bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of Corynebacterium Pseudotuberculosis Infection on Serologic Surveillance for Johne's Disease in Goats
Authors: Manning Elizabeth J. B. , Cushing Heather F. , Hietala Sharon , Wolf Cynthia B.
Published: May 2011

False-positive results on serologic assays for Mycobacterium avium subsp. paratuberculosis (MAP) are believed to occur due to cross-reacting antibody produced by Corynebacterium pseudotuberculosis (C. pstb) infection in goats. This issue of compromised specificity was evaluated by testing 771 adult goats from 10 Midwestern goat herds in 2004. Assays for MAP infection included radiometric fecal culture and 2 enzyme-linked immunosorbent assays (ELISAs); ELISA-positive samples were tested by agar gel immunodiffusion (AGID). A synergistic hemolysin inhibition assay (SHI) was used to detect C. pstb antibody. Four infection status categories were evaluated. Category 1 goats (free of both MAP and C. pstb infection) tested negative on all MAP fecal cultures and SHI tests. Five of 181 goats were positive in both ELISAs, and 2 more were positive in ELISA-1 only. For Category 2 (MAP infected; no C. pstb infection), all animals were SHI negative. Six goats were fecal culture positive and strongly positive in both ELISAs; 2 more goats were positive only in ELISA-1. For Category 3 ( C. pstb infected or vaccinated; no history of MAP infection), all fecal cultures were negative and 91% were SHI test-positive. In this population, only 2 goats were positive in both MAP ELISAs, while 84 additional goats were test-positive only on ELISA-1. In the absence of C. pstb infection, both ELISAs performed comparably, but when C. pstb infection was present the performance of ELISA-1 was significantly perturbed. Use of the ELISA-2 for goats is an effective and efficient method for Johne's disease surveillance in any goat herd.
Toxoplasma Gondii in an African Crested Porcupine (Hystrix Cristata)
Authors: Harrison Tara M. , Moorman Jamee Black , Bolin Steven R. , Grosjean Nicole L. , Lim Ailam , Fitzgerald Scott D.
Published: May 2011

An adult female crested porcupine ( Hystrix cristata) was evaluated for acute onset of neurologic signs including head tilt, circling, and ataxia. She was found dead in her holding area 2 days after initially exhibiting clinical signs. Necropsy was unremarkable. Histopathology of brain tissue revealed the presence of protozoal cysts associated with inflammation as the underlying cause of clinical signs and death. Immunohistochemical staining of brain tissue for Toxoplasma gondii was strongly positive. PCR on fresh brain confirmed T. gondii as the causative organism. An adult male in the same enclosure has demonstrated similar neurologic signs over the past 3 years and has failed to respond to various medical treatments. Clinical disease associated with T. gondii has not been previously reported in this porcupine species or any other Old World porcupines, although there are several reports of clinical toxoplasmosis involving New World porcupine species.
Taxus sp. Intoxication in Three Francois' Langurs (Trachypithecus Francoisi)
Authors: Lacasse Claude , Gamble Kathryn C. , Poppenga Robert H. , Farina Lisa L. , Landolfi Jennifer , Terio Karen
Published: May 2011

Three Francois' langurs ( Trachypithecus francoisi) were found dead, without previous clinical signs, over a 48-hour period at a zoological institution after transfer to a new exhibit. A hybrid yew shrub ( Taxus baccata × T. cuspidata) was found in close proximity to the exhibit perimeter fence. Despite clinical suspicion of yew intoxication, thin-layer chromatography performed on gastric contents was negative. However, microscopic examination of gastric contents revealed multiple yew fragments, and taxine alkaloids were detected by gas chromatography and mass spectrometry of the gastric contents to confirm yew intoxication. Acute death of the animals prevented treatment. The fourth langur in the collection survived, most likely because of its low rank in the troop's hierarchy, with a suspected small amount or none of the plant ingested. To the authors' knowledge, this case report is the first yew intoxication documented in a nonhuman primate species. Taxus spp. intoxication is an often fatal condition reported in domestic animals and humans. In comparison with these species, mortality appeared delayed in the Francois' langurs, most likely because of their unique gastrointestinal anatomy, with both foregut and colonic fermentation. Plant intoxication should be a differential diagnosis when multiple acute deaths are observed after recent introduction to a new enclosure.
Development of a 1-Step Enzyme-Linked Immunosorbent Assay for the Rapid Diagnosis of Bovine Respiratory Syncytial Virus in Postmortem Specimens
Authors: Quinting Birgit , Robert Béatrice , Letellier Carine , Boxus Mathieu , Kerkhofs Pierre , Schynts Frédéric , Collard Alfred
Published: May 2011

Bovine respiratory syncytial virus (BRSV) is associated with severe respiratory disease in cattle. BRSV infection frequently leads to the death of young infected animals. The presence of BRSV in postmortem specimens is routinely detected using indirect immunofluorescence (IIF). However, this technique requires special equipment and considerable expertise. The present paper describes the development of a 1-step ELISA for rapid (1.5 hours) detection of BRSV antigen in organ homogenates. The performance of the new 1-step ELISA was evaluated using bovine postmortem specimens ( n = 108) in comparison with 3 other BRSV diagnostic techniques: indirect immunofluorescence, the Clearview respiratory syncytial virus (RSV) test, and real-time reverse transcriptase polymerase chain reaction (RT-PCR). The relative sensitivity, specificity, and the kappa coefficient of 1-step ELISA, the Clearview RSV electroimmunoassay (EIA), and IIF were calculated, using real-time RT-PCR as the reference test. The new 1-step ELISA was the most sensitive and specific of the 3 tests. Thus, the new 1-step ELISA is a reliable test for detecting BRSV antigen in organ homogenates.
An Indirect Enzyme-Linked Immunosorbent Assay for Detection of Antibodies to Actinobacillus Pleuropneumoniae Serovar 7 in Pig Serum
Authors: Klausen Joan , Ekeroth Lars , Grøndahl-Hansen Jan , Andresen Lars Ole
Published: May 2011

Lipopolysaccharide (LPS) antigen was purified from Actinobacillus pleuropneumoniae serovar 7 by phenol-water extraction and fractionated on a, S-100 Sephacryl column. High molecular weight fractions of LPS purified from the S-100 column were pooled and used as antigen in an indirect serovar 7 ELISA. The ELISA was evaluated with sera from pigs experimentally infected with 11 different A. pleuropneumoniae serovars of biotype 1. Estimation of sensitivity and specificity of the A. pleuropneumoniae serovar 7 ELISA was performed using pig sera from herds naturally infected with A. pleuropneumoniae serovar 7 as well as sera from herds free of infection with A. pleuropneumoniae serovar 7. When compared to the complement fixation test (CFT) as a reference test, the ELISA showed much higher sensitivity and statistically equivalent specificity.
Growth and Metabolic Characterization of Macrorhabdus Ornithogaster
Authors: Hannafusa Yasuko , Bradley Allison , Tomaszewski Elizabeth E. , Libal Melissa C. , Phalen David N.
Published: May 2011

Macrorhabdus ornithogaster ( M. ornithogaster) is an anamorphic ascomycetous yeast found only in the stomach of birds. Infection is often benign but has also been associated with disease in some species of birds under some circumstances. In vitro efforts to grow M. ornithogaster have been largely unsuccessful. In this report, multiple liquid and solid media of varying pH, sugar concentration, and fetal bovine serum (FBS) concentrations, incubated at various temperatures in room air or microaerophilic conditions, were examined for their ability to support the growth of M. ornithogaster, obtained from a budgerigar ( Melopsittacus undulatus). Optimum growth conditions were found to be Basal Medium Eagle's, pH 3 to 4, containing 20% FBS, and 5% glucose or sucrose under microaerophilic conditions at 42°C. Using these conditions, M. ornithogaster was repeatedly passaged without loss of viability. Polyclonal isolates of M. ornithogaster consistently assimilated glucose, sucrose, and trehalose. M. ornithogaster did not grow with prolonged exposure to atmospheric oxygen, but growth in microaerophilic conditions was moderately enhanced by preincubation with atmospheric oxygen for 24 hours. An isolate of M. ornithogaster was found to be infective to day-old chickens, reduce their rate of weight gain, and induce a mild to moderate heterophilic inflammation of the isthmus. M. ornithogaster was reisolated from the chicks 7 days after infection, fulfilling Koch's postulates. A 761-bp sequence of 18S rDNA from this isolate was compared to the originally reported M. ornithogaster sequence and was found to be 97% identical.
Characterization of Rupture of Abdominal Artery Aneurysm in Dairy Cattle
Authors: Lamm Catherine G. , Guard Charles L. , Erb Hollis N. , Njaa Bradley L.
Published: May 2011

In this retrospective study, the authors describe the gross and histologic changes associated with rupture of an abdominal artery aneurysm in 33 mature female Holstein cattle between January 1980 and June 2005 from 29 farms in upstate New York and northern Pennsylvania. Over this period, there was an increase in the number of cases submitted for necropsy per year, and a seasonal trend did not exist. Affected animals ranged from 2.5 to 5.5 years of age. Grossly affected cattle exhibited marked hemoabdomen. There was marked dilation and rupture of the abdominal aorta or one of its branches, including the mesenteric, left gastric, celiac artery, right ruminal artery, or left ruminal artery. Histologically, the tunica media of the affected arteries was often thin and irregular in width with disrupted, fragmented, and coiled elastin. Granulation tissue and hemorrhage was localized at the site of rupture. There was mild-to-marked hyperplasia of the tunica intima that was irregular and disorderly with adjacent smooth-muscle hyperplasia within the tunica media. In addition, in a fraction of cases, there was mild-to-moderate mucinous change (48%) and mineralization (30%) within the tunica media. To the authors' knowledge, this is the first description of the gross and histologic changes in Holstein cows with abdominal artery aneurysm and rupture.
Sodium Fluoride/Copper Naphthenate Toxicosis in Cattle
Authors: DeBey Brad M. , Jacob Binod , Oehme Fred W. , Imerman Paula
Published: May 2011

Fourteen cattle on a Kansas pasture died from ingestion of a wood preservative compound containing sodium fluoride and copper naphthenate. Clinical signs included depression, anorexia, ataxia, diarrhea, and recumbency. Grossly visible lesions included perirenal edema, pale kidneys, and forestomach ulceration. All 3 cows that had postmortem evaluations had extensive renal cortical tubular necrosis. Tissue concentrations of fluoride were slightly elevated above expected background levels, while copper tissue concentrations were not elevated. The findings indicated that the sodium fluoride caused renal tubular necrosis leading to renal failure. Copper naphthenate may have contributed to abomasal ulceration; however, tissue copper concentrations indicated that copper from the formulation was not appreciably absorbed from the gastrointestinal tract.
Infections Caused by Pathogenic Free-Living Amebas (Balamuthia Mandrillaris and Acanthamoeba sp.) in Horses
Authors: Kinde Hailu , Read Deryck H. , Daft Barbara M. , Manzer Michael , Nordhausen Robert W. , Kelly Daryl J. , Fuerst Paul A. , Booton Gregory , Visvesvara Govinda S.
Published: May 2011

This article describes amebic infections in 4 horses: granulomatous amebic encephalitis caused by Balamuthia mandrillaris and Acanthamoeba culbertsoni and systemic infections caused by Acanthamoeba sp. The former infection occurred in 1 of 4 horses spontaneously without any underlying conditions; the latter amebic infection was perhaps “opportunistic” considering the visceral involvement by this protozoan in association with Aspergillus sp. and/or Escherichia coli and Pseudomonas sp. The clinicopathologic findings and demonstration of the amebic organisms using immunohistochemical techniques, culture, polymerase chain reactions, and electron microscopy are presented.
Comparison of Tests for Detection of Bovine Viral Diarrhea Virus in Diagnostic Samples
Authors: Edmondson Misty A. , Givens M. Daniel , Walz Paul H. , Gard Julie A. , Stringfellow David A. , Carson Robert L.
Published: May 2011

Currently, a variety of tests are used to detect bovine viral diarrhea virus (BVDV) in persistently infected (PI) cattle. These tests include immunohistochemical staining (IHC), antigen capture enzyme-linked immunosorbent assay (ACE), virus isolation (VI), and reverse transcription-polymerase chain reaction (RT-PCR). However, a lack of methods standardization could compromise the ability to consistently identify animals infected with BVDV. This study evaluated the diagnostic proficiency of current methods for detecting BVDV in infected cattle using intra- and interlaboratory comparisons. Samples were collected from 4 animals more than 7 months of age (2 BVDV negative animals, a PI animal, and a PI animal that previously lacked detectable virus in serum as determined by VI). Samples were submitted to 23 participating diagnostic laboratories using the respective laboratory's standard submission protocol. Samples collected for submission included: 1) serum for ACE, RT-PCR, and VI; 2) whole blood for RT-PCR and VI; and 3) skin biopsies for ACE and IHC. The ACE performed on skin provided the greatest consistency in detecting positive samples and a perfect level of agreement among laboratories. Reverse transcription-polymerase chain reaction and IHC performed well by correctly identifying ≤85% of samples positive for BVDV. Virus isolation performed on serum yielded the lowest consistency in detecting positive samples and the lowest level of agreement. The level of agreement between laboratories for detecting BVDV in persistently infected cattle ranged from perfect to less than expected by chance. The variation between laboratories suggests a need for training opportunities in standardized laboratory protocols and proficiency testing.
Validation of Nonnested and Real-Time PCR for Diagnosis of Sheep-Associated Malignant Catarrhal Fever in Clinical Samples
Authors: Traul Donald L. , Taus Naomi S. , Oaks J. Lindsay , Toole Donal O' , Rurangirwa Fred R. , Baszler Timothy V. , Li Hong
Published: May 2011

Sheep-associated malignant catarrhal fever (SA-MCF), a frequently fatal disease primarily of certain ruminants, is caused by ovine herpesvirus 2 (OvHV-2). Molecular diagnosis of SA-MCF in affected animals has relied on detection of OvHV-2 DNA using a nested PCR, which has significant potential for amplicon contamination as a routine method in diagnostic laboratories. In this report, a nonnested and a previously developed real-time PCR were validated for detection of OvHV-2 DNA in samples from clinically affected animals. Three sets of blood or tissue samples were collected: 1) 97 samples from 97 naturally affected animals with evidence of clinical SA-MCF; 2) 200 samples from 8 animals with experimentally induced SA-MCF; and 3) 100 samples from 100 animals without any evidence of clinical SA-MCF. Among 97 positive samples defined by nested PCR from clinically affected animals, 95 (98%) were positive by nonnested PCR and 93 (96%) were positive by real-time PCR, respectively. One hundred percent of the samples from the animals with experimentally induced MCF were positive by real-time PCR, while 99% were positive by nonnested PCR. Neither nonnested PCR nor real-time PCR yielded a positive result on any of the 100 nested PCR-negative samples from animals without evidence of clinical MCF. The data confirmed that both nonnested and real-time PCR maintained high specificity and sensitivity for the detection of OvHV-2 DNA in clinical samples.
Use of Formalin-Fixed Tissues to Determine Fumonisin B1-Induced Sphingolipid Alterations in Swine
Authors: Hsiao Shih-Hsuan , Tumbleson Mike E. , Constable Peter D. , Haschek Wanda M.
Published: May 2011

Fumonisin B1 is a mycotoxin that causes lethal pulmonary edema in swine. Sphinganine, sphingosine, and the sphinganine to sphingosine ratio are important biomarkers for fumonisin B1 exposure. Currently, tissues selected for sphinganine and sphingosine analyses are frozen at −80°C until analyses take place. However, for diagnostics and some research projects, formalin is used more routinely as a preservative for long-term storage of tissues. To determine whether formalin-fixed tissues could be used for sphinganine and sphingosine analyses, sphinganine and sphingosine concentrations were quantified in both frozen and formalin-fixed lung, liver, kidney, and heart from fumonisin B1-treated and control pigs. Tissues were evaluated 3 months after freezing and 3, 6, and 12 months after formalin fixation. Sphinganine, sphingosine, and the sphinganine to sphingosine ratio of both frozen and formalin-fixed lung and liver from fumonisin B1-treated pigs were elevated. Formalin-fixed tissues had lower sphinganine and sphingosine concentrations but higher sphinganine to sphingosine ratios than the corresponding frozen tissues. Storage in formalin for up to 12 months did not affect the results. Sphingosine analysis could not be performed in formalin-fixed heart and kidney because of noninterpretable chromatograms. Therefore, formalin-fixed lung and liver can be used to determine fumonisin B1-induced sphinganine and sphingosine alterations in swine, with the sphinganine to sphingosine ratio being the most useful.
Development and Validation of Monoclonal and Polyclonal Antibodies for the Detection of Immunoglobulin G of Bottlenose Dolphins (Tursiops Truncatus)
Authors: Nollens Hendrik H. , Green Linda G. , Duke Diane , Walsh Michael T. , Chittick Beth , Gearhart Scott , Klein Paul A. , Jacobson Elliott R.
Published: May 2011

Antibodies directed against species-specific immunoglobulin G (IgG) have a broad range of applications in serologic and immunologic research and in the development of clinical assays. Validated anti-IgG antibodies for marine mammal species are in short supply. The objective of this study was to produce and validate antibodies with specificity for IgG of the common bottlenose dolphin ( Tursiops truncatus). Bottlenose dolphin IgG was purified using protein G. Two mouse monoclonal antibodies and a rabbit polyclonal antibody were developed from mice and rabbits immunized with bottlenose dolphin IgG. The specificity of the monoclonal antibodies and the polyclonal antibody for bottlenose dolphin IgG was first verified by Western blot analysis and enzyme-linked immunosorbent assay (ELISA). For further validation, both monoclonal antibodies and the polyclonal antibody were incorporated in an indirect ELISA for the detection of the immune response of bottlenose dolphins to a vaccine antigen. Three bottlenose dolphins were immunized with a commercial Erysipelothrix rhusiopathiae vaccine, and serial blood samples were collected from all dolphins for measurement of levels of circulating antibodies. Seroconversion was observed in all 3 dolphins by use of both monoclonal antibodies and the polyclonal antibody. Circulating antibodies were detectable as early as 6 days after immunization in 1 dolphin. Peak antibody levels were detected 14 days after the immunization. The ability to detect seroconversion in all 3 immunized bottlenose dolphins firmly establishes the specificity of the monoclonal antibodies and the polyclonal antibody for IgG of the common bottlenose dolphin.
Osteocartilaginous Metaplasia in the Right Atrial Myocardium of Healthy Adult Sheep
Authors: Gopalakrishnan Gopakumar , Blevins William E. , Van Alstine William G.
Published: May 2011

Among 172 hearts from clinically normal adult sheep aged 1.5 to 7 years evaluated for the presence of cartilage and/or bone in the right atrial myocardium, 3.49% (6/172) had palpable evidence of osteocartilaginous foci. An additional 8% prevalence was estimated based on radiographs of hearts that contained ≤1 mm foci of nonpalpable, radiographically dense bone. Microscopically, the nodules in the atria were characterized by mature lamellar bone enclosing adipose tissue, with occasional new bone formation by endochondral ossification. No degenerative changes were evident in the affected atrial myocardium, suggesting that these lesions were clinically insignificant background changes.
Occurrence of Canine Parvovirus Type 2c in the United States
Authors: Hong Charles , Decaro Nicola , Desario Costantina , Tanner Patrick , Pardo M. Camila , Sanchez Susan , Buonavoglia Canio , Saliki Jeremiah T.
Published: May 2011

Canine parvovirus (CPV) type 2 (CPV-2) emerged around 1978 as a major pathogen of dogs worldwide. In the mid-1980s, the original CPV-2 had evolved and was completely replaced by 2 variants, CPV-2a and CPV-2b. In 2000, a new variant of CPV (named CPV-2c) was detected in Italy and now cocirculates with types 2a and 2b in that country. The CPV-2c has also been reported from single outbreaks in Vietnam and Spain. This study was conducted to determine if CPV-2c occurs in the United States. Thirty-three fecal samples were collected from dogs in 16 states between April 2006 and April 2007 and were tested for CPV using real-time polymerase chain reaction (PCR). Positive samples were further tested using conventional PCR and minor-groove binding TaqMan PCR assays to determine the viral type and to differentiate vaccine strains from field strains. Twenty-seven samples were positive for CPV, 7 of which were CPV-2c from 5 states: Arizona, California, Georgia, Oklahoma, and Texas. Of the 7 isolates, 4 differed from European CPV-2c isolates by 2 additional single-nucleotide mutations at positions 4076 and 4104, the latter of which produces a ThrAla change at residue 440 located near a major antigenic site. The coast-to-coast geographic distribution of the states in which CPV-2c was detected strongly suggests that this new CPV variant is probably widespread in the United States. The continuous evolution of CPV requires that monoclonal antibody-based and nucleic acid-based diagnostic assays should be periodically checked for sensitivity on prevalent CPV strains.
Comparison of the in Vitro Susceptibility of Rodent Isolates of Pseudomonas Aeruginosa and Pasteurella Pneumotropica to Enrofloxacin
Authors: Sasaki Hiraku , Kawamoto Eiichi , Kunita Satoshi , Yagami Ken-ichi
Published: May 2011

The objectives of this study were to determine and compare the in vitro enrofloxacin susceptibility of 94 Pseudomonas aeruginosa isolates obtained from enrofloxacin-treated and untreated mice and that of 40 Pasteurella pneumotropica strains and also to assess the efficacy and effects of enrofloxacin treatment of laboratory mice. The minimum inhibitory concentrations (MICs) of enrofloxacin against all the Ps. aeruginosa isolates were in the range of 1 to 4 μg/ml, whereas those against all the P. pneumotropica strains were less than 0.5 μg/ml. The mutation frequency in 54% of the Ps. aeruginosa isolates on treatment with enrofloxacin ranged from 10−6 to 10−8; however, none of the P. pneumotropica strains could grow on medium containing more than 3 μg/ml enrofloxacin. Comparison of in vitro enrofloxacin susceptibilities suggested that enrofloxacin was effective for eliminating P. pneumotropica but not for eliminating Ps. aeruginosa for which the MIC of enrofloxacin was more than 1 μg/ml. These results indicated that the enrofloxacin susceptibility of P. pneumotropica was higher than that of Ps. aeruginosa, and that the enrofloxacin treatment might not affect the susceptibility of Ps. aeruginosa.
Nocardia Tenerifensis Genome Identification in a Cutaneous Granuloma of a Cat
Authors: Ramos-Vara José A. , Wu Ching Ching , Lin Tsang Long , Miller Margaret A.
Published: May 2011

An adult Domestic Longhair cat developed a subcutaneous mass in its tail. Histologically, this mass consisted of ill-defined pyogranulomas centered around aggregates of gram-positive, acid-fast filamentous bacteria, consistent with Nocardia. Due to the lack of fresh samples, DNA was extracted from formalin-fixed paraffin-embedded tissue sections and subjected to polymerase chain reaction amplification and DNA sequencing of 16S ribosomal RNA gene encompassing Nocardia sp.-specific sequences. Sequences analyzed using the GenBank database revealed 99.5% homology with Nocardia spp. and had the highest sequence homology of 98.2% with Nocardia tenerifensis among Nocardia spp. To the authors’ knowledge, this is the first report of detection of N. tenerifensis genome associated with cutaneous nocardiosis in an animal.
Assessment of Melamine and Cyanuric Acid Toxicity in Cats
Authors: Puschner Birgit , Poppenga Robert H. , Lowenstine Linda J. , Filigenzi Michael S. , Pesavento Patricia A.
Published: May 2011

The major pet food recall associated with acute renal failure in dogs and cats focused initially on melamine as the suspect toxicant. In the course of the investigation, cyanuric acid was identified in addition to melamine in the offending food. The purpose of this study was to characterize the toxicity potential of melamine, cyanuric acid, and a combination of melamine and cyanuric acid in cats. In this pilot study, melamine was added to the diet of 2 cats at 0.5% and 1%, respectively. Cyanuric acid was added to the diet of 1 cat at increasing doses of 0.2%, 0.5%, and 1% over the course of 10 days. Melamine and cyanuric acid were administered together at 0%, 0.2%, 0.5%, and 1% to 1 cat per dose group. No effect on renal function was observed in cats fed with melamine or cyanuric acid alone. Cats dosed with a combination were euthanized at 48 hours after dosing because of acute renal failure. Urine and touch impressions of kidneys from all cats dosed with the combination revealed the presence of fan-shaped, birefringent crystals. Histopathologic findings were limited to the kidneys and included crystals primarily within tubules of the distal nephron, severe renal interstitial edema, and hemorrhage at the corticomedullary junction. The kidneys contained estimated melamine concentrations of 496 to 734 mg/kg wet weight and estimated cyanuric acid concentrations of 487 to 690 mg/kg wet weight. The results demonstrate that the combination of melamine and cyanuric acid is responsible for acute renal failure in cats.
Development of a Sensitive and Specific Indirect Enzyme-Linked Immunosorbent Assay Based on a Baculovirus Recombinant Antigen for Detection of Specific Antibodies against Ehvlichia Canis
Authors: López Lissett , Venteo Angel , Aguirre Enara , García Marga , Rodríguez MaJosé , Amusátegui Inmaculada , Tesouro Miguel A. , Vela Carmen , Sainz Angel , Rueda Paloma
Published: May 2011

An indirect enzyme-linked immunosorbent assay (ELISA) based on baculovirus recombinant P30 protein of Ehrlichia canis and the 1BH4 anticanine IgG monoclonal antibody was developed and evaluated by examining a panel of 98 positive and 157 negative sera using the indirect fluorescent antibody (IFA) test as the reference technique. The P30-based ELISA appeared to be sensitive and specific (77.55% and 95.54%, respectively) when qualitative results (positive/negative) were compared with those of the IFA test; the coefficient of correlation (R) between the 2 tests was 0.833. Furthermore, it was possible to establish a mathematical formula for use in comparing the results of both techniques. These results indicate that recombinant P30 antigen-based ELISA is a suitable alternative of the IFA test for simple, consistent, and rapid serodiagnosis of canine ehrlichiosis. Moreover, the use of this recombinant protein as antigen offers a great advantage for antigen preparation in comparison with other techniques in which the whole E. canis organism is used as antigen.
Toxoplasmosis in a Woodchuck (Marmota Monax) and Two American Red Squirrels (Tamiasciurus Hudsonicus)
Authors: Bangari Dinesh S. , Mouser Pamela , Miller Margaret A. , Stevenson Gregory W. , Vemulapalli Ramesh , Thacker H. Leon
Published: May 2011

Toxoplasmosis was diagnosed in a woodchuck ( Marmota monax) and 2 American red squirrels ( Tamiasciurus hudsonicus). The woodchuck was euthanized by a wildlife rescue organization in New York after progressive clinical signs of head tilt, circling, and rapid weight loss. Necropsy examination revealed acute subdural hemorrhage over the right cerebral hemisphere. Histologic lesions included meningoencephalitis, myocarditis, and hepatitis. Protozoal cysts were present in affected and unaffected neuroparenchyma. The squirrels were found dead, emaciated, and moderately infested with fleas near a park in northern Indiana. In both squirrels, the lungs were consolidated with numerous nodules up to 2 mm in diameter. Histologically, pneumonia and encephalitis were associated with intracellular and free protozoa. Additional histologic lesions included multifocal lymphoplasmacytic encephalitis with intralesional protozoa in both squirrels. The protozoa were positive with Toxoplasma gondii-specific immunohistochemistry and had ultrastructural features consistent with T. gondii in both squirrels. A real-time polymerase chain reaction test using T. gondii–specific probes demonstrated protozoal DNA in the lung, brain, and kidney of the squirrels and in the brain and heart of the woodchuck. To the authors' knowledge, this is the first report of toxoplasmosis in woodchucks or American red squirrels. Because rodents are common near urban settlements, this finding underscores their role as important intermediate hosts for T. gondii.
Peribiliary Cysts Associated with Severe Liver Disease: A Previously Unrecognized Tumor in a Lion (Panthera Leo)
Authors: Yu Chi-Ho , Kim Kyoo-Tae , Hwang Du-Na , Yhee Ji-Young , Moon Chang-Taek , Hur Tai-Young , Sur Jung-Hyang
Published: May 2011

A 13-year-old male lion ( Panthera leo) from Dae Jeon Zoo, Republic of Korea, presented with clinical signs of lethargy and anorexia. Despite treatment with antibiotics and fluid therapy, the lion died 6 days after initial presentation. Postmortem examination revealed multiple masses measuring 5–10 cm in diameter and cysts throughout the liver. A diagnosis of spontaneous peribiliary cysts was made on the basis of microscopic lesions as well as special staining and immunohistochemical characteristics. Histologically, the neoplasm was surrounded and composed of compact collagenous tissue. The inner cystic single layer resembled biliary mucosa and was composed of cuboidal or flattened epithelial lining that was strongly immunopositive for cytokeratin AE1/AE3. This layer was surrounded by fibrous tissue that stained blue by Masson's trichrome staining. Given the presence of multiple organized cysts in the liver, the lesion was consistent with peribiliary cysts. To the authors' knowledge, this is the first report of peribiliary cysts in an animal.
Investigation into the Effectiveness of Pooled Fecal Samples for Detection of Verocytotoxin-Producing Escherichia Coli O157 in Cattle
Authors: Arnold Mark E. , Ellis-Iversen Johanne , Cook Alasdair J. C. , Davies Robert H. , McLaren Ian M. , Kay Anthony C. S. , Pritchard Geoff C.
Published: May 2011

Verocytotoxin-producing Escherichia coli O157 (VTEC O157) may cause severe illness in people. Cattle are regarded as an important source of VTEC O157, and in an outbreak investigation, there is a necessity to establish whether or not the putative contact herd shares infection with the human case. The effectiveness of a herd investigation is impacted by the number of samples required, which will influence the time taken to collect samples and then process these in the laboratory. The objective of this study was to investigate the effectiveness of pooled sampling for detecting VTEC O157 in cattle herds in the United Kingdom. On farm 1, 150 individual fecal samples were collected during the course of a VTEC O157 outbreak investigation. One-gram and 10-g subsamples were tested from each individual sample. Once the culture results of the individual sample were known, pools comprising 5 and 10 individual samples were formed, with each pool containing a known number of positive samples. This data showed that the sensitivity of pooled sampling depended upon the proportion of positive samples in the pool. Further samples were collected from 2 more infected farms (2 and 3). Each individual sample was tested in duplicate. Pools of 5 feces were formed on-farm, and half the number of pooled feces were tested as individual feces. There was no significant difference between the number of cultures required for pooled sampling, as was the same for individual sampling, and therefore pooling did not improve the effectiveness of detection of VTEC O157.
Sensitivity of Polymerase Chain Reaction for Detection of Bovine Viral Diarrhea Virus in Pooled Serum Samples and Use of Pooled Polymerase Chain Reaction to Determine Prevalence of Bovine Viral Diarrhea Virus in Auction Market Cattle
Authors: Smith Rebecca L. , Sanderson Michael W. , Walz Paul H. , Givens M. Daniel
Published: May 2011

Two reverse transcription-nested polymerase chain reaction tests, 1 quantitative (qRT-nPCR) and 1 standard (RT-nPCR), were evaluated to assess sensitivity for detection of bovine viral diarrhea virus (BVDV) of a single positive serum sample in a pool of 30. The RT-nPCR and qRT-nPCR each detected 95 of 100 known positives. The RT-nPCR was used to estimate the prevalence of BVDV in adult beef cows. Serum samples were obtained from the US Department of Agriculture brucellosis testing laboratories in 3 Midwestern states. Samples originated from auction markets and private treaty sales throughout the 3 states. A total of 2,990 serum samples were collected and randomly pooled into 100 pools for testing. Two of the 100 pools of field samples were positive, and each positive pool had a single positive individual sample upon confirmation. The estimate of BVDV prevalence in adult cows in this study was 0.07%. This study estimates the diagnostic sensitivity of RT-nPCR for BVDV and confirms that it is a useful diagnostic tool for pools of 30 serum samples and that prevalence of BVDV in adult cattle from auction markets is low.
Malignant Melanoma of the Horn Base in a Pygora Goat
Authors: Mavangira Vengai , Hughes Jennifer M. , Middleton John R. , Mitchell William J. , Kim Dae Young
Published: May 2011

A case of malignant melanoma, originating at the base of the left horn, was diagnosed in a white 11-year-old Pygora doe. At necropsy, the goat had an ulcerated, black-pigmented, 2.5-cm mass at the base of the left horn. Firm masses diffusely black on the cut surface were present at the left horn base, subcutaneously over the frontal bones, and infiltrating into the frontal sinuses, the submandibular lymph node, and left parotid salivary gland. The left maxillary premolars and molars were loosened from the periodontia. Multiple black foci of metastasis were observed in the liver. Histologically, the masses consisted predominantly of moderately pleomorphic, polyhedral to spindle cells that contained variable amounts of dark brown intracytoplasmic pigment granules. The tumor cells were positive for Melan A by immunohistochemical staining.
Adaptation and evaluation of a rapid test for the diagnosis of sheep scrapie in samples of rectal mucosa
Authors: González Lorenzo , Horton Robert , Ramsay Drew , Toomik Reet , Leathers Valerie , Tonelli Quentin , Dagleish Mark P. , Jeffrey Martin , Terry Linda
Published: May 2011

In recent publications, it was shown that disease-associated prion protein (PrPd) accumulates in the lymphoid tissue of the rectal mucosa of a high proportion of scrapie-infected sheep at clinical and preclinical stages, regardless of several host factors; PrPd can also be detected in biopsy specimens of rectal mucosa, with an increased probability proportional to age or incubation period and with an efficiency almost identical to that of tonsil biopsies. Rectal biopsies have the advantages of providing higher numbers of lymphoid follicles and of being simpler to perform, which makes them suitable for scrapie screening in the field. In biopsy samples, PrPd could be demonstrated by immunohistochemical (IHC) and Western immunoblotting methods, and the purpose of the present study was to optimize and evaluate a “rapid test” for the diagnosis of scrapie in rectal biopsy samples. The HerdChek CWD (chronic wasting disease) antigen EIA (enzyme immunoassay) test was chosen and, once optimized, provided specificity and sensitivity figures of 99.2% and 93.5%, respectively, compared with IHC results in the same samples obtained at a postmortem. The sensitivity of the assay increased from 82.1%, when a single rectal mucosa sample was tested to 99.4% for those sheep in which 3 or more samples were analyzed. Similarly, sensitivity values of the HerdChek CWD antigen EIA test on biopsy samples increased from 95% to 100% for sheep subjected to 1 or 2 sequential biopsies 4 months apart, respectively. Thus, a preclinical diagnosis of scrapie in live sheep can be achieved by a combination of a simple sampling procedure, which can be repeated several times with no detrimental effect for the animals, and a rapid and efficient laboratory method.
The effects of formalin fixation and tissue embedding of bovine liver on copper, iron, and zinc analysis
Authors: Bischoff Karyn , Lamm Catherine , Erb Hollis N. , Hillebrandt Joseph R.
Published: May 2011

Veterinary analytical chemistry laboratories might be called upon to analyze formalin-fixed or paraffin-embedded tissue samples for trace minerals. The purpose of this study was to determine whether concentrations of copper (Cu), iron (Fe), and zinc (Zn) are comparable among fresh or frozen, formalin-fixed, and paraffin-embedded bovine liver samples on an as-received basis. Three liver sample subtypes (fresh or frozen, formalin-fixed, and paraffin-embedded) from 12 cows were collected and analyzed for Cu, Fe, and Zn concentrations. Concentrations were measured by using inductively coupled argon plasma atomic-emission spectroscopy. There was no significant difference in mineral measurements between fresh or frozen and formalin-fixed samples for Cu and Zn (both P ≥ 0.052). The median concentration of Fe was lower in the fresh or frozen samples than in the formalin-fixed samples. However, for every pair of fresh or frozen and paraffin-embedded samples for all 3 minerals, the fresh or frozen sample had a lower measurement than the paraffin-embedded sample (all P = 0.005). Differences in mineral measurements associated with tissue processing did not result in differences in classification (within or outside the reference range) for Fe. However, the classification of Cu and Zn was different up to 25% of the time with fresh or frozen versus formalin-fixed or embedded liver. Although Cu, Fe, and Zn concentrations attained from processed tissue may be useful, they must be evaluated with caution.
Quality Documentation Challenges for Veterinary Clinical Pathology Laboratories
Authors: Sacchini Federico , Freeman Kathleen P.
Published: May 2011

An increasing number of veterinary laboratories worldwide have obtained or are seeking certification based on international standards, such as the International Organization for Standardization/International Electrotechnical Commission 17025. Compliance with any certification standard or quality management system requires quality documentation, an activity that may present several unique challenges in the case of veterinary laboratories. Research specifically addressing quality documentation is conspicuously absent in the veterinary literature. This article provides an overview of the quality system documentation needed to comply with a quality management system with an emphasis on preparing written standard operating procedures specific for veterinary laboratories. In addition, the quality documentation challenges that are unique to veterinary clinical pathology laboratories are critically evaluated against the existing quality standards and discussed with respect to possible solutions and/or recommended courses of action. Documentation challenges include the establishment of quality requirements for veterinary tests, the use or modification of human analytic methods for animal samples, the limited availability of quality control materials satisfactory for veterinary clinical pathology laboratories, the limited availability of veterinary proficiency programs, and the complications in establishing species-specific reference intervals.
The Correlation of Potassium Content and Moisture in Bovine Liver Samples Analyzed for Trace Mineral Concentrations
Authors: Ludwick Taylor P. , Poppenga Robert H. , Green Peter G. , Puschner Birgit , Melton Larry A. , Hoar Bruce R. , Nyberg Nicole L. , Maas John
Published: May 2011

Beef cattle in the United States are often found to be deficient in essential trace minerals such as copper and zinc. Established reference ranges for mineral concentrations exist and usually designate a concentration as adequate, marginal, deficient, or excessive. This research investigates a new method of interpreting detected elemental concentrations in bovine liver that will add confidence to the final diagnosis. This is based on the hypothesis that a correlation exists between potassium concentration and moisture in a bovine liver sample. This relationship between potassium and moisture content enables the diagnostician to more accurately predict mineral concentrations and wet weight regardless of sample moisture loss. Correlations were found between potassium content and percentage of moisture in experimental samples, clinical biopsies, and a validation study, to a statistical significance of P < 0.001. Experimental samples had a correlation coefficient of R 2 = 0.95 and the mathematical relationship y = 2513.2 x−1.0662. Clinical biopsies had a correlation of R 3 = 0.83 and the mathematical relationship y = 2203.4 x−0.991. The validation study had a correlation of R 2 = 0.55 and a mathematical relationship y = 2321.4 x−0.952. An exponent of −1 is predicted by conservation of potassium mass. These findings have practical significance in maintaining and improving cattle growth, health, reproduction, and food safety.
Characterization of Fluoroquinolone Resistance in Escherichia Coli Strains from Ruminants
Authors: Jurado Sonia , Orden José A. , Horcajo Pilar , De La Fuente Ricardo , Ruiz-Santa-Quiteria José A. , Martínez-Pulgarín Susana , Domínguez-Bernal Gustavo
Published: May 2011

Thirty-seven fluoroquinolone-resistant Escherichia coli strains from ruminants (according to Clinical and Laboratory Standards Institute guidelines) were screened by molecular methods for mutations in the quinolone resistance-determining region (QRDR) of the gyrA and parC genes and for the presence of the qnrA gene. One of the strains studied was an enterohemorrhagic E. coli (EHEC) strain potentially pathogenic for humans. Three E. coli strains resistant to enrofloxacin (minimal inhibitory concentration [MIC] = 2 μg/ml) but not to ciprofloxacin (MIC = 1 μg/ml) presented single mutations in the gyrA and parC genes, while 34 strains resistant to both fluoroquinolones presented double and single mutations in gyrA and parC, respectively (31 strains), or double mutations in gyrA and parC (3 strains). The EHEC strain presented a double amino acid substitution in the GyrA protein (Ser-83→Leu and Asp-87→Gly) and a double amino acid substitution in the ParC protein (Gly-78→Cys and Ser-80→Arg), one of which has not been previously described. The present study shows that most of the mutations in the QRDR of the gyrA and parC genes of fluoroquinolone-resistant E. coli strains from ruminants are the same as those seen in E. coli strains from other animal species and humans and that there are no differences in mutation patterns in the QRDR of E. coli strains from healthy ruminants and those with diarrhea. No strains carried qnrA, which indicates that this gene does not play an important role in the selection of fluoroquinolone-resistant E. coli strains from ruminants.
Fatal Yersiniosis in Farmed Deer Caused by Yersinia Pseudotuberculosis Serotype O:3 Encoding a Mannosyltransferase-Like Protein WbyK
Authors: Zhang Shuping , Zhang Zhenyu , Liu Shiliang , Bingham Willie , Wilson Floyd
Published: May 2011

Sudden death of 9 deer occurred in a large enclosed deer farm with approximately 400 heads of cervids. Fatal yersiniosis was diagnosed in 2 deer that were submitted for laboratory diagnosis. Histopathologically, the disease was characterized by multifocal pulmonary hemorrhage and mild interstitial pneumonia, marked diffuse cholangiohepatitis, minimal myocarditis with mild myocardial degeneration, and mild multifocal suppurative cystic colitis. Yersinia pseudotuberculosis was isolated from the lungs and colon of the affected animals. The isolates were PCR-positive for genes virF, inv, yopB, and yopH, which are essential for invasion and colonization of host intestine and lung. The isolates reacted with polyclonal antibodies against serotype O:3 antigen. The O-genotyping patterns of the isolates were identical with each other, but different from those of the 21 O-genotypes (or serotypes) reported previously. In addition to the O-antigen genes possessed by classical serotype O:3, a gene ( wbyK) encoding a mannosyltransferase-like protein was detected in these isolates. The wbyK gene of the isolates showed 94% of DNA sequence homology with the wbyK gene harbored by Y. pseudotuberculosis O:1b. On the basis of pathology, bacteriology, and serology, the authors concluded that the acute deaths of these deer were caused by Y. pseudotuberculosis infection. Molecular characterization of the isolate revealed a genetic heterogeneity in the O-antigen gene cluster of Y. pseudotuberculosis serotype O:3.
Suggested Guidelines for Immunohistochemical Techniques in Veterinary Diagnostic Laboratories
Authors: Ramos-Vara José A. , Kiupel Matti , Baszler Timothy , Bliven Laura , Brodersen Bruce , Chelack Brian , West Keith , Czub Stefanie , Del Piero Fabio , Dial Sharon , Ehrhart E. J. , Graham Tanya , Manning Lisa , Paulsen Daniel , Valli Victor E.
Published: May 2011

This document is the consensus of the American Association of Veterinary Laboratory Diagnosticians (AAVLD) Subcommittee on Standardization of Immunohistochemistry on a set of guidelines for immunohistochemistry (IHC) testing in veterinary laboratories. Immunohistochemistry is a powerful ancillary methodology frequently used in many veterinary laboratories for both diagnostic and research purposes. However, neither standardization nor validation of IHC tests has been completely achieved in veterinary medicine. This document addresses both issues. Topics covered include antibody selection, fixation, antigen retrieval, antibody incubation, antibody dilutions, tissue and reagent controls, buffers, and detection systems. The validation of an IHC test is addressed for both infectious diseases and neoplastic processes. In addition, storage and handling of IHC reagents, interpretation, quality control and assurance, and troubleshooting are also discussed. Proper standardization and validation of IHC will improve the quality of diagnostics in veterinary laboratories.
Canine Parvovirus 2c Infection in Central Portugal
Authors: Vieira Maria João , Silva Eliane , Oliveira João , Vieira Ana Luísa , Decaro Nicola , Desario Costantina , Muller Alexandra , Carvalheira Júlio , Buonavoglia Canio , Thompson Gertrude
Published: May 2011

Canine parvovirus (CPV) has been evolving, generating new genetic and antigenic variants throughout the world. This study was conducted to determine the types of CPV circulating in dogs in Figueira da Foz, Portugal. Thirty fecal samples, collected between 2006 and 2007 from dogs with clinical signs of CPV infection, were tested for CPV by a rapid, in-clinic, enzyme-linked immunosorbent assay (ELISA)/immunomigration test, by conventional real-time polymerase chain reaction (PCR), and by minor-groove binding TaqMan PCR. Of the 29 PCR-positive samples, 15 were identified as CPV-2b and 14 as CPV-2c. No CPV-2a was detected. The sensitivity of the ELISA test was 82.76% compared with the PCR assays. No significant associations were found between CPV type, clinical outcome, breed, vaccination status, or age.