bluuebunny/crossref_metadata_2025 huggingface dataset:
Small Intestine Adenocarcinoma in Conjunction with Multiple Adenomas Causing Acute Colic in a Horse
Authors: Muñoz Moran Juan A. , Lemberger Karin , Cadoré Jean-Luc , Lepage Olivier M.
Published: May 2011

An 11-year-old Andalusian stallion developed marked signs of colic associated with an acute small intestine obstruction. Exploratory laparotomy revealed a distal jejunum full-thickness wall induration and multiple small adherent intraluminal masses. Fifteen centimeters of jejunum, including the induration, and several intraluminal masses were resected. Histologic examination revealed an adenocarcinoma and multiple polypoid adenomas. The horse was discharged, and no complications were reported 12 months postoperatively. Colic was considered secondary to partial jejunal lumen obstruction by the adenocarcinoma. Adenocarcinoma recurrence or transformation from remaining adenomas into an adenocarcinoma is still a major risk.
Assessing the Effect of Sample Handling on the Performance of a Commercial Bovine Viral Diarrhea Virus Antigen-Capture Enzyme-Linked Immunosorbent Assay
Authors: Reed Matthew C. , O'Connor Annette M. , Yoon Kyoung-Jin , Cooper Vickie L.
Published: May 2011

Handling practices of specimens may affect the sensitivity or specificity of diagnostic tests. In this study, as part of the Voluntary Iowa Bovine Viral Diarrhea Virus Screening Project held in 2006, 2 sample-handling practices were evaluated to determine how they affect the sensitivity and specificity of the antigen-capture enzyme-linked immunosorbent assay (ACE) for bovine viral diarrhea virus (BVDV). The null hypotheses investigated were 1) that maintenance of samples at room temperature would not be associated with decreased sensitivity, and 2) that continued use of a single pair of ear notchers would not be associated with cross-contamination of virus from 1 notch to another and reduce specificity. These hypotheses were tested in 2 studies by collecting known positive and negative samples and giving groups of samples different treatments. The first study used ACE on 4 groups of skin samples, all from a known-positive animal. Each group was subjected to different lengths of time at room temperature, from 24 to 96 hours at 24–hour intervals. No difference in test results was found between specimens subjected to different lengths of time at room temperature. The second study tested the effects of giving 3 different treatments to an ear notcher in between sample collecting (water rinse, Nolvasan solution rinse, or no treatment) on ACE results. No effect on sensitivity or specificity of ACE was observed. No difference in test results was found between the 3 ear-notcher treatment groups. The sample handling practices evaluated appeared to have little impact on test sensitivity or specificity of ACE for BVDV.
Comparison of histologic techniques for the diagnosis of bovine tuberculosis in the framework of eradication programs
Authors: Varello Katia , Pezzolato Marzia , Mascarino Daniela , Ingravalle Francesco , Caramelli Maria , Bozzetta Elena
Published: May 2011

Rapid diagnosis of tuberculosis in cattle reacting positive in antemortem assays is crucial in countries where eradication programs are operated to confirm the presence of the infection in tuberculosis-free herds. This study evaluated the accuracy of histopathologic examination by hematoxylin and eosin and Ziehl-Neelsen (ZN) staining applied in this framework, when suspected lesions are caused by low infectious doses and are detected in early stages of the disease. For this purpose, histologic methods were compared with mycobacterial culture as reference test on suspected lymph node samples from 173 cattle reacting positive in antemortem tests. Histopathology demonstrated high sensitivity (93.4%) and specificity (92.3%), while ZN sensitivity and specificity were respectively 33.9% and 100%. There was good agreement between histopathology and bacterial culture, suggesting that histopathologic examination is a reliable tool for rapid diagnosis in countries where active tuberculosis eradication programs allow the prompt identification and elimination of reactor cattle. Histopathology permits identification of typical mycobacterial lesions and its differentiation from other causes.
Development and validation of an enzyme-linked immunosorbent assay for the diagnosis of porcine proliferative enteropathy
Authors: Wattanaphansak Suphot , Asawakarn Tanong , Gebhart Connie J. , Deen John
Published: May 2011

The objective of this study was to develop an indirect enzyme-linked immunosorbent assay (ELISA) using a sonicated pure culture of Lawsonia intracellularis as the antigen (So-ELISA). A total of 332 serum samples, consisting of 232 experimentally infected animals and 100 animals naturally infected with L. intracellularis, were used to assess the diagnostic sensitivity. Three hundred and fifty-five sera from uninfected animals were used to determine the diagnostic specificity. The receiver operating characteristic and mean +3 standard deviation of optical density (OD) values from uninfected animals were used for selecting cut-off points. The diagnostic accuracy of So-ELISA was considered to be high as the area under the curve index was 0.991 with 0.0029 standard error. The optimal cut-off for So-ELISA was set at 0.45 OD with 89.8% sensitivity and 99.4% specificity based on a combination of good sensitivity and high specificity. No cross-reactivity was found in sera from pigs exposed to Brachyspira pilosicoli, B. hyodysenteriae, Campylobacter mucosalis, C. jejuni, or C. coli. Inter- and intracoefficient of variation of all control sera tested with So-ELISA was less than 10%. The observed agreements between So-ELISA and the immunoperoxidase monolayer assay tested with experimental challenge animals and field samples were 95.08% with 0.88 kappa and 90.65% with 0.74 kappa value, respectively. So-ELISA was able to detect the seroconversion of infected animals at 2 to 4 weeks after exposure to L. intracellularis. Based on the validation results, So-ELISA could be used as an alternative serology for proliferative enteropathy diagnosis.
Antemortem versus postmortem methods for detection of betanodavirus in Senegalese sole (Solea senegalensis)
Authors: Olveira José Gabriel , Soares Florbela , Engrola Sofia , Dopazo Carlos P. , Bandín Isabel
Published: May 2011

The suitability of nested reverse transcription polymerase chain reaction (nRT-PCR) to detect betanodavirus in blood samples from naturally infected Senegalese sole (Solea senegalensis) was evaluated in comparison with other diagnostic methods. Results indicated that histologic examination of brain lesions could be regarded as the most consistent indicator of nodavirus infection in this species. The nRT-PCR showed low to moderate levels of detection; the best values were obtained in brain samples followed by blood samples. Inoculation of SSN-1 and SAF-1 cells with fish samples did not cause cytopathic effect, although virus was detected by reverse transcription polymerase chain reaction in approximately 25% of the SSN-1 inoculated wells. The efficiency of detection of the viral genome was dramatically increased by the use of nRT-PCR, reaching 90.6% of positives in brain samples and 84.4% in blood samples. The sensitivity and the negative predictive value of nRT-PCR in blood samples were slightly lower than those obtained using brain samples. Nevertheless, it is suggested that the advantage of being able to perform diagnosis on live fish adequately counterbalances the slightly lower sensitivity of nRT-PCR on blood samples. This technique is proposed as a useful tool, not only for the selection of nodavirus-free breeders but also to check the fish status during ongrowing.
Hemolytic-uremic syndrome in a postpartum mare concurrent with encephalopathy in the neonatal foal
Authors: Dickinson Charles E. , Gould Daniel H. , Davidson Ann H. , Avery Paul R. , Legare Marie E. , Hyatt Doreene R. , DebRoy Chitrita
Published: May 2011

A postpartum mare and foal were presented for evaluation of fever and lethargy in the mare. The mare was diagnosed with endometritis and initially responded well to treatment. On the second day of hospitalization, the mare developed renal insufficiency characterized by oliguria, azotemia, hemolysis, and thrombocytopenia. Concurrently, the foal developed rapidly progressive central nervous system signs culminating in refractory seizures. Both animals failed to respond to treatment and were euthanized. Thrombotic microangiopathy involving glomeruli was evident on microscopic examination of the mare's kidneys. Microscopic evidence of brain edema was the principal postmortem finding in the foal. No specific etiology was confirmed in either case. Notably, Escherichia coli 0103:H2 was isolated from the mare's uterus and the gastrointestinal tracts of both animals. To the authors' knowledge, this is the first report in which an organism implicated as a cause of hemolytic-uremic syndrome was isolated from an animal with clinical signs and postmortem findings consistent with the disease.
Hematogenous metastasis of embryonal rhabdomyosarcoma originating from skeletal muscle in a young dog
Authors: Yhee Ji-Young , Kim Dae-Hyun , Hwang Du-Na , Moon Chang-Taek , Yu Chi-Ho , Kim Hwi-Yool , Doster Alan R. , Sur Jung-Hyang
Published: May 2011

An 8-month-old, intact male Golden Retriever with a history of left forelimb lameness for 2 months was presented to the Veterinary Medical Teaching Hospital of Konkuk University (Seoul, Korea). Results of a physical examination revealed a mass in the left axillary region. A thoracic radiography showed an osteolytic lesion in the scapula and the presence of a soft tissue density from the thoracic wall to the scapula. A computerized tomography revealed a mass invading into the scapula, and small nodules in the lung that suggested metastasis. At necropsy, a pale-yellow, irregular, firm, 8 × 10 × 5 cm mass extended from axillary region and destroyed the scapular. In addition, small nodules were noted in the lung. On microscopic examination, the mass consisted of round-to-oval cells, with eccentrically located hyperchromatic nuclei and eosinophilic cytoplasm in fibromyxoid stroma. Tumor cells were observed in blood vessels in the primary mass. Tumor cells strongly expressed vimentin, desmin, and myoglobin. In phosphotungstic acid-hematoxylin staining, cross-striations were detected in rhabdomyoblasts. In periodic acid-Schiff reaction, only a few cells were detected. The diagnosis was primary rhabdomyosarcoma of the appendicular muscle of a young dog. The tumor presumably originated in the skeletal muscle of the limb, invaded into the adjacent scapular bone, and metastasized to the lung.
Histopathologic and Immunohistochemical Findings in Two White-Tailed Deer Fawns Persistently Infected with Bovine Viral Diarrhea Virus
Authors: Duncan Colleen , Ridpath Julia , Palmer Mitchell V. , Driskell Elizabeth , Spraker Terry
Published: May 2011

Bovine viral diarrhea virus (BVDV) is an important pathogen of domestic cattle. Serologic, experimental, and individual case studies explored the presence and pathogenesis of the virus in wild ungulates; however, there remain large gaps in knowledge regarding BVDV infection in nonbovine species. Live twins were born from a white-tailed deer ( Odocoileus virginianus) doe infected with noncytopathic BVDV during its first trimester of pregnancy. The twins died at 1 day of age from trauma unrelated to the infection, and tissues were collected for histologic and immunohistochemical examination. The most significant histologic abnormality was diffuse depletion of B-lymphocytes in both fawns. The BVDV antigen was distributed widely throughout many tissues and cell types, most notably epithelium and vascular endothelium, consistent with that reported in cattle. In contrast to cattle, lymphocytes exhibited only very rare positive staining.
Evaluation of Four Commercial Enzyme-Linked Immunosorbent Assays for the Diagnosis of Bovine Paratuberculosis in Chilean Dairy Herds
Authors: Fry M. Paulina , Kruze Juan , Collins Michael T.
Published: May 2011

The accuracy of 4 commercial enzyme-linked immunosorbent assays (ELISAs) for diagnosis of bovine paratuberculosis was compared using sera from 53 Mycobacterium avium subsp. paratuberculosis (MAP) fecal culture–positive dairy cows (cases) and sera from 345 dairy cattle resident in 11 fecal culture–negative herds on 2 consecutive occasions 1 year apart (controls). The specificity of all 4 ELISA kits was >99%, and their diagnostic sensitivity ranged from 30.2% to 41.5%. Pairwise comparison of ELISAs found no significant differences (McNemar's chi-square test > 0.05), and assay agreement for categorical assay interpretation (positive or negative) was high (>98%) with κ values ranging from 0.84 to 0.95. Receiver operating characteristic (ROC) curve analysis and the corresponding area under the ROC curves indicate that kit B had the highest overall accuracy. Thus, all 4 ELISA kits for bovine paratuberculosis had comparable accuracy when tested on Chilean dairy cattle, with kit B having a slight statistical advantage based on ROC area under the curve analysis. This suggests that any of the 4 kits could be appropriate for herd certification and for paratuberculosis control programs on Chilean dairy cattle.
Comparison of Two Commercial Radial Immunodiffusion Assays for Detection of Bovine Immunoglobulin G in Newborn Calves
Authors: Ameri Mehrdad , Wilkerson Melinda J.
Published: May 2011

Bovine failure of passive transfer (FPT), defined as inadequate transfer of colostral immunoglobulins from the dam to the calf, has been associated with increased risk in neonatal mortality. Currently, radial immunodiffusion (RID) assay is considered to be the gold standard in determining FPT in serum samples from calves. There are 2 commercial RID assays routinely used for serodiagnosis of FPT in calves: VET-RID and SRID. Discrepancies between results of these RID assays were observed in the authors' laboratory. The objective of this study was to compare 2 commercial RID assays by testing a paired panel of 30 blood samples collected from newborn Holsteins at birth before, and 24 hr after, ingestion of colostrum, a commercial bovine reference serum, and a panel of different concentrations of 2 purified bovine immunoglobulin G (IgG) products. Overall, the results of this study showed a high level of discrepancy and poor agreement between the 2 RID kits. The interassay precision study revealed lower between-run coefficients of variation for the VET-RID kit compared with the SRID kit. The spiking and recovery study using purified bovine IgG products demonstrated that the VET-RID kit more closely approximates the expected concentrations of the purified bovine IgG products, whereas the SRID kit consistently overestimates the concentration of purified bovine IgG products. It was concluded that this may be due to inaccuracies in the internal standards of the SRID kit.
Differentiation of Erysipelothrix Rhusiopathiae Strains by Nucleotide Sequence Analysis of a Hypervariable Region in the spaA Gene: Discrimination of a Live Vaccine Strain from Field Isolates
Authors: Nagai Shinya , To Ho , Kanda Akira
Published: May 2011

Erysipelothrix rhusiopathiae causes erysipelas in swine and is considered a reemerging disease contributing substantially to economic losses in the swine industry. Since an attenuated live vaccine was commercialized in 1974 in Japan, outbreaks of acute septicemia or subacute urticaria of erysipelas have decreased dramatically. In contrast, a chronic form of erysipelas found during meat inspections in slaughterhouses has been increasing. In this study, a new strain-typing method was developed based on nucleotide sequencing of a hypervariable region in the surface protective antigen ( spaA) gene for discrimination of the live vaccine strain from field isolates. Sixteen strains isolated from arthritic lesions found in slaughtered pigs were segregated into 4 major patterns: 1) identical nucleotide sequence with the vaccine strain: 3 isolates; 2) 1 nucleotide substitution (C to A) at position 555: 5 isolates; 3) 1 nucleotide substitution at various positions: 5 isolates; and 4) 2 nucleotide substitutions: 3 isolates. Isolates with the same nucleotide sequence as the vaccine strain were further characterized by other properties, including the mouse pathogenicity test. One strain isolated from pigs on a farm where the live vaccine had been used was found to be closely related to the vaccine strain. The phylogenetic tree constructed based on the spaA sequence suggests that the evolutionary distance of the isolates is related to the pathogenicity in mice. The new strain-typing system based on nucleotide sequencing of the spaA region is useful to discriminate the vaccine strain from field isolates.
Hemagglutinating Activity of Serovar Reference Strains of Ornithobacterium Rhinotracheale
Authors: Vega Vicente , Zepeda Andrea , Ramírez Saúl , Morales Vladimir , Fernóndez Pomposo , de Oca Roberto Montes , Guerra-Infante Fernando M. , de Haro-Cruz María de Jesús , Blackall Patrick J. , Soriano Edgardo V.
Published: May 2011

In the present study, the hemagglutinating activity of 9 reference strains (serovars A–I) of Ornithobacterium rhinotracheale was investigated by using fresh erythrocytes from 15 different species: chicken (broiler, rooster, hen), turkey, pigeon, quail, duck, Harris hawk ( Parabuteo unicinctus), house finch ( Carpodacus mexicanus), cow, sheep, horse, dog, rabbit, pig, human (groups A, B, AB, and O), and rainbow trout ( Oncorhynchus mykiss). All 9 strains agglutinated rabbit erythrocytes. None of the strains was able to agglutinate hen, cow, horse, or rainbow trout erythrocytes. The number of positive reactions among the remaining species varied. Results indicate that the use of rabbit erythrocytes is better suited for testing the hemagglutinating activity of O. rhinotracheale.
Clinical and Histopathological Features of a Thymolipoma in a Dog
Authors: Ramírez Gustavo A. , Spattini Giliola , Altimira Jaume , García Beatríz , Vilafranca Miquel
Published: May 2011

This study presents a case of a canine thymolipoma, which is a rare, slow-growing, benign tumor of the thymus composed of mature adipose tissue and thymic tissue. A 9-year-old spayed, female miniature pinscher presented with chronic cough and dyspnoea. Radiology revealed pleural effusion and a mediastinal mass with a fatty appearance. The mass was attached to, and silhouetted, the adjacent pericardium. Microscopically, the mass was composed of adipose tissue with numerous cords and nests of thymic tissue without corticomedullary arrangement. To the authors' knowledge, this is the first description of this uncommon neoplasm in a dog. The gross and histological findings are similar to those described for thymolipomas in humans. The possible histogenesis for this neoplasia in a dog is also discussed.
Detection of Feline Leukemia Virus in the Endangered Iberian Lynx (Lynx Pardinus)
Authors: Luaces Inés , Doménech Ana , García-Montijano Marino , Collado Victorio M. , Sónchez Celia , Tejerizo J. German , Galka Margarita , Fernóndez Pilar , Gómez-Lucía Esperanza
Published: May 2011

Feline retroviruses are rarely reported in lynx species. Twenty-one Iberian lynx ( Lynx pardinus) blood and tissue samples collected from Doñana National Park and Los Villares (Sierra Morena) in southern Spain during 1993–2003 were analyzed by polymerase chain reaction to amplify nucleic acids from feline retroviruses. Six samples were positive for Feline leukemia virus (FeLV), but no samples tested positive for Feline immunodeficiency virus. The BLAST analysis indicated that 5 of the 6 sequences were closely related to FeLV strain Rickard subgroup A, whereas 1 sequence was identical to FeLV. To the authors' knowledge, this is the first report of FeLV in the endangered Iberian lynx.
Comparison of Three Serological Assays to Determine the Cross-Reactivity of Antibodies from Eight Genetically Diverse U.S. Swine Influenza Viruses
Authors: Leuwerke Brad , Kitikoon Pravina , Evans Richard , Thacker Eileen
Published: May 2011

Swine influenza virus is an economically important pathogen to the U.S. swine industry. New influenza subtypes and isolates within subtypes with different genetic and antigenic makeup have recently emerged in U.S. swineherds. As a result of the emergence of these new viruses, diagnosticians' ability to accurately diagnose influenza infection in pigs and develop appropriate vaccine strategies has become increasingly difficult. The current study compares the ability of subtype-specific commercial enzyme-linked immunosorbent assays (ELISA), hemagglutination inhibition (HI), and serum neutralization (SN) assays to detect antibodies elicited by multiple isolates within different subtypes of influenza virus. Pigs were infected with genetically and antigenically different isolates of the 3 major circulating subtypes within populations of swine (H1N1, H1N2, and H3N2). Serum was collected when all pigs within a group collectively reached HI reciprocal titers ≥160 against that group's homologous challenge virus. The antibody cross-reactivity of the sera between isolates was determined using ELISA, HI, and SN assays. In addition, the correlation between the 3 assays was determined. The assays differed in their ability to detect antibodies produced by the viruses used in the study. The results provide important information to diagnostic laboratories, veterinarians, and swine producers on the ability of 3 common serological assays used in identifying infection with influenza in pigs.
Pathology and Toxicology Findings for Search-and-Rescue Dogs Deployed to the September 11, 2001, Terrorist Attack Sites: Initial Five-Year Surveillance
Authors: Fitzgerald Scott D. , Rumbeiha Wilson K. , Braselton W. Emmett , Downend Amanda B. , Otto Cynthia M.
Published: May 2011

A long-term surveillance study was conducted on 95 search-and-rescue (S&R) dogs deployed to the September 11, 2001, terrorist attack sites; an additional 55 nondeployed S&R dogs served as controls. After 5 years of surveillance, 32% of the deployed dogs have died and 24% of the nondeployed dogs. The mean age at the time of death in these 2 groups of dogs is not significantly different. Causes of death in both groups of dogs include inflammatory, degenerative, and proliferative conditions. No primary pulmonary tumors have been identified to date nor has any significant level of toxicant been found in the tissues from these dogs using assays for general organic compounds and metals or, specifically, for polychlorinated biphenyls. However, significant numbers of both deployed and nondeployed dogs have evidence of inhaled matter as demonstrated by the presence of anthracotic pigments or refractile particulate matter in pulmonary tissue. Although S&R activities in response to the 9/11 terrorist attacks exposed dogs to a wide variety of potentially toxic compounds, to date, these dogs do not appear to suffer from higher mortality or increased pulmonary disease compared with nondeployed dogs. To the authors' knowledge, the current survey represents the first long-term and large-scale survey of the pathology and toxicology of S&R dogs deployed to a major disaster site.
Detection of Persistent Cytauxzoon Felis Infection by Polymerase Chain Reaction in Three Asymptomatic Domestic Cats
Authors: Brown Holly M. , Latimer Kenneth S. , Erikson Laura E. , Cashwell Marsha E. , Britt James O. , Peterson David S.
Published: May 2011

Repeated polymerase chain reaction (PCR) testing of 3 asymptomatic domestic cats were positive for Cytauxzoon felis DNA, suggesting persistent infection. Two cats initially presented with clinical signs consistent with acute cytauxzoonosis and, in both cases, signs of illness resolved after treatment. Parasitemia was detected in peripheral blood smears from these cats upon presentation with illness and, at subsequent follow-up appointments, in the absence of clinical illness. Polymerase chain reaction analysis was positive for C. felis from blood sampled at each time point. A third cat, a housemate of a cat fatally infected with C. felis, was preventatively treated for infection at the time of the housemate cat's death. This contact cat, having never shown signs of clinical illness consistent with cytauxzoonosis infection, had no detectable parasitemia but was positive for C. felis on repeated PCR testing. Detection of asymptomatically infected cats allows for the possibility of a yet unrecognized population of infected domestic cats that may have the capacity to serve as an additional reservoir host for C. felis, altering the currently accepted paradigm of C. felis transmission to domestic cats through bobcats as the reservoir host. In cases of very low parasitemia, more sensitive means of parasite detection, such as PCR testing, may be necessary to detect infected cats. Increased detection of asymptomatically infected cats will aid in understanding the epidemiology of C. felis infection and enhance the ability to prevent this highly fatal infectious disease of domestic cats.
Development of Multiplex Polymerase Chain Reaction Assays for Detecting Enterotoxigenic Escherichia Coli and their Application to Field Isolates from Piglets with Diarrhea
Authors: Lee Su In , Kang Sang Gyun , Kang Mi Lan , Yoo Han Sang
Published: May 2011

Fimbriae and enterotoxins are major virulence factors associated with enterotoxigenic Escherichia coli (ETEC). In this study, 3 sets of multiplex polymerase chain reaction (mPCR) assays targeting fimbriae, enterotoxins, and other adherence factors were developed for detecting ETEC. A total number of 188 E. coli field isolates were examined, and percentages of E. coli strains carrying each virulence factors were as follows: F4 (7.45%), F5 (29.79%), F6 (6.38%), F18 (15.43%), F41 (3.72%), STa (10.11%), STb (20.74%), LT (9.57%), Stx2e (2.13%), EAST1 (42.02%), F1 (67.55%), AIDA-I (2.66%), and pAA (7.45%). Of the 188 E. coli field isolates examined, 25.53% were found to be pathogenic ETEC, having both fimbriae and enterotoxins. However, the ratio increased to 44.68% when the presence of other adhesins was considered as criteria for virulence. Among the adherence factors, F1 was found to be the most prevalent. AIDA-I and pAA were also found with similar ratio as compared with other virulence factors. In addition, virulence patterns carrying these alternate adhesive genes with enterotoxins were detected with significant ratio. Therefore, it is desirable that alternate adhesins be considered as markers for diagnosis of ETEC.
A Collaborative Canadian-United Kingdom Evaluation of an Immunohistochemistry Protocol to Diagnose Bovine Spongiform Encephalopathy
Authors: Manning Lisa , O'Rourke Katherine I. , Knowles Donald P. , Marsh Sarah A. , Spencer Yvonne I. , Moffat Estella , Wells Gerald A. H. , Czub Stefanie
Published: May 2011

Collaboration was established in 2001 to evaluate a commercially available immunohistochemistry assay kit for the detection of bovine spongiform encephalopathy (BSE) disease-associated prion protein in formic acid-treated formalin-fixed samples of bovine brain. The kit protocol was evaluated at the National Centre for Foreign Animal Diseases (Winnipeg, Canada) and the Veterinary Laboratories Agency (Weybridge, U.K.). The U.K. laboratory provided paraffin-embedded blocks of brainstem (medulla oblongata at the level of the obex) from 100 positive cases defined by clinical signs and histopathology, and 100 clinically suspect but BSE-negative samples defined by histopathology and immunohistochemistry with anti-PrP monoclonal antibody R145. The Canadian laboratory provided 400 blocks from surveillance cases defined as clinically suspect but negative by histopathology and immunohistochemistry with anti-PrP antibody 6H4. Consecutive sections from each block were cut and coded. Each set of 600 slides was immunolabeled and read in each laboratory. Evaluation parameters included estimates of diagnostic sensitivity and specificity and reproducibility of the results. The kit performed with 100% sensitivity, specificity, and reproducibility in spite of minor differences between the laboratories in brain sample areas, fixation and processing, and in the immunolabeling protocol. Although enzyme linked immunosorbent assays are widely used in high throughput surveillance programs, standardized protocols and reagents for manual immunohistochemistry provide a useful adjunct to surveillance efforts, particularly in laboratories testing small numbers of samples or using immunohistochemistry for confirmation and characterization of BSE cases.
Western Blot Detection of PrPSc in Archived Paraffin-Embedded Brainstem from Scrapie-Affected Sheep
Authors: Kunkle Robert A. , Nicholson Eric M. , Lebepe-Mazur Semakaleng , Orcutt Dennis L. , Srinivas Megan L. , Greenlee Justin J. , Alt David P. , Hamir Amir N.
Published: May 2011

Scrapie is a naturally occurring fatal neurodegenerative disease of adult sheep and goats, one of a group of mammalian diseases known as transmissible spongiform encephalopathies (TSE) or prion diseases. Immunoassays that identify disease-associated prion protein (PrPSc) are integral to the diagnosis of scrapie and other prion diseases. Results obtained by either immunohistochemistry (IHC) or Western blot (WB) assay are generally adequate for the definitive diagnosis. Approved or accepted methods for WB diagnosis of TSEs requires the use of fresh or frozen nonfixed tissue samples, whereas formalin-fixed, paraffin-embedded tissue is required for the localization of PrPSc by IHC. Because disparate processing methods are used for these accepted diagnostic techniques, separate tissue samples are collected from the same animal. Occasions arise in which there is either insufficient quantity of tissue available to complete analysis by both techniques or initial tissue processing is incompatible with one of the assays. Also, results between the assays may differ because of the vagaries of sampling, especially in case material that contains moderate-to-low levels of PrPSc. The present article describes a method to conduct a WB assay from the same paraffin-embedded brainstem sample used for the IHC diagnosis of experimentally induced sheep scrapie.
Development and Use of a Multiplex Real-Time Quantitative Polymerase Chain Reaction Assay for Detection and Differentiation of Porcine Circovirus-2 Genotypes 2a and 2b in an Epidemiological Survey
Authors: Gagnon Carl A. , del Castillo Jerome R. E. , Music Nedzad , Fontaine Guy , Harel Josëe , Tremblay Donald
Published: May 2011

By the end of 2004, the Canadian swine population had experienced a severe increase in the incidence of Porcine circovirus-associated disease (PCVAD), a problem that was associated with the emergence of a new Porcine circovirus-2 genotype (PCV-2b), previously unrecovered in North America. Thus, it became important to develop a diagnostic tool that could differentiate between the old and new circulating genotypes (PCV-2a and PCV-2b, respectively). Consequently, a multiplex real-time quantitative polymerase chain reaction (mrtqPCR) assay that could sensitively and specifically identify and differentiate PCV-2 genotypes was developed. A retrospective epidemiologic survey that used the mrtqPCR assay was performed to determine if cofactors could affect the risk of PCVAD. From 121 PCV-2-positive cases gathered for this study, 4.13%, 92.56%, and 3.31% were positive for PCV-2a, PCV-2b, and both genotypes, respectively. In a data analysis using univariate logistic regressions, the PCVAD-compatible (PCVAD/c) score was significantly associated with the presence of Porcine reproductive and respiratory syndrome virus (PRRSV), PRRSV viral load, PCV-2 viral load, and PCV-2 immunohistochemistry (IHC) results. Polytomous logistic regression analysis revealed that PCVAD/c score was affected by PCV-2 viral load ( P = 0.0161) and IHC ( P = 0.0128), but not by the PRRSV variables ( P > 0.9), which suggests that mrtqPCR in tissue is a reliable alternative to IHC. Logistic regression analyses revealed that PCV-2 increased the odds ratio of isolating 2 major swine pathogens of the respiratory tract, Actinobacillus pleuropneumoniae and Streptococcus suis serotypes 1/2, 1, 2, 3, 4, and 7, which are serotypes commonly associated with clinical diseases.
Use of Dried Blood Smears for Detection of Feline Hemoplasmas Using Real-Time Polymerase Chain Reaction
Authors: Sykes Jane E. , Owens Sean D. , Terry Jeralyn C. , Lindsay LeAnn L. , Pusterla Nicola
Published: May 2011

The objective of the current study was to determine the sensitivity and specificity of real-time polymerase chain reaction (real-time PCR) for feline hemoplasmas when applied to DNA extracted from dried whole-blood smears in comparison to that for DNA extracted from liquid whole blood. Blood samples were collected into ethylenediamine tetra-acetic acid tubes from 305 cats with possible or suspected hemoplasmosis, and dried blood smears from each sample were prepared. DNA was extracted from blood smears and a 160-μl aliquot of each liquid blood sample by using a robotic extractor and was subjected to real-time PCR for feline glyceraldehyde-3-phosphate dehydrogenase (liquid blood), 18S ribosomal RNA (rRNA; dried blood), and “ Candidatus Mycoplasma haemominutum”, Mycoplasma haemofelis, and “ Candidatus Mycoplasma turicensis” DNA. When using the results for liquid whole blood as the gold standard, the sensitivity of each assay for “ Ca. M. haemominutum”, M. haemofelis, and “ Ca. M. turicensis” was 49 of 66 (74%), 11 of 13 (85%), and 11 of 20 (55%), respectively. The specificity of each assay was 224 of 234 (96%), 287 of 287 (100%), and 280 of 280 (100%), respectively. When possible, liquid blood samples should be submitted for detection of feline hemoplasmas by using real-time PCR. The improved sensitivity of real-time PCR on blood smears for M. haemofelis compared with that of the other hemoplasma species may reflect the higher organism burdens associated with infection with this species.
Homocysteine Measurement by an Enzymatic Method and Potential Role of Homocysteine as a Biomarker in Dogs
Authors: Rossi Silvia , Rossi Gabriele , Giordano Alessia , Paltrinieri Saverio
Published: May 2011

In humans, homocysteine (Hcy) is employed to monitor renal, cardiovascular, and other diseases and their complications. The aim of the current study was to define the analytical performances of an enzymatic method not yet validated in dogs for measuring homocysteine, and to assess the possible clinical usefulness of Hcy measurement. Using conventional approaches, the analytical performances were investigated by assessing, imprecision, inaccuracy, and interference of hemoglobin, triglycerides, and bilirubin. The possible clinical usefulness of Hcy determination was assessed by comparing the results of healthy dogs ( n = 8) with those of dogs with heart disease ( n = 10), inflammation ( n = 6), gastrointestinal disorders ( n = 7), neoplasia ( n = 8), renal failure ( n = 4), trauma ( n = 7), and other miscellaneous diseases ( n = 6). Preliminary evaluation of this enzymatic method showed it to be precise at Hcy levels close to or higher than the values in dogs with renal or cardiac disorders that had the highest Hcy levels. By contrast, at low Hcy levels, which were recorded basically in control dogs, the method suffers from high imprecision. The sample of choice is serum. The use of icteric samples should be avoided, while hemoglobin and lipids have only a minor effect on Hcy measurement. In conclusion, the enzymatic method employed in the current study provides useful information in dogs and could be used to monitor cardiac and renal disorders, in which Hcy concentrations are elevated.
Pathologic Lesions of Mycotic Pneumonia in an Alpaca following Third Compartment Ulceration
Authors: Hughes Katherine , Mueller Karin
Published: May 2011

A 4-year-old female alpaca was referred to The Queen's Veterinary School Hospital (Cambridge, United Kingdom) with a history of abortion 6 days previous, at approximately 245 days in gestation. The patient had developed watery diarrhea 24 hr after abortion and exhibited bruxism, progressive depression, and ultimately collapse. On presentation to the referral institute, the alpaca was unable to stand and exhibited severe tachypnea, dyspnea, and tachycardia. The patient was considered to be moribund and was euthanized. Macroscopic postmortem assessment and subsequent histological investigations revealed multifocal ulceration of the third gastric compartment, with vascular mycotic invasion and disseminated multifocal embolic mycotic pneumonia. Real-time polymerase chain reaction targeting the large ribosomal subunit (28S) performed on paraffin-embedded lung tissue successfully amplified 28S RNA specific for Aspergillus species. The primary cause of the depression and collapse of this animal was considered to be a combination of respiratory compromise due to severe embolic mycotic pneumonia and endotoxic shock associated with the ulcerative gastritis of the third gastric compartment. To the authors' knowledge, this is the first report of ulcerative gastritis of the third gastric compartment histologically illustrated as a portal of entry for Aspergillus species causing embolic mycotic pneumonia in an alpaca.
A Case of Coccygeal Chondroid Chordoma in a Cat: Morphological and Immunohistochemical Features
Authors: Carminato Antonio , Marchioro Wendy , Melchiotti Erica , Vascellari Marta , Mutinelli Franco
Published: May 2011

In August 2007, a 4-year-old, intact, female Domestic Shorthair cat was examined for a mass on the tip of the tail. Histological examination performed after apical caudectomy revealed a neoplasm affecting the distal part of the last coccygeal vertebra. The neoplasm consisted of lobules of physaliferous cells surrounding cartilaginous tissue and a central core of trabecular bone. A diagnosis of chondroid chordoma was made based on histomorphological features and immunohistochemical results. Chondroid chordoma has been previously reported in humans, rats, ferrets, and mink. To the authors' knowledge, chondroid chordoma has not been reported in cats. Neither recurrence nor metastasis was reported 7 months after surgery.
Characterization of Shedding Patterns of Porcine Circovirus Types 2a and 2b in Experimentally Inoculated Mature Boars
Authors: Madson Darin M. , Ramamoorthy Sheela , Kuster Chris , Pal Narinder , Meng Xiang-Jin , Halbur Patrick G. , Opriessnig Tanja
Published: May 2011

Porcine circovirus-2 (PCV-2) is an economically important swine pathogen and causes PCV-associated disease (PCVAD) in pigs worldwide. Currently, 2 genotypes of PCV-2, PCV-2a and −2b, are circulating in U.S. swine herds. The objectives of the current study were to evaluate the amount of PCV-2 DNA present in semen over time, compare and correlate incidence and amount of PCV-2 present in semen samples to that present in serum samples and blood swabs, and determine if there are differences in shedding patterns between PCV-2a and −2b. Fifteen 7-month-old PCV-2-naïve Landrace boars ( Sus scrofa) were randomly allocated to 3 treatment groups. The boars in group 1 ( n = 3) served as negative controls, and those in groups 2 ( n = 6) and 3 ( n = 6) were intranasally and intramuscularly inoculated with PCV-2a and −2b, respectively. Semen, serum, and blood swab samples were collected up to 90 days postinoculation (DPI), and necropsies were performed on DPI 23,48, and 90. Larger quantities of both PCV-2a and − 2b DNA were detected earlier in serum and blood swab samples than in raw semen of experimentally inoculated boars. The incidence and duration of presence of PCV-2 DNA in semen varied among boars; however, intermittent shedding was not observed. In all sex glands, PCV-2 DNA was detected by polymerase chain reaction; however, PCV-2 antigen was not detected by immunohistochemistry, and PCV-2 had no effect on sperm morphology. Differences in shedding patterns between PCV-2a and −2b were not observed under the study conditions.
Detection of Helicobacter in Gastric Washing of Cats
Authors: Pregel Paola , Rota Ada , Palmerini Domenico , Guarda Franco , Appino Simonetta
Published: May 2011

The aim of the current study was to evaluate the efficacy of gastric washing for detecting the presence of Helicobacter spp. in feline gastric mucus. Gastric fluids were collected from 11 cats undergoing routine surgical procedures. The fluids were centrifuged and the pellets were subjected to cytological examination by May-Grünwald/Giemsa (MGG) staining and testing by polymerase chain reaction (PCR) for the presence of Helicobacter after DNA extraction. Helicobacter spp. were detected in 8 of 11 samples by MGG staining and in 10 of 11 samples by PCR. Gastric washing proved to be a useful sampling technique and a valuable alternative to the gold-standard method of gastric biopsy, which has the risk of missing colonized spots in the case of patchy Helicobacter colonization. Gastric washing, a noninvasive way of sampling, could be applied to cats undergoing general anesthesia for different causes. Detection of Helicobacter in cats could allow epidemiological investigations, and sequencing of samples could assist in assessing the distribution of various Helicobacter species.
Genetic Diversity of Bovine Viral Diarrhea Virus 1: Italian Isolates Clustered in at Least Seven Subgenotypes
Authors: Giammarioli Monica , Pellegrini Claudia , Casciari Cristina , Rossi Elisabetta , De Mario Gian Mia
Published: May 2011

Bovine viral diarrhea virus (BVDV) is an economically important pathogen of cattle. Two approved species are recognized, namely BVDV-1 and BVDV-2. To date, only 4 subgenotypes of BVDV-2 are known, and at least 11 distinct subgenotypes have been detected for BVDV-1. In a previous study, the genetic characteristics of 38 field isolates of BVDV from northern Italy were investigated, and all 38 isolates were classified as BVDV-1 and could be assigned to 5 different subgenotypes, namely BVDV-1b, BVDV-1d, BVDV-1e, BVDV-1h, and BVDV-1f. However, the circulation of BVDV-2 has been reported in Italy as well. The aim of the current study was to type 88 BVD viruses found throughout Italy. Genetic study was based on the 5′-UTR, supported by select comparison within the Npro coding region. Phylogenetic analysis showed that 5 isolates could be typed as BVDV-2a. The remaining 83 isolates were typed as BVDV-1 and were found to belong to 7 distinct subgenotypes, namely BVDV-1a ( n = 8), BVDV-1b ( n = 37), BVDV-1d ( n = 3), BVDV-1e ( n = 22), BVDV-1f ( n = 4), BVDV-1g ( n = 4), and BVDV-1h ( n = 5). The majority of cattle farms in the current study were predominately infected by BVDV-1b and BVDV-1e isolates, whereas the other BVDV subgenotypes occurred only sporadically. The results also provided evidence for circulation of additional subgenotypes BVDV-1a and BVDV-1g. The occurrence of BVDV-2 was also reconfirmed.
Daily Rhythm of Serum Lipase and α-Amylase Activity in Fed and Fasted Dogs
Authors: Piccione Giuseppe , Giannetto Claudia , Fazio Francesco , Giudice Elisabetta
Published: May 2011

The aim of the current study was to investigate the influence of feeding and fasting on the daily rhythm of serum lipase and α-amylase activity in dogs. Fourteen purebred Beagle dogs were housed in individual pens lined with wood shavings under a 12:12 light:dark cycle. The dogs were divided into 2 groups. Group A received normal feeding once a day, and group B was fasted starting 12 hr prior to the first blood collection. Water was available ad libitum. Blood samples were collected every 4 hr for 48 hr. Serum lipase and α-amylase activity were analyzed with standard kits by means of an ultraviolet spectrophotometer. Statistical analysis of the data was performed by 2-way analysis of variance and by single cosinor method. Results showed no statistical influence of feeding schedule on lipase and α-amylase and a robust daily rhythmicity of lipase and α-amylase in fed and fasted dogs. The current study could provide additional documentation of the structure of the dog circadian timing system and increase the necessary information related to the clinical approach to pancreatic diseases and to the therapeutic efficacy of timed administration of drugs or rations.
Immunohistochemical Detection of Brucella Melitensis Antigens in Cases of Naturally Occurring Abortions in Sheep
Authors: Ilhan Fatma , Yener Zabit
Published: May 2011

Brucella melitensis, a worldwide zoonotic pathogen, is a significant cause of abortion in sheep and goats in some countries. The present study was carried out to determine, by immunohistochemistry, the presence of B. melitensis antigens in 110 naturally occurring aborted sheep fetuses. Sections of lung, liver, kidney, and spleen of each fetus were stained with immunoperoxidase to detect Brucella antigens. Brucella melitensis antigens were detected in 33 of 110 fetuses (30%). In the 33 positive cases, Brucella antigens were found in lung (25 [22.7%]), liver (21 [19%]), spleen (13 [11.8%]), and kidney (6 [5.4%]). Microscopic studies demonstrated that Brucella antigens were mainly located in the cytoplasm of macrophages and neutrophils of the lung, and in the cytoplasm of macrophages in the portal infiltrates and Kupffer cells of the liver. It was concluded that immunohistochemistry in formalin-fixed, paraffin-embedded tissues is a useful tool for the diagnosis of spontaneous ovine abortion caused by B. melitensis.
Malignant Catarrhal Fever in a Captive American Bison (Bison Bison) in Italy
Authors: Campolo Marco , Lucente Maria Stella , Mari Viviana , Elia Gabriella , Tinelli Antonella , Laricchiuta Pietro , Caramelli Maria , Nava Donatella , Buonavoglia Canio , Decaro Nicola
Published: May 2011

Malignant catarrhal fever (MCF) is a fatal, systemic disease of cattle and other domestic and wild ruminants that, in Europe, is caused by Ovine herpesvirus 2 (OvHV-2). American bison ( Bison bison) are highly susceptible to the disease. An adult American bison, housed in a zoo in southern Italy in close cohabitation with a group of domestic sheep ( Ovis aries aries) displayed clinical signs that resembled the acute form of MCF. By real-time polymerase chain reaction, OvHV-2 DNA was detected intravitam in blood, in nasal and ocular swabs, and postmortem in tissue samples of the bison. By indirect fluorescent antibody test, high MCF antibody titers were found in the bison serum. Ovine herpesvirus 2 DNA and antibodies were also found in blood samples from the domestic sheep, thus suggesting a potential role of these animals as a source of the infection. To the authors' knowledge, this is the first MCF case in captive ruminants in Italy and the second confirmed case in captive bison of European zoos.
Antemortem Detection of PrPCWD in Preclinical, Ranch-Raised Rocky Mountain Elk (Cevvus Elaphus Nelsoni) by Biopsy of the Rectal Mucosa
Authors: Spraker Terry R. , VerCauteren Kurt C. , Gidlewski Thomas , Schneider David A. , Munger Randy , Balachandran Aru , O'Rourke Katherine I.
Published: May 2011

Antemortem biopsy of the rectal mucosa was evaluated as a method for the preclinical diagnosis of chronic wasting disease (CWD) in a herd of ranch-raised Rocky Mountain elk ( Cervus elaphus nelsoni) quarantined because of exposure to CWD. Biopsy samples were obtained from 41 elk during the winter of 2005–2006 and from 26 elk from that herd still alive and available for testing during the winter of 2006–2007. Samples were examined for PrPCWD, the protein marker for CWD infection, by immunohistochemistry. PrPCWD was detected in follicles of the rectoanal mucosa–associated lymphoid tissue in biopsy samples from 1 elk with clinical signs of chronic wasting disease and 5 clinically normal elk. The diagnosis was confirmed in all 6 animals by postmortem analysis of brain and peripheral lymph nodes. PrPCWD was also observed in the submucosal plexus and myenteric plexus of the enteric nervous system, and in close association with nonmyelinated mucosal and submucosal nerve fibers. In antemortem rectal biopsy samples from positive animals, immunostaining was consistently observed in approximately 60% of the mucosa-associated lymphoid tissue follicles if 10 or more total follicles per biopsy were present for evaluation. Most antemortem biopsy samples obtained from elk younger than 6.5 years contained at least 10 follicles per rectal mucosal biopsy. These findings support the analysis of antemortem biopsy of the rectal mucosa samples as part of an integrated strategy to manage chronic wasting disease in Rocky Mountain elk.