bluuebunny/crossref_metadata_2025 huggingface dataset:
Histological and Immunohistochemical Detection of Different Helicobacter Species in the Gastric Mucosa of Cats
Authors: Scanziani Eugenio , Simpson Kenneth W. , Monestiroli Silvia , Soldati Sabina , Strauss-Ayali Dalit , Piero Fabio Del
Published: May 2011

Detailed histopathological evaluation of the gastric mucosa of Helicobacter-infected cats is complicated by the difficulty of recognizing Helicobacter organisms on hematoxylin and eosin (HE)-stained sections and the ability of multiple Helicobacter species to infect cats. In this study, the presence and localization of different species of Helicobacter in the stomachs of cats was investigated using silver staining and immuno-histochemistry. Five groups containing 5 cats each were established (group 1: urease negative and Helicobacter free; groups 2, 3, 4, and 5: urease positive and infected with Helicobacter heilmannii, unclassified Helicobacter spp., Helicobacter felis, and Helicobacter pylori, respectively). Gastric samples were evaluated by HE and silver staining and by immunohistochemistry with 3 different anti- Helicobacter primary antibodies. Helicobacter were detected by Steiner stain in all infected cats at the mucosal surface, in the lumen of gastric glands, and in the cytoplasm of parietal cells. In silver-stained sections, H. pylori was easily differentiated from H. felis, H. heilmannii, and unclassified Helicobacter spp., which were larger and more tightly coiled. No organisms were seen in uninfected cats. Helicobacter antigen paralleled the distribution of organisms observed in Steiner-stained sections for 2 of the 3 primary antibodies tested. The antisera were not able to discriminate between the different Helicobacter species examined. A small amount of Helicobacter antigen was present in the lamina propria of 3 H. pylori-, 3 H. felis-, and 1 H. heilmannii-infected cat. Minimal mononuclear inflammation was present in uninfected cats and in those infected with unclassified Helicobacter spp. and H. heilmannii cats. In H. felis-infected cats, lymphoid follicular hyperplasia with mild pangastric mononuclear inflammation and eosinophilic infiltrates were present. The H. pylori-infected cats had severe lymphoid follicular hyperplasia and mild to moderate mononuclear inflammation accompanied by the presence of neutrophils and eosinophils. These findings indicate that Steiner staining and immunohistochemistry are useful for detecting Helicobacter infections, particularly when different Helicobacter species can be present. Monoclonal antibodies specific for the different Helicobacter species could be important diagnostic aids. There appear to be differences in the severity of gastritis in cats infected with different Helicobacter species.
Cutaneous Neosporosis in Two Adult Dogs on Chronic Immunosuppressive Therapy
Authors: la Perle Krista M. D. , Del Piero Fabio , Carr Ruthann F. , Harris Cheryl , Stromberg Paul C.
Published: May 2011

Antemortem diagnosis of generalized ulcerative and pyogranulomatous dermatitis with numerous intralesional tachyzoites was made from skin biopsy specimens from 2 adult dogs on chronic immunosuppressive therapy. A 9-year-old Italian Greyhound was on long-term corticosteroid therapy for the treatment of a lupus-like systemic autoimmune disorder, and a 7-year-old Labrador Retriever had received several months of chemotherapy for lymphosarcoma. The tachyzoites were identified as Neospora caninum by immunoperoxidase immunohistochemistry. Both dogs were treated with clindamycin. Lesions in the Greyhound resolved; however, the Labrador Retriever was euthanized because of evidence of neuromuscular disease, despite improvement of the skin lesions. These 2 cases indicate that cutaneous neosporosis can occur in adult dogs on chronic immu-nosuppressive therapy. The disease may result from reactivation of a congenital infection and/or a recently acquired primary infection.
Application of a Direct Flow Cytometric Erythrocyte Immunofluorescence Assay in Dogs with Immune-Mediated Hemolytic Anemia and Comparison to the Direct Antiglobulin Test
Authors: Quigley Kelley A. , Chelack Brian J. , Haines Deborah M. , Jackson Marion L.
Published: May 2011

A direct flow cytometric erythrocyte immunofluorescence assay (FC) was developed and compared with the direct antiglobulin test (DAT) for detection of erythrocyte-bound immunoglobulin (IgG and IgM) and complement (C3) in dogs with immune-mediated hemolytic anemia (IMHA). Tests were performed on erythrocytes from 13 healthy nonanemic dogs and from 13 anemic dogs with IMHA. The FC and DAT were negative for erythrocyte-bound immunoglobulin in all healthy dogs. The FC was negative for erythrocyte-bound C3 in 12 healthy dogs and positive in 1 healthy dog, and the DAT was negative for C3 in all healthy dogs. Of the 13 IMHA dogs tested for erythrocyte-bound IgG, 12 were positive using the FC and 7 were positive using the DAT. Sensitivity for the detection of erythrocyte-bound IgG in the 26 dogs was 92% for FC and 53% for DAT. Specificity for detection of erythrocyte bound IgG for FC and DAT was 100%. The addition of IgM and/ or C3 did not increase the sensitivity for FC or DAT. In this group of dogs, the FC provided a more rapid, cost-effective, sensitive, objective method to quantitate erythrocyte-bound immunoglobulin and/or complement compared with the currently used DAT.
Digital Papillomatosis in a Confined Beagle
Authors: DeBey Brad M. , Bagladi-Swanson Mary , Kapil Sanjay , Oehme Frederick W.
Published: May 2011

Papillomavirus-induced papillomas were diagnosed on multiple digits of all 4 feet of a young Beagle. No other cutaneous or oral involvement was identified. Papillomavirus antigen was confirmed by im-munoperoxidase localization within keratinocyte nuclei. In addition to the typical basophilic intranuclear inclusions associated with papillomavirus infections, keratinocytes within the papillomas contained large, eosino-philic cytoplasmic inclusions that previously have been described in a Boxer with cutaneous lesions associated with a papillomavirus infection. The papillomas in this Beagle regressed completely within 2 months of the initial diagnosis.
An Immunoassay for Canine Pancreatic Elastase 1 as an Indicator for Exocrine Pancreatic Insufficiency in Dogs
Authors: Spillmann T. , Wittker A. , Teigelkamp S. , Eim C. , Burkhardt E. , Eigenbrodt E. , Sziegoleit A.
Published: May 2011

The detection of pancreatic elastase 1 in stool samples has become the noninvasive gold standard for the diagnosis of pancreatic insufficiency in humans. Accordingly, the development of a sandwich-ELISA specific for canine pancreatic elastase 1, based on monoclonal antibodies, is presented here. The test has a detection range of 4–240 μg canine pancreatic elastase 1/g feces. The intraassay coefficient of variation is 7.4%, and the interassay coefficient of variation is 7.7%. Spiking experiments show that canine elastase 1 is quantitatively detectable in fecal samples. Interestingly, the range of the elastase 1 concentration in canine feces within several days is higher as compared with humans. As the proposed cutoff of 10 μg/g is below this variation range in 96.1% of the tested samples, the effect on the test specificity is negligible. Because the test detects neither human nor bovine and porcine elastase 1, pancreatic function can be monitored without interrupting an enzyme replacement therapy.
Relationship between Sample Weight, Homogeneity, and Sensitivity of Fecal Culture for Salmonella Enterica
Authors: Cannon Rob M. , Nicholls Terry J.
Published: May 2011

When testing for organisms such as Salmonella enterica in an aliquot taken from a sample of feces, the sensitivity of the procedure is considerably higher if the organisms are randomly distributed in the feces than if the same concentration of organisms is in random clusters. However, dispersing the clusters (by homogenizing the sample before taking the aliquot) increases the sensitivity. The increase depends on the weight of the fecal sample taken.
Isolation of Arcanobacterium (Actinomyces) Pyogenes from Cases of Feline Otitis Externa and Canine Cystitis
Authors: Billington S. J. , Post K. W. , Jost B. H.
Published: May 2011

Arcanobacterium pyogenes is a normal inhabitant of the mucous membranes of domestic animals, such as cattle, sheep, swine, and goats. It is also an opportunistic pathogen in these animals, where it causes a variety of purulent infections involving the skin, joints, and visceral organs. Two recent cases of isolation of A. pyogenes from companion animals are reported. In the first case, a cat presented with a chronic otitis externa, from which A. pyogenes was isolated in pure culture. The second case involved a dog with a urinary tract infection, where A. pyogenes was isolated from urine as the predominant bacterial species. In both cases, the A. pyogenes isolates were presumptively identified by macrobiochemical tests, and then their identities were confirmed by polymerase chain reaction analysis and 16S rRNA gene sequencing.
Visceral Nodular Cartilaginous Metaplasia in Rockfishes (Sebastes Spp.) in the Eastern North Pacific Ocean
Authors: Heidel Jerry R. , Nowak Barbara F. , Fischer Kay A. , Watral Virginia G. , Kent Michael L.
Published: May 2011

Multiple, discrete, nodular foci of cartilaginous metaplasia were found in the spleens and kidneys of rockfishes taken from the northeastern Pacific Ocean during a survey to determine the incidence and the nature of diseases in these animals. These nodules sometimes occurred in association with granulomatous inflammation and distinct granulomas. Many of these fish were infected by Ichythophonus spp. or acid-fact bacteria (presumably Mycobacteria spp.). Some of the metaplastic foci contained encapsulated accumulations of eosinophilic vesicles and basophilic granular debris, described by other authors as “cysts of unknown etiology,” which have been observed at different sites in a variety of temperate and tropical fish species.
Use of a Japanese Quail Fibrosarcoma Cell Line (QT-35) in Serologic Assays to Determine the Antigenic Relationship of Avian Metapneumoviruses
Authors: Sabara Marta I. , Larence June E. , Halayko Stacey
Published: May 2011

The ability of a Japanese quail fibrosarcoma cell line (QT-35) to support the replication of avian metapneumoviruses belonging to the 3 subgroups A (14/1 virus), B (Colorado virus), and C (Hungary virus) enabled the development of assays for the detection and evaluation of virus-specific antibodies. On the basis of the results of enzyme-linked immunosorbent assay (ELISA), plaque reduction neutralization assay (PRNA), immunofluorescent assay (IFA), and Western blot analysis, some degree of antigenic cross-reactivity was observed between prototype viruses belonging to each of the 3 subgroups A, B, and C. The antigen produced in QT-35 cells was found to be superior with respect to its reactivity with virus-specific antibodies, as determined when used in ELISA and IFA. Standardization of both the input virus and the virus-specific antibodies in PRNA enabled a more detailed analysis of the antigenic relationship between these viruses. Specifically, it was observed that 14/1 virus shared more neutralizing regions with Hungary and Colorado viruses than did either of these viruses with 14/1 virus. In addition, Hungary virus shared comparatively fewer neutralizing epitopes with the Colorado virus than did 14/1 virus. Western blot analysis of the reactivity patterns of virus antigen, produced in QT-35 cells, with subgroup-specific antibodies identified a cross-reactive protein migrating at approximately 18 kD. These assays and the information from the Western blot will enable further analysis of avian metapneumovirus isolates to determine antigenic relationships.
Use of Immunohistochemical Marker Calretinin in the Diagnosis of a Diffuse Malignant Metastatic Mesothelioma in an Equine
Authors: Stoica G. , Cohen N. , Mendes O. , Kim Hun-Taek
Published: May 2011

Mesotheliomas are rarely reported in animal species. In this report, the occurrence of a diffuse, metastatic mesothelioma in a 6-year-old gray Arabian mare is described. The mare was presented on clinical examination with ascites, bilateral pleural effusion, and pleural roughening. Necropsy revealed abundant fluid in the abdominal and thoracic cavities. The surface of all organs was thick and fibrosed with multiple raised nodules and hemorrhages. Histology was characteristic of a generalized, biphasic mesothelioma with vascular and lymph nodes metastases. It is believed that the primary tumor developed in the pericardium and spread through lymphatics. In this report, calretinin was used as an immunohistochemical marker in the diagnosis of mesothelioma in an equine species for the first time.
Biology of PrPsc Accumulation in Two Natural Scrapie-Infected Sheep Flocks
Authors: Caplazi Patrick , O'Rourke Katherine , Wolf Cynthia , Shaw Daniel , Baszler Timothy V.
Published: May 2011

Sheep scrapie is a prion disease that requires interaction of exogenous prions with host prion protein (PrP) supporting prion formation. Disease is associated with deposition of a host-generated conformational variant of PrP, PrPsc, in a variety of tissues, including brain, resulting in fatal spongiform encephalopathy. Efficiency of PrPsc formation is determined by polymorphisms in the PrP-coding sequence. This article adds to previous data of natural sheep scrapie, concentrating on the effect of host genotype and age on PrPsc accumulation patterns during preclinical and clinical disease. Two entire scrapie-infected, predominantly Suffolk-cross, sheep flocks euthanized for regulatory purposes were genotyped and analyzed for PrPsc deposition in various tissues using single- and dual-label immunohistochemistry. Scrapie, as defined by PrPsc deposition, occurred in 13/80 sheep. Preclinical disease was evident in nearly 70% of infected sheep, ranging in age from 14 months to 7 years. PrPsc accumulated systemically in the nervous tissue, various lymphoid tissues, both alimentary tract related and non–alimentary tract related, and the placenta. Clinical neurological illness was always associated with spongiform encephalopathy and PrPsc deposition in the brain. Only 6 of 9 sheep with preclinical scrapie had PrPsc deposition in the brain but widespread PrPsc deposition in peripheral lymphoid tissue, supporting previous data showing peripheral PrPsc accumulation preceding deposition in the brain. PrPsc colocalized with a marker for follicular dendritic cells throughout the lymphoid system. PrPsc also accumulated in the peripheral nervous system, particularly the nervous supply of the gastrointestinal tract. Abundant PrPsc was evident in trophoblast cells of placentomes but not in the endometrium, myometrium, or associated nervous plexus. PrPsc deposits were not observed in the mammary parenchyma or bone marrow. Scrapie susceptibility was defined genetically by PrP codon 171: PrPsc deposition was restricted to PrP genotype AA136RR154QQ171 in 12/13 cases or AV136RR154QQ171 in 1/13 cases. The earliest accumulation was observed in the single VRQ/ARQ heterozygous animal, consistent with the reported high scrapie susceptibility and brief incubation period observed in breeds with predominance of the V136R154Q171 allele. Disease occurred within, as well as independent of, mother–daughter lines, suggesting both maternal and nonmaternal transmission in the flocks.
Prunus Spp. Intoxication in Ruminants: A Case in a Goat and Diagnosis by Identification of Leaf Fragments in Rumen Contents
Authors: Radi Zaher A. , Styer Eloise L. , Thompson Larry J.
Published: May 2011

Prunus serotina Ehrh. (black cherry) intoxication was diagnosed on postmortem examination of a goat. The clinical signs were weakness, depression, seizure-like activity, and lateral recumbency. Natural cases of black cherry intoxication have not been reported in goats in the United States. In the absence of a history of access to black cherry or the ability to detect cyanide or cyanogenic glycosides in blood or tissues, black cherry intoxication may be diagnosed in ruminants by the identification of black cherry leaves in rumen contents. Three distinctive features facilitate identification of black cherry leaves or leaf fragments: 1) a pair of small glands that protrude from the sides of the petiole just below the base of the blade, 2) incurved, gland-tipped (callous) teeth along the margins of the leaf, and 3) a band of hairs to each side of the lower half of the midvein on the surface of the leaf. Shape of the marginal teeth, presence or absence of glands at the tips of these teeth, the morphology of these glands, and presence or absence of petiolar glands and their morphology may allow identification and differentiation of small fragments of leaves from the 6 most important cyanogenic Prunus spp. in eastern North America: black cherry, Carolina laurel cherry, peach, English laurel cherry, choke cherry, and fire cherry.
Evaluation of a New Antibody-Based Enzyme-Linked Immunosorbent Assay for the Detection of Bovine Leukemia Virus Infection in Dairy Cattle
Authors: Monti Gustavo E. , Frankena Klaas , Engel Bas , Buist Willem , Tarabla Héctor D. , de Jong Mart C. M.
Published: May 2011

The objective of this study was to validate a new blocking enzyme-linked immunosorbent assay (ELISA) (designated M108 for milk and S108 for serum samples) for detecting bovine leukemia virus (BLV) infection in dairy cattle. Milk, serum, and ethylenediaminetetraacetic acid–blood samples were collected from 524 adult Holstein cows originating from 6 dairy herds in Central Argentina. The M108 and S108 were compared with agar gel immunodiffusion (AGID), polymerase chain reaction and a commercial ELISA. Because there is currently no reference test capable of serving as a gold standard, the test sensitivity (SE) and specificity (SP) were evaluated by the use of a latent class model. Statistical inference was performed by classical maximum likelihood and by Bayesian techniques. The maximum-likelihood analysis was performed assuming conditional independence of tests, whereas the Bayesian approach allowed for conditional dependence. No clear conclusion could be drawn about conditional dependence of tests. Results with maximum likelihood (under conditional independence) and posterior Bayes (under conditional dependence) were practically the same. Conservative estimates of SE and SP (with 95% confidence intervals) for M108 were 98.6 (96.7; 99.6) and 96.7 (92.9; 98.8) and for S108 99.5 (98.2; 99.9) and 95.4 (90.9; 98.1), respectively. The ELISA 108 using either milk or serum to detect BLV-infected animals had comparable SE and SP with the official AGID and a commercial ELISA test, which are currently the most widely accepted tests for the serological diagnosis of BLV infection. Therefore, ELISA 108 can be used as an alternative test in monitoring and control programs.
Investigation of Falsely Reported Resistance of Streptococcus Equi Subsp. Zooepidemicus Isolates from Horses to Trimethoprim–Sulfamethoxazole
Authors: Feary Darien J. , Hyatt Doreene , Traub-Dargatz Josie , Roach Susan , Jones Robert L. , Wu Ching Ching , Morley Paul S.
Published: May 2011

The objective of this study was to investigate the perceived increase in resistance of Streptococcus equi subsp. zooepidemicus ( S. zooepidemicus) isolated from the lower respiratory tract of horses to trimethoprim–sulfamethoxazole (SXT). The recorded SXT-susceptibility results of 50 S. zooepidemicus isolates from the tracheal wash fluid of equine patients examined at Colorado State University Veterinary Teaching Hospital from each of 2 time periods (1987–1990 and 1997–2001) were compared and statistically analyzed using a cross-sectional study design. There was a statistically significant difference between the documented resistance of S. zooepidemicus isolated in the 1987–1990 time period (8%), using quantitative microbroth dilution, and the resistance reported for isolates from the 1997–2001 time period (42%), using Kirby–Bauer agar disk diffusion. Laboratory investigation revealed inadequate quality control of media and subsequent falsely reported resistance of S. zooepidemicus from 1997 to 2001 time period. This study demonstrates how minor deviations from prescribed laboratory-testing guidelines can have a major effect on antimicrobial susceptibility test results. The study also underscores the need for regular surveillance and monitoring of trends in antimicrobial susceptibility to detect and correct such problems. In addition, epidemiologists and others collecting data from laboratories should be cautioned to interact with the laboratory regarding interpretation of results of various testing methods to ensure accurate analysis and conclusions.
Occurrence of Piscirickettsiosis-Like Syndrome in Tilapia in the Continental United States
Authors: Mauel Michael J. , Miller Debra L. , Styer Eloise , Pouder Deborah B. , Yanong Roy P. E. , Goodwin Andrew E. , Schwedler Thomas E.
Published: May 2011

From 2001 to 2003, tilapia ( Oreochromis sp.) farms in Florida, California, and South Carolina experienced epizootics of a systemic disease causing mortality. The fish exhibited lethargy, occasional exophthalmia, and skin petechia. The gills were often necrotic, with a patchy white and red appearance. Grossly, the spleen and kidneys were granular with whitish irregular nodules throughout. Granulomatous infiltrates were observed in kidney, spleen, testes, and ovary tissues, but not in the liver. The granulomas contained pleomorphic coccoid bacteria, measuring 0.57 ± 0.1 × 0.8 ± 0.2 μm, that were Giemsa-positive, acid-fast-negative, and Gram-negative. The bacteria had a double cell wall, variable electron-dense and -lucent areas, and were present in the cytoplasm and within phagolysosomes. The syndrome was associated with cold stress and poor water conditions. These findings are consistent with an infectious process caused by a Piscirickettsia-like bacterium described previously in tilapia in Taiwan and Hawaii. This report involves the first identified cases of a piscirickettsiosis-like syndrome affecting tilapia in the continental United States.
Rapid, Multiwell Colorimetric Assay for Measuring Neutrophil Chemoattractant Activity in Bronchoalveolar Lavage Fluid of Horses with Recurrent Airway Obstruction
Authors: Hall Jean A. , Hoyt Dana , Zuver Clarissa , Skinner Monica M. , Schlipf John W.
Published: May 2011

The criteria used to diagnose recurrent airway obstruction (RAO) in affected horses include demonstration of reversible lower airway obstruction and greater than 25% neutrophils in bronchoalveolar lavage fluid (BALF). Additional objective laboratory tests are needed to improve diagnostic accuracy and to monitor response to treatment. The goal of this study was to determine if neutrophil chemoattractant activity of BALF could be measured by using a previously described, rapid, multiwell colorimetric assay for chemotaxis. In this assay, neutrophils that have migrated through a membrane filter are collected into the bottom well of a disposable chemotaxis–cell migration chamber. The number of viable cells collected in the bottom well is quantified by measurement of the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenol tetrazolium bromide (MTT), which is reduced by dehydrogenase in mitochondria of live cells. The number of migrating cells corresponds to the amount of MTT reduced, which is measured with an enzyme-linked immunosorbent assay plate reader. Fourteen adult horses were enrolled in this study, 7 of which had owner histories consistent with RAO. Each horse was sedated, a bronchoalveolar lavage tube was passed, and saline was infused and immediately aspirated. An aliquot of BALF was used for differential cell count, and BALF supernatant was harvested to assess neutrophil chemoattractant activity. Normal control horses and RAO-affected horses were distinguished according to clinical signs and percent neutrophils in BALF. Neutrophil chemoattractant activity of BALF was significantly greater in RAO-affected horses ( P = 0.001) compared with control horses. This assay may be useful in future studies for monitoring response to therapy in RAO-affected horses.
Vitamin a Deficiency in Turkey Poults
Authors: Cortes P. L. , Tiwary A. K. , Puschner B. , Crespo R. M. , Chin R. P. , Bland M. , Shivaprasad H. L.
Published: May 2011

Vitamin A deficiency was diagnosed in a commercial flock of 13,000 4–6-week-old turkey poults in the summer of 2004. The birds were initially submitted for examination because of a 3% increase in the reported daily mortality of the flock. Clinically, affected birds had stunted growth and ruffled feathers, showed signs of incoordination, and were depressed. At necropsy, pale white pseudomembranous to mucoid material was observed on the mucosal surface of the tongue, oral cavity, portions of the esophagus, and the crop of some birds. Histologically, there was squamous metaplasia of the mucosal epithelium of the oral mucosa, esophagus, sinuses, nasal glands, bronchi, proventriculus, and the bursa of Fabricius. Vitamin A was not detected in the feed sample at a detection limit of 0.5 mg/kg. Serum vitamin A concentrations in 7 birds were very low and ranged from 0.05 to 0.1 mg/L. Vitamin A concentrations in livers were extremely low (0.1 mg/kg wet weight, 1/7 poults) or undetectable (<0.1 mg/kg wet weight, 6/7 poults). A diagnosis of vitamin A deficiency was made based on gross and microscopic lesions and vitamin A concentrations in serum, liver, and feed. To the authors' knowledge, this is the first documented case of vitamin A deficiency in poults submitted from a commercial meat turkey producer comparatively depicting the gross and microscopic lesions with those found in other species of birds and mammals.
Skin Test and Gamma Interferon Enzyme-Linked Immunosorbent Assay Results in Sheep Exposed to Dead Mycobacterium Avium Subspecies Paratuberculosis Organisms
Authors: Robbe-Austerman Suelee , Stabel Judith R. , Morrical Daniel G.
Published: May 2011

Cell-mediated immunity (CMI) diagnostic tests, such as the gamma interferon enzyme-linked immunosorbent assay (IFN-γ ELISA) and the Johnin skin test, have the potential to detect animals infected with Mycobacterium avium subspecies paratuberculosis (MAP) early in the course of the disease. While these CMI tests tend to be relatively specific in noninfected flocks, in MAP-infected flocks, these tests often identify animals that cannot be confirmed infected by any other reference test, including necropsy and culture. The aim of this study was to determine if antigen exposure by inhalation or oral ingestion of killed MAP organisms would cause a detectable CMI response in sheep. Forty-eight lambs 4 months of age were randomly divided into a control group, an orally exposed group (dosed with 1 × 10 10 autoclaved MAP organisms 3 times), and an inhalation-exposed group (dosed once with 1 × 10 5 dead organisms). Lambs were skin tested and/or bled pre-exposure and 1, 2, 3, 4, and 12 months postexposure. No significant difference was seen with either the oral- or inhalation-exposed groups of lambs versus controls with either the IFN-γ ELISA or the skin test at any time pre- or postexposure. These results suggest that infection/invasion of MAP organisms must occur in order to have a positive skin test or IFN-γ ELISA beyond the false-positive rate. Simple exposure is not enough to elicit a detectable CMI response.
Congenital Lobar Emphysema and Tension Pneumothorax in a Dog
Authors: Gopalakrishnan Gopakumar , Stevenson Gregory W.
Published: May 2011

Congenital lobar emphysema (CLE) and tension pneumothorax (TPT) are rarely reported in dogs. A case of CLE of the right middle lung lobe predisposing to air trapping, alveolar hyperinflation, and pleural rupture resulting in fatal spontaneous TPT in a 6-month-old mixed breed dog is described. The unique alteration of “bloat line” was observed in this case in addition to compressive atelectasis of all other lung lobes and lack of negative pressure within the thoracic cavity, signifying markedly elevated intrathoracic pressure. Bronchial cartilage hypoplasia and bronchiectasis were confirmed microscopically, which likely led to abnormal dynamic collapse of bronchi during expiration, consequentially leading to increased intrapulmonary pressure, bullous emphysema, and pleural rupture resulting in TPT. TPT consequent to CLE may therefore be considered one of the potential causes of sudden death in young dogs without overt clinical illness.
Light-Chain Multiple Myeloma in a Cat
Authors: Yamada Osamu , Tamura Kyoichi , Yagihara Hiroko , Isotani Mayu , Azakami Mari , Sawada Satoko , Ono Kenichiro , Washizu Tsukimi , Bonkobara Makoto
Published: May 2011

A diagnosis of light-chain multiple myeloma was made in an 11-year-old male American Shorthair cat. The cat showed atypical plasma cell infiltration in the bone marrow, biclonal gammopathy caused by polymerization of myeloma protein (M-protein), and Bence-Jones proteinuria. The M-protein in the serum of the cat was analyzed by using 12% sodium dodeyl sulfate (SDS) polyacrylamide gel electrophoresis with Coomassie brilliant blue staining. An intense band with a size of 27 kDa, the size of the immunoglobulin light chain, was clearly observed, whereas the band corresponding to the immunoglobulin heavy chain (59 kDa) was undetectable. The 27-kDa band was confirmed to be an immunoglobulin light chain by Western blotting by using antibodies for feline immunoglobulin. These data suggested that the neoplastic plasma cells produce light chain only, leading to the diagnosis of light-chain multiple myeloma in the cat.
Detection of Leptospira Interrogans DNA and Antigen in Fixed Equine Eyes Affected with End-Stage Equine Recurrent Uveitis
Authors: Pearce Jacqueline W. , Galle Laurence E. , Kleiboeker Steve B. , Turk James R. , Schommer Susan K. , Dubielizig Richard R. , Mitchell William J. , Moore Cecil P. , Giuliano Elizabeth A.
Published: May 2011

Equine recurrent uveitis (ERU) is the most frequent cause of blindness in horses worldwide. Leptospira has been implicated as an etiologic agent in some cases of ERU and has been detected in fresh ocular tissues of affected horses. The objective of this study was to determine the presence of Leptospira antigen and DNA in fixed equine ocular tissues affected with end-stage ERU. Sections of eyes from 30 horses were obtained. Controls included 1) 10 normal equine eyes and 2) 10 equine eyes with a nonrecurrent form of uveitis. The experimental group consisted of 10 eyes diagnosed with ERU based on clinical signs and histologic lesions. Sections were subjected to immunohistochemical staining with an array of rabbit anti- Leptospira polyclonal antibodies. DNA extractions were performed by using a commercial kit designed for fixed tissue. Real-time PCR analysis was completed on extracted DNA. The target sequence for PCR was designed from alignments of available Leptospira 16S rDNA partial sequences obtained from GenBank. Two of 10 test samples were positive for Leptospira antigen by immunohistochemical assay. Zero of 20 controls were positive for Leptospira antigen. All test samples and controls were negative for Leptospira DNA by real-time PCR analysis. Leptospira was detected at a lower frequency than that previously reported for fresh ERU-affected aqueous humor and vitreous samples. Leptospira is not frequently detectable in fixed ocular tissues of horses affected with ERU when using traditional immunohistochemical and real-time PCR techniques.
Elimination of False-Positive Polymerase Chain Reaction Results Resulting from Hole Punch Carryover Contamination
Authors: Bonne Nicolai , Clark Phillip , Shearer Patrick , Raidal Shane
Published: May 2011

The collection of biological material (e.g., blood) directly onto filter paper for subsequent use in laboratory assays such as polymerase chain reaction (PCR), has become a common practice. Dried cells or fluid on the paper can be readily rehydrated and retrieved into a standard volume of an appropriate elution buffer but introduces a dilution factor to the sample. The use of a common cutting instrument for excising a standard-sized piece of paper that contains the material also introduces the potential for transferring biological material from one sample to subsequent samples, causing false-positive results by PCR. In the present study, filter-paper-collected blood that contained beak and feather disease virus was used to determine if viral DNA could be transferred between samples by a hole punch used to excise sequential filter papers. It was determined that false-positive results could be obtained at least 13 times after a positive sample. Subsequently, the efficacy of 4 methods of hole punch disinfection, flaming, VirkonS, bleach, and a bleach-ethanol combination, was assessed. The only effective and practical method to destroy DNA was a method where the hole punch was agitated in commercial bleach, rinsed in water, the water was displaced with 100% ethanol and air-dried. This method was simple, cheap, and relatively rapid, and allowed for the use of a single hole punch for a series of samples, without carryover contamination and consequent false-positive results.
Development and field validation of a Mycoplasma iowae real-time polymerase chain reaction assay
Authors: Cai Hugh Y. , Bell-Rogers Patricia , Parker Lois , Ferencz Arpad , Pozder Peter
Published: May 2011

A Mycoplasma iowae real-time polymerase chain reaction (PCR) assay using primers and probes targeting the 16S rRNA gene was developed and field-validated in this study. The assay specifically identified M. iowae with a detection limit of 80 colony-forming units (cfu) per turkey cloacal swab sample (3.2 cfu per PCR reaction). It was validated by testing 154 field turkey cloacal swab samples in parallel with culture isolation. The diagnostic sensitivity of the PCR was 97.6%, and the specificity was 95.5%. The real-time PCR developed in this study is a rapid, sensitive, and cost-effective alternative to culture isolation for detecting M. iowae from cloacal swab samples.
Detection of a Broad Range of Class I and II Newcastle Disease Viruses Using a Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay
Authors: Kim L. Mia , Suarez David L. , Afonso Claudio L.
Published: May 2011

Prompt detection of virulent strains of Newcastle disease virus (vNDV) using real-time reverse transcription polymerase chain reaction (RT-PCR) is challenging because of the broad genetic variability across 2 clades comprising 18 recognized genotypes. A large proportion of class I low virulence ND viruses recently identified in samples recovered from wild birds and from poultry in live bird markets are not detected by the validated real-time RT-PCR assay that targets the matrix gene (M-gene assay). This study describes the identification and sequencing of a conserved region from the polymerase gene of class I NDV and the design and evaluation of a real-time RT-PCR assay (L-TET assay) that identifies a broad range of NDV, demonstrates a 10-fold increase in sensitivity over a previously reported L-gene assay, and works in conjunction with the existing M-gene assay using the same protocol. The L-TET assay detects ≤1 fg of homologous transcribed RNA from genotypes 5, 7, and 8 of class I, and from class II genotype II in either single- or multiplex format. Differential detection of mixed class I and II viruses down to 100 fg is possible because L-TET uses an alternate fluorophore from the M-gene assay. The multiplexed assay is capable of detecting a broad range of class I and II ND viruses with <1 threshold cycle decrease in sensitivity compared to the single probe. A total of 140 class I ( n = 108, genotypes 1–2 and 4–9) and class II ( n = 32, genotypes I–VII) were correctly identified by both the single- and multiplex formats.
Cholangiectasis and Duodenal Diverticulosis in a Farm-Raised Elk (Cervus Elaphus)
Authors: Owston Michael A. , Van Alstine William G. , Couetil Laurent L. , Miskovic Marybeth
Published: May 2011

A 6-year-old male elk presented in August with a 4-month history of weight loss and a 1-day history of being recumbent and unable to rise. The elk's body condition score was 1.5/5. Hematology and blood chemistry showed an inflammatory leukogram, mild electrolyte abnormalities, and elevated liver enzyme activities. Because of poor prognosis, the owner elected euthanasia. On necropsy, 8 diverticula were in the duodenum distal to the common bile duct, which was dilated and filled with feed material. Many intrahepatic bile ducts were dilated up to 5 cm, were filled with ingesta and gritty material, and had walls thickened up to 1.5 mm. On histologic examination, mural thickening of dilated bile ducts was because of increased fibrous connective tissue with inflammatory cell infiltration and hyperplastic and hypertrophic epithelial lining. Portal tracts diffusely contained hyperplastic bile ducts and had increased amounts of fibrous connective tissue. Abscesses that contained bacteria were scattered throughout the liver. Duodenal diverticula lacked the tunica muscularis, and the muscularis mucosa was hypertrophied. Caudal to some diverticula, the duodenal lumen was narrowed. An additional finding was embolic pneumonia with fungi morphologically and immunohistochemically consistent with Aspergillus spp.
A Real-Time Polymerase Chain Reaction Assay for the Detection of the Myxozoan Parasite Henneguya Ictaluri in Channel Catfish
Authors: Griffin Matt J. , Wise David J. , Camus Alvin C. , Mauel Michael J. , Greenway Terrence E. , Pote Linda M.
Published: May 2011

Proliferative gill disease (PGD), caused by the myxozoan parasite Henneguya ictaluri, is the most prevalent parasitic infection affecting commercial channel catfish ( Ictalurus punctatus) aquaculture. There are currently no effective chemotherapeutic or biological control measures for PGD, which often peaks during the spring and fall when water temperatures are between 16–25°C. The current diagnostic techniques of gross examination of gill clip wet mounts and histopathology are subject to false-negatives during the early stages of infection, and the quantifiable nature of end-point polymerase chain reaction (PCR) is subjective. Consequently, a rapid and more sensitive quantitative real-time PCR assay was developed for the detection of H. ictaluri during the early stages of infection in channel catfish. A 23 base-pair TaqMan probe was designed based on previously published H. ictaluri PCR protocols. The sensitivity of the assay was the equivalent of a single H. ictaluri actinospore, and in a pond challenge study, quantitative real-time PCR proved to be more sensitive than gross examination, microscopic examination of gill clip wet mounts, and histopathologic examination of gill tissue sections. Future applications of this assay will focus on developing methodologies to be used in conjunction with current pond-monitoring protocols to evaluate potential treatments and better manage this significant seasonal disease.
Evaluation of Two Avian Influenza Type A Rapid Antigen Tests under Indonesian Field Conditions
Authors: Loth Leo , Prijono Walujo Budi , Wibawa Hendra , Usman Tri Bhakti
Published: May 2011

The current study evaluated the test characteristics of 2 commercially available rapid antigen tests for highly pathogenic avian influenza. Diagnostic specimens were collected from free-ranging village chickens in Indonesia. A total of 174 healthy, sick, and dead birds were included in the study. The relative diagnostic test sensitivity and the relative diagnostic test specificity were calculated by using real-time reverse transcription polymerase chain reaction (RT-PCR) as the confirmatory diagnosis. The AnigenR Rapid AIV Ag test had a relative diagnostic sensitivity of 0.69 (95% confidence interval [CI]: 0.56–0.80) and a relative diagnostic specificity of 0.98 (95% CI: 0.93–0.99). The Flu DetectTM Antigen Capture test strip had a relative diagnostic sensitivity of 0.71 (95% CI: 0.58–0.82) and a relative diagnostic specificity of 0.98 (95% CI: 0.93–0.99). These tests are a valuable tool for the Indonesian avian influenza control program by reliably and quickly detecting Influenza A virus from oropharyngeal swabs from sick or dying chickens.
Stability of Bovine Viral Diarrhea Virus Antigen in Ear Punch Samples Collected from Bovine Fetuses
Authors: Ridpath Julia F. , Chiang Yu-Wei , Waldbillig Jill , Neill John D.
Published: May 2011

Fourteen first-calf heifers were tested free of antibodies against Bovine viral diarrhea viruses (BVDV) by serum neutralization and free of BVDV by polymerase chain reaction. Twelve were exposed to BVDV-1b strain CA0401186a at 84–86 days of gestation, and 2 were exposed to mock inoculum and served as negative controls. Fetuses were harvested by cesarean section at 115–117 days of gestation. The 12 fetuses removed from the BVDV-exposed heifers were BVDV positive based on virus isolation from kidney, thymus, cerebellum, and spleen. It can be assumed that these fetuses would have developed into persistently infected calves had they been allowed to go to term. Virus was not isolated from the fetuses of control animals. Ear punch samples were collected from all fetuses at time of harvest. Antigen capture enzyme-linked immunosorbent assay (ACE), using a commercial kit, was performed on ear punch samples that were frozen within 5 hr of collection and stored at −20°C until tested, tested after storage for 7 days at room temperature (18–25°C), or tested after storage for 7 days at 37°C. Samples stored for 7 days at room temperature or 37°C lost an average of 34% of their starting weight. All samples from BVDV isolation–positive fetuses tested positive by ACE, whereas samples from nonexposed fetuses tested negative, regardless of storage conditions. These results suggest that ACE testing of skin samples collected from aborted fetuses and stillborn calves found in the field may represent a practical surveillance method for BVDV-induced reproductive disease.
Comparison of Genotypes of Escherichia Coli Strains Carrying F18ab and F18ac Fimbriae from Pigs
Authors: DebRoy Chitrita , Roberts Elisabeth , Scheuchenzuber William , Kariyawasam Subhashinie , Jayarao Bhushan M.
Published: May 2011

Escherichia coli carrying the F18 fimbriae colonize the small intestine of pigs and cause postweaning diarrhea and edema disease. There are 2 closely related antigenic variants of F18, F18ab, and F18ac. While F18ab-positive strains are known to be associated with edema disease, E. coli–carrying F18ac are known to cause diarrhea. One hundred ninety-eight E. coli isolates obtained from cases of diarrhea and edema disease in pigs isolated from feces or intestine were screened for the presence of the fedA gene encoding for F18 fimbriae. To distinguish between F18ab and F18ac, the fedA gene was sequenced in 69 F18-positive isolates/strains. The translated protein sequences of the fedA gene in the 2 variants differ; F18ac protein carries proline at amino acid residue 121, which is substituted or missing in F18ab. The F18ab- and F18ac-positive E. coli strains were compared for the presence of virulence attributes, serotypes of the isolates, and relatedness between the strains. Contrary to earlier reports that E. coli F18ab-positive strains mostly express Shiga toxin and F18ac-positive strains generally express enterotoxins, the current report shows conclusively for the first time that both variant types may carry genes for Shiga toxins and/or enterotoxins. Monoclonal antibodies produced against F18ab or F18ac fimbriae could not distinguish the strains carrying the 2 variants. Therefore, it was concluded that either of the 2 F18 variants, F18ab or F18ac, may be involved in causing postweaning diarrhea or edema disease in pigs.
Pathology of Natural Highly Pathogenic Avian Influenza H5N1 Infection in Wild Tufted Ducks (Aythya Fuligula)
Authors: Bröjer Caroline , Ågren Erik O. , Uhlhorn Henrik , Bernodt Karin , Mörner Torsten , Désirée  , Jansson S. , Mattsson Roland , Zohari Siamak , Thorén Peter , Berg Mikael , Gavier-Widén Dolores
Published: May 2011

Highly pathogenic avian influenza (HPAI) subtype H5N1 is an infectious systemic viral disease that results in high morbidity and mortality in poultry, and has been reported in a wide range of wild bird species during the last few years. An outbreak of HPAI H5N1 occurred in wild birds in Sweden in 2006 that affected several duck species, geese, swans, gulls, and raptors. Tufted ducks ( Aythya fuligula) accounted for the largest number of positive cases and, therefore, were selected for more in-depth histologic and immunohistochemical evaluations. The main histologic lesions associated with the presence of avian influenza antigen were found in the brain, pancreas, and upper respiratory tract. Other tissues in which influenza antigen was variably found included liver, lung, adrenal glands, kidneys, and peripheral nerve ganglia. The current study describes the pathology and viral tissue targeting of H5N1 by using histology, polymerase chain reaction, and immunohistochemistry, and highlights the range and variation in the presentation of the natural disease in tufted ducks.
Quadrigeminal Arachnoid Cysts in a Kitten and a Dog
Authors: Reed Scott , Cho Doo Youn , Paulsen Dan
Published: May 2011

Two quadrigeminal arachnoid cysts with different pathogenesis are described in 2 different species. A 10-week-old male Persian kitten with a progressively decreasing level of consciousness died spontaneously. At necropsy, mild internal hydrocephalus, caudal cerebellar coning, and cerebellar herniation through the foramen magnum were associated with a congenital quadrigeminal arachnoid cyst compressing the rostral cerebellum and shifting the entire cerebellum caudally. In contrast, a possibly acquired quadrigeminal cyst was observed in a 2-year-old male neutered Yorkshire Terrier in association with necrotizing encephalitis. Quadrigeminal arachnoid cysts have been rarely reported in dogs and humans.
Congenital Infiltrative Lipomas in a Calf
Authors: Sickinger Marlene , von Erichsen Jasmin , Koehler Kernt , Doll Klaus , Reinacher Manfred
Published: May 2011

The current report describes a case of congenital subcutaneous and intramuscular tumors of the neck and tail base in a 4-week-old female Angus-Charolais crossbred calf. Results of clinical and ultrasound examination are summarized. Biopsy and necropsy findings indicated an infiltrative lipoma. Congenital lipomas are uncommon tumors in bovids. Clinical and morphologic differentials, as well as classification and the possible pathogenesis of congenital neoplasms, are discussed.
A Case of Interscapular Fibrosarcoma in a Dwarf Rabbit (Oryctolagus Cuniculus)
Authors: Petterino Claudio , Modesto Paola , Strata Daniela , Vascellari Marta , Mutinelli Franco , Ferrari Angelo , Ratto Alessandra
Published: May 2011

A 1-year-old, intact, male dwarf rabbit ( Oryctolagus cuniculus) was vaccinated against myxomatosis and rabbit viral hemorrhagic disease in February 1999, and a localized reaction appeared in the same anatomic site within a few days. No regression was observed after subcutaneous antibiotic treatment. The rabbit was kept under observation, and the swelling apparently disappeared in 3 months. The owner then decided to avoid any further subcutaneous drug administration. The referring veterinarian examined the animal on July 2006 for the sudden appearance of a nodular, 4.5 cm × 3.5 cm × 2.0 cm, subcutaneous mass located over the interscapular space. Fine-needle aspiration was performed, and a population of neoplastic spindle cells, rare pleomorphic multinucleated cells, and rare leukocytes were observed. The mass was surgically removed, fixed in 10% neutral buffered formalin, and routinely processed for histologic, histochemical, and immunohistochemical diagnostic investigation. The neoplastic tissue exhibited fascicles composed of malignant spindle-shaped cells with elongated to oval hyperchromatic nuclei and scant cytoplasm. Occasional multinucleated cells were also observed. The neoplastic cells were immunoreactive for vimentin but did not stain for smooth muscle actin, desmin, myoglobin, and cytokeratins (AE1/AE3). Moreover, the histochemical stain for aluminum was positive. The diagnosis was fibrosarcoma based on morphologic and immunohistochemical results. The histologic features of this neoplasm were remarkably similar to feline injection-site sarcoma.