bluuebunny/crossref_metadata_2025 huggingface dataset:
Recombinant K39 Dipstick Immunochromatographic Test: A New Tool for the Serodiagnosis of Canine Leishmaniasis
Authors: Otranto Domenico , Paradies Paola , Sasanelli Mariateresa , Leone Nicola , de Caprariis Donato , Chirico Jan , Spinelli Rosa , Capelli Gioia , Brandonisio Olga
Published: May 2011

The spread of human leishmaniasis has prompted the scientific community to study dogs as reservoirs for Leishmania infantum. Canine leishmaniasis (CanL) is widespread in the Mediterranean area with a prevalence of up to 50%. The first step toward controlling the disease is to monitor its distribution, mainly in stray dogs. The validity of a recombinant K39 (rK39) dipstick test,a commercially available for the serodiagnosis of human leishmaniasis, was evaluated using sera from 165 dogs selected on the basis of positive or negative lymph node smears at parasitological examination. The results were compared with the indirect fluorescent antibody test (IFAT) (cutoff 1:80). Sera from a group of dogs with other diagnosed diseases but negative for leishmaniasis were also tested to evaluate any cross-reactivity. Various procedures were used for testing whole blood samples. The relative specificity of the rK39 dipstick and IFAT was 100% (97 of 97) and 98.97% (96 of 97), whereas the relative sensitivity was 97.06% (66 of 68) and 98.53% (67 of 68), respectively. The results of the dipstick and IFAT corresponded except for 2 sera (k = 0.987). This data confirm the usefulness of rK39 antigen for diagnosing CanL both in symptomatic and asymptomatic dogs. The rK39 dipstick proved to be a rapid, sensitive, and specific test that may be very useful in the field for large-scale screening and also in veterinary practice, requiring minimal equipment and operator expertise.
Microbiologic and Pathologic Findings in an Epidemic of Equine Pericarditis
Authors: Bolin David C. , Donahue J. Mike , Vickers Mary L. , Harrison Lenn , Sells Steve , Giles Ralph C. , Hong C. B. , Poonacha K. B. , Roberts John , Sebastian Manu M. , Swerczek Tom W. , Tramontin Robert , Williams Neil M.
Published: May 2011

During the spring and summer of 2001 and in association with the mare reproductive loss syndrome, 22 terminal and 12 clinical cases of equine pericarditis were diagnosed in central Kentucky. Actinobacillus species were the principal isolates from 8 of 10 nontreated, terminally affected and 3 of 10 clinically affected horses. Enterococcus faecalis and Streptococcus zooepidemicus were cultured from the remaining 2 nontreated terminal cases. No viruses were isolated in tissue culture. Nucleic acid of equine herpesvirus-2 was detected in pericardial and tracheal wash fluids of 3 and 1 individuals, respectively. Microscopic alterations in sections of heart and parietal pericardium were consistent with chronic fibrinous bacterial pericarditis. This report confirms a significant role of Actinobacillus species in equine pericarditis and describes an epidemic of this infrequently observed syndrome in the horse.
Development and Evaluation of an Experimental Model of Cutaneous Columnaris Disease in Koi Cyprinus Carpio
Authors: Tripathi Niraj K. , Latimer Kenneth S. , Gregory Christopher R. , Ritchie Branson W. , Wooley Richard E. , Walker Randal L.
Published: May 2011

A reproducible, experimental model of columnaris disease was developed to study the pathogenesis of cutaneous disease associated with Flavobacterium columnare infection in koi ( Cyprinus carpio). In experimental infections, lesions were usually restricted to skin and fins; gill necrosis was not a consistent finding. Cytologic and histopathologic examinations provided a presumptive diagnosis of columnaris disease. Specific detection of F. columnare was done using the polymerase chain reaction and DNA in situ hybridization (ISH). Polymerase chain reaction allowed the detection of F. columnare in fresh biological material and in formalin-fixed, paraffin-embedded tissues. The DNA ISH technique allowed the identification and localization of F. columnare in formalin-fixed, paraffin-embedded tissues. Using these molecular techniques, F. columnare was readily detected in skin specimens from infected fish; however, the bacterium was infrequently detected in specimens of liver, kidney, and spleen. These observations suggest that columnaris disease generally presents as a cutaneous disease that is unassociated with systemic infection in koi. Hematologic studies indicated that most infected koi developed microcytic, normochromic, nonregenerative anemia and leukopenia characterized by lymphopenia, mild neutrophilia, and monocytosis. Biochemical changes in diseased fish included significant hyperglycemia, hyponatremia, and hypochloridemia.
An Outbreak of Sheep-Associated Malignant Catarrhal Fever in Bison (Bison Bison) after Exposure to Sheep at a Public Auction Sale
Authors: Berezowski John Andrew , Appleyard Greg D. , Crawford Timothy B. , Haigh Jerry , Li Hong , Middleton Dorothy M. , O'Connor Brendan P. , West Keith , Woodbury Murray
Published: May 2011

An outbreak of malignant catarrhal fever (MCF) among bison sold at an auction market was studied for an 18-month period. Forty-five of 163 bison submitted for sale from 8 different bison farms died on 7 other destination farms. The outbreak began on day 50 after the sale, peaked between days 60 and 70, and ended on day 220. Twenty-one dead bison were confirmed to be MCF cases by clinical histories, pathology, and detection of ovine herpesvirus-2 DNA in their tissues with polymerase chain reaction assays. Twenty-four dead bison were classified as suspect MCF cases from clinical histories. No cases of MCF were observed among bison remaining on originating farms or resident bison mixed with sale bison on destination farms. There were no sheep reported within 3 km of originating or destination farms, limiting bison exposure to sheep to the auction facility, where sheep were present for less than 1 day. The outbreak provides an illustration of the temporal distribution of MCF mortality expected in bison and an estimate of the time from exposure until death from MCF after a single short exposure to sheep. The study provides evidence that bison with MCF do not transmit MCF to other bison.
Diagnosis of Infectious Canine Hepatitis Virus (CAV-1) Infection in Puppies with Encephalopathy
Authors: Caudell D. , Confer A. W. , Fulton R. W. , Berry A. , Saliki J. T. , Fent G. M. , Ritchey J. W.
Published: May 2011

Nine weaned Labrador Retriever puppies from a litter of 11 were presented with signs of acute central nervous system (CNS) disease that included ataxia and blindness. All puppies died. Gross examination of tissues from 2 puppies revealed regionally diffuse hemorrhages in the brain stem and swollen hemorrhagic lymph nodes. Light microscopic examination of hematoxylin and eosin-stained tissues showed numerous large, basophilic intranuclear inclusion bodies within CNS vascular endothelium and occasionally in individual hepatocytes. Immunohistochemical staining of the tissue was positive using an antibody against canine adenovirus-1. Virus isolation for infectious canine hepatitis virus was achieved using inoculated cell cultures. Polymerase chain reaction amplification of DNA from cell culture material revealed shared homology with other mammalian adenoviruses.
Detection of the saa Gene in Verotoxin-Producing Escherichia coli from Ruminants
Authors: Orden José A. , Cortés Carmen , Ruiz-Santa-Quiteria José A. , Martínez Susana , de la Fuente Ricardo
Published: May 2011

A total of 163 verotoxin-producing Escherichia coli (VTEC) strains isolated from diarrheic and healthy cattle, sheep, and goats were analyzed for the presence of the saa gene by polymerase chain reaction. Seventeen (45.9%) and 5 (29.4%) of the VTEC isolated from healthy cattle and diarrheic calves, respectively, had the saa gene. None of the saa-positive strains carried the eae gene, but 20 of the 22 saa positive were ehxA positive. In contrast with cattle VTEC, none of the VTEC isolated from small ruminants were saa positive. These results show that the saa gene is commonly associated with bovine eae-negative VTEC strains but not with ovine or caprine VTEC strains.
Nutritional Degenerative Myopathy in a Population of Captive Bred Uroplatus Phantasticus (Satanic Leaf-Tailed Geckoes)
Authors: Gabor Les J.
Published: May 2011

Severe generalized degenerative myopathy was diagnosed in a population of captive bred satanic leaf-tailed geckoes ( Uroplatus phantasticus). The diagnosis was based on characteristic histological changes and response to dietary therapy. This is the first reported case of nutritional myopathy in the satanic leaf-tailed gecko.
Whole-Blood and Plasma Cholinesterase Levels in Normal Koi (Cyprinus Carpio)
Authors: Murphy Lisa A. , Lewbart Gregory A. , Meerdink Gavin L. , Bargren Gerald L.
Published: May 2011

Organophosphate and carbamate insecticides produce their toxic effects by binding with cholinesterase enzymes, resulting in the inhibition of acetylcholine catabolism. Excessive acetylcholine levels cause excessive synaptic neurotransmitter activity. In most animal species, whole blood is the preferred sample for evaluation of cholinesterase activity. Previous reports of cholinesterase activity in carp have involved sampling of serum, plasma, brain, liver, muscle, and heart. Whole-blood samples were obtained from 12 healthy koi ( Cyprinus carpio). Whole-blood cholinesterase activity was measured in all 12 samples. Plasma cholinesterase activity was also measured in 4 samples. The overall average of the whole-blood cholinesterases was 0.08 μmol/ml/minute (SD = 0.024). The results for the plasma cholinesterases were similar, with an average of 0.08 μmol/ml/minute (SD = 0.012). Because these reference values are below the minimum detection limit for the method of analysis used, results from blood samples from koi suspected to be suffering from anticholinesterase toxicosis may be uninterpretable.
Actinobacillus Capsulatus Septicemia in a Domestic Rabbit (Oryctolagus Cuniculus)
Authors: Meyerholz David K. , Haynes Joseph S.
Published: May 2011

A 5-year-old pet rabbit ( Oryctolagus cuniculus) died after a 3-day history of anorexia and depression. At necropsy, the stomach was distended with dough-like ingesta and hair consistent with gastric stasis syndrome. The lungs had multifocal, raised red nodules with circumferential hemorrhage. Microscopic examination showed pulmonary hemorrhage with intravascular fibrin thrombi and bacterial colonies, which were present in lesser amounts in the kidney, heart, and liver. Bacterial culture of the lung produced a heavy pure growth of Actinobacillus capsulatus. Acute septicemia is a novel presentation for this pathogen. This is the first documented case of A. capsulatus disease in the contiguous United States and may represent an underdiagnosed to emerging disease of lagomorphs.
Epithelioid Leiomyosarcoma in the Visceral Peritoneum of an American Badger (Taxidea Taxus)
Authors: Wang Fun-In , Chang Chih-Hua , Liu Chen-Hsuan , Jeng Chian-Ren
Published: May 2011

A 12-year-old female American badger was presented to the Taipei city zoo veterinary ward with anorexia and weakness. Treatments were ineffective, and the badger died of chronic interstitial nephritis and uremia. At necropsy, numerous firm white nodules, measuring 0.5–2.0 cm, were present on the surface of the liver, stomach, spleen, small intestine, pancreas, and diaphragm. Most nodules were encapsulated and well demarcated from the organs to which they were attached. A poorly demarcated mass, measuring 0.5 cm in diameter, had invaded the hepatic parenchyma and appeared to be the origin of all the nodules derived by transcavitary implantation. Histologically, the nodules contained primarily oval or spindle-shaped cells, typical of smooth muscle cells, forming alternating bundles attached to the surface of the various organs. In some nodules, aggregates of individual polyhedral to round cells with round to oval centrally located nuclei and abundant eosinophilic cytoplasm, typical of smooth muscle origin, were noted. Zones of subcapsular necrosis and multifocal necrosis were also observed in some nodules. Tumor cells stained positively for α-smooth muscle actin and vimentin and negatively for desmin, cytokeratin, estrogen, and progesterone receptors. This tumor is similar to but distinguishable from the “disseminated peritoneal leiomyomatosis (DPL)” found in women.
Comparison of Ear Notch Immunohistochemistry, Ear Notch Antigen-Capture ELISA, and Buffy Coat Virus Isolation for Detection of Calves Persistently Infected with Bovine Viral Diarrhea Virus
Authors: Cornish Todd E. , van Olphen Alberto L. , Cavender Jacqueline L. , Edwards Joan M. , Jaeger Paula T. , Vieyra Leslie L. , Woodard Lynn F. , Miller Dan R. , O'Toole Donal
Published: May 2011

Two techniques performed on skin biopsy samples (ear notches), immunohistochemistry (IHC) and antigen-capture ELISA (AgELISA), were compared for detection of bovine viral diarrhea virus (BVDV) persistent infection (PI) in 559 Angus calves between the ages of 1 and 5 months. The calves also were tested for BVDV infection using virus isolation (VI) and reverse transcription (RT)-PCR on buffy coat samples and for antibodies to BVDV types 1a and 2 by serum neutralization (SN). Sixty-seven of 559 (12.0%) calves tested positive at initial screening by IHC, AgELISA, or VI, and all 67 were kept for a minimum of 3 months and retested monthly by IHC, AgELISA, VI, RT-PCR, and SN. Of the calves positive at initial screening, 59/67 (88.1%) were determined PI and 8/67 (11.9%) were determined acutely infected. Both IHC and AgELISA detected 100% of PI calves; however, IHC and AgELISA also detected 6 and 8 acutely infected calves, respectively, at initial screening. Furthermore, IHC and AgELISA continued to detect 3 and 4 acutely infected calves, respectively, 3 months after initial screening. Three acutely infected calves had IHC staining indistinguishable from PI calves at initial screening. Both IHC and AgELISA are accurate at detecting BVDV-infected calves, but veterinarians and producers should be advised that both tests detect some calves acutely infected with BVDV in addition to PI animals. Repeat testing using VI or RT-PCR on buffy coat samples should be performed at 30 days after initial screening to conclusively discriminate between acute and PI.
Evaluation of a PCR to Detect Salmonella in Fecal Samples of Horses Admitted to a Veterinary Teaching Hospital
Authors: Ward Michael P. , Alinovi Catherine A. , Couëtil Laurent L. , Wu Ching Ching
Published: May 2011

The diagnostic accuracy of a PCR used to identify horses shedding Salmonella spp. in their feces during hospitalization was estimated, relative to bacterial culture of serially collected fecal samples, using longitudinal data. Five or more fecal samples were collected from each of 116 horses admitted as inpatients, for reasons other than gastrointestinal disease, between July 26, 2001 and October 25, 2002. All 873 fecal samples collected were tested with a PCR based on oligonucleotide primers defining a highly conserved segment of the histidine transport operon gene of Salmonella typhimurium, and each sample was cultured for Salmonella spp. One or more samples from 87 (75%) horses were PCR positive, and Salmonella was cultured from 1 or more samples from 11 (9.5%) horses. All culture-positive horses had at least 1 PCR-positive result, whereas only 29 (28%) culture-negative horses were PCR negative on all fecal samples tested. The PCR was most specific, relative to bacterial culture of serially collected fecal samples, when used to test samples from Quarterhorse or breeds other than Thoroughbred or Standardbred, or from clinical (vs. healthy, accompanying horses) cases. Overall, the PCR had the greatest agreement (70%), compared with bacterial culture of serially collected fecal samples, using a cutoff of 2 or more positive PCR test results to define a Salmonella-positive horse. The reasons why some fecal samples, from which Salmonella organisms cannot be isolated, are PCR positive need to be determined before the PCR can be incorporated into Salmonella surveillance programs for hospitalized equine populations.
Clinical and Virological Findings in Pups Naturally Infected by Canine Parvovirus Type 2 Glu-426 Mutant
Authors: Decaro Nicola , Desario Costantina , Campolo Marco , Elia Gabriella , Martella Vito , Ricci Dominga , Lorusso Eleonora , Buonavoglia Canio
Published: May 2011

An outbreak of canine parvovirus type 2 infection caused by the Glu-426 mutant in 2 litters of pups is reported. The infected pups ( n = 6) were monitored daily for evidence of clinical signs and hematological changes and for the evaluation of viral shedding in the feces. The disease induced by the Glu-426 mutant was mild in all the infected pups. Vomiting and hemorrhagic diarrhea were not observed; however, the pups developed mucoid diarrhea (3.5 median days), depression (1.5 median days), and relative leukopenia and lymphopenia (2.5 median days). Fever and loss of appetite were observed only in 2 pups. Virus was detected in the feces for 4.5, 6.5, and 46 median days by hemagglutination, virus isolation on cell cultures, and real-time polymerase chain reaction (PCR), respectively. By real-time PCR, the highest viral DNA titers were detected in the feces of both litters at day 10, reaching median values of more than 1010 DNA copies/mg of feces.
Combined Use of the ASK and SHK-1 Cell Lines to Enhance the Detection of Infectious Salmon Anemia Virus
Authors: Rolland Jill B. , Bouchard Deborah , Coll John , Winton James R.
Published: May 2011

Infectious salmon anemia (ISA) is a severe disease primarily affecting commercially farmed Atlantic salmon ( Salmo salar) in seawater. The disease has been reported in portions of Canada, the United Kingdom, the Faroe Islands, and the United States. Infectious salmon anemia virus (ISAV), the causative agent of ISA, has also been isolated from several asymptomatic marine and salmonid fish species. Diagnostic assays for the detection of ISAV include virus isolation in cell culture, a reverse transcriptase–PCR, an enzyme-linked immunosorbent assay, and an indirect fluorescent antibody test. Virus isolation is considered the gold standard, and 5 salmonid cell lines are known to support growth of ISAV. In this study, the relative performance of the salmon head kidney 1 (SHK-1), Atlantic salmon kidney (ASK), and CHSE-214 cell lines in detecting ISAV was evaluated using samples from both experimentally and naturally infected Atlantic salmon. Interlaboratory comparisons were conducted using a quality control–quality assurance ring test. Both the ASK and SHK-1 cell lines performed well in detecting ISAV, although the SHK-1 line was more variable in its sensitivity to infection and somewhat slower in the appearance of cytopathic effect. Relative to the SHK-1 and ASK lines, the CHSE-214 cell line performed poorly. Although the ASK line appeared to represent a good alternative to the more commonly used SHK-1 line, use of a single cell line for diagnostic assays may increase the potential for false-negative results. Thus, the SHK-1 and ASK cell lines can be used in combination to provide enhanced ability to detect ISAV.
Comparative Analysis of Expert and Machine-Learning Methods for Classification of Body Cavity Effusions in Companion Animals
Authors: Hotz Christine S. , Templeton Steven J. , Christopher Mary M.
Published: May 2011

A rule-based expert system using CLIPS programming language was created to classify body cavity effusions as transudates, modified transudates, exudates, chylous, and hemorrhagic effusions. The diagnostic accuracy of the rule-based system was compared with that produced by 2 machine-learning methods: Rosetta, a rough sets algorithm and RIPPER, a rule-induction method. Results of 508 body cavity fluid analyses (canine, feline, equine) obtained from the University of California–Davis Veterinary Medical Teaching Hospital computerized patient database were used to test CLIPS and to test and train RIPPER and Rosetta. The CLIPS system, using 17 rules, achieved an accuracy of 93.5% compared with pathologist consensus diagnoses. Rosetta accurately classified 91% of effusions by using 5,479 rules. RIPPER achieved the greatest accuracy (95.5%) using only 10 rules. When the original rules of the CLIPS application were replaced with those of RIPPER, the accuracy rates were identical. These results suggest that both rule-based expert systems and machine-learning methods hold promise for the preliminary classification of body fluids in the clinical laboratory.
First Outbreak of Respiratory Disease Associated with Swine Influenza H1N2 Virus in Pigs in Korea
Authors: Jung Kwonil , Chae Chanhee
Published: May 2011

This report describes the first diagnosis of swine influenza H1N2 virus infection in growing pigs in Korea by virus isolation, reverse transcription–polymerase chain reaction (RT-PCR), histopathology, and in situ hybridization. The subtype of swine influenza virus isolates was determined to be H1N2 by RT-PCR. The most consistent and predominant histological feature was bronchointerstitial pneumonia. Lung tissues from these pigs were hybridized with the nonradioactive digoxigenin-labeled complementary DNA (cDNA) probe from H1 HA and N2 NA genes but not from H3 HA and N1 NA genes. A strong hybridization signal was seen in bronchial- and bronchiolar-lining epithelial cells and in alveolar and interstitial macrophages.
Validation of a Human Immunoturbidimetric Assay to Measure Canine Albumin in Urine and Cerebrospinal Fluid
Authors: Gentilini Fabio , Dondi Francesco , Mastrorilli Cinzia , Giunti Massimo , Calzolari Claudia , Gandini Gualtiero , Mancini Danilo , Bergamini Paolo Famigli
Published: May 2011

The aim of this study was to validate an automated immunoturbidimetric assay used to quantify human albumin in urine and to accurately measure canine albumin concentrations in both urine and cerebrospinal fluid. The partial homology existing between human and canine albumin limited the accuracy of the human assays in measuring canine albumin without method modifications. Thus, the assay was modified by calibrating the analyzer with calibrators made in the laboratory containing known concentrations of canine albumin. To prepare the set of calibrators, the albumin concentration of pooled sera of healthy dogs was assessed in 5 replicates using the BromocresolGreen assay. Pooled samples were aliquoted and serially diluted to obtain the expected concentrations of albumin (0.5, 1, 5, 13, and 30 mg/dl) for establishing the canine calibration curve. Thereafter, the performance was assessed by analyzing canine urine and CSF. The modified assay accurately quantified canine albumin in both specimens, as indicated by the following. Intra- and interassay variability was 0.92% and 2.74%, respectively; recovery was 99.66% and 99.07% in urine and 105.02% in CSF. No interference was detected when hemolysate and glucose were added to urine. The test was linear within the verified range (0–225 mg/dl). These results demonstrate that the modified human albumin immunoturbidimetric assay can be a useful tool in the veterinary diagnostic laboratory. It is accurate and tends itself to automatization on chemistry analyzers.
Enterotoxemia Associated with Beta2 Toxin–Producing Clostridium Perfringens Type A in Two Asiatic Black Bears (Selenarctos Thibetanus)
Authors: Greco Grazia , Madio Anna , Martella Vito , Campolo Marco , Corrente Marialaura , Buonavoglia Domenico , Buonavoglia Canio
Published: May 2011

Beta2 (β2) toxin–producing Clostridium perfringens type A strains were found to be associated with necrotic and hemorrhagic intestinal lesions in 2 Asiatic black bears ( Selenarctos thibetanus) that died suddenly. Ten isolates were obtained from the liver, lungs, heart, and small and large intestine of the animals and were examined by multiplex polymerase chain reaction for the genes encoding the 4 lethal toxins (alpha, beta, epsilon, and iota) for classification into toxin types as well as for the genes encoding enterotoxin and the novel β2-toxin for subclassification. In addition, the cpb2 sequence of the 10 isolates was different from the published sequence of cpb2 of pig type C isolate CWC245, whereas it was highly similar to the cpb2 sequence of the C. perfringens type A strain 13. This finding suggests the existence of 2 cpb2 subtypes. This is the first report of enterotoxemia associated with the presence of C. perfringens producing β2-toxin in the tissues and intestinal content of Asiatic black bears.
Detection of Bovine Leukemia Virus by in Situ Polymerase Chain Reaction in Tissues from a Heifer Diagnosed with Sporadic Thymic Lymphosarcoma
Authors: Duncan Robert B. , Scarratt William K. , Buehring Gertrude C.
Published: May 2011

An 18-month-old bovine heifer was presented for clinical evaluation after a sudden onset of ventral edema. Clinical and pathological evaluations were consistent with thymic lymphosarcoma, a sporadic form of lymphosarcoma in cattle, which is not generally considered to be associated with bovine leukemia virus (BLV). This heifer was seropositive for BLV at 6 and 18 months of age. Tissues obtained at necropsy were evaluated using in situ polymerase chain reaction. The BLV proviral DNA was detected in lymphocytes of the thymus as well as in epithelial cells of the liver and kidney. This report presents evidence that thymic lymphosarcomas can be associated with BLV infection and that BLV may have a broader cellular tropism than was supposed previously.
Neuronal Ceroid-Lipofuscinosis in a Holstein Steer
Authors: Hafner S. , Flynn S. , Harmon B. G. , Hill J. E.
Published: May 2011

A young, partially blind Holstein steer was affected by mild cerebral atrophy. Formalin-fixed cerebral gray matter was diffusely yellow brown. Microscopically, there were eosinophilic, autofluorescent granules primarily in the cytoplasm of cerebral neurons. There was also extensive retinal atrophy with complete loss of the rod and cone layers. Ultrastructural examination of affected cerebral neurons revealed a mixture of granular osmiophilic and lamellar patterns in the cytoplasmic storage bodies. This suggests the existence of neuronal ceroid-lipofuscinosis in the Holstein breed.
Fatal Mycobacteriosis with Hepatosplenomegaly in a Young Dog Due to Mycobacterium Avium
Authors: O'Toole D. , Tharp S. , Thomsen B. V. , Tan E. , Payeur J. B.
Published: May 2011

Cases of disseminated Mycobacterium avium infections in dogs are rare because it appears that the species is innately resistant to infection. A 2-year-old, castrated, 5 kg Shih Tzu-Poodle-cross developed anemia, abdominal pain, lethargy, and splenomegaly. Histological examination of surgically removed spleen indicated marked granulomatous splenitis with myriad intracytoplasmic acid-fast bacterial rods. Ultrastructural examination revealed the presence of 3–4-μm-long mycobacteria in phagolysosomes of epithelioid macrophages. Tissue extract of lightly fixed spleen was positive for M. avium 16S ribosomal RNA and negative for M. tuberculosis complex IS 6110 DNA by polymerase chain reaction testing. Anemia was associated with the presence of mycobacteria-infected macrophages in bone marrow. The animal's condition deteriorated, and euthanasia was performed after a clinical course of 2 months. The principal morphological findings at necropsy were severe diffuse granulomatous hepatitis, enteric lymphadenomegaly, and segmental granulomatous enteritis with intralesional mycobacteria present. Mycobacterium avium was cultured from enteric lymph nodes sampled at necropsy. The source of infection was not established but was presumed to be environmental with an enteric portal of entry.
Use of Touch-Down Polymerase Chain Reaction to Enhance the Sensitivity of Mycobacterium Bovis Detection
Authors: Zumárraga Martín J. , Meikle Virginia , Bernardelli Amelia , Abdala Alejandro , Tarabla Hector , Romano María I. , Cataldi Angel
Published: May 2011

The confirmatory diagnosis of Mycobacterium bovis ( M. bovis) in animal samples is carried out by culture in Stonebrink media. However, culture is very slow because of the extremely long duplication time of the bacillus and difficult because of the scarcity of bacilli in diagnostic samples. This study describes the development of a single-tube touch-down polymerase chain reaction (PCR) protocol for the detection of M. bovis using primers that target the IS 6110 element. Spiked water and milk as well as routine diagnostic samples (milk and nasal swabs) from M. bovis–positive cattle were tested. This protocol allows the rapid and sensitive detection of M. bovis in bovine samples by enhancing the sensitivity of standard PCR amplification.
Diagnosis of Taxus (Yew) Poisoning in a Horse
Authors: Tiwary Asheesh K. , Puschner Birgit , Kinde Hailu , Tor Elizabeth R.
Published: May 2011

A 2-year-old bay Thoroughbred colt was found dead overnight in its stall without a known history of any illness, existing disease, or toxicant exposure. No information on the clinical signs before this animal's death was reported. A full necropsy was performed the next morning and revealed a mild to moderate degree of endocardial hemorrhages in both ventricles. Microscopic examination of the heart showed an acute mild mutifocal necrosis of papillary muscles and ventricles. The stomach content contained approximately 2% Taxus alkaloids as determined by gas chromatography/mass spectrometry. In the past, diagnosis of Taxus poisoning has been mainly based on history of exposure and the presence of plant parts in the gastrointestinal tract. Pathological lesions associated with Taxus poisoning have not been published for horses. Therefore, this is the first report of cardiac lesions in a horse after lethal exposure to Taxus. On the basis of these findings, it is suggested that Taxus exposure needs to be considered in the differential diagnosis of horses that die suddenly or have cardiac lesions suggestive of Taxus exposure, even if intact plant parts are not identified in the stomach by the naked eye.
Pneumatosis Cystoides Intestinalis in a Rabbit Doe (Oryctolagus Cuniculus)
Authors: Peris B. , García V. , Blas E. , Segura P. , Martínez J. , Corpa J. M.
Published: May 2011

Pneumatosis cystoides intestinalis (PCI) is an infrequent condition of animals characterized by the existence of numerous thin-walled, gas-filled cystic structures within the intestinal wall and adjacent lymph nodes. Microscopically, the cystic structures appear to be dilated lymphatics located in the lamina propria, submucosa, muscularis, subserosa, mesentery, and mesenteric lymph nodes. This report describes a case of pneumatosis cystoides intestinalis in a rabbit doe from an organic farm where 20 rabbit does were fed ad libitum with a natural diet consisting of whole barley, pea beans, alfalfa hay, and a pelleted vitamin–mineral blend. A combination of nutritional, bacterial, and other factors are hypothesized as possible predisposing factors in the development of PCI.
Experimental Transmission of Chronic Wasting Disease Agent from Mule Deer to Cattle by the Intracerebral Route
Authors: Hamir Amir N. , Kunkle Robert A. , Cutlip Randall C. , Miller Janice M. , O'Rourke Katherine I. , Williams Elizabeth S. , Miller Michael W. , Stack Mick J. , Chaplin Melanie J. , Richt Jürgen A.
Published: May 2011

This communication reports final observations on experimental transmission of chronic wasting disease (CWD) from mule deer to cattle by the intracerebral route. Thirteen calves were inoculated intracerebrally with brain suspension from mule deer naturally affected with CWD. Three other calves were kept as uninoculated controls. The experiment was terminated 6 years after inoculation. During that time, abnormal prion protein (PrPres) was demonstrated in the central nervous system (CNS) of 5 cattle by both immunohistochemistry and Western blot. However, microscopic lesions suggestive of spongiform encephalopathy (SE) in the brains of these PrPres-positive animals were subtle in 3 cases and absent in 2 cases. Analysis of the gene encoding bovine PRNP revealed homozygosity for alleles encoding 6 octapeptide repeats, serine (S) at codon 46, and S at codon 146 in all samples. Findings of this study show that although PrPres amplification occurred after direct inoculation into the brain, none of the affected animals had classic histopathologic lesions of SE. Furthermore, only 38% of the inoculated cattle demonstrated amplification of PrPres. Although intracerebral inoculation is an unnatural route of exposure, this experiment shows that CWD transmission in cattle could have long incubation periods (up to 5 years). This finding suggests that oral exposure of cattle to CWD agent, a more natural potential route of exposure, would require not only a much larger dose of inoculum but also may not result in amplification of PrPres within CNS tissues during the normal lifespan of cattle.
Disseminated Liposarcoma in a Dog
Authors: Wang Fun-In , Liang Sao-Ling , Eng Hock-Liew , Jeng Chian-Ren , Pang Victor Fei
Published: May 2011

A 9-year-old, female Mongrel dog was presented for posterior hindlimb weakness, inability to stand, and pain in the lumbosacral and pelvic regions. Radiography revealed a lytic lesion extending from L5 to L6 to the ilium. At necropsy, an 8 − 2 to 3.2 − 3 cm, irregular, white, firm mass was identified extending from the left dorsolateral aspect of the L6 vertebrae to the sacrum, crossing the sacroiliac joint to the ilium, and reaching the acetabulum without affecting the joint cartilage. Tumor masses were also present bilaterally near the costochondral junction of several ribs. White, soft nodules were present in the heart, pericardium, lungs, spleen, and kidneys as well. Histologically, osteolysis with disruption of the cortical bone and reactive bone with the presence of multinucleated osteoclasts was noted. Neoplastic cells consisted of variable, small basophilic round cells (SBRC) with very scant cytoplasm, larger polygonal cells with abundant eosinophilic cytoplasm, and vacuolated cells resembling adipocytes. Within the marrow cavity, vacuolated cells with necrosis predominated, whereas in periosteal areas, polygonal and vacuolated cells that were mixed with a lower percentage of SBRC were more common. In the lungs and heart, SBRC predominated, and in the spleen, polygonal cells were more numerous. Tumor cells stained positive for vimentin and S-100 and stained negative for CD99, neuron-specific enolase, synaptophysin, chromogranin A, cytokeratins, desmin, myoglobin, and actin. This tumor most likely arose from the marrow cavity of the L6 and later invaded through the vertebral body into adjacent vertebrae and various visceral sites.
Comparison of Individual and Pooled Sampling Methods for Detecting Bacterial Pathogens of Fish
Authors: Mumford Sonia , Patterson Chris , Evered Joy , Brunson Ray , Levine Jay , Winton Jim
Published: May 2011

Examination of finfish populations for viral and bacterial pathogens is an important component of fish disease control programs worldwide. Two methods are commonly used for collecting tissue samples for bacteriological culture, the currently accepted standards for detection of bacterial fish pathogens. The method specified in the Office International des Epizooties Manual of Diagnostic Tests for Aquatic Animals permits combining renal and splenic tissues from as many as 5 fish into pooled samples. The American Fisheries Society (AFS) Blue Book/US Fish and Wildlife Service (USFWS) Inspection Manual specifies the use of a bacteriological loop for collecting samples from the kidney of individual fish. An alternative would be to more fully utilize the pooled samples taken for virology. If implemented, this approach would provide substantial savings in labor and materials. To compare the relative performance of the AFS/USFWS method and this alternative approach, cultures of Yersinia ruckeri were used to establish low-level infections in groups of rainbow trout ( Oncorhynchus mykiss) that were sampled by both methods. Yersinia ruckeri was cultured from 22 of 37 groups by at least 1 method. The loop method yielded 18 positive groups, with 1 group positive in the loop samples but negative in the pooled samples. The pooled samples produced 21 positive groups, with 4 groups positive in the pooled samples but negative in the loop samples. There was statistically significant agreement (Spearman coefficient 0.80, P < 0.001) in the relative ability of the 2 sampling methods to permit detection of low-level bacterial infections of rainbow trout.
Use of Fine Needle Aspirates and Flow Cytometry for the Diagnosis, Classification, and Immunophenotyping of Canine Lymphomas
Authors: Sözmen Mahmut , Tasca Silvia , Carli Erika , Lorenzi Davide De , Furlanello Tommaso , Caldin Marco
Published: May 2011

Fifty canine lymphomas were classified cytomorphologically using the updated Kiel classification scheme. Aspirates of lymph nodes from dogs with lymphoma were stained using 5 canine-specific antibodies and 3 human-specific antibodies that cross-react with canine lymphocytes. The antibody-stained aspirates were analyzed by flow cytometry. A total of 32 (64%) of the 50 lymphomas were characterized as B-cell origin and 18 (36%) were of T-cell origin. B-cell lymphomas were identified in 12 females and 20 males with a mean age of 8.35 years. T-cell lymphomas were identified in 8 females and 10 males with a mean age of 7.9 years. A minority of the lymphomas were low-grade B-cell and T-cell lymphomas (6/50, 12% and 4/50, 8%, respectively). The most common morphologic types were high-grade centroblastic and unclassifiable plasmacytoid for B- and T-cell lymphomas (18/50, 36% and 7/50, 14%, respectively).
Comparison of Selenium Determination in Liver Samples by Atomic Absorption Spectroscopy and Inductively Coupled Plasma–Mass Spectrometry
Authors: Miksa Irina Rudik , Buckley Carol L. , Carpenter Nancy P. , Poppenga Robert H.
Published: May 2011

Selenium (Se) is an essential trace element that is often deficient in the natural diets of domestic animal species. The measurement of Se in whole blood or liver is the most accurate way to assess Se status for diagnostic purposes. This study was conducted to compare hydride generation atomic absorption spectroscopy (HG-AAS) with inductively coupled plasma–mass spectrometry (ICP-MS) for the detection and quantification of Se in liver samples. Sample digestion was accomplished with magnesium nitrate and nitric acid for HG-AAS and ICP-MS, respectively. The ICP-MS detection was optimized for 82Se with yttrium used as the internal standard and resulted in a method detection limit of 0.12 μg/g. Selenium was quantified by both methods in 310 samples from a variety of species that were submitted to the Toxicology Laboratory at New Bolton Center (Kennett Square, PA) for routine diagnostic testing. Paired measurements for each sample were evaluated by a mean difference plot method. Limits of agreement were used to describe the maximum differences likely to occur between the 2 methods. Results suggest that under the specified conditions ICP-MS can be reliably used in place of AAS for quantitation of tissue Se at or below 2 μg/g to differentiate between adequate and deficient liver Se concentrations.
Comparison of Rabbit Monoclonal and Mouse Monoclonal Antibodies in Immunohistochemistry in Canine Tissues
Authors: Vilches-Moure José G. , Ramos-Vara José A.
Published: May 2011

Rabbit monoclonal (RM) antibodies appear to have higher affinity for antigens than mouse monoclonal (MM) antibodies. However, RM antibodies have not been used in veterinary diagnostic immunohistochemistry. The authors compared reactivities of RM and MM antibodies on formalin-fixed, paraffin-embedded canine tissues, targeting 11 different antigens: CD3, CD79a, calcitonin, calretinin, chromogranin A, COX-2, estrogen receptor, Ki67, progesterone receptor, synaptophysin, and vimentin. Paraffin-embedded tissue sections were processed by 1 of 2 antigen-retrieval methods: 1) proteinase K digestion or 2) steam heat in citrate buffer. An additional set of slides did not receive antigen retrieval. Immunostaining was performed using an automated stainer, and scores were assigned to the different dilutions and antigen-retrieval methods on the basis of staining intensity and number of positive cells. Steam heat was usually the best antigen-retrieval method. The optimal dilution for each antibody was that which resulted in the highest specific staining and the lowest nonspecific (background) staining. The RM or MM antibodies yielded a specific reaction for all antigens examined except calretinin. The RM and MM antibodies yielded a specific reaction for 4 antigens only: COX-2, Ki67, synaptophysin, and vimentin. Three antigens (CD3, chromogranin A, and progesterone receptor) were detected only with RM antibodies, whereas the other 3 (CD79a, calcitonin, estrogen receptor) were detected only with MM antibodies. The results of this study differed from those reported for human tissues by the manufacturers of the antibodies. These results emphasize that, regardless of manufacturers' recommendations, each antibody must be individually standardized and validated before routine use in canine tissues.
Performance of a Johne's Disease Enzyme-Linked Immunosorbent Assay Adapted for Milk Samples from Goats
Authors: Salgado Miguel , Manning Elizabeth J. B. , Collins Michael T.
Published: May 2011

Antibody detection–based tests for paratuberculosis offer speed and economy, 2 diagnostic test attributes important to animal industries with narrow profit margins. Application of such tests to individual milk samples instead of serum samples can further improve testing efficiency and decrease testing cost. Accuracy of a commercial bovine paratuberculosis enzyme-linked immunosorbent assay (ELISA) adapted for use on goat serum and milk samples was determined. Fecal, blood, and milk samples were collected from 159 goats belonging to 2 Wisconsin goat herds with a prior history of paratuberculosis and 1 herd of 50 goats from a paratuberculosis-free Wisconsin herd. Fecal samples were cultured using the modified BACTEC 12B media. Sera were tested according to the manufacturer's instructions for bovine samples. Milk samples were centrifuged and mixed with the ELISA kit's Mycobacterium phlei–containing diluent at a ratio of 1:2. Using fecal culture as the “gold standard,” the sensitivity of the ELISA on goat serum was 64% and the sensitivity of the ELISA on goat milk was 48%. The milk ELISA had higher agreement with fecal culture results (kappa = 0.525) than the serum ELISA (kappa = 0.425). ELISA specificity was 100% on both serum and milk. Regression analysis also showed good correlation between serum and milk S/P values ( r2 = 0.67). Although less sensitive, the ELISA on goat milk samples appears to offer a useful, low-cost alternative for detection of goats with paratuberculosis that have progressed to the stage of shedding M. paratuberculosis in their feces.
Comparison of Blood Polymerase Chain Reaction and Enzyme-Linked Immunosorbent Assay for Detection of Mycobacterium Avium Subsp. Paratuberculosis Infection in Cattle and Sheep
Authors: Juste Ramon A. , Garrido Joseba M. , Geijo Marivi , Elguezabal Natalia , Aduriz Gorka , Atxaerandio Raquel , Sevilla Iker
Published: May 2011

A study was carried out to compare the performance of enzyme-linked immunosorbent assay (ELISA) and blood polymerase chain reaction (PCR) for diagnosis of paratuberculosis in cattle and sheep. For cattle, a set of 278 samples from 1 paratuberculosis-affected Friesian farm was used; it included 80 ELISA-positive samples and 198 ELISA-negative samples from an age-matched group. Ninety-four samples were from heifers and 184 were from 2–5-year-old cows. The overall analysis showed a clear association (Fisher exact test [FET] P = 0.0049) but a weak negative agreement (45.3%, kappa = −0.1665 ± 0.0994) between the 2 tests. It reflected a moderate agreement among heifers (87.7%, kappa = 0.4471 ± 0.2435) and a moderate disagreement among cows (62.7%, kappa = −0.3670 ± 0.1057). For sheep, 496 blood samples from 53 Latxa dairy flocks were used; 180 of the blood samples were from dam/offspring pairs. The overall association between the 2 tests on ovine samples was strong (FET, P = 0.0005), whereas the agreement was low (kappa = 0.1622 ± 0.1188). There was slightly better agreement for ewes (kappa = 0.2135 ± 0.1992) than for lambs (kappa = 0.1193 ± 0.1301). There was also a highly unlikely proportion of dam/offspring positive results (FET, P < 0.0001, kappa = 0.6269 ± 0.1854). Four of 6 lambs that were necropsied 1 year after testing had paratuberculosis microscopic lesions in the ileocecal valve (3 lambs) or a PCR-positive result (4 lambs). These results suggest that blood PCR testing might be a potentially useful new approach in paratuberculosis diagnosis, especially in young animals.
Development and Use of a Multiplex Polymerase Chain Reaction Assay Based on Apx Toxin Genes for Genotyping of Actinobacillus Pleuropneumoniae Isolates
Authors: Rayamajhi Nabin , Shin Sung Jae , Kang Sang Gyun , Lee Deog Yong , Ahn Jeong Min , Yoo Han Sang
Published: May 2011

Actinobacillus pleuropneumoniae ( A. pleuropneumoniae) is the etiological agent of a porcine pleuropneumonia that threatens the global swine industry. The major pathogenic toxins of A. pleuropneumoniae include ApxI, ApxII, ApxIII, and ApxIV, which are serotype or serovar specific. Several techniques have been developed for the identification and typing of A. pleuropneumoniae. Serological assays are used to identify and serotype A. pleuropneumoniae, but factors such as cross-reactivity limit their specificity. Labor, time, and the requirement for specific antibodies are also drawbacks of these assays. Multistep polymerase chain reaction (PCR) techniques based on apx genes have been reported for the identification and typing of A. pleuropneumoniae. This study developed multiplex PCR for the identification and genotyping of A. pleuropneumoniae based on apx genes. This multiplex PCR technique was successful in differentiating 11 of 15 reference serotypes. Five different primer sets were used to amplify the 4 apx genes from each serotype in a single-step reaction. The multiplex PCR reported in this study was further used in genotyping 51 field isolates of A. pleuropneumoniae from different regions of Korea. The concomitant amplification of all 4 apx genes makes multiplex PCR more specific and convenient for the diagnosis and genotyping of A. pleuropneumoniae.
Characterization of 26 Isolates of Staphylococcus Aureus, Predominantly from Dairy Sheep, Using Four Different Techniques of Molecular Epidemiology
Authors: Vautor Eric , Jay Corinne , Chevalier Nicolas , Visomblin Nathalie , Vernet Guy , Pépin Michel
Published: May 2011

Little information is available regarding the molecular epidemiology of Staphylococcus aureus–induced mastitis in dairy sheep. In this study, 4 different typing techniques were compared in typing 26 S. aureus isolates, predominantly from cases of subclinical mastitis in dairy ewes. The 4 techniques were pulsed-field gel electrophoresis (PFGE), restriction fragment length polymorphism (RFLP) on 2 genes (coagulase and clumping factor B), randomly amplified polymorphic DNA–polymerase chain reaction (PCR) (RAPD-PCR), and multilocus sequence typing (MLST). On the basis of discriminatory power as the key parameter of typing systems, MLST and PFGE were found to be the most powerful techniques. The MLST and PFGE could contribute to epidemiological surveillance and evaluation of mastitis control programs, by documenting prevalence and dissemination of endemic clones in infected populations. The results of this study show that a single clone of S. aureus is widely distributed in infected ewe mammary glands.