxin has potential value for use both in vaccines and in diagnostic tests.
A New Serotype Adenovirus Isolated from a Goat in the United States
Authors: Lehmkuhl H. D. , DeBey B. M. , Cutlip R. C.
Published: May 2011

A virus (T94-0353) isolated from the small intestine of a 3-week-old kid with diarrhea and serous ocular and nasal discharge was identified as an adenovirus based on morphologic and physicochemical characteristics. Neutralization tests and restriction endonuclease analysis comparing the caprine adenovirus with the prototype bovine and ovine adenovirus serotypes and a previously isolated caprine adenovirus showed that the caprine isolate was antigenically distinct, produced a unique restriction pattern compared with currently recognized bovine, caprine, and ovine adenoviruses, and represents a new adenovirus type. The role and significance of naturally acquired adenovirus infection in respiratory and enteric disease in goats has not been established. Isolation of adenovirus from goats with disease coupled with seroepidemiologic and pathogenicity studies will help define the role of the adenoviruses in disease production.
PCR-Based Detection of an Emerging Avian Pneumovirus in US Turkey Flocks
Authors: Dar Arshud M. , Tune Kathy , Munir Shirin , Panigrahy Brundaban , Goyal Sagar M. , Kapur Vivek
Published: May 2011

Avian pneumovirus (APV) or turkey rhinotracheitis virus (TRTV) is an important respiratory pathogen of domesticated poultry in many countries in Europe, Africa, and Asia. Until recently, the United States was considered free of APV. In late 1996, an atypical upper respiratory tract infection appeared in turkey flocks in Colorado and shortly thereafter in turkey flocks in Minnesota. An avian pneumovirus (APV-US) that was serologically distinct from the previously described TRTV was isolated as the primary cause of the new syndrome. The nucleotide sequence of a fragment of the APV-US fusion gene was determined and used to develop a polymerase chain reaction–based assay that specifically detects APV-US viral nucleic acid sequences in RNA extracts of tracheal swabs and turbinate homogenates. The assay is highly sensitive in that it can detect <0.01 TCID50 of APV. The availability of this assay enables the rapid and accurate determination of APV-US in infected poultry flocks.
Spatial and Temporal Clustering of Salmonella Serotypes Isolated from Adult Diarrheic Dairy Cattle in California
Authors: Sato K. , Carpenter T. E. , Case J. T. , Walker R. L.
Published: May 2011

Electronic records from the California Animal Health and Food Safety Laboratory System between January 1, 1991, and June 30, 1998, were reviewed, and data on 593 Salmonella serotypes isolated from feces or intestinal contents of adult dairy cattle with diarrhea were analyzed to determine spatial and temporal clustering. A statistically significant clustering in space was detected with 10 (Typhimurium, Montevideo, Muenster, Anatum, Give, Menhaden, Kentucky, Agona, Derby, and Newport) of the 13 serotypes examined. Significant temporal clustering was also detected with 10 serotypes (Typhimurium, Montevideo, Muenster, Anatum, Give, Newbrunswick, Menhaden, Kentucky, Derby, and Newport). Six serotypes (Anatum, Menhaden, Montevideo, Muenster, Newbrunswick, and Newport) were significantly spatially and temporally clustered. A difference in temporal and spatial distribution patterns of some serotypes associated with diarrhea in adult dairy in California was found. Knowledge of the specific type of clustering, if present, should improve our understanding of the transmission and control of salmonellosis in the field.
Foxtail-Induced Ulcerative Stomatitis Outbreak in a Missouri Stable
Authors: Turnquist S. E. , Ostlund E. N. , Kreeger J. M. , Turk J. R.
Published: May 2011

Twenty of 25 horses in a well-managed Missouri boarding stable were diagnosed with gingivitis/stomatitis. Gross examination of the affected horses revealed varying degrees of gingivitis ranging from mild periodontal swelling to marked swelling and erythema with ulceration and hemorrhage. Fine hair-like material was embedded within the intensely affected areas. Gingival biopsies from 4 affected horses contained pyogranulomatous inflammation with, in some cases, numerous eosinophils and several grass awns in cross and longitudinal section. Numerous foxtail seed heads were identified in hay samples. Examination of the records revealed that all of the affected horses had been fed the suspect hay, with the exception of 1 horse. Although not deliberately fed the suspect hay, this horse did have access to the hay when turned out into the exercise paddock. The lesions resolved following a change in hay source.
An Outbreak of Toxoplasmosis in Farmed Mink (Mustela Vison S.)
Authors: Frank Rodney K.
Published: May 2011

A large Wisconsin mink ( Mustela vison) farm experienced an outbreak of toxoplasmosis in the spring of 1999 following an outbreak of canine distemper during the previous fall. Major clinical signs for pregnant females included reduced feed consumption, abortions, and stillborn kits; kits <3 weeks old experienced ataxia and mortality. Of 7,800 females, 1,976 (26%) lost their entire litter either from abortion or neonatal mortality. Kit mortality from 7 days to 3 weeks of age was 3,300, and overall kit mortality attributed to the toxoplasmosis outbreak was 10,408. Six neonatal mink kits, 12 3-week-old kits, and 2 adult female mink were submitted to the University of Minnesota Veterinary Diagnostic Laboratory for diagnostic workup. Gross postmortem lesions were limited to empty stomachs (12 of 12 kits) and pale livers (4 of 12 kits) for the 3-week-old kits. Major microscopic lesions included interstitial pneumonia, encephalitis, encephalomalacia, and myocarditis. Toxoplasmosis was diagnosed by microscopic lesions, microscopic lesion distribution, and the detection of Toxoplasma gondii tachyzoites by immunohistochemistry. This is the first detailed report of an outbreak of toxoplasmosis in mink in the United States.
Evaluation of a New Sandwich Enzyme-Linked Immunosorbent Assay for Detection of Bovine Viral Diarrhea Virus in Unprocessed Fetal Bovine Serum
Authors: Plavsic Mark Z. , Prodafikas Gilda
Published: May 2011

A new sandwich enzyme-linked immunosorbent assay (S-ELISA) kit that uses raw (unprocessed) fetal bovine serum (FBS) as the testing sample was evaluated for upstream bovine viral diarrhea virus (BVDV) testing. Pooled FBS samples ( n = 84) were tested using the S-ELISA. Thirty serum samples originating from persistently infected (PI) calves that had been confirmed by virus isolation (VI) as BVDV positive and another 30 samples previously confirmed by VI as BVDV negative were also evaluated. Of the 84 field samples, the S-ELISA detected 13 (15.5%) BVDV-positive specimens. When these 13 positive samples were tested by VI and immunofluorescent assay, 11 (84.6%) were positive and 2 (15.4%) were negative. The S-ELISA was positive for all 30 PI samples (100%) and negative for all 30 negative samples (100%). These data indicate that the new kit is a relatively reliable diagnostic tool and can be considered for upstream detection of BVDV-contaminated raw FBS pools.
Hepatic Hyaline Globules in an Eclectus Parrot (Eclectus Roratus)
Authors: Boyd Kelli L. , Latimer Kenneth S.
Published: May 2011

Hepatic hyaline globules, similar to those reported in some human livers, were observed in liver tissue from an Eclectus parrot ( Eclectus roratus). The cytoplasmic inclusions were periodic acid-Schiff positive and diastase resistant and failed to stain by acid-fast or Congo red techniques. Ultrastructurally, the hepatic globules were composed of granular amorphous material with small peripheral striations that extended into the cytoplasm.
Complex Vertebral Malformation in Holstein Calves
Authors: Agerholm Jorgen S. , Bendixen Christian , Andersen Ole , Arnbjerg Jens
Published: May 2011

A recently observed lethal congenital defect of purebred Holstein calves is reported. Eighteen genetically related calves were necropsied. One calf had been aborted on gestation day 159, and the others were delivered between day 250 and day 285. Birth weights were reduced. The defect was characterized by shortening of the cervical and thoracic parts of the vertebral column due to multiple hemivertebrae, fused and misshaped vertebrae, and scoliosis. Symmetrical flexures of the carpal joints and the metacarpophalangeal joint in combination with a slight lateral rotation of the phalanges also were present. Similar low-grade arthrogryposis was present in the posterior limbs. Fifty percent of the calves had heart malformation. Other malformations occurred in a few calves. Complex vertebral malformation (CVM) is proposed as the designation for this defect. A genetic etiology is indicated because cases occurred following breeding between genetically related individuals. Two common ancestors were found; both were former elite sires of US Holstein origin. Because of the widespread international use of semen from sires occurring in the pedigrees of affected calves, CVM is expected to occur in several countries.
Early Detection of Maedi-Visna (Ovine Progressive Pneumonia) Virus Seroconversion in Field Sheep Samples
Authors: Varea R. , Monleón E. , Pacheco C. , Luján L. , Bolea R. , Vargas M. A. , van Eynde G. , Saman E. , Dickson L. , Harkiss G. , Amorena B. , Badiola J. J.
Published: May 2011

The aim of this work was to investigate whether an enzyme-linked immunosorbent assay (ELISA) was useful for early detection of maedi-visna virus (MVV) infection in sheep under field conditions. An ELISA based on p25 recombinant protein and a gp46 synthetic peptide was used. Sequentially obtained serum samples ( n = 1,941) were studied for 4 years. ELISA results were compared with those of the agar gel immunodiffusion (AGID) test, and results of both tests were compared with a reference result established using consensus scores for at least 2 of 3 serologic techniques (AGID, ELISA, and western blotting, which was used to resolve result discrepancies between the other 2 techniques). A total of 247 discrepancies were observed between ELISA and AGID. Of these, 131 were due to an earlier detection of 120 sera by the ELISA and 11 sera by AGID. The remaining discrepancies (116) were due to the presence of false reactions in both tests. Fewer false-negative results were found by ELISA than with AGID (6 vs. 69 sera, respectively), whereas the number of false-positive results was virtually the same for ELISA and AGID (21 vs. 20, respectively). In relation to the reference result, ELISA sensitivity and specificity were 97.8% and 98.2%, respectively, whereas values for AGID were 76.3% and 98.3%, respectively. The agreement between ELISA and the reference result was higher than that between AGID and the reference result (κ value: 0.96 and 0.77, respectively). A variation in the ELISA signal (based on optical density) was observed during the study period, suggesting different antibody levels throughout the animal's life. The ELISA was useful for detecting MVV-infected sheep in field conditions and has potential for use in control and eradication programs.
Evaluation and Use of a Nested Polymerase Chain Reaction Assay in Cats Experimentally Infected with Bartonella Henselae Genotype I and Bartonella Henselae Genotype II
Authors: Roy Alma F. , Corstvet Richard E. , Tapp Ronald A. , O'Reilly Kathy L. , Cox Hollis U.
Published: May 2011

Cats have been shown to be infected with Bartonella henselae genotype I, B. henselae genotype II, and B. clarridgeiae. Feline bartonellosis infections and the strains involved in these infections are important in both veterinary and human medicine. Nucleic acid amplification methods such as polymerase chain reaction (PCR) are being used in both research and diagnostics as tools for understanding many infectious diseases. Bartonella bacteremia in cats is detected by blood culture; however, because of the limitations of culture (delayed turnaround time and sensitivity limits), PCR may be a more efficient method for identifying infected cats. Three distinct PCR assays that could differentiate among B. henselae genotype I, B. henselae genotype II, and B. clarridgeiae were developed and used to detect as few as 3.2 organisms. Fourteen cats experimentally infected with B. henselae genotype I and B. henselae genotype II were followed by bacterial culture and PCR through the course of infection, including periods of primary and relapsing bacteremia. The PCR assay was positive in 11 of the 14 cats for periods of 1–9 weeks after culture became negative. Of the 223 blood specimens that were culture negative, the PCR assay was positive in 38 (17%) of the specimens. Two of the 14 cats developed relapsing bacteremia. The 2 B. henselae genotypes were amplified in the cats and the bacteremic phase of these infections as determined by PCR lasted for a longer period than previously determined by culture. Using laboratory assays such as PCR to understand the strains involved in feline bartonellosis and the course of the infection is important in the understanding of these zoonotic agents.
Correlation of Ergovaline and Lolitrem B Levels in Endophyte-Infected Perennial Ryegrass (Lolium Perenne)
Authors: Hovermale Jeannette T. , Craig A. Morrie
Published: May 2011

The varieties of perennial ryegrass ( Lolium perenne) infected with the endophytic fungus Neotiphodium lolii contain several classes of toxic alkaloids, including ergopeptide alkaloids and lolitrem alkaloids. Lolitrem B, a potent tremorgen, is generally considered to be the predominant alkaloid in endophyte-infected perennial ryegrass. Ergovaline, a vasoconstrictor normally associated with endophyte-infected tall fescue ( Festuca arudinacea), is also present in endophyte infected perennial ryegrass. Clinical signs of animals ingesting endophyte-infected perennial ryegrass are consistent with the presence of lolitrem B. However, clinical signs normally associated with ergovaline poisoning are not usually observed in animals ingesting endophyte-infected perennial ryegrass. A survey was conducted to quantitate both lolitrem B and ergovaline in 459 perennial ryegrass straw samples received at the Oregon State University College of Veterinary Medicine. Samples were analyzed for each alkaloid using separate high-performance liquid chromatography analyses. A strong positive correlation between the 2 alkaloids ( r2 = 0.7335) was observed, especially in the samples containing <3,000 ppb (ng/g) lolitrem B. The threshold levels above which clinical signs typically occur are 2,000 ppb lolitrem B and 300–400 ppb ergovaline. All of the samples analyzed contained <425 ppb ergovaline.
Immunoreactivity of A103, An Antibody to Melan A, in Canine Steroid-Producing Tissues and their Tumors
Authors: Ramos-Vara José A. , Beissenherz Marilyn E. , Miller Margaret A. , Johnson Gayle C. , Kreeger John M. , Pace Lanny W. , Turk James R. , Turnquist Susan E. , Watson Gary L. , Yamini Ben
Published: May 2011

The monoclonal antibody A103 to the melanocytic differentiation antigen Melan A stains human steroid-producing cells and their tumors. A total of 200 formalin-fixed, paraffin-embedded canine normal tissues and hyperplastic and neoplastic lesions of the adrenal gland, testis, and ovary were immunohistochemically tested for Melan A with antibody A103. Leydig cell tumors (23/23, 100%), Sertoli cell tumors (14/15, 93%), and adrenocortical adenomas (12/13, 92%) were consistently positive. Adrenocortical carcinomas (23/35, 65%) and granulosa cell tumors (10/17, 59%) were less frequently positive. All pheochromocytomas, seminomas, and dysgerminomas were negative. The pattern of staining was cytoplasmic, but nuclear staining was also frequently seen in normal Leydig cells and their tumors. As in human tumors, immunohistochemistry for Melan A stains many canine steroid-producing tumors and can be used to distinguish these tumors from those of nonstereidogenic cells.
Nucleotide Sequence and Polymerase Chain Reaction/Restriction Fragment Length Polymorphism Analyses of Aleutian Disease Virus in Ferrets in Japan
Authors: Murakami Masaru , Matsuba Chikako , Une Yumi , Nomura Yasuo , Fujitani Hideo
Published: May 2011

Two ferrets with spontaneous Aleutian disease (AD) were found in Japan. The diagnosis was verified by polymerase chain reaction (PCR) amplification of part of the capsid gene specific to AD virus (ADV). The nucleotide sequences (365 bp in length) of the amplified fragments from the 2 ferrets differed by a single nucleotide, producing an amino acid alteration. Compared with other types of ADV, these isolates had 96% sequence similarity to a published ferret ADV (FADV) in contrast to <91% homology to various types of mink ADV (MADV). The phylogenetic tree of ADVs indicates that these 2 isolates and the published FADV belong to the same genetic group and definitely are divergent from MADVs. The predicted amino acid sequence of the hypervariable segment in the capsid gene was conserved among the 3 types of FADV. These results indicated that the 2 isolates found in Japan were new DNA types of FADV and could have been derived from FADV(s). A restriction fragment length polymorphism (RFLP) method to distinguish the ferret types of ADV from the mink types of ADV was developed on the basis of differences in their nucleotide sequences. Digestion of the PCR products with AfaI or ScaI provided different cleavage patterns for FADV and MADV. This PCR/RFLP analysis of the ADV capsid gene will be a valuable asset for diagnosis of this virus infection in ferrets.
Idiopathic Disseminated Intracytoplasmic Neuronal Vacuolation in a Neonatal Holstein Calf Born in the USA
Authors: Hamir Amir N. , Habecker Perry , Jenny Allen , Hutto David , Stack Mick J. , Chaplin Melanie J. , Stasko Judi
Published: May 2011

Histopathologic, immunohistochemical, and ultrastructural evaluations were made of a 6-day-old Holstein calf with severe vacuolation of the neuronal perikarya that was widely distributed throughout the central nervous system. No evidence of storage material within the vacuoles was revealed by histopathologic and ultrastructural examinations. Immunohistochemical and electron microscopic examinations were negative for protease-resistant prion protein and scrapie-associated fibrils, respectively. These results indicate that the clinical signs in this calf were not associated with transmissible spongiform encephalopathy. Neuronal vacuo-lation has not previously been documented in calves.
Rhinosporidiosis in a Cat
Authors: Moisan Peter G. , Baker Stephanie V.
Published: May 2011

A polypoid nasal mass from an adult cat was submitted for routine biopsy examination and was found to contain sporangia and sporangiospores consistent with Rhinosporidium seeberi. Inflammatory infiltrates were moderate and pyogranulomatous to lymphohistiocytic and were associated with hyperplasia of the transitional nasal epithelium. Apparently, this is the first reported case of rhinosporidiosis in a cat.
A Simpler, More Sensitive Competitive Inhibition Enzyme-Linked Immunosorbent Assay for Detection of Antibody to Malignant Catarrhal Fever Viruses
Authors: Li Hong , McGuire Travis C. , Müller-Doblies Uwe U. , Crawford Timothy B.
Published: May 2011

An earlier competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) was developed for detection of specific antibody against malignant catarrhal fever (MCF) viruses (MCFV) in ruminants. In this study, the indirect CI-ELISA was improved by conjugating the monoclonal antibody 15-A directly with horseradish peroxidase and by developing a method of producing precoated, dried antigen plates. This new test is referred to as a direct CI-ELISA. The reformatted test yielded a significantly improved sensitivity, and the time required was reduced to about one-sixth of the previous time. Of 37 MCF cases in cattle that were confirmed by histopathology and polymerase chain reaction (PCR) assay, 37 (100%) were positive by the new test, whereas the indirect CI-ELISA detected only 23 (62%). The direct CI-ELISA detected antibody to MCFV in 100% of 48 sheep that had been defined as infected with ovine herpesvirus 2 (OvHV-2) by PCR, whereas the indirect CI-ELISA detected only 41 (85%). Comparison of antibody titers measured by the 2 assays for sera collected from OvHV-2-infected sheep and from cattle, bison, and deer with clinical sheep-associated MCF revealed that the direct CI-ELISA offered a 4-fold increase in analytical sensitivity over the indirect format. The number of seropositive animals detected by the direct CI-ELISA among apparently normal cattle and bison was 2–3 times greater than the number detected by the indirect CI-ELISA, indicating that a significant percentage of normal cattle and bison are subclincally infected with MCFV.