bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of the Usefulness of the Portable Device Lactate Pro for Measurement of Lactate Concentrations in Equine Whole Blood
Authors: van Oldruitenborgh-Oosterbaan Marianne M. Sloet , van den Broek Erica T. W. , Spierenburg Astrid J.
Published: May 2011

Blood lactate measurements are commonly used in exercising horses to determine the onset of lactate accumulation and in colic patients to assess clinical status and to indicate prognosis. To study the usability of a portable blood lactate meter based on dry chemistry (Lactate Pro), the data from this instrument were compared to data from a laboratory-used lactate meter based on wet chemistry (ABL 605 blood gas analyzer [ABL]). Heparinized blood samples were obtained from horses participating in a jumping experiment (n = 9), from horses cantering at maximal speed on a racetrack (n = 7), and from patients admitted to the Department of Equine Sciences at Utrecht University for severe colic (n = 13). Seventeen of these samples were tested in duplicate on both instruments to determine the repeatability of the measurements. Blood lactate concentrations measured with the Lactate Pro ranged from 0.8–17.6 mmol/liter and with the ABL from 1.0–18.6 mmol/liter. The correlation between lactate concentrations obtained using the Lactate Pro and values from the ABL was 0.90, and the relationship was represented by the following formula: y = 0.90 · x + 0.36, indicating a linear relationship between values produced by the ABL and Lactate Pro. The repeatability for the Lactate Pro was high (0.997), which is comparable to the ABL (0.999).
Benign Peripheral Nerve Sheath Tumor of the Perianal Region in a Young Pony
Authors: Sturgeon Ben P. R. , Milne Elspeth M. , Smith Kenneth C.
Published: May 2011

A 20 × 10 cm, lobulated mass was present in the perianal region of a 4-year-old Dales pony. Histopathology revealed an unencapsulated, loose arrangement of sheets and whorls of narrow mesenchymal cells, situated in the deep dermis. Intervening areas had a myxomatous appearance. The whorls were often arranged around a central structure resembling an axon or a vascular structure. Immunohistochemistry revealed that the elongated mesenchymal cells and central axon-like structures expressed vimentin, S-100, and neuron-specific enolase, but not pancytokeratin, glial fibrillary acid protein, and the neurofilament markers. NR4 and 2F11. On the basis of the histopathology and immunohistochemistry, a diagnosis of benign peripheral nerve sheath tumor (schwannoma type) was made. This case was unusual in that the concentric laminations of Schwann cells were very loosely arranged, with an intervening myxomatous stroma (Antoni type B appearance) and despite its benign histological appearance, the mass extended deeply to the proximal sacral vertebrae. Its exact origin was unclear; it may have arisen from cutaneous nerves with deep extension or from neural structures in the sacral region. Due to the incomplete surgical removal, regrowth of the mass occurred.
Pooled Polymerase Chain Reaction to Detect Tritrichomonas Foetus in Beef Bulls
Authors: Kennedy James A. , Pearl Dayla , Tomky Linda , Carman Jane
Published: May 2011

Preputial scraping samples from 305 mixed breed beef bulls were examined for the detection of Tritrichomonas foetus infection. All samples were collected by veterinarians and transported in commercial media to an accredited lab. Upon arrival samples underwent microscopic examination for the presence of Tritrichomonas foetus and were then incubated until 5 days postcollection before final microscopic examination. Culture detected 14 samples with Trichomonad spp.; all were confirmed to be Tritrichomonas foetus by polymerase chain reaction (PCR). After final examination samples were randomly placed in groups of 5 samples; technicians were blinded as to culture results of the individual samples constituting each pool. From each sample within a group, a portion of the fluid sediment was removed and pooled with the other samples of the group to form 61 pools. From each of the formed pools an aliquot was removed for PCR. PCR detected 16 positive pools; an additional 2 positive samples were then identified on individual PCR on samples previously diagnosed as culture negative. Relative to culture, the 95% confidence intervals for sensitivity and specificity of PCR pools to detect Tritrichomonas foetus were 76.8% to 100% (mean value: 100%) and 85.5 to 99.5% (mean value: 93.4%), respectively.
Lymphangiosarcoma in the Nictitating Membrane of a Horse
Authors: Puff Christina , Herder Vanessa , Philipp Andrea , Baumgärtner Wolfgang
Published: May 2011

A 15-year-old Haflinger gelding presented with a mass in the left nictitating membrane. Two biopsies and the excised nictitating membrane were taken at different time points as a result of reoccurrence of the mass and submitted for histopathologic evaluation. The horse was euthanized as a result of poor prognosis following the reoccurrence of the mass after surgical removal. Histologically, the mass consisted of dilated, thin-walled vascular clefts and channels, lined by flattened to cuboidal endothelial cells with moderate cellular pleomorphism. There was up to 1 mitotic figure per high power field. The channels were empty or contained few erythrocytes. In the collagen-rich stroma, few lymphocytes, focal follicular lymphoid aggregations, and marked lymphangiectasia were observed. Immunohistochemically, the neoplastic cells stained positive for vimentin and partially positive for factor VIII-related antigen. Ultrastructural analysis revealed discontinuous endothelial lining vascular channels that partially lacked a basal membrane. Based on the histopathologic, immunohistochemical, and ultrastructural features lymphangiosarcoma was diagnosed.
Immunohistochemical Characterization of a Hepatic Neuroendocrine Carcinoma in a Cat
Authors: Ferreira-Neves Patricia , Lezmi Stéphane , Lejeune Typhaine , Rakotovao Farasao , Dally Claire , Fontaine Jean-Jacques , Bernex Florence , Cordonnier Nathalie
Published: May 2011

A hepatic mass was identified in a 5-year-old, female mixed-breed cat that died spontaneously after a clinical history of progressive emaciation, ptyalism, and persistent coryza. At necropsy, a 7-cm-diameter, yellow-brown, firm, multilobulated tumor was identified in the liver. Microscopically, the mass consisted of neoplastic cells arranged in small, closely packed nests within a thin fibrovascular stroma. These cells were of medium sized and polygonal, with fine argyrophilic cytoplasmic granules. Nuclei were predominantly round with finely stippled chromatin and indistinct nucleoli. Mitotic figures were numerous. Immunohistochemically, most of the neoplastic cells were immunoreactive for chromogranin A, neuron-specific enolase (NSE), and cytokeratin AE1/AE3 and weakly labeled for synaptophysin. The tumor was negative for glial fibrillary acidic protein (GFAP), vimentin, and cytokeratins 5, 6, 8, and 17. Vascular emboli and intrahepatic micrometastasis were also identified with chromogranin A. All these features were consistent with a hepatic neuroendocrine carcinoma and emphasized the importance of using a panel of antibodies to diagnose such rare tumors.
Prion protein in sheep urine
Authors: Andrievskaia Olga , Algire James , Balachandran Aru , Nielsen Klaus
Published: May 2011

The misfolded form of cellular prion protein (PrPc) is the main component of the infectious agent of transmissible spongiform encephalopathies and the validated biomarker for these diseases. The expression of PrPc is highest in the central nervous system and has been found in peripheral tissues. Soluble PrPc has been detected in cerebrospinal fluid, urine, serum, milk, and seminal plasma. In this study, attempts were made to characterize prion protein in urine samples from normal and scrapie-infected sheep. Urine samples from scrapie-infected sheep and age-matched healthy sheep were collected and analyzed by Western blot following concentration. A protease K-sensitive protein band with a molecular weight of approximately 27–30 kDa was visualized after immunoblotting with anti-PrP monoclonal antibodies to a C-terminal part of PrPc, but not after immunoblotting with monoclonal antibodies to an N-terminal epitope of PrPc or with secondary antibodies only. The amount of PrPc in the urine of 49 animals (control group: n = 16; naturally scrapie-infected group: n = 33) was estimated by comparison with known amounts of ovine recombinant PrP in the immunoblot. Background concentration of PrPc in urine was found to be 0–0.16 ng/ml (adjusted to the initial nonconcentrated volume of the urine samples). Seven out of 33 naturally scrapie-infected animals had an elevated level (0.3–4.7 ng/ml) of PrPc in urine. The origin of PrPc in urine and the reason for the increased level of PrPc in scrapie-infected sheep urine has yet to be explored.
Comparison of two real-time reverse transcription polymerase chain reaction assays for the detection of Equine arteritis virus nucleic acid in equine semen and tissue culture fluid
Authors: Lu Zhengchun , Branscum Adam J. , Shuck Kathleen M. , Zhang Jianqiang , Dubovi Edward J. , Timoney Peter J. , Balasuriya Udeni B. R.
Published: May 2011

Two previously developed TaqMan fluorogenic probe-based 1-tube real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assays (T1 and T2) were compared and validated for the detection of Equine arteritis virus (EAV) nucleic acid in equine semen and tissue culture fluid (TCF). The specificity and sensitivity of these 2 molecular-based assays were compared to traditional virus isolation (VI) in cell culture. The T1 real-time RT-PCR had a higher sensitivity (93.4%) than the T2 real-time RT-PCR (42.6%) for detection of EAV RNA in semen. However, the T1 real-time RT-PCR was less sensitive (93.4%) than the World Organization for Animal Health (OIE)-prescribed VI test (gold standard). The sensitivity of both PCR assays was high (100.0% [T1] and 95.2% [T2]) for detecting EAV RNA in TCF. In light of the discrepancy in sensitivity between either real-time RT-PCR assay and VI, semen that is negative for EAV nucleic acid by real-time RT-PCR that is from an EAV-seropositive stallion should be confirmed free of virus by VI. Similarly, the presence of EAV in TCF samples that are VI-positive but real-time RT-PCR-negative should be confirmed in a 1-way neutralization test using anti-EAV equine serum or by fluorescent antibody test using monoclonal antibodies to EAV. If the viral isolate is not identified as EAV, such samples should be tested for other equine viral pathogens. The results of this study underscore the importance of comparative evaluation and validation of real-time RT-PCR assays prior to their recommended use in a diagnostic setting for the detection and identification of specific infectious agents.
Detection of Porcine reproductive and respiratory syndrome virus infection in porcine oral fluid samples: a longitudinal study under experimental conditions
Authors: Prickett John , Simer Robert , Christopher-Hennings Jane , Yoon Kyoung-Jin , Evans Richard B. , Zimmerman Jeffrey J.
Published: May 2011

Isolation of Porcine reproductive and respiratory syndrome virus (PRRSV) from oral fluids was first reported in 1997. The objective of the present study was to determine whether PRRSV and/or anti-PRRSV antibodies were present in oral fluids at diagnostic levels. The level and duration of PRRSV and anti-PRRSV antibodies in serum and oral fluids was evaluated in 3 age groups of pigs (4,8, or 12 weeks of age) inoculated with a type 2 (North American) PRRSV isolate. Serum, buccal swabs, and pen-based oral fluid samples were collected for 63 days following inoculation. Specimens were assayed for PRRSV by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), and for anti-PRRSV antibodies by enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT). Porcine reproductive and respiratory syndrome virus was detected by real-time qRT-PCR in serum for approximately 5 weeks and in oral fluids for approximately 4 weeks postinoculation. Pig age at the time of inoculation had no effect on the quantity or duration of virus in oral fluid samples. Low levels of anti-PRRSV antibody were detected in oral fluid samples by ELISA and IFAT. Although the approach remains to be validated in the field, the results of this experiment suggest that pen-based oral fluid sampling could be an efficient, cost-effective approach to PRRSV surveillance in swine populations.
Comparison of flow cytometry and immunofluorescence microscopy for the detection of Giardia duodenalis in bovine fecal samples
Authors: Uehlinger Fabienne D. , Barkema Herman W. , O'Handley Ryan M. , Parenteau Monique , Parrington Lorna J. , VanLeeuwen John A. , Dixon Brent R.
Published: May 2011

The performance of flow cytometry (FC) was compared with immunofluorescence microscopy (IM) for detection of Giardia duodenalis in bovine feces. Samples from 36 adult dairy cows and 208 dairy calves were collected. Flow cytometry test characteristics were calculated using continuous, ordinal, and dichotomized results. Spearman correlation coefficients comparing the results of the 2 tests were 0.47 and 0.68 for cows and calves, respectively. Using IM as indicative of presence or absence of G. duodenalis cysts in each sample, likelihood ratios of FC results with 0, 1, and ≥2 gated events indicated that samples with 1 gated event were likely to be positive in the cows but not in the calves. Immunofluorescence microscopy detected G. duodenalis in 69.7% and 48.1% of cows and calves, respectively. When dichotomizing the FC results at a cutoff point of 1 or 2 gated events, 46.3% and 19.9% of the cow and 51.9% and 35.1% of the calf samples, respectively, were classified as G. duodenalis-positive. Relative to IM, the sensitivity in the cows was 0.59 and 0.28, respectively, and 0.76 and 0.64, respectively, in the calves. At a cut-off point of 1, 65.7% and 73.1% of the cow and calf samples, respectively, were correctly classified in FC, and at a cut-off point of 2, 49.3% and 78.4% were correctly classified in the cows and calves, respectively. Flow cytometry was less sensitive than IM. Possible reasons and research needed to improve FC for G. duodenalis detection are discussed.
Investigation of characteristics and factors associated with avian mycobacteriosis in zoo birds
Authors: Witte Carmel L. , Hungerford Laura L. , Papendick Rebecca , Stalis Ilse H. , Rideout Bruce A.
Published: May 2011

The objective of the current study was to identify factors associated with avian mycobacteriosis in zoo birds. Inventory data, population health records, and necropsy data from eligible birds in the Zoological Society of San Diego's (ZSSD) collection from 1991–2005 ( n = 13,976) were used to describe disease incidence, prevalence, and postmortem findings. A matched case-control study was then conducted to identify factors describing demographic, temporal, and enclosure characteristics, along with move and exposure histories. Cases (disease-positive birds; n = 167) were matched in a 1:7 ratio with controls (disease-negative birds; n = 1169) of similar age and taxonomic grouping. Potential risk factors were evaluated using univariate and multivariable conditional logistic regression. Disease prevalence and incidence were estimated for the study period at 1.2% and 3 cases/(1,000 bird-years at risk), respectively. Lesion characteristics and order prevalence are described. In the multivariable model, case birds were more likely to have been previously housed with a bird with mycobacterial disease involving the intestinal tract (odds ratio [OR] = 5.6, P < 0.01) or involving only nonintestinal sites (OR = 2.0, P < 0.01). Cases were more likely to have been imported into the collection than hatched at the ZSSD (OR = 4.2, P < 0.01). Cases were moved among ZSSD enclosures more than controls (OR = 1.1 for each additional move, P < 0.01). Findings will help guide future management of this disease for zoo bird populations.
Frequency of yeasts and dermatophytes from healthy and diseased dogs
Authors: Prado Marilena R. , Brilhante Raimunda S. N. , Cordeiro Rossana A. , Monteiro André J. , Sidrim José J. C. , Rocha Marcos F. G.
Published: May 2011

The aim of this study was to investigate the presence of dermatophytes and yeasts in healthy and diseased dogs. A total of 633 samples were collected from 26 healthy animals (104 samples), 131 with dermatitis (343 samples), 74 with otitais (148 samples), and 19 with ocular diseases (38 samples). Cultures from healthy animals were positive for Malassezia pachydermatis in 13.5% (7/52) of samples from skin, 42.3% (11/26) from ear, and 3.8% (1/26) from eye. Fungal growth was observed in 20.4% (70/343) samples from animals with dermatitis. Microsporum canis was the most isolated fungus ( n = 39), followed by M. pachydermatis ( n = 30) and Malassezia sp. ( n = 3). Of the 148 samples from dogs with otitis, 90 (60.8%) were positive for M. pachydermatis, and of the clinical specimens from the conjunctiva of animals with ophthalmic disease, 2.6% (1/38) presented positive cultures for M. pachydermatis. Only 14.3% (2/14) of the positive cultures for M. pachydermatis and 40.9% (9/22) of those for M. canis were positive in the direct exam. Direct exams were positive in 84.3% (70/83) of the culture positive samples from affected ears of dogs with otitis. Malassezia pachydermatis may act as an aggravating factor in the occurrence of cutaneous diseases, or the isolation of M. canis may be associated with the onset of dermatophytosis. Fungal culture, rather than microscopic examination, should be used as the definitive diagnostic test for dermatomycoses and otitis.
A real-time polymerase chain reaction assay to detect single nucleotide polymorphisms at codon 171 in the prion gene for the genotyping of scrapie susceptibility in sheep
Authors: McKay Jerome T. , Brigner Tiffany A. , Caplin Brian E. , McCurdy Kimberly S. , Forde Richard L.
Published: May 2011

The objective of this study was to report a reliable real-time polymerase chain reaction assay compatible with the Roche LightCycler 2.0 capable of genotyping sheep for scrapie susceptibility at codon 171. The single nucleotide polymorphisms (SNPs) in the prion protein gene in sheep that may govern resistance to scrapie at codon 171 encode for lysine (K), histidine (H), glutamine (Q), and arginine (R). A modified proteinase K method for leukocytes or whole blood was used to isolate genomic DNA from sheep blood. Fluoresentric developed and optimized primers and probes for the codon 171 SNP assay. The assay was initially validated using 218 determinations from whole blood of known genotypes with 100% correct identity. The assay was further validated through a whole-blood check test provided annually by the National Veterinary Services Laboratory with a correct identification rate of 100%. From January 2005 to December 2006, 3,672 samples from blood were genotyped at codon 171. The genotypes were QR171 ( n = 1,838, 50.05%). RR171 ( n = 1,423, 38.75%), QQ171 ( n = 407, 11.08%), HR171 ( n = 2, 0.05%), and HQ171 ( n = 2, 0.05%). The combination of this simple extraction method and the novel Fluoresentric assay is very accurate, is capable of identifying all 4 SNPs at codon 171 in one reaction, and has proven to be a useful tool for producers in their selective breeding programs.
Culture of Piscirickettsia salmonis on enriched blood agar
Authors: Mauel Michael J. , Ware Cynthia , Smith Pedro A.
Published: May 2011

Piscirickettsia salmonis is the etiologic agent of piscirickettsiosis, an economically significant disease offish. Isolation of P. salmonis by culturing on fish cell lines has been the standard technique since the initial isolation of the organism. The ability to grow P. salmonis on artificial media would relieve facilities of the cost of maintaining cell lines, permit isolation at fish culture sites with fewer contamination problems, and allow easier transport of isolates to diagnostic facilities for confirmation assays. This report describes the successful culture of P. salmonis on enriched blood agar.
Association of canine obesity with reduced serum levels of C-reactive protein
Authors: Veiga Angela P. M. , Price Christopher A. , de Oliveira Simone T. , dos Santos Andréa P. , Campos Rómulo , Barbosa Patricia R. , González Félix H. D.
Published: May 2011

The prevalence of obesity is increasing in dogs as well as in humans. C-reactive protein (CRP) is an important tool for the detection of inflammation and/or early tissue damage and is linked to obesity in humans. The objective of the present study was to determine if serum CRP levels are altered in obese dogs. Fifteen lean (control group) and 16 overweight (obese group) dogs were examined. Blood samples were collected under fasted conditions for serum determination of CRP, glucose, insulin, cholesterol, triglyceride, and fructosamine. Results indicated that obese dogs were insulin resistant because serum insulin and insulin/glucose ratios were higher than in lean dogs ( P ≤ 0.05). Serum CRP concentrations were lower in obese dogs than in controls ( P ≤ 0.001). C-reactive protein was negatively correlated with insulin/glucose ratio ( R = −0.42) and cholesterol ( R = −0.39; P ≤ 0.05). Furthermore, levels of cholesterol, triglycerides, and fructosamine were increased in the obese group compared with the control group. Based on these results, it can be postulated that CRP production is inhibited by obesity and insulin resistance in dogs.
Identification of complex vertebral malformation carriers in Chinese Holstein
Authors: Chu Qin , Sun Dongxiao , Yu Ying , Zhang Yi , Zhang Yuan
Published: May 2011

Complex vertebral malformation (CVM) is a monogenic autosomal recessive hereditary defect of Holstein dairy cattle. It is caused by a point mutation from G to T at the nucleotide position 559 in bovine solute carrier family 35, member 3 gene (SLC35A3), which changes the amino acid sequence of uridine 5′-diphosphate-N-acetylglucosamine transporter protein from a valine to a phenylalanine in position 180. The elite U.S. Holstein sire Penstate Ivanhoe Star was identified as the common ancestor of the current CVM carriers. Because his offspring, mainly those of Carlin-M Ivanhoe Bell, were used in many countries, CVM has potentially spread into China. In the present study, using the polymerase chain reaction-single-stranded conformational polymorphism (PCR-SSCP) technique, 10 CVM carriers were found among 68 at-risk Chinese Holstein bulls, and 282 carriers were found among 602 at-risk cows. The results of this study indicate that the CVM gene exists in the Chinese Holstein population.
Respiratory herpesvirus infection in two Indian Ringneck parakeets
Authors: Lazic Tatjana , Ackermann Mark R. , Drahos Jo M. , Stasko Judith , Haynes Joseph S.
Published: May 2011

A flock of Indian Ringneck parakeets ( Psittacula krameri manillensis) was imported to the United States from Australia. Soon after, 1 parakeet suddenly died, and a second parakeet died after a 2-day course of illness, which consisted of anorexia, lethargy, emaciation, and dyspnea. At necropsy, the affected birds had diffuse consolidation and red discoloration of the lungs, as well as thickened, congested air sacs. The microscopic examination revealed multifocal, necrotizing bronchitis, parabronchitis, and interstitial pneumonia. The lumen of the affected airways contained numerous, large syncytial cells with up to 15 nuclei. The nuclei of these syncytial cells often contained large, eosinophilic inclusion bodies, consistent with herpesvirus. The epithelium of the trachea and air sacs was hypertrophied and contained syncytial cells with intranuclear inclusion bodies similar to the bronchi. In addition, a few intranuclear inclusion bodies were also present in the epithelial cells that line the air capillaries. On ultrastructural examination, the nuclei of degenerating epithelial cells contained clusters of viral nucleocapsid proteins and unenveloped, icosahedral, viral particles that were approximately 90 nm in diameter. In addition, some epithelial cells contained clusters of enveloped viral particles approximately 105 nm in diameter, within the cytocavitary network. These lesions are characteristic of those caused by respiratory herpesvirus of parakeets.
Diagnosis of Clostridium Perfringens Intestinal Infections in Sheep and Goats
Authors: Uzal Francisco A. , Songer J. Glenn
Published: May 2011

Clostridium perfringens produces enteric diseases, generically called enterotoxemias, in sheep, goats, and other animals. This microorganism can be a normal inhabitant of the intestine of most animal species, including humans, but when the intestinal environment is altered by sudden changes in diet or other factors, C. perfringens proliferates and produces potent toxins that act locally or are absorbed into the general circulation with usually devastating effects on the host. History, clinical signs, and gross postmortem findings are useful tools for establishing a presumptive diagnosis of clostridial enterotoxemia in sheep and goats. Definitive diagnosis requires laboratory confirmation. Isolation of some types of C. perfringens (e.g., B and C) can be of diagnostic value, but other types (e.g., A) are so commonly found in the intestine of normal animals that isolation is meaningless from a diagnostic point of view. The most accepted criterion in establishing a definitive diagnosis of enterotoxemia is detection of C. perfringens toxins in intestinal contents. Also, histopathological examination of brain is very useful for diagnosis of type D disease, as lesions produced by epsilon toxin in the brains of sheep and goats are pathognomonic for type D enterotoxemia. Ancillary tests, such as measuring urine glucose or observing Gram-stained smears of intestinal mucosa, can be used. However, although such tests have a presumptive diagnostic value when positive, they cannot be used to rule out a diagnosis of enterotoxemia when negative.
Porcine Circovirus-2 DNA Concentration Distinguishes Wasting from Nonwasting Pigs and is Correlated with Lesion Distribution, Severity, and Nucleocapsid Staining Intensity
Authors: Harding John C. S. , Baker Crissie D. , Tumber Anju , McIntosh Kathleen A. , Parker Sarah E. , Middleton Dorothy M. , Hill Janet E. , Ellis John A. , Krakowka Steven
Published: May 2011

The emergence of severe porcine circoviral disease in North America is associated with Porcine circovirus-2 genotype b (PCV-2b), which has led to speculation that PCV-2b is more virulent than PCV-2a. The objectives of this study were to 1) correlate the PCV-2 DNA concentration and lesions in wasting (WST) and age-matched healthy (HLTH) pigs from 2 clinically affected farms, and unaffected (UNFCT) pigs from a farm with no prior clinical or diagnostic history of PCVD; and 2) to determine the initial estimates of sensitivity and specificity of PCV-2 quantitative polymerase chain reaction (qPCR). PCV-2b was confirmed in all 3 farms. Compared with HLTH pigs, WST pigs demonstrated significantly more prevalent thymic atrophy, failure of normal pulmonary collapse, and ascites ( P < 0.017 for all). The HLTH and UNFCT pigs had significantly more pronounced lymphoid germinal centers and proliferative paracortical T-dependent zones, compared with WST pigs ( P < 0.017). Across all tissues, PCV-2 DNA concentrations were significantly higher in WST compared with HLTH and UNFCT pigs ( P < 0.017 for all). The PCV-2 DNA concentrations were strongly correlated with PCV-2 nucleocapsid staining intensity in lymph node, spleen, Peyer's patches, lung, liver, and kidney (0.60 ≤ r ≤ 0.84). In the current study, the PCV-2 DNA log10 cutoff concentrations best able to distinguish WST from HLTH and UNFCT pigs were between 7.0 and 8.0 per gram for tissues, and between 4.0 and 5.0 per milliliter for sera. The presence of PCV-2b in UNFCT pigs is evidence that PCV-2b by itself is not sufficient to induce severe disease.
Evaluation of a Nested Polymerase Chain Reaction Assay to Differentiate Between Two Genotypes of Porcine Circovirus-2
Authors: Lyoo Kwang Soo , Kim Hyeun Bum , Joo Han Soo
Published: May 2011

Porcine circovirus-2 (PCV-2) is associated with several diseases in pigs, including postweaning multisystemic wasting syndrome (PMWS). A new genotype of PCV-2 was isolated from swine farms with and without clinical PMWS in North America. The new genotype was differentiated in a separate cluster by phylogenetic analyses and is now named PCV-2b compared with PCV-2a for the previously known genotype. The purpose of this study was to develop and evaluate a nested polymerase chain reaction (nPCR) assay to detect and differentiate between PCV-2a and PCV-2b. Genotype-specific primer sets were designed by using sequence data published for different PCV-2 strains. Specificity and sensitivity of the nPCR were examined by using PCV-2 isolates with known genotype. Nested PCR was found to be highly specific and sensitive for detecting and differentiating between the PCV-2 genotypes compared with the conventional 1-step PCR assay. Nested PCR was applied to detect PCV-2 and to identify the genotype in serum samples from swine farms with and without a clinical history of PMWS. Of 60 serum samples collected from 4 farms during clinical PMWS outbreaks, PCV-2a and PCV-2b were detected in 6 and 49 samples, respectively. Six of the 10 samples from one of the 4 farms had both PCV-2a and PCV-2b. Of 20 serum samples from 2 farms without PMWS, 11 were positive for PCV-2a only. These results suggest that the differential nPCR can be used to detect PCV-2 and to differentiate the 2 genotypes from field samples.
Encephalitis in Aborted Bovine Fetuses Associated with Bovine Herpesvirus 1 Infection
Authors: Brower Alexandra , Homb Kelly M. , Bochsler Philip , Porter Robert , Woods Karen , Ubl Sue , Krueger David , Cigel Francine , Toohey-Kurth Kathy
Published: May 2011

Brain tissue from 12 aborted bovine fetuses submitted to the Wisconsin Veterinary Diagnostic Laboratory revealed histologic lesions that consisted of glial nodules and variable degrees of mononuclear inflammation, microhemorrhage, neuronal necrosis, and cerebral cortical cavitation. A diagnosis of Bovine herpesvirus 1 (BHV-1) abortion had been made in all of these cases through multiple testing modalities. Brain tissue from 8 of the 12 fetuses was immunohistochemically stained with a monoclonal antibody specific to BHV-1, and, in 5 fetuses, there was positive intralesional staining of neurons, glial cells, and endothelial cells. This preliminary data suggested that herpesviral infection of brain tissue led to the described neurologic lesions. BHV-1 was then amplified from brain tissue in all 12 of the fetuses and was confirmed by partial sequencing of the thymidine kinase and glycoprotein C genes. To the authors' knowledge, neurologic lesions have not previously been described in BHV-1–infected fetuses, nor has BHV-1 previously been identified in bovine fetal brain tissue. The neurologic histopathology attributed to BHV-1 infection in these cases overlaps with the neurologic lesions produced by Neospora caninum, a common etiologic agent of bovine abortion. Therefore, when bovine fetal neurologic lesions are found, both etiologies should be considered and then distinguished by using additional diagnostic tools.
Hepatic Hemorrhage, Hemocoelom, and Sudden Death due to Haemoproteus Infection in Passerine Birds: Eleven Cases
Authors: Donovan Taryn A. , Schrenzel Mark , Tucker Tammy A. , Pessier Allan P. , Stalis Ilse H.
Published: May 2011

Haemoproteus spp. are ancient apicomplexan hemoparasites that have undergone extensive coevolution with their natural hosts and are typically species specific, with inapparent or minimal pathogenicity. A promiscuous genotype of Haemoproteus capable of undergoing host switching on a familial level was identified. This protozoan caused severe disease with high mortality in 6 species of exotic passerine birds housed in California at the San Diego Zoo's Wild Animal Park: Surinam crested oropendola ( Psarocolius decumanus decumanus), Guianan turquoise tanager ( Tangara mexicana mexicana), blue-necked tanager ( Tangara cyanicollis caeruleocephala, Guianan red-capped cardinal ( Paroaria gularis gularis), magnificent bird of paradise ( Diphyllodes magnificus hunsteini), and superb bird of paradise ( Lophorina superba). The birds had few or no clinical signs. Necropsy findings consisted of hemocoelom and irregularly scattered areas of hemorrhage and hepatocellular necrosis. Affected areas of liver contained solitary protozoal megaloschizonts in varied states of degeneration and peripheral nonsuppurative inflammation. No other parasite life stages were found in parenchymal organs or blood smears. Polymerase chain reaction using consensus primers for an avian malarial mitochondrial cytochrome B gene segment was positive in all cases. Sequencing and BLAST analysis identified the protozoan as a Haemoproteus sp. related to Haemoproteus spp. found in asymptomatic passerine birds native to North America. In situ hybridization was performed in 3 animals with a mitochondrial cytochrome B probe and was positive only in megaloschizonts. These findings suggest the recognition of a genotype of Haemoproteus that exhibits high levels of host infidelity and causes severe disease in captive birds exotic to North America.
Validation of an In-Clinic Enzyme-Linked Immunosorbent Assay Kit for Diagnosis of Borrelia Burgdorferi Infection in Horses
Authors: Johnson Amy L. , Divers Thomas J. , Chang Yung-Fu
Published: May 2011

Confirmation of Borrelia burgdorferi infection in horses has required enzyme-linked immunosorbent assay (ELISA) or Western blot tests performed by reference laboratories. An in-clinic C6 ELISA SNAP kit has been marketed for dogs. This canine kit was evaluated for horses using serum from experimentally infected ponies. Serum samples originated from 2 previous studies. In the first study, 7 ponies were exposed to B. burgdorferi–infected ticks; 4 ponies served as uninfected controls. Serum samples were obtained bimonthly for 9 months. In the second study, 16 ponies were exposed to B. burgdorferi–infected ticks. After confirmation of infection by skin culture, polymerase chain reaction (PCR), and serology, the ponies were allocated to 4 groups that received tetracycline, doxycycline, ceftiofur, or no treatment. Serum samples were obtained monthly, both before and after antibiotic treatments, for 11 months. For the current study, selected samples ( n = 220) from both studies were tested with IDEXX SNAP Heartworm Ab/ Borrelia burgdorferi Ab/ Ehrlichia canis Ab Test Kits. Tested samples included samples taken before infection, from various times postinfection, and after antibiotic treatments. Results from confirmed positive or negative samples were used to determine sensitivity and specificity of the assay. Results indicate that the test kits have fair sensitivity (63%) and very high specificity (100%) for horses recently infected with B. burgdorferi. Validation of this test provides equine practitioners with an inexpensive, in-clinic method to confirm infection, although its moderate sensitivity may result in a moderate chance of a false negative test.
Real-Time Fluorogenic Reverse Transcription Polymerase Chain Reaction Assay for Detection of African Horse Sickness Virus
Authors: Agüero Montserrat , Gómez-Tejedor Concepción , Cubillo Ángeles María , Rubio Consuelo , Romero Esther , Jiménez-Clavero Miguel Angel
Published: May 2011

African horse sickness is an arthropod-borne disease of the equine included in the World Organization for Animal Health (OIE) list with important economic consequences for horse trade. The disease is caused by African horse sickness virus (AHSV; family Reoviridae, genus Orbivirus), which is transmitted by Culicoides midges. It is endemic in sub-Saharan Africa, spreading occasionally outside this area where the occurrence of Culicoides vectors allows virus transmission. Currently, only conventional (gel-based) reverse transcription polymerase chain reaction (RT-PCR) protocols are available for its detection; however, these methods are cumbersome and difficult to apply when large numbers of samples are to be tested, as in the case of epizootics. To overcome this problem, a real-time RT-PCR method has been developed, based on a 5'-Taq nuclease-3′-minor groove binder-DNA probe (TaqMan MGB) for detection of a wide range of AHSV serotypes and strains designed to the highly conserved region of the VP7 gene (segment 7). The method was able to detect all prototype strains from the 9 known serotypes of the virus, with a high analytical sensitivity; no cross-reactions were observed with other orbiviruses or with other viruses affecting horses. The diagnostic sensitivity was assessed using a panel of AHSV-positive tissue samples from an epizootic that occurred in Spain between 1987 and 1990. This method, which can be performed in 96-well format, is suitable for large-scale surveillance of AHSV in areas where it can potentially spread.
Neurological Disease in Cattle in Southern Brazil Associated with Bovine Herpesvirus Infection
Authors: Rissi Daniel Ricardo , Pierezan Felipe , Silva Mariana Só e , Flores Eduardo Furtado , de Barros Claudio Severo Lombardo
Published: May 2011

The occurrence of neurological disease in cattle caused by Bovine herpesvirus in 11 farms from southern Brazil between 1987 and 2007 is described. Twenty-two animals were necropsied. Major clinical signs included excessive salivation, nasal and ocular discharge, circling, recumbency, depression, incoordination, grinding of teeth, and paddling movements. Necropsy findings in 10 of 22 cattle included hyperemia and softening of the rostral portions of the telencephalic cortex, with flattening of gyri, and malacia. Cattle in 10 cases did not show any gross lesions. Histological examination in most cases revealed nonsuppurative and necrotizing meningoencephalitis with acute neuronal necrosis, edema, eosinophilic intranuclear inclusion bodies in astrocytes and neurons, and infiltration of gitter cells. No histologic lesions could be detected in 4 cases. The initial diagnosis was based upon the clinical, epidemiological, and pathological findings. The diagnosis was confirmed by virus isolation in cell culture followed by virus identification by a glycoprotein C–based polymerase chain reaction. Seven isolates were identified as Bovine herpesvirus 5, and 4 were identified as Bovine herpesvirus 1.
Pseudomonas-Associated Discospondylitis in a Two-Month-Old Llama
Authors: Sura Radhakrishna , Creden Anne , Van Kruiningen Herbert J.
Published: May 2011

A 2-month-old female llama with a history of tetraparesis was presented for necropsy. This cria was apparently normal until it became ataxic during its second week of life. It had diminished flexor reflexes of the left forelimb and both hind limbs. Hematology and serum biochemistry revealed neutrophilia, elevated alkaline phosphatase, elevated phosphorus, and modest hyperglycemia. Radiography of the cervical spine demonstrated an abnormal C3–C4 intervertebral disc and bony proliferation of the vertebral end plates. The llama was treated with antibiotics and corticosteroids but failed to respond. Postmortem examination revealed cervical intervertebral disc disease and vertebral exostosis. Microscopically, there was necrosis of intervertebral connective tissue and focal malacia of the spinal cord. Pseudomonas aeruginosa was isolated from the intervertebral disc space and from cerebrospinal fluid. To the authors' knowledge, this is the first description of Pseudomonas-associated discospondylitis in a llama.
A Case of Two Different Tumors in the Heart of a Dog
Authors: Grieco Valeria , Locatelli Chiara , Riccardi Elena , Brambilla Paola
Published: May 2011

A 9-year-old, spayed, female Maremmano shepherd had a bilateral mastectomy for multiple mammary adenocarcinomas 2 years previous and was referred to the Cardiology Service of the School of Veterinary Medicine of Milan after an acute episode of cardiogenic collapse. Because of severe cardiovascular symptoms and poor prognosis, the dog was euthanized. Necropsy examination revealed the presence of multiple firm grayish neoplastic nodules in the myocardium of the left ventricle and scattered in the pulmonary parenchyma. Neoplastic nodules were also detected in the spleen, pancreas, liver, kidneys, and omentum. Histological examination revealed the coexistence of tubular adenocarcinoma and an undifferentiated sarcoma in the myocardium. Immunohistochemical staining of the sarcoma cells was negative for cytokeratin, desmin, and smooth muscle myosin, thus excluding their epithelial or myoepithelial origin, as well as an origin from smooth muscles cells. These findings, together with the coexpression of vimentin and α-smooth muscle actin, suggested that the sarcoma was derived from myofibroblasts. To the authors' knowledge, this is the first report describing cardiac sarcoma of presumptive myofibroblastic origin in a dog with simultaneous occurrence of cardiac metastasis of mammary gland adenocarcinoma.
An Outbreak of Chronic Pneumonia and Polyarthritis Syndrome Caused by Mycoplasma Bovis in Feedlot Bison (Bison Bison)
Authors: Dyer Neil , Hansen-Lardy Lynae , Krogh Darlene , Schaan Lynn , Schamber Evelyn
Published: May 2011

Mycoplasma bovis was identified by a specific lesion, conventional bacterial culture, immunohistochemistry, and polymerase chain reaction in 2 feedlot bison found dead with severe, chronic, caseonecrotic pneumonia; polyarthritis; and laryngitis. On microscopic examination, pulmonary lesions were characterized by prominent, well-defined areas of caseous necrosis and bronchiectasis. Immunohistochemical analysis of lung exhibited staining in bronchiolar epithelium and in random areas of caseous necrosis. On gross examination, the laryngeal lesion observed in 1 animal was typical of changes seen in cases of calf diphtheria. Nasal swabs taken from 6 clinically ill bison from the same feedlot revealed 1 animal shedding M. bovis by the nasal route. No other pathogens were recovered from the pulmonary or laryngeal lesions; however, Mannheimia haemolytica was cultured from the nasal swabs of 2 clinically ill bison, although not from the animal found to be shedding M. bovis. Several other affected bison had swollen joints and exhibited lameness and a reluctance to move. Changes observed in dead and clinically ill bison from this feedlot are similar to what has been described in the literature as chronic pneumonia and polyarthritis syndrome in feedlot cattle caused by M. bovis. Based on the severity of the lesions, and the number of dead and affected animals, bison in a feedlot setting appear to exhibit sensitivity to infection with M. bovis.
Malignant Mediastinal Extra-Adrenal Paraganglioma with Spinal Cord Invasion in a Dog
Authors: Rizzo Scott A. , Newman Shelley J. , Hecht Silke , Thomas William B.
Published: May 2011

A 1.5-year-old castrated, male, mixed-breed dog was evaluated because of a 2-week history of lethargy, stiffness, and progressive paraparesis. Spinal radiographs, myelography, and computed tomography of the region showed a locally invasive mass involving the thoracic wall. Upon necropsy, an encapsulated, fluctuant mass was noted attached to the right dorsal body wall in the region of the fifth to seventh thoracic vertebra. Churukian-Schenk staining revealed positive granules within the neoplastic cell cytoplasm and immunohistochemistry was positive for expression for cytoplasmic neuron-specific enolase and synaptophysin. Chromogranin A and S100 expression were found to be negative. Immunohistochemistry and silver staining did not allow further differentiation of the tumor, and the diagnosis remains consistent with either a chromaffin paraganglioma or a nonchromaffin paraganglioma (chemodectoma) with some production of catecholamines. Extra-adrenal paragangliomas are rare neoplasms in dogs but should be considered as a possible differential diagnosis for a primary paraspinal thoracic mass.
Hepatic Intranuclear Glycogen Inclusions in Western Barred Bandicoots (Perameles Bougainville)
Authors: Bennett Mark D. , Woolford Lucy , Nicholls Philip K. , Warren Kristin S. , O'Hara Amanda J.
Published: May 2011

The western barred bandicoot, Perameles bougainville, is an endangered Australian marsupial species. Routine histology of liver samples collected at necropsy from 19 of 20 (95%) western barred bandicoots revealed the sporadic to common occurrence of abnormal hepatocyte nuclei characterized by margination of chromatin and concomitant central pallor. Some abnormal hepatocyte nuclei were mildly to markedly enlarged and irregularly shaped. Periodic acid-Schiff reagent stained 131 of 142 (92%) of these abnormal hepatocyte nuclei. Positive staining was completely eliminated by diastase pretreatment. Transmission electron microscopy revealed that abnormal hepatocyte nuclei with marginated chromatin did not contain viral particles. Rather, glycogen β-particles and α-rosettes were identified within some abnormal hepatocyte nuclei. Glycogen intranuclear inclusions were an incidental finding in western barred bandicoot hepatocytes.
Fonsecaea Pedrosoi Skin Infection in a Dog
Authors: Rajeev Sreekumari , Clifton Gail , Watson Cindy , Miller Debra
Published: May 2011

Fonsecaea pedrosoi is the most common fungal agent associated with human chromoblastomycosis. In the current study, a phaeohyphomycotic condition of the skin caused by Fonsecaea pedrosoi is described in a dog.
Variation in the VP1 Gene of Foot-And-Mouth Disease Virus Serotype a Associated with Epidemiological Characteristics of Outbreaks in the 2001 Epizootic in Argentina
Authors: Perez Andres M. , König Guido , Späth Ernesto , Thurmond Mark C.
Published: May 2011

A mixed binomial Bayesian regression model was used to quantify the relation between nucleotide differences in the VP1 gene of Foot-and-mouth disease virus (FMDV) serotype A, and epidemiologic characteristics of the outbreaks from which the viruses were obtained between January and December 2001 in Argentina. An increase in the probability of different nucleotides between isolates was associated with a longer time between isolation dates, a greater distance between isolation locations, an increase in the difference between attack rates, and an increase in the difference in outbreak durations. The farther apart the outbreak herds were in the southerly and easterly directions, the greater the difference in nucleotide changes. The model accurately predicted genetic distances of isolates obtained in 2001 compared with a 2002 isolate (P < 0.01), which suggested that the predictive modeling approach applied in the present study may be useful in understanding the epidemiology of evolution of FMDV and in forensic analysis of disease epidemics.