bluuebunny/crossref_metadata_2025 huggingface dataset:
Field Evaluation of a Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay for Detection of Vesicular Stomatitis Virus
Authors: Wilson William C. , Letchworth Geoffrey J. , Jiménez Carlos , Herrero Marco V. , Navarro Roberto , Paz Pedro , Cornish Todd E. , Smoliga George , Pauszek Steven J. , Dornak Carrie , George Marcos , Rodriguez Luis L.
Published: May 2011

Sporadic outbreaks of vesicular stomatitis (VS) in the United States result in significant economic losses for the U.S. livestock industries because VS is a reportable disease that clinically mimics foot-and-mouth disease. Rapid and accurate differentiation of these 2 diseases is critical because their consequences and control strategies differ radically. The objective of the current study was to field validate a 1-tube multiplexed real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assay for the rapid detection of Vesicular stomatitis New Jersey virus and Vesicular stomatitis Indiana virus strains occurring in Mexico and North and Central America. A comprehensive collection of 622 vesicular lesion samples obtained from cattle, horses, and swine from throughout Mexico and Central America was tested by the real-time RT-PCR assay and virus isolation. Overall, clinical sensitivity and specificity of the real-time RT-PCR were 83% and 99%, respectively. Interestingly, VS virus isolates originating from a specific region of Costa Rica were not detected by real-time RT-PCR. Sequence comparisons of these viruses with the real-time RT-PCR probe and primers showed mismatches in the probe and forward and reverse primer regions. Additional lineage-specific primers and a probe corrected the lack of detection of the missing genetic lineage. Thus, this assay reliably identified existing Mexican and Central American VS viruses and proved readily adaptable as new VS viruses were encountered. An important secondary result of this research was the collection of hundreds of new VS virus isolates that provide a foundation from which many additional studies can arise.
Immunophenotyping and Gene Rearrangement Analysis in Dogs with Lymphoproliferative Disorders Characterized by Small-Cell Lymphocytosis
Authors: Yagihara Hiroko , Uematsu Yosuke , Koike Ayumi , Tamura Kyoichi , Isotani Mayu , Yamaguchi Tomohiro , Ono Kenichiro , Washizu Tsukimi , Bonkobara Makoto
Published: May 2011

Lymphocytosis caused by neoplastic proliferation of small lymphocytes is occasionally difficult to distinguish by morphological examination from nonneoplastic lymphocytosis. To examine the clinical utility of gene rearrangement analysis for demonstrating neoplastic proliferation of small lymphocytes, gene rearrangement analysis was performed in comparison with immunophenotyping using peripheral lymphocytes in dogs with small lymphocytosis. Thirty-one dogs with small-cell lymphocytosis (8,100–884,300/μl) were enrolled. By immunophenotyping, lymphocytosis of all dogs was suggested to be neoplastic in nature based on the detection of marked expansion of phenotypically homogeneous lymphocytes or the presence of an aberrant antigen-expressing population of lymphocytes. In contrast, gene rearrangement analysis represented clonality in 27 dogs (detection rate of 87%). From the present study, gene rearrangement analysis was considered to be worthwhile to strengthen the evidence of neoplastic proliferation of small lymphocytes when coupled with immunophenotyping and to be a suitable diagnostic substitute if immunophenotyping is not available in clinical practice.
Chromatographic Analysis of Lipid Fractions in Healthy Dogs and Dogs with Obesity or Hyperadrenocorticism
Authors: Jericó Márcia Marques , De Chiquito Fernanda Camargo , Kajihara Kazuo , Antonio Márcio , Moreira Bastistela , Gonzales Rodrigo , Machado Fabricio Lorenzini Aranha , Nunes Valéria Sutti , Catanozi Sergio , Nakandakare Edna Regina
Published: May 2011

Obesity and endogenous hyperadrenocorticism (HAC) are common clinical conditions in veterinary practice, and both conditions have clinical and laboratory similarities, such as weight gain and dyslipidemia. The objective of the present study was to characterize and compare the lipid profiles and plasma lipoprotein fractions in healthy dogs ( n = 10), in obese dogs ( n = 10), and in dogs with HAC ( n = 6). All of the dogs were client owned. The lipoproteins were separated by fast protein liquid chromatography, and the plasma concentrations of total cholesterol and total triacylglycerol (TAG) were determined by enzymatic methods. When compared with the healthy and obese groups, dogs with HAC had a significant increase ( P < 0.01) in the total concentrations of TAGs and cholesterol (CHOL), with higher distribution in the very low-density lipoprotein (VLDL)–CHOL fractions. In addition, the distributions of the high-density lipoprotein (HDL)–CHOL and HDL–TAG fractions were significantly lower ( P < 0.01) in dogs with HAC than in healthy dogs. Considering the animals in this study, it was determined that the dogs with HAC differed significantly from the healthy and obese dogs regarding the metabolism of CHOL and TAG, as well as their VLDL and HDL fractions. Similar laboratory findings could allow veterinarians to distinguish obese dogs from those with HAC. In addition, dogs with HAC may be at higher risk for developing metabolic and atherosclerotic complications.
Baccharis Pteronioides Toxicity in Livestock and Hamsters
Authors: Stegelmeier Bryan L. , Sani Yulvian , Pfister James A.
Published: May 2011

Baccharis pteronioides DC has been intermittently associated with livestock poisoning in the southwestern United States. In 2004, nearly 100 cows were reported poisoned by B. pteronioides in southern New Mexico. Initial field studies and postmortem examinations found drought conditions, evidence of B. pteronioides consumption, and a reported mortality of nearly 40%. Because postmortem materials were unsuitable for further examination, plant samples were collected for feeding trials and chemical evaluation. Forty-eight Syrian hamsters (8 weeks old) were randomly divided into 4 groups and dosed with 0, 50, 100, and 200 mg of B. pteronioides for 10 days. After dosing, the hamsters were necropsied; sera were analyzed biochemically; and tissues were collected and evaluated histologically. The hamsters treated with 200 mg and several of the 100-mg animals developed anorexia and diarrhea. These animals developed multiple hemorrhagic infarcts in the liver and kidney, with severe hemorrhagic enteritis. Histologically, the higher-dosed animals had severe necrotizing vasculitis with vascular thrombosis of hepatic and renal vessels. Many glomerular capillaries contained fibrin thrombi. The superficial intestinal and colonic mucosa was necrotic, with extensive hemorrhage and proliferation of luminal bacteria. Lower-dosed animals had mild hepatocellular swelling, with proliferation of intestinal and gastric bacteria and yeast. The findings indicate that at high doses, B. pteronioides is toxic to hamsters and produces lesions that are very similar to bacterial endotoxin–produced vasculitis and infarction. Research to purify and identify the toxin, the toxic dose, and mechanism of toxicity are ongoing.
Detection of All Eight Serotypes of Epizootic Hemorrhagic Disease Virus by Real-Time Reverse Transcription Polymerase Chain Reaction
Authors: Wilson William C. , O'Hearn Emily S. , Tellgren-Roth Christian , Stallknecht David E. , Mead Daniel G. , Mecham James O.
Published: May 2011

Epizootic hemorrhagic disease virus (EHDV) has been associated with bluetongue-like disease in cattle. Although U.S. EHDV strains have not been experimentally proven to cause disease in cattle, there is serologic evidence of infection. Differentiation of Bluetongue virus (BTV) and EHDV is necessary because diagnosis of infection caused by these viruses is often confused. The previously developed nested reverse transcription polymerase chain reaction (nRT-PCR) test for indigenous EHDV disease is sensitive and specific, but it is prone to contamination problems. Additionally, the EHDV nRT-PCR only detects 7 of the 8 serotypes. To develop an improved diagnostic test, sequence analysis was performed on 2 conserved target genes; one is highly expressed in infected mammalian cells, whereas the other is highly expressed in infected insect cells. This information was used to develop a rapid EHDV real-time PCR that detects all 8 EHDV serotypes. The EHDV assay did not cross-react with BTV strains and performed similarly to the nRT-PCR tests with archived clinical samples. In addition, it is superior to the nRT-PCR, not only because it is a closed system with fewer cross-contamination problems, but also because it detects all 8 serotypes and is less labor and time intensive.
A Polymerase Chain Reaction Strategy for the Diagnosis of Camelpox
Authors: Balamurugan Vinayagamurthy , Bhanuprakash Veerakyathappa , Hosamani Madhusudhan , Jayappa Kallesh Danappa , Venkatesan Gnanavel , Chauhan Bina , Singh Raj Kumar
Published: May 2011

Camelpox is a contagious viral skin disease that is mostly seen in young camels. The disease is caused by the Camelpox virus (CMLV). In the present study, a polymerase chain reaction (PCR) assay based on the C18L gene (encoding ankyrin repeat protein) and a duplex PCR based on the C18L and DNA polymerase ( DNA pol) genes were developed. The former assay yields a specific amplicon of 243 bp of the C18L gene, whereas the duplex PCR yields 243- and 96-bp products of the C18L and DNA pol genes, respectively, in CMLV, and only a 96-bp product of the DNA pol gene in other orthopoxviruses. The limit of detection was as low as 0.4 ng of viral DNA. Both PCR assays were employed successfully for the direct detection and differentiation of CMLV from other orthopoxviruses, capripoxviruses, and parapoxviruses in both cell culture samples and clinical material. Furthermore, a highly sensitive SYBR Green dye–based, realtime PCR was optimized for quantitation of CMLV DNA. In the standard curve of the quantitative assay, the melting temperature of the specific amplicon at 77.6°C with peak measured fluorescence in dissociation plot was observed with an efficiency of 102%. To the authors' knowledge, this is the first report to describe a C18L gene–based PCR for specific diagnosis of camelpox infection.
Field Estimation of the Flock-Level Diagnostic Specificity of an Enzyme-Linked Immunosorbent Assay for Avian Metapneumovirus Antibodies in Turkeys
Authors: Muñoz-Zanzi Claudia , Trampel Darrell , Hanson Tim , Harrison Kristen , Goyal Sagar , Cortinas Roberto , Lauer Dale
Published: May 2011

Routine serologic testing for Avian metapneumovirus (AMPV) infection of turkey flocks at slaughter is currently being used to monitor changes in the occurrence of AMPV infection in endemic areas and can also be used to detect the emergence of infection in currently unaffected areas. Because of the costs associated with false-positive results, particularly in areas that are free of AMPV infection, there is a need to obtain improved estimates of flock-level specificity (SP). The objective of this study was to estimate flock-level SP of a program to monitor AMPV infection in turkey flocks at processing using a standard enzyme-linked immunosorbent assay (ELISA). A study was carried out in which 37 AMPV-free flocks from 7 Midwest operations were followed serologically. Six percent, 3%, and 0.2% of total samples tested AMPV positive at 8 weeks, 12 weeks, and at processing, respectively. Overall, flock-level SP increased as the cutoff increased and as age increased. Flock-level SP at processing was 97%, if a cutoff of 1 was used (the flock was classified as positive if at least 1 sample tested positive), and 100%, if any other cutoff was used. Administration of antibiotics ( P = 0.02) and vaccination for Bordetella avium (P = 0.08) were positively associated with the probability of (false) positive test results. These findings suggest possible cross-reactions with other infections and highlight the need to consider variable diagnostic performance depending on farm conditions.
Systemic Toxoplasmosis and Gram-Negative Sepsis in a Southern Chamois (Rupicapra Pyrenaica) from the Pyrenees in Northeast Spain
Authors: Marco Ignasi , Velarde Roser , López-Olvera Jorge R. , Cabezón Oscar , Pumarola Martí , Lavín Santiago
Published: May 2011

A 6-year-old, male southern chamois ( Rupicapra pyrenaica) had an absence of flight response and was captured by hand in the Catalan Pyrenees in northeast Spain. On clinical examination, the animal was in good body condition, and only atrophy of the right eye was observed. Blood samples were collected and hematologic analysis performed, but no alterations were observed. The animal was sent to a Wildlife Rescue Centre, where it developed chronic wasting and died after 32 days in captivity. At necropsy, the animal was cachectic and had edematous, mottled lungs. Histopathologic examination revealed systemic toxoplasmosis and acute Gram-negative septicemia. The protozoan organisms were identified as Toxoplasma gondii based on immunohistochemistry. An indirect fluorescent antibody test was performed, and the animal was positive with an antibody titer of 1:50.
Cotoneaster sp. Poisoning in a Llama (Lama Glama)
Authors: Grüss Aurélie , Priymenko Nathalie
Published: May 2011

A case of poisoning with a Cotoneaster sp. is described in a llama ( Lama glama). The clinical signs were dyspnea, with congested mucous membranes, chewing, recumbency, and opisthotonos. Several hours after ingestion of the plant, the llama died, despite treatment with sodium thiosulfate and nitrite. Necropsy revealed approximately 676 g of leaves and fruit of Cotoneaster in the first compartment of the stomach. The blood was light red and did not clot. Because Cotoneaster sp. contains low concentrations of cyanogenic glycosides, toxicosis occurs only following massive ingestion of plant material.
Infection with Ovine Herpesvirus 2 in Norwegian Herds with a History of Previous Outbreaks of Malignant Catarrhal Fever
Authors: Løken Torleiv , Bosman Anna-Marì , van Vuuren Moritz
Published: May 2011

Infection with Ovine herpesvirus 2 (OvHV-2) in healthy cattle, swine, sheep, and goats was investigated on 43 selected Norwegian farms; of which, 41 (95%) had experienced outbreaks of malignant catarrhal fever (MCF) in cattle and/or swine during the preceding 5 years. Two of the farms had no history of MCF and were included for control purposes. Blood samples from 384 cattle, 40 sows, 75 sheep, and 4 goats were examined for OvHV-2 by polymerase chain reaction assay (PCR) and for antibodies using a competitive inhibition enzyme-linked immunosorbent assay (ciELISA). All samples were also tested for antibodies reactive to Alcelaphine herpesvirus 1 with an indirect fluorescent antibody test (IFAT). All but 4 of the sheep and all 4 goats tested positive with 1 or more of the tests. Eighty-nine (25%) of the cattle and 17 (43%) of the swine on the farms with previous MCF outbreaks tested positive with 1 or more of the tests. On 22 of the farms, at least 1 bovine tested positive with ciELISA and/or PCR, whereas 8 other farms had test-positive cattle with IFAT only. The 2 control farms yielded no positive results with any of the tests. Four of the farms had swine that tested positive with PCR, but none with ciELISA, whereas 4 other farms had test-positive swine with IFAT only. The prevalence of infection in cattle and swine seemed not to be influenced either by their age or the degree of contact with the sheep and goats.
Pituitary Acidophil Macroadenoma in a Pygmy Goat (Capra Hircus Hircus)
Authors: Reed Scott D. , Bauer Rudy W.
Published: May 2011

This report describes a pituitary acidophil macroadenoma in a goat. Antemortem clinical findings included hypothermia and rumen stasis. Clinicopathologic findings included refractory hypoglycemia, low total thyroxin and insulin concentrations, elevated bile acid concentration, and hyposthenuria. In addition to the pituitary macroadenoma, bilateral atrophy of the zona reticularis of the adrenal glands was observed histologically.
Isolation of Campylobacter Fetus Subspecies Fetus in a Two-Year-Old Quarterhorse with Chronic Diarrhea of an Undetermined Etiology
Authors: Hurcombe Samuel D. A. , Fox James G. , Kohn Catherine W.
Published: May 2011

A 2-year-old Quarterhorse was evaluated for chronic diarrhea and weight loss of 5 weeks duration after numerous diagnostic tests failed to identify an underlying cause. Historically, the horse was housed at pasture where human household waste vehicles were routinely cleaned and the effluent could run onto the field. Physical examination revealed poor body condition and frequent high-volume diarrhea. Diagnostic testing for Salmonella spp., endoparasites, Cryptosporidium spp., Clostridium spp., and diffuse infiltrative bowel disease were negative. Rectal tissue histopathology failed to identify Mycobacterium spp., spirochetal organisms, or submucosal infiltration with cells. Rectal tissue biopsy and a fresh fecal sample identified numerous Campylobacter organisms with microaerobic culture. Molecular testing revealed the species as Campylobacter fetus subsp. fetus as a possible organism associated with large bowel disease in this filly. The organism was sensitive to fluoroquinolone antimicrobials in vitro. The filly responded transiently to therapy, forming discrete fecal balls after 72 hr of treatment. At 5 months follow-up, the horse had gained weight, was alert and responsive, but reverted back to having soft “cow-pie” feces. Reculture of the feces at 9 months failed to identify any Campylobacter organisms. To the authors' knowledge, this is the first report to identify C. fetus subsp. fetus from feces and tissue of a horse with the use of molecular methods. This organism could play a role in the etiology of chronic diarrhea in horses.
Pneumonia from Angiostrongylus Vasorum Infection in a Red Panda (Ailurus Fulgens Fulgens)
Authors: Patterson-Kane Janet C. , Gibbons Lynda M. , Jefferies Ryan , Morgan Eric R. , Wenzlow Nanny , Redrobe Sharon P.
Published: May 2011

A 9-year-old, male, captive red panda ( Ailurus fulgens fulgens) in an urban zoo in the United Kingdom presented with respiratory distress and weight loss. The animal was euthanatized, and a postmortem examination was performed. The lungs were diffusely consolidated with extensive mineralization. Microscopically, there was extensive obliteration of normal pulmonary architecture by sheets and coalescing nodules of partially mineralized fibrous tissue and granulomatous inflammation centered on large numbers of nematode larvae and eggs. First stage nematode larvae were isolated from lung tissue and were characterized as Angiostrongylus vasorum on the basis of their morphology and sequencing of the 18S ribosomal RNA gene and the entire second internal transcribed spacer. Although A. vasorum has previously been reported in red pandas in a zoological collection in Denmark, this study is the first reported case in the United Kingdom and occurs against a background of geographical spread and increased incidence of disease in domestic and wild canids. Angiostrongylus vasorum should be considered a differential diagnosis for respiratory disease in the red panda and taken into account when planning parasite and pest control programs for zoological collections.
Isolation of Propionibacterium Acnes from a Case of Placentitis and Abortion in a Cow
Authors: Lyons Jeremiah A. , Bemis David A. , Bryant Mary Jean , Kania Stephen A. , Abd-Eldaim Mohamed , Newman Shelley J.
Published: May 2011

On the basis of the scarcity of reports in the veterinary literature, it appears that Propionibacterium spp. are rarely associated with disease or isolated from cattle tissues. Recently, Propionibacterium spp. has been associated with multifocal abscessation in cattle. This report describes a case of necrosuppurative placentitis and abortion in an adult Holstein cow. Numerous colonies of small, pleomorphic, Gram-positive, rod-shaped bacteria were observed within the fibrin lattice associated with placental lesions and within the fetal atelectatic lung. Propionibacterium acnes was isolated in high numbers from the placenta, fetal lung, and stomach contents. To the authors' knowledge, this is the first report of placentitis associated with propionibacteria in a cow.
Feline Intestinal T-Cell Lymphoma: Assessment of Morphologic and Kinetic Features in 30 Cases
Authors: Cesari Alessandro , Bettini Giuliano , Vezzali Enrico
Published: May 2011

In this study, 30 feline intestinal T-cell lymphomas (ITCLs) from 77 cats with gastrointestinal lymphoma were evaluated. Neoplastic lesions were composed predominantly of small ( n = 21) or medium to large ( n = 9) anaplastic cells. Different patterns of tumor growth were observed. A starry-sky pattern was evident in 7 cases (23.3%), fibrosis in 18 cases (60%), and neovascularization in 19 cases (63.3%). The cell proliferation index (assessed by MIB1 [mindbomb homolog 1] immunohistochemistry) ranged from 0.21% to 66.91%. Mean MIB1 index was 3.49% within the first 33rd percentile, 18.31% within the second 33rd percentile, and 40.16% within the third 33rd percentile ( P < 0.000001). Microscopic and kinetic features provided evidence that ITCL in cats exhibits a spectrum in cytological composition and growth patterns that could putatively reflect differences in biologic behavior.
Metastatic Uterine Adenocarcinoma and Hepatic Lipomatosis in a Llama (Lama Glama)
Authors: Klopfleisch Robert , van der Grinten Elisabeth , Gruber Achim D.
Published: May 2011

A 6-year-old, female llama ( Lama glama) presented with progressive anorexia and ascites. Postmortem examination revealed an infiltrative uterine adenocarcinoma with widespread metastases. The neoplasm completely replaced and infiltrated the myometrium of the uterine body and cervix and metastasized largely to the serosal surfaces of the peritoneal cavity. Histopathology identified a highly invasive growth of neoplastic cells in solid packets or tubular arrangements and marked fibroplasia. No bovine or llama papillomavirus DNA was detected intralesionally by polymerase chain reaction.
Sarcocystis Falcatula—Associated Encephalitis in a Free-Ranging Great Horned Owl (Bubo Virginianus)
Authors: Wünschmann Arno , Rejmanek Daniel , Cruz-Martinez Luis , Barr Bradd C.
Published: May 2011

A great horned owl ( Bubo virginianus) was admitted to a rehabilitation clinic with severe neurologic signs that were unresponsive to supportive care. The animal was euthanatized because of a poor prognosis. Marked granulomatous encephalitis with focal brainstem malacia was detected microscopically. The brainstem was the most severely affected brain location and the only place in which schizonts and merozoites, morphologically compatible with Sarcocystis spp., were detected. Immunohistochemistry with the use of polyclonal antisera indicated the presence of Sarcocystis falcatula. The species identification of the protozoa as S. falcatula was confirmed by polymerase chain reaction. To the author's knowledge, this is the first report of spontaneous S. falcatula–associated encephalitis in a great horned owl.
Systemic Iridovirus Infection in the Banggai Cardinalfish (Pterapogon Kauderni Koumans 1933)
Authors: Weber E. Scott , Waltzek Thomas B. , Young Devon A. , Twitchell Erica L. , Gates Amy E. , Vagelli Alejandro , Risatti Guillermo R. , Hedrick Ronald P. , Salvatore Frasca
Published: May 2011

Iridoviruses infect food and ornamental fish species from a wide range of freshwater to marine habitats across the globe. The objective of the current study was to characterize an iridovirus causing systemic infection of wild-caught Pterapogon kauderni Koumans 1933 (Banggai cardinalfish). Freshly frozen and fixed specimens were processed for histopathologic evaluation, transmission electron microscopic examination, virus culture, molecular virologic testing, microbiology, and in situ hybridization (ISH) using riboprobes. Basophilic granular cytoplasmic inclusions were identified in cytomegalic cells often found beneath endothelium, and hexagonal virus particles typical of iridovirus were identified in the cytoplasm of enlarged cells by transmission electron microscopy. Attempts at virus isolation in cell culture were unsuccessful; however, polymerase chain reaction (PCR)-based molecular testing resulted in amplification and sequencing of regions of the DNA polymerase and major capsid protein genes, along with the full-length ATPase gene of the putative iridovirus. Virus gene sequences were then used to infer phylogenetic relationships of the P. kauderni agent to other known systemic iridoviruses from fishes. Riboprobes, which were transcribed from a cloned PCR amplification product from the viral genome generated hybridization signals from inclusions within cytomegalic cells in histologic sections tested in ISH experiments. To the authors' knowledge, this is the first report of a systemic iridovirus from P. kauderni. The pathologic changes induced and the genomic sequence data confirm placement of the Banggai cardinalfish iridovirus in the genus Megalocytivirus family Iridoviridae. The ISH provides an additional molecular diagnostic technique for confirmation of presumptive infections detected in histologic sections from infected fish.
Detection of Border Disease Virus in Fetuses, Stillbirths, and Newborn Lambs from Natural and Experimental Infections
Authors: García-Pérez Ana L. , Minguijón Esmeralda , Barandika Jesús F. , Aduriz Gorka , Povedano Inés , Juste Ramón A. , Hurtado Ana
Published: May 2011

The purpose of the present study was to evaluate the use of enzyme-linked immunosorbent assay (ELISA) antigen detection in blood or fetal fluids and reverse transcription polymerase chain reaction (RT-PCR) amplification in tissues for routine laboratory diagnosis of Border disease virus (BDV) infection. Samples from 67 fetuses, 6 stillbirths, and 11 lambs from 25 commercial flocks with suspicion of BDV abortion and 3 fetuses, 7 stillbirths, and 15 lambs obtained from an experimental infection with a local isolate (BDV genotype 4) were investigated. Presence of BDV was detected by RT-PCR in 7.9% of fetuses, 50% of stillbirths, and 50% of lambs from the commercial flocks analyzed, corresponding to 8 of the 25 farms (32%). A similar percentage of the lambs and stillbirths from the experimental infection were positive by RT-PCR of tissue samples (54.5%), and the highest positivity was detected in lymph node, thyroid gland, and kidney. The current study revealed that RT-PCR analysis of stillbirths and lambs with clinical symptoms is more suitable than the analysis of fetuses to confirm the presence of BDV in a flock. Pestiviral antigen was detected by antigen ELISA in a high proportion of fetuses (24/58) and stillbirths (3/4) from commercial flocks, but in lambs, the presence of colostral antibodies masked the detection of the antigen by ELISA. Nevertheless, in lambs from the experimental infection that were not fed colostrum, antigen ELISA was less efficient than RT-PCR in detecting viral presence in stillbirths and lambs. Antigen ELISA is therefore recommended for fetuses with advanced autolysis that can adversely affect RNA integrity.
Comparison of Three DNA Extraction Methods for Detection of Erysipelothrix Rhusiopathiae in Chicken Blood by Polymerase Chain Reaction
Authors: Harada Kazuki , Uchiyama Mariko , Hoshi Teruyuki , Takahashi Toshio
Published: May 2011

A previously reported Erysipelothrix-specific polymerase chain reaction (PCR) was used to detect Erysipelothrix bacteremia in chickens. The sensitivity of PCR using 3 DNA extraction methods (boiling method, commercial gene matrix, and DNA extractor kit) was compared by using a serial 10-fold dilution of a chicken isolate of Erysipelothrix rhusiopathiae strain in chicken blood. Of the techniques used, the DNA extractor kit, followed by PCR, provided the most sensitive method for the detection of the E. rhusiopathiae strain in chicken blood (approximately 100 CFU/0.1 ml of blood). Two E. rhusiopathiae infection experiments were then attempted. In a total of 10 inoculated chickens, bacteremia developed in 9 chickens, consisting of all 5 chickens used in the first trial (ranging from 5.1 × 101 to 2.0 × 103 CFU/0.1 ml of blood) and 4 of the 5 chickens used in the second trial (ranging from 1.0 × 100 to 3.3 × 102 CFU/0.1 ml of blood). In the second trial, the 3 detection techniques were applied to the chickens with bacteremia, and the organism could be detected by using the DNA extractor kit in blood specimens from the 3 chickens exhibiting bacteremia of ≥4.2 × 101 CFU/0.1 ml of blood. This observation suggests that most E. rhusiopathiae–infected chickens develop more critical bacteremia than the detectable level by PCR with the DNA extractor kit, and the PCR detection method can be used as a first-line screening of avian erysipelas.
Validation of the Medonic CA620/530 Vet 20-μ1 Microcapillary Sampler System for Hematology Testing of Feline Blood
Authors: Nathalie H. Bourgès-Abella , Reynolds Brice S. , Anne Geffré , Braun Jean-Pierre , Trumel Catherine
Published: May 2011

The aim of the current study was to compare feline hematologic variables in blood collected in microcapillary tubes (20 μl) and conventional blood tubes with the Medonic CA620/530 Vet in-house hematologic analyzer. A comparison of results obtained in 60 cats presented at the clinics of the veterinary school showed that the correlations between the 2 methods were 0.97 for white blood cell, 0.95 for red blood cell, and 0.93 for platelet counts; 0.92 for hemoglobin concentration; and 0.99 for mean corpuscular volume. No clinically relevant differences between the 2 blood sampling techniques were observed for any variable, which suggests that both techniques are interchangeable in cats. Moreover, microcapillary tubes would allow easier repeated sampling in the same cat and would likely be useful in other small species.
Quantitative Analysis of Telomerase in Feline Mammary Tissues
Authors: Fusaro Laura , Panarese Serena , Brunetti Barbara , Zambelli Daniele , Benazzi Cinzia , Sarli Giuseppe
Published: May 2011

The purpose of this study was to validate immunohistochemistry (IHC) as an alternative to telomerase repeat amplification protocol (TRAP) analysis to detect telomerase activity. TRAP–enzyme-linked immunosorbent assay (ELISA) reactivity was compared with telomerase reverse transcription (TERT) IHC staining in 22 feline mammary tissues (6 normal mammary glands, 2 dysplastic mammary glands, 1 fibroadenoma, and 13 malignant neoplasms [6 solid mammary carcinomas, 2 squamous-cell carcinomas, 4 tubulopapillary mammary carcinomas, and 1 mammary carcinosarcoma]). TERT IHC staining revealed enzymatic expression in nuclear, nucleolar, cytoplasmic, and combined nuclear and nucleolar staining patterns that were separately quantified by image analysis and expressed as the absolute number (average) of positive cells or percentage of positive cells with respect to overall cellularity. With TERT IHC staining, the absolute number and percentage of cells with positive nuclei and nucleoli within the same cell were the variables with the greatest discrimination between benign and malignant mammary lesions (analysis of variance [ANOVA], average P < 0.0001; percentage P < 0.001). For TRAP-ELISA–positive versus TRAP-ELISA–negative tissues, a positive test result provided greater differentiation between malignant versus benign mammary lesions (ANOVA, average P = 0.00038; percentage P = 0.0022). The same IHC pattern of expression showed a proportional and significant (average P = 0.004; percentage P = 0.002) but low (average R = 0.60; percentage R = 0.63) correlation with TRAP-ELISA by the Pearson test. The correlation coefficients obtained show that IHC and TRAP cannot be considered interchangeable because the 2 methods are more complementary than exclusive.
Equine Colitis X Associated with Infection by Clostridium Difficile NAP1/027
Authors: Songer J. Glenn , Trinh H. T. , Dial Sharon M. , Brazier Jon S. , Glock Robert D.
Published: May 2011

A 14-year-old Quarter Horse with a 48-hr history of colic was euthanized after failure to respond to treatment. At necropsy, cecal and colonic mucosae were congested throughout, and there was segmental edema and significant thickening of the intestinal wall. Excessive numbers of mononuclear cells were found in mucosal lamina propria. Submucosal hemorrhage was diffuse and extensive, and Clostridium difficile toxins A and B were detected. Large numbers of C. difficile were isolated, and genetic characterization revealed them to be North American pulsed-field gel electrophoresis type 1, polymerase chain reaction ribotype 027, and toxinotype III. Genes for the binary toxin were present, and toxin negative–regulator tcdC contained an 18-bp deletion. This genotype comprises the current human “epidemic strain,” which is associated with human C. difficile–associated disease of greater than historical severity. The diagnosis was peracute typhlocolitis, with lesions and history typical of those attributed to colitis X.
An Atypical Equine Gastrointestinal Stromal Tumor
Authors: Muravnick Kathleen B. , Parente Eric J. , Del Fabio Piero
Published: May 2011

A 17-year-old, gelded Quarter Horse cross was found to have a large, intra-abdominal mass. Clinical signs included infrequent mild colic, weight loss, and chronic anemia. Surgery revealed a very large, discrete, hemorrhagic, multilobular mass with vascular attachments to the transverse colon, mesocolon, jejunal mesentery, and omentum; the site of origin was the transverse colon. Histologic examination demonstrated dense sheets, fascicles, palisades, and interconnecting streams of neoplastic spindle cells with lesser numbers of admixed multinucleated giant cells. Based on morphology alone, this neoplasm might have been misdiagnosed as a peripheral nerve sheath tumor because many of the morphologic features were suggestive of neural differentiation. Neoplastic cells expressed cluster of differentiation (CD)117 (c-kit), vimentin, desmin, smooth muscle actin, neuron-specific enolase, and S-100 protein and did not express cytokeratin. Based predominantly on the immunohistochemical profile, especially the CD117 positivity, this neoplasm was diagnosed as a gastrointestinal stromal tumor with both myogenic and neurogenic differentiation. The morphology and immunohistochemical profile of this neoplasm were different from published cases of equine gastrointestinal stromal tumors. Unusual aspects included the large size of this neoplasm, the neuroid rather than myxomatous morphology, the presence of multinucleated giant cells, and the expression of desmin.
Copper Toxicosis with Hemolysis and Hemoglobinuric Nephrosis in Three Adult Boer Goats
Authors: Bozynski Chantelle C. , Evans Tim J. , Dae Young Kim , Johnson Gayle C. , Hughes-Hanks Jennifer M. , Mitchell William J. , Rottinghaus George E. , Perry Jeanette , Middleton John R.
Published: May 2011

Acute and, particularly, chronic copper exposures, along with defects in hepatic copper metabolism, altered excretion of copper, and/or nutritional imbalances between copper and other trace elements, can lead to hepatic accumulation of copper and primary copper toxicosis. There is interspecies variation in susceptibility to copper toxicosis, with sheep being the species most likely to develop this condition. Adult dairy goats and Boer crosses are generally considered resistant to chronic copper toxicosis, especially the hemolytic stage of this disease. The current report is rather unusual in that it describes instances of naturally occurring copper toxicosis with hemolysis and hemoglobinuric nephrosis in 3 adult Boer goats. In 2 of these goats, a possible source of excessive dietary copper was investigated but not definitively identified. In the third goat, the etiologic factors associated with the copper toxicosis were not determined. It appears that mature Boer goats are susceptible to the hemolytic stage of chronic copper toxicosis, which was not observed in a recent, large-scale copper intoxication involving lactating dairy goats. Copper analyses on both liver and kidney samples were necessary to confirm the diagnosis of copper toxicosis in all 3 goats. All feedstuffs associated with instances of copper toxicosis should be analyzed for iron, molybdenum, sulphur, and zinc as well as copper to determine what nutritional factors are contributing to the pathogenesis of this disease. Consideration also should be given to the ingestion of hepatotoxic plants and other toxic exposures, which could predispose an animal to secondary chronic copper toxicosis.
Verminous Myelitis in a Pit Bull Puppy
Authors: Snook Eric R. , Baker David G. , Bauer Rudy W.
Published: May 2011

A 10-week-old, male pit bull dog presented to the referring veterinarian with hind limb paresis and epaxial muscle atrophy. No spinal lesions were identified at gross necropsy; however, histologically there was marked granulomatous myelitis in the spinal cord between T13 and L2 with occasional, intralesional nematode larvae. Based on morphologic characteristics, the nematode larvae were identified as Strongyloides spp., possibly Strongyloides stercoralis.
Fatal Onion (Allium Cepa) Toxicosis in Water Buffalo (Bubalus Bubalis)
Authors: Borelli Vanessa , Lucioli Joelma , Fernando Henrique Furlan , Patrícia Giovana Hoepers , Juliano Fleck Roveda , Sandra Davi Traverso , Gava Aldo
Published: May 2011

Toxicosis caused by the ingestion of onion ( Allium cepa) by 5 water buffalo ( Bubalus bubalis) occurred in the district of Caçador, Santa Catarina, Brazil. The water buffalo died after ingestion of a large quantity of onion that had been left in the pasture. Clinical signs started 8 days postingestion and were characterized by pale mucous membranes, lethargy, and dark urine. At necropsy, pieces of onions were found in the rumen of 1 animal. The carcass smelled strongly of onion, and the kidneys and urine were dark brown. Microscopic renal lesions included tubular degeneration and necrosis with deposits of eosinophilic material in the cytoplasm of renal tubular epithelial cells and tubular lumina. These changes were consistent with hemoglobinuric nephrosis. Centrilobular coagulation necrosis was observed in the liver accompanied by hemorrhage and macrophages containing brown cytoplasmic pigment. A diagnosis of hemolytic anemia caused by onion toxicosis was based on the epidemiological data, clinical signs, macroscopic changes, and histological lesions.
Diagnosis of Intramammary Infections at Dry-off Based on Sampling Strategy, Epidemiology of Pathogens, and Agreement Beyond Chance
Authors: Torres Audrey H. , Rajala-Schultz Päivi J. , DeGraves Fred J.
Published: May 2011

Isolation of pathogens from duplicate or multiple milk samples is currently considered the gold standard in diagnosis of bovine intramammary infections (IMI). However, in large field studies and especially in normal dairy production conditions, collection of single samples is often the most practical option to determine the causal agents of mastitis in a herd. The objective of the present study was to determine how well results between the first and the second sample in pairs of duplicate and successive quarter milk samples agree, using 5 different IMI definitions, based on the number of colony forming units (CFU) per milliliter of milk and epidemiology of the pathogens isolated. Agreement between microbiologic results from the first and the second sample of a pair was assessed by calculating the percentage of agreement and kappa coefficient. Milk samples collected at dry-off from 561 Holstein cows in 4 Ohio dairy herds were included in the analyses. Results of the study indicate that the agreement between the first and the second sample of a duplicate pair was high when criteria to call a sample positive was adjusted for the number of CFU/ml of milk by considering the epidemiology of different mastitis organisms. This finding suggests that an IMI can be accurately diagnosed with single samples. For contagious pathogens (Staphylococcus aureus) a cutoff of 100 CFU/ml and a cutoff of 1,000 CFU/ml for major environmental and minor pathogens will serve as a sensible approach to diagnose bovine IMI with single milk samples.
Bayesian Estimation of the Performance of Using Clinical Observations and Harvest Lung Lesions for Diagnosing Bovine Respiratory Disease in Post-weaned Beef Calves
Authors: White Brad J. , Renter David G.
Published: May 2011

Bovine respiratory disease (BRD) diagnosis during the postweaning phase of beef production is an important component of effective preventive health and treatment programs. Although identification of diseased animals based on signs of clinical illness (CI) is a common method in the beef industry for identifying BRD, very little information is available on the accuracy of this method. Previous investigators hypothesized that monitoring pulmonary lesions at harvest (LU) could be a more reliable indicator of disease status during the postweaning phase. A structured literature review was conducted to identify research that compared CI and LU. Because there is no true gold standard for diagnosing BRD, Bayesian methods were used to estimate the sensitivity and specificity of each diagnostic method relative to a BRD diagnosis at any time during the postweaning phase. Results from the current study indicate that the estimated diagnostic sensitivity and specificity of CI were 61.8% (97.5% probability interval [PI]: 55.7, 68.4) and 62.8% (97.5% PI: 60.0, 65.7), respectively. Use of LU for a BRD diagnosis was estimated to have a sensitivity of 77.4% (97.5% PI: 66.2, 87.3) and a specificity of 89.7% (97.5% PI: 86.0, 93.8). Further analysis revealed that the probabilities of LU having higher sensitivity and specificity than CI were 99.4% and 100%, respectively. The present research indicates that neither method was perfect, and both methods were relatively poor at correctly classifying truly diseased animals (sensitivity) but that LU was more accurate than CI for BRD diagnosis. Results from the present study should be considered when these diagnostic methods are used to evaluate BRD outcomes in clinical and research settings.
Nor98 Scrapie Identified in the United States
Authors: Loiacono Christie M. , Thomsen Bruce V. , Hall S. Mark , Kiupel Matti , Sutton Diane , O'Rourke Katherine , Barr Bradd , Anthenill Lucy , Keane Delwyn
Published: May 2011

A distinct strain of scrapie identified in sheep of Norway in 1998 has since been identified in numerous countries throughout Europe. The disease is known as Nor98 or Nor98-like scrapie, among other names. Distinctions between classic scrapie and Nor98 scrapie are made based on histopathology and immunodiagnostic results. There are also differences in the epidemiology, typical signalment, and likelihood of clinical signs being observed. In addition, sheep that have genotypes associated with resistance to classic scrapie are not spared from Nor98 disease. The various differences between classic and Nor98 scrapie have been consistently reported in the vast majority of cases described across Europe. The current study describes in detail the pathologic changes and diagnostic results of the first 6 cases of Nor98 scrapie disease diagnosed in sheep of the United States.
Axonal Degeneration in Sheep Caused by the Ingestion of Halimium Brasiliense
Authors: Riet-Correa Franklin , Barros Severo S. , Méndez Maria C. , Gevehr-Fernandes Cristina , Neto Pereira Otaviano A. , Soares Marcio P. , McGavin M. Donald
Published: May 2011

Nervous system disease is reported in sheep from 2 farms in southern Brazil and in 33 farms in Uruguay. The illness was seasonal, occurring from May to November, during the growing season of Halimium brasiliense, and primarily affected sheep older than 3 years of age. Clinical signs included transient seizures that occurred mainly when sheep were disturbed or frightened. Most affected sheep recovered when removed to other pastures. Feeding trials produced clinical signs in 1 sheep after the ingestion of 2,117 g/kg of body weight of H. brasiliense over 142 days. Two sheep that had previously recovered from spontaneous toxicosis developed clinical signs after the ingestion of 263 g and 565 g of H. brasiliense per kg body weight given over 36 and 31 days, respectively. The main histologic lesion was vacuolation of the brain and spinal cord, with rare axonal spheroid formation. Transmission electron microscopy revealed segmental axonal swelling with degeneration and disappearance of the axonal organelles and vacuolation of the axoplasm. A pigment identified as ceroid was also present in neurons, astrocytes, and macrophages. These lesions suggested a novel morphologic manifestation of a toxic axonopathy.
Characterization of Cervidpoxvirus Isolates from Oregon, California, and Eastern Canada
Authors: Moerdyk-Schauwecker Megan , Eide Kathleen , Bildfell Robert , Baker Rocky J. , Black Wendy , Graham Daniel , Thompson Kim , Crawshaw Graham , Rohrmann George F. , Jin Ling
Published: May 2011

The present report describes the analysis of 4 Deerpox virus isolates from California, Oregon, and Ontario, Canada. All 4 isolates were associated with cutaneous crusting lesions. Examination of selected samples by electron microscopy demonstrated that the viruses were morphologically similar to orthopoxviruses. Phylogenetic analysis of the A21 gene, which is found in all poxviruses, indicated that the 4 isolates form a lineage distinct from other members except for those belonging to the genus Cervidpoxvirus of the subfamily Chordopoxvirinae. Members of the Cervidpoxvirus lineage encode a set of genes not found in other poxviruses. These include homologs of genes encoding interleukin 1 receptor antagonists (IL-1Ra) and C-type lectin-like receptors (CTLR). In the current investigation, genes encoding homologs of IL-1Ra and CTLR were amplified from all the isolates and were found to be closely related to orthologs found in the Cervidpoxvirus genus, which further supports the inclusion of these isolates in the Cervidpoxvirus genus.
Genetic Variability of Archived Cytauxzoon Felis Histologic Specimens from Domestic Cats in Georgia, 1995–2007
Authors: Brown Holly M. , Modaresi Shirin M. , Cook Jessica L. , Latimer Kenneth S. , Peterson David S.
Published: May 2011

Cytauxzoon felis infection in domestic cats has historically been nearly 100% fatal. However, increasing reports of domestic cats that survive cytauxzoonosis and reports of asymptomatic cats with C. felis infections suggest the existence of different parasite strains that vary in pathogenicity. The objective of the current study was to obtain epidemiologic information about cytauxzoonosis through genotypic characterization of archived histologic specimens from domestic cats with C. felis infections that were diagnosed in Georgia between 1995 and 2007. Such retrospective data on genetic variability will provide an historic context for current studies of C. felis genotype frequencies. Cytauxzoon felis DNA was obtained from formalin-fixed, paraffin-embedded tissues from infected cats diagnosed with cytauxzoonosis at necropsy. Genetic characterization of C. felis was performed using sequence analysis of the polymerase chain reaction–amplified ribosomal internal transcribed spacer regions 1 and 2 (ITS1, ITS2). Eleven different combined ITS1 and ITS2 sequences were identified, the majority of which were identical to those previously reported in fatally infected cats from Georgia. The findings of the current study document the existence of genetically distinct C. felis populations in historical samples and, together with data from contemporary samples, demonstrate a diverse population structure for C. felis.