bluuebunny/crossref_metadata_2025 huggingface dataset:
Experimental Reproduction of Tumbleweed Shield Lichen (Xanthoparmelia Chlorochroa) Poisoning in a Domestic Sheep Model
Authors: Dailey Rebecca N. , Montgomery Donald L. , Ingram James T. , Siemion Roger , Raisbeck Merl F.
Published: May 2011

Xanthoparmelia chlorochroa, commonly called tumbleweed lichen, is found throughout the Rocky Mountain region. This particular species of lichen was incriminated in the poisoning of cattle and sheep in Wyoming during the 1930s. More than 70 years elapsed before another case was reported. There is virtually no information in the veterinary literature regarding toxicity of this lichen. This report describes X. chlorochroa poisoning in domestic sheep fed lichen collected from different locales and at different times of the year. Affected animals voided red urine and displayed incoordination. A transient spike in serum creatine kinase activity occurred in all ewes during the course of the feeding trial. Histologically, necrosis of a few individual appendicular skeletal myocytes was observed in 1 ewe, but grossly discernible myonecrosis was absent. The severity of clinical signs varied depending on the location and/or time of year the lichen was collected, indicating that toxicity of the lichen may be influenced by environmental conditions. Results demonstrate that domestic sheep are a useful model for further investigation of X. chlorochroa intoxication. The current study should act as a starting point for elucidating the pathogenesis of X. chlorochroa poisoning and aid in the development of a diagnostic assay to confirm lichen poisoning.
Immunohistochemical Characterization of Mammary Squamous Cell Carcinoma of the Dog
Authors: Sassi Francesco , Sarli Giuseppe , Brunetti Barbara , Morandi Federico , Benazzi Cinzia
Published: May 2011

Squamous cell carcinoma of the mammary gland is rare in both veterinary and human medicine. Whereas human metaplastic and squamous variants are known, the objectives of the current study were to ascertain the presence of such entities in canine mammary tumors and to distinguish them from other (epidermal, sweat gland) squamous tumors that may develop in the same area. A panel of antibodies (anticytokeratin [CK] 19, CK 14, CK 5/6, pancytokeratin, and vimentin) was used on 18 mammary gland malignancies with squamous features and 16 malignant skin tumors (11 squamous cell carcinomas of the skin and 5 sweat glands). Fifteen of the 18 mammary carcinomas were classified as metaplastic carcinomas, and the remaining 3 were classified as squamous cell carcinomas. The 2 most useful markers to establish the histogenesis of mammary tumors were pancytokeratin and CK 19. All other antibodies were equally expressed (CK 14 and 5/6) in all histotypes. The antibody panel discriminated primary epidermal squamous tumors (pancytokeratin positive and CK 19 negative) from gland-derived squamous neoplasms (pancytokeratin positive and CK 19 positive) but failed to distinguish primary mammary tumors from other squamous tumors of glandular origin.
Effect of Anticoagulant and Storage Conditions on Platelet Size and Clumping in Healthy Dogs
Authors: Mylonakis Mathios E. , Leontides Leonidas , Farmaki Rania , Kostoulas Polychronis , Koutinas Alexander F. , Christopher Mary
Published: May 2011

The potential impact of preanalytical factors, such as type of anticoagulant, storage temperature, and time, on the formation of macroplatelets and platelet aggregates (platelet clumping) in dogs is largely elusive. The objective of the current study was to assess the effect of anticoagulant, temperature, and blood storage time in the light microscopy-generated macroplatelet percentages and the frequency of visually inspected platelet aggregates in clinically healthy dogs. Giemsa-stained blood smears from 70 healthy dogs were reviewed after exposure to different anticoagulants (ethylenediamine tetra-acetic acid [EDTA] vs. citrate), temperatures (25°C vs. 4°C), and storage times (up to 24 hr postsampling). The mean percentage of macroplatelets (platelets with diameter or length ≥ μm) was higher ( P = 0.0006) when EDTA was used as the anticoagulant. For either anticoagulant, the mean percentage of macroplatelets was higher ( P < 0.0001) at 25°C than at 4°C. Platelet clumping was 1.9 times ( P < 0.0001) more likely to occur when citrate- rather than EDTA-anticoagulated blood was examined; regardless of the anticoagulant used, clumping occurred 3 times ( P < 0.0001) more often when samples were preserved at 4°C than when they were preserved at 25°C. Storage time did not significantly influence the macroplatelet percentages or the frequency of platelet clumping. The results of this study indicate that macroplatelet percentages in the canine blood should be interpreted in relation to anticoagulant- and temperature-specific reference intervals and that future studies are warranted in order to investigate the clinical relevance of this calculation. In addition, the significant association of citrate with the formation of platelet aggregates may preclude its use for platelet enumeration in the dog.
Evaluation of an Indirect Enzyme-Linked Immunosorbent Assay for Routine Screening of Theiler's Murine Encephalomyelitis Virus Antibodies in Mice Colonies
Authors: Laborde Juan M. , Carbone Cecilia , Corva Santiago G. , Galosi Cecilia M.
Published: May 2011

The current study demonstrates the ability of an indirect enzyme-linked immunosorbent assay (iELISA) to detect antibodies against Theiler's murine encephalomyelitis virus in mice colonies. The antigen was produced from infected baby hamster kidney (BHK)-21 cells and treated with 1% Nonidet P40 in saline buffer. Control antigen was prepared following the same procedure using uninfected BHK-21 cells. The optimal antigen and serum dilutions were established. The reaction was revealed using an anti-mouse-horseradish peroxidase conjugate and 2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). Optimized iELISA was validated by detection of antibodies in known positive and negative serum samples before testing the samples of unknown status. Performance of the iELISA was compared with the indirect fluorescent antibody test, and the cutoff value was determined by receiver operating curve. Indirect ELISA showed 100% sensitivity, 99.38% specificity, and 97.78% predictive positive value. The antigen used is easy to produce, and no special equipment is required. The iELISA developed is simple and provides a rapid and less costly tool for diagnosis and research.
Predictive Values of Aspartate Aminotransferase and Gamma-Glutamyl Transferase for the Hepatic Accumulation of Copper in Cattle and Buffalo
Authors: Minervino Antonio H. H. , Barrêto Raimundo A. , Queiroz Genilson F. , Headley Selwyn A. , Ortolani Enrico L.
Published: May 2011

Ten cattle and 10 buffalo were divided into 2 groups (control [ n = 8] and experimental [ n = 12]) that received daily administration of copper. Three hepatic biopsies and blood samples were performed on days 0, 45, and 105. The concentration of hepatic copper was determined by spectrophotometric atomic absorption, and the activities of aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) were analyzed. Regression analyses were done to verify the possible existing relationship between enzymatic activity and concentration of hepatic copper. Sensitivity, specificity, accuracy, and positive and negative predictive values were determined. The serum activities of AST and GGT had coefficients of determination that were excellent predictive indicators of hepatic copper accumulation in cattle, while only GGT serum activity was predictive of hepatic copper accumulation in buffalo. Elevated serum GGT activity may be indicative of increased concentrations of hepatic copper even in cattle and buffalo that appear to be clinically healthy. Thus, prophylactic measures can be implemented to prevent the onset of a hemolytic crisis that is characteristic of copper intoxication.
Hepatic Encephalopathy in Two Goat Kids with Common Paternity
Authors: Wilkerson Gregory K. , Bera Monali M. , Holt Timothy N. , Callan Robert J. , Pandher Karamjeet
Published: May 2011

Two juvenile, intact, female mixed-breed goats from a common sire were presented for periodic neurologic deficits, seizures, and a generalized loss of body condition that occurred over a 4–6-week period. On physical examination, both goats were thin, obtunded, blind, and ataxic. Laboratory diagnostics revealed increased serum bile acids (95 μmol/l; reference interval: 0–50 μmol/l) in one of the goats. Both goats exhibited progressive physical and mental deterioration, and were eventually euthanized. Upon necropsy, no significant macroscopic lesions were noted. Microscopic examination, however, demonstrated hepatocellular atrophy and anomalies in the hepatic microvasculature, including duplication of hepatic arteries, small-to-indistinct portal veins, and oval cell hyperplasia. In addition, spongiform change was microscopically identified throughout the parenchyma of the brain, most notably within the white matter and along the junction of gray and white matter. The diagnosis of congenital portal vein hypoperfusion (suggestive of a portosystemic shunt) with resultant hepatic encephalopathy was proposed in each case based on the characteristic microscopic lesions in conjunction with the signalment and history of the goats. The observation that the affected kids were sired by the same buck suggests a hereditary basis for the condition in these animals as well.
Equid Herpesvirus 2-Associated Oral and Esophageal Ulceration in a Foal
Authors: Vengust Modest , Baird John D. , van Dreumel Tony , Ackerley Cameron , Bienzle Dorothee
Published: May 2011

A case of a 1-month-old Thoroughbred foal with dysphagia, salivation, pyrexia, oral mucosal pustules, and esophageal ulceration is reported. Swabs from the ulcerated lesions yielded Equid herpesvirus 2 (EHV-2) in virus isolation assays, and histopathology of a biopsy from the esophageal lesion identified nuclear inclusions suggestive of herpesviruses. Immunohistochemical staining with antibodies specific for EHV-2 was positive for epithelial cells in the vicinity of the ulcer but not in more distant mucosa. Electron microscopic evaluation of the biopsy showed herpesviral particles in epithelial cells. The foal recovered over 5 days of supportive and gastroprotective therapy, and the esophageal ulcers healed. Serology and immunohistochemistry indicated that this foal likely had lesions associated with EHV-2 and not EHV-1, −4, or −5.
Fibropapilloma of the Glans Penis in a Horse
Authors: Gardiner David W. , Teifke Jens P. , Podell Brendan K. , Kamstock Debra A.
Published: May 2011

An 18-year-old Arabian stallion was presented for recent onset of stranguria. Physical examination of the distal portion of the glans penis revealed multiple, smooth, glistening, grayish-pink, variably sized, exophytic, nodular masses circumferentially surrounding the external urethral orifice. Partial penile amputation was performed, and the entire specimen was submitted for histological evaluation. Microscopically, the masses consisted of abundant amounts of loosely arranged fibrovascular stroma with low numbers of spindloid to stellate fibrocytes. The overlying epithelium was mildly to moderately hyperplastic with short anastomosing rete ridges (pseudoepitheliomatous hyperplasia). The lesion was diagnosed as fibropapilloma because of features similar to bovine penile fibropapilloma including anatomical location, gross appearance, and histological characteristics. A sarcoid was considered but negated as the lesion lacked the classical streaming and interlacing spindle cell population, “picket-fence” appearance at the epithelial interface, and long, thin, dissecting rete ridges typical of most equine sarcoids. Polymerase chain reaction for the Bovine papillomavirus-1 and Bovine papillomavirus-2 E5 gene and for Equine herpesvirus 1, 3, and 4 was negative on formalin-fixed tissue specimens.
Herpesvirus-Associated Neurological Disease in a Donkey
Authors: Vengust Modest , Wen Xin , Bienzle Dorothee
Published: May 2011

A 4-year-old donkey was evaluated for progressive neurological abnormalities consisting of depression, stupor, weakness, and recumbency. Diagnostic evaluation for viral involvement identified an asinine herpesvirus in DNA extracted from deep pharyngeal swabs. Specific primers were designed based on comparison with equine herpesviral DNA polymerase sequences and yielded an 875-base pair product from the donkey. This sequence had complete identity with short sequences of asinine herpesvirus previously identified in donkeys with interstitial pneumonia. Amino acid analysis of the entire sequence indicated high similarity with Equid herpesvirus 7 (91%), Zebra herpesvirus 1 (90%), and Equid herpesvirus 2 (89%). With supportive treatment and physical therapy, the donkey gradually recovered over 5 days of hospitalization and returned to normal function. The current case illustrates the potential of a novel asinine herpesvirus to induce neurological disease in donkeys and provides a large viral sequence allowing confident assignment of this virus to the subfamily Gammaherpesvirinae.
Perforin Expression in Feline Epitheliotropic Cutaneous Lymphoma
Authors: Neta Michal , Naigamwalla Dinaz , Bienzle Dorothee
Published: May 2011

Cutaneous lymphomas are uncommon in people and companion animals. The tumors can be broadly categorized into epitheliotropic and nonepitheliotropic forms, which appear to have different biological behaviors. The present case describes a feline cutaneous epitheliotropic lymphoma. Masses in a 9-year-old cat were first identified on the tail. The cat was treated with chemotherapy, but additional skin masses developed on the flank, face, and ears. Local radiation induced transient tumor regression, but eventual dissemination prompted euthanasia 13 months after initial tumor appearance. Granular lymphocytes were consistently detected on blood smears, and histologically, the tumor involved the skin and superficial subcutis. Tumor lymphocytes expressed cluster of differentiation 3 (CD3) and perforin molecules, suggestive of a cytotoxic phenotype. Location, histopathological features, and perforin expression were similar to a distinct entity in human medicine designated primary cutaneous, CD8-positive, epidermotropic, cytotoxic, T-cell lymphoma.
A Nonhealing Ulcerative Skin Lesion Associated with Trichinella Nativa Infection in a Cat
Authors: Saari Seppo , Airas Niina , Näreaho Anu , Vihma Veikko , Autio Karoliina , Sankari Satu , Sukura Antti
Published: May 2011

Trichinella spp. can infect various domestic and wild species, including companion animals. Infection occurs because of the ingestion of raw meat (e.g., infected prey). In experimental studies, cats have been found to be a very susceptible host to infection by Trichinella spp.; naturally occurring feline infections have also been reported. However, clinically apparent disease seems to be a rare manifestation of this infection in cats. The skin biopsy of an 8-year-old, neutered, male, domestic cat revealed an inflammatory granulation tissue that surrounded a well-preserved cyst that contained a Trichinella sp. larva. Distinct seropositive reaction against Trichinella spp. antigens was demonstrated by enzyme-linked immunosorbent assay and Western blot. Immunohistochemistry, by using serum from the infected cat as the source of antibody, showed strong immunostaining of Trichinella spp. larvae. During a 1-year follow-up, a postexcisional local tissue reaction was observed. This manifested as a firm, poorly circumscribed subcutaneous mass adjacent to the eye, which demonstrated clinical features and histopathologic findings indicative of chronic inflammation associated with granulation tissue and fibrodysplasia. Digestion of the muscle biopsy revealed one Trichinella sp. larva, which was identified by multiplex polymerase chain reaction as Trichinella nativa. To the authors' knowledge, this is the first documented case of trichinellosis in a cat with a nonhealing ulcerative skin lesion as the main clinical manifestation of the infection.
Hypertrophic Osteopathy Associated with Mycotic Pneumonia in Two Juvenile Elk (Cervus Elaphus)
Authors: Ferguson Nicole M. , Lévy Michel , Ramos-Vara José A. , Baird Debra K. , Wu Ching Ching
Published: May 2011

Two yearling bull elk ( Cervus elaphus) from the same farm developed anorexia, weight loss, and lameness. On physical examination, both elk were thin and showed diffuse swelling of all lower limbs. Radiographs of the lower limbs showed periosteal thickening of the distal extremities, consistent with hypertrophic osteopathy. Thoracic radiographs indicated the presence of pulmonary nodules. Cytologic evaluations of tracheal washes on both elk were consistent with inflammation. Acid-fast stains on both samples were negative. Because of the poor prognosis, both elk were euthanized. At necropsy, the carpal, metacarpal, tarsal, and metatarsal bones, as well as the radius, ulna, and tibia had thickening of cortical bone. There were multiple encapsulated nodules throughout the lungs, lymph nodes, and kidney, and smaller nodules in the myocardium. On microscopic examination, these nodules contained myriads of hyphae, and immunohistochemistry for Aspergillus sp. was strongly positive. Aspergillus fumigatus was isolated from affected tissue in 1 elk. Necropsy findings in both elk were consistent with disseminated fungal granulomas and periosteal hyperostosis. This case presents the first description of hypertrophic osteopathy in elk. The source of infection was undetermined, but inhalation of spores from contaminated feed or bedding was suspected.
An Outbreak of Acute Aflatoxicosis on a Chinchilla (Chinchilla Lanigera) Farm in Argentina
Authors: Pereyra María L. González , Carvalho Eulogio C. Q. , Tissera Jorge L. , Keller Kelly M. , Magnoli Carina E. , Rosa Carlos A. R. , Dalcero Ana M. , Cavaglieri Lilia R.
Published: May 2011

Chinchillas ( Chinchilla lanigera) are known to be very sensitive to aflatoxins, and often a large number of animals die if toxicosis occurs. An outbreak of acute aflatoxicosis on a chinchilla farm in Argentina is described in the present study. A commercial feed suspected of causing the death of 200 animals was sampled. Livers from 9 dead chinchillas were analyzed for their macroscopic and microscopic characteristics via necropsy and histopathology. Aflatoxins B1, B2, G1, and G2 were determined, by thin layer chromatography, to be in the feed. Macroscopic inspection of livers revealed general enlargement, pale-yellowish coloration, hypertrophy, rounded borders, and increased friability. Size and color were remarkably different from a healthy organ. Histopathologic analyses of hepatic parenchyma showed severe, diffuse cytoplasmic vacuolation of hepatocytes. Sudan III staining confirmed the presence of lipid within the vacuoles. The feed was positive for aflatoxin B1 in quantities that exceeded the recommended levels. Histologic lesions were typical of aflatoxin intake. Monitoring feed for mycotoxins is crucial to prevent outbreaks of toxicosis, to improve management practices, and to diminish exposure risk of animals and humans to these harmful toxins.
Practical Sample Size Calculations for Surveillance and Diagnostic Investigations
Authors: Fosgate Geoffrey T.
Published: May 2011

The likelihood that a study will yield statistically significant results depends on the chosen sample size. Surveillance and diagnostic situations that require sample size calculations include certification of disease freedom, estimation of diagnostic accuracy, comparison of diagnostic accuracy, and determining equivalency of test accuracy. Reasons for inadequately sized studies that do not achieve statistical significance include failure to perform sample size calculations, selecting sample size based on convenience, insufficient funding for the study, and inefficient utilization of available funding. Sample sizes are directly dependent on the assumptions used for their calculation. Investigators must first specify the likely values of the parameters that they wish to estimate as their best guess prior to study initiation. They further need to define the desired precision of the estimate and allowable error levels. Type I (alpha) and type II (beta) errors are the errors associated with rejection of the null hypothesis when it is true and the nonrejection of the null hypothesis when it is false (a specific alternative hypothesis is true), respectively. Calculated sample sizes should be increased by the number of animals that are expected to be lost over the course of the study. Free software routines are available to calculate the necessary sample sizes for many surveillance and diagnostic situations. The objectives of the present article are to briefly discuss the statistical theory behind sample size calculations and provide practical tools and instruction for their calculation.
Design and Evaluation of a Multiplex Polymerase Chain Reaction Assay for the Simultaneous Identification of Genes for Nine Different Virulence Factors Associated with Escherichia Coli that Cause Diarrhea and Edema Disease in Swine
Authors: Casey Thomas A. , Bosworth Brad T.
Published: May 2011

A multiplex polymerase chain reaction (mPCR) assay was developed for detection and characterization of pathogenic Escherichia coli that cause diarrhea and edema disease in swine. The mPCR assay was designed as a single reaction for detecting 5 different adhesins (K88, K99, 987P, F41, and F18), 3 enterotoxins (LT, STaP, and STb), and the Shiga toxin (Stx2e) associated with porcine pathogenic E. coli. The specificity of the mPCR assay was evaluated by comparison with results from previous analysis of 100 porcine isolates characterized by colony blot hybridization with DNA probes for the 5 adhesins and 4 toxin genes. There was complete agreement between the 2 methods. The mPCR assay for E. coli pathogens isolated from swine was further evaluated by examination of strains containing virulence factors that are known to have different antigenic subtypes or DNA sequence variations. It was found that the mPCR assays targeting genes encoding for K88 and F18 amplified products with the appropriate sizes from strains containing genes for different K88 and F18 antigenic subtypes; mPCR assays targeting the gene encoding for STaP amplified product from only STaP-positive but not STaH-positive isolates; and mPCR assays targeting the gene encoding for the Stx2 amplified products from only Stx2-positive and not Stx1-positive isolates. Similarly, mPCR assays targeting the gene encoding for LTI did not produce the appropriate product from strains containing genes for LTII. The mPCR assays are simple to perform, and they should be useful for diagnosis of porcine colibacillosis, including the genotypic characterization of E. coli isolates from pigs with diarrhea or edema disease.
Phenotypic and Molecular Typing of Tuberculous and Nontuberculous Mycobacterium Species from Slaughtered Pigs in Egypt
Authors: Mohamed Amr M. , El-Ella Ghada A. Abou , Nasr Essam A.
Published: May 2011

A total of 745 slaughtered pigs were examined during routine meat inspection for suspected tuberculous lesions. Specimens from suspected lesions were collected for conventional mycobacteriologic examinations. Suspected mycobacterial colonies were subjected to molecular typing based on the Mycobacterium species-specific intergenic spacer (IGS) target. The study resulted in detection of suspected lesions in 110 (14.8%) carcasses, from which only 67 specimens produced suspected mycobacterial colonies. Conventional examination was only able to identify 56 isolates as Mycobacterium species, which was confirmed by polymerase chain reaction amplification of the IGS target. Interestingly, out of these, 18 and 12 isolates were Mycobacterium tuberculosis and Mycobacterium bovis, respectively. Sequence analysis of IGS resolved the identities of 10 of the 11 conventionally unidentified isolates as being 4 different nontuberculous Mycobacterium species. The last isolate was proposed as a non -Mycobacterium species and was confirmed by its identification as Rhodococcus equi based on the 16S ribosomal DNA sequence analysis. The study described the isolation of Mycobacterium tuberculosis from pigs and revealed high burden of infection with both tuberculous and nontuberculous mycobacterial species among pigs in Egypt. In addition, the study showed the usefulness of IGS sequence analysis as a reliable molecular tool that would be useful for further epidemiologic and public health studies.
Evaluation of Susceptibility Test Breakpoints Used to Predict mecA-Mediated Resistance in Staphylococcus Pseudintevmedius Isolated from Dogs
Authors: Bemis David A. , Jones Rebekah D. , Frank Linda A. , Kania Stephen A.
Published: May 2011

Clinical and Laboratory Standards Institute interpretive breakpoints for in vitro susceptibility tests that predict mecA-mediated oxacillin resistance in Staphylococcus pseudintermedius isolates from animals have been changed twice in the past decade. Moreover, there are no counterpart recommendations for human isolates of S. pseudintermedius. Individual medical and veterinary laboratories variably use interpretive breakpoints identical to those recommended for use with Staphylococcus aureus or identical to those recommended for use with coagulase-negative staphylococci. The purpose of the current study was to examine correlations between oxacillin disk diffusion, oxacillin gradient diffusion, oxacillin microbroth dilution, and cefoxitin disk diffusion tests used to predict mecA-mediated resistance in S. pseudintermedius and to retrospectively estimate, from disk diffusion zone diameter measurements, the prevalence and rate of increase of oxacillin resistance among canine S. pseudintermedius isolates submitted to a veterinary teaching hospital laboratory. Oxacillin disk diffusion zone diameters of ≤17 mm and oxacillin minimum inhibitory concentrations of ≥0.5 μg/ml were highly correlated with detection of mecA in canine S. pseudintermedius isolates by polymerase chain reaction. MecA-mediated resistance among S. pseudintermedius isolates from dogs increased from less than 5% in 2001 to near 30% in 2007. More than 90% of the methicillin-resistant S. pseudintermedius isolates in 2006 and 2007 were also resistant to representatives of ≥4 additional antimicrobial drug classes. Cefoxitin disk diffusion with the resistance breakpoint set at ≤24 mm significantly underestimated the presence of mecA in S. pseudintermedius.
Genetic Variability of Cytauxzoon Felis from 88 Infected Domestic Cats in Arkansas and Georgia
Authors: Brown Holly M. , Berghaus Roy D. , Latimer Kenneth S. , Britt James O. , Rakich Pauline M. , Peterson David S.
Published: May 2011

Although cytauxzoonosis has historically been nearly 100% fatal in domestic cats, increasing number of reports of infected cats that demonstrate less-severe disease suggest the existence of different strains of Cytauxzoon felis. To test this hypothesis, the genetic variability of C. felis was examined in blood samples from naturally infected domestic cats from Arkansas and Georgia by using the first and second ribosomal internal transcribed spacer regions (ITS1, ITS2) as markers to assess genotypic variability. In addition, the clinical outcome of infection (survival vs. fatal disease) was analyzed. Within the C. felis ITS1 region, there were a total of 8 single nucleotide polymorphisms (SNP) and a single nucleotide insertion. Within the ITS2 region, there were a total of 4 SNPs and a single 40 base pair insertion. When taken together, the ITS1 and ITS2 sequence data defined a total of 11 different sequences and 3 unique genotypes. One unique ITS1-ITS2 genotype was detected in samples submitted exclusively from Arkansas, and a second unique genotype was submitted exclusively from Georgia. There was a significant association between infection with C. felis that contained particular ITS genotypes and survival of the infected domestic cat. The identification of unique C. felis genotypes obtained from different geographic areas and the association of particular ITS genotypes with the outcome of infection suggest the existence of parasite strains that may vary in pathogenicity to the domestic cat and offer an explanation for the survival of some infected cats in more recent case studies.
Disseminated Phaeohyphomycosis in Weedy Seadragons (Phyllopteryx Taeniolatus) and Leafy Seadragons (Phycodurus Eques) Caused by Species of Exophiala, Including a Novel Species
Authors: Nyaoke Akinyi , Weber E. Scott , Innis Charles , Stremme Donald , Dowd Cynthia , Hinckley Lynn , Gorton Timothy , Wickes Brian , Sutton Deanna , de Hoog Sybren , Frasca Salvatore
Published: May 2011

During the period from January 2002 to March 2007, infections by melanized fungi were identified with greater frequency in aquarium-maintained leafy seadragons ( Phycodurus eques)and weedy seadragons ( Phyllopteryx taeniolatus), pivotal species to the educational and environmental concerns of the aquarium industry and conservation groups. The objective of this study was to characterize the pathology and identify fungi associated with phaeohyphomycotic lesions in these species. Samples from 14 weedy and 6 leafy seadragons were received from 2 institutions and included fresh, frozen, and formalin-fixed tissues from necropsy and biopsy specimens. Fresh and frozen tissues were cultured for fungi on Sabouraud dextrose agar only or both Sabouraud dextrose agar and inhibitory mold agar with gentamicin and chloramphenicol at 30°C. Isolates were processed for morphologic identification and molecular sequence analysis of the internal transcribed spacer region and D1/D2 domains of the large subunit ribosomal RNA gene. Lesions were extensive and consisted of parenchymal and vascular necrosis with fungal invasion of gill (11/20), kidney (14/20), and other coelomic viscera with or without cutaneous ulceration (13/20). Exophiala sp. isolates were obtained from 4 weedy and 3 leafy seadragons and were identified to species level in 6 of 7 instances, namely Exophiala angulospora (1) and a novel species of Exophiala (5), based on nucleotide sequence comparisons and phylogenetic analyses. Disseminated phaeohyphomycosis represents an important pathologic condition of both weedy and leafy seadragons for which 2 species of Exophiala,1a novel species, have been isolated.
Plasma Progesterone Analysis by a Time-Resolved Fluorescent Antibody Test to Monitor Estrous Cycles in Goats
Authors: Blaszczyk Barbara , Stankiewicz Tomasz , Udala Jan , Gączarzewicz Dariusz
Published: May 2011

The objective of the current study was to evaluate whether blood plasma progesterone (P4) measurements with a time-resolved fluorescent antibody test (TR-FAT) kit designed for humans was applicable for goats. The first experiment was designed to verify whether the concentrations of P4 measured by TR-FAT can be used to monitor the estrous and ovarian activity in goats ( n = 14). Blood samples (322) were collected, and the ovaries were scanned using ultrasonography. The second experiment was carried out on 4 goats (60 samples) and designed to compare the TR-FAT with radioimmunoassay (RIA). The time interval between the lowest concentrations of P4 assayed by TR-FAT was 21 ± 0.3 days and did not differ significantly from the length of the interestrous interval. The highest concentrations of P4 were confirmed by detection of corpus luteum. During estrus, the mean concentration did not differ significantly between both methods. Significant differences were present during the luteal phases; however, the profiles of P4 assayed by both methods followed a similar pattern. Regression analysis showed a correlation between the 2 methods ( r = 0.98; r 2 = 0.96; P < 0.0001). The Bland-Altman plot showed that all averages were within the 95% limits of agreement; however, the differences between both methods tend to be greater as the average increases. The results demonstrated that the TR-FAT method can be applied to monitor estrous cycles in goats through measurements of plasma P4 concentrations. Moreover, not only does the TR-FAT meet the requirements for safety, but it is also a method of high throughput, rapidity, and simplicity.
Performance of a Commercial Swine Influenza Virus H1N1 and H3N2 Antibody Enzyme-Linked Immunosorbent Assay in Pigs Experimentally Infected with European Influenza Viruses
Authors: Barbé Filip , Labarque Geoffrey , Pensaert Maurice , Van Reeth Kristien
Published: May 2011

The IDEXX Swine influenza virus H1N1 and H3N2 enzyme-linked immunosorbent assays (ELISAs) are used worldwide, but their capacity to detect antibodies to European Swine influenza viruses (SIVs) has not been documented. A total of 313 well-defined sera from SIV seronegative pigs and pigs experimentally infected with European SIVs were used to compare the performance of both ELISAs and the hemagglutination inhibition (HI) test. The ELISAs largely failed to detect pigs that had been infected with H1N1 (0/42 positive in H1N1 ELISA) or H3N2 only (9/18 positive in H3N2 ELISA; group 1). Higher ELISA detection rates were found after consecutive infection of pigs with either H1N1 or H3N2 and 1 other subtype (7/40 and 11/22 positive in H1N1 and H3N2 ELISA, respectively; group 2). Of 39 pigs that had been vaccinated twice with 1 of 4 commercial SIV vaccines (group 3), 25 tested positive in the H1N1 and 4 in the H3N2 ELISA. Pigs that had received a single vaccination after a prior infection with H1N1 and/or H3N2 (group 4) were more frequently positive than group 1 or 3 pigs (23/24 and 15/24 positive in H1N1 and H3N2 ELISA, respectively). Both the H1N1 and H3N2 ELISA showed a low sensitivity (39% and 35%, respectively) relative to the HI test. Because pigs in the field are frequently infected and/or vaccinated with multiple SIV subtypes and variants, they are more likely to test positive in the ELISAs. However, the interpretation of ELISA results will be difficult, and HI remains the method of choice.
Field Performance of Two Rapid Screening Tests in Active Surveillance of Transmissible Spongiform Encephalopathies in Small Ruminants
Authors: Seuberlich Torsten , Doherr Marcus G. , Botteron Catherine , Nicolier Alexandra , Schwermer Heinzpeter , Brünisholz Hervé , Heim Dagmar , Zurbriggen Andreas
Published: May 2011

Recently, screening tests for monitoring the prevalence of transmissible spongiform encephalopathies specifically in sheep and goats became available. Although most countries require comprehensive test validation prior to approval, little is known about their performance under normal operating conditions. Switzerland was one of the first countries to implement 2 of these tests, an enzymelinked immunosorbent assay (ELISA) and a Western blot, in a 1-year active surveillance program. Slaughtered animals ( n = 32,777) were analyzed in either of the 2 tests with immunohistochemistry for confirmation of initial reactive results, and fallen stock samples ( n = 3,193) were subjected to both screening tests and immunohistochemistry in parallel. Initial reactive and false-positive rates were recorded over time. Both tests revealed an excellent diagnostic specificity (> 99.5%). However, initial reactive rates were elevated at the beginning of the program but dropped to levels below 1% with routine and enhanced staff training. Only those in the ELISA increased again in the second half of the program and correlated with the degree of tissue autolysis in the fallen stock samples. It is noteworthy that the Western blot missed 1 of the 3 atypical scrapie cases in the fallen stock, indicating potential differences in the diagnostic sensitivities between the 2 screening tests. However, an estimation of the diagnostic sensitivity for both tests on field samples remained difficult due to the low disease prevalence. Taken together, these results highlight the importance of staff training, sample quality, and interlaboratory comparison trials when such screening tests are implemented in the field.
Comparison of Three Immunological Diagnostic Tests for the Detection of Avian Tuberculosis in Naturally Infected Red Deer (Cervus Elaphus)
Authors: Fernández Jorge González , Fernández-de-Mera Isabel , Reyes Luís E. , Ferreras María C. , Pérez Valentín , Gortazar Christian , Fernández Marta , García-Marín Juan F.
Published: May 2011

Cases of tuberculosis due to Mycobacterium avium subsp. avium in 52 adult red deer ( Cervus elaphus) from a farm were studied using different diagnostic techniques. Immunological probes consisted of the comparative cervical tuberculin (CCT) skin test, the interferon-γ (IFN-γ) assay, and 2 enzyme-linked immunosorbent assays (ELISAs) employing either avian purified protein derivatives or protoplasmatic antigen (PPA-3) as antigens. Three of the animals were euthanized due to severe weakness, loss of weight, and emaciation. Macroscopically, the 3 animals showed tuberculous lesions located mainly in lymph nodes of the digestive system and small intestine but also in other organs and lymph nodes. Polymerase chain reaction was carried out on samples from the 3 deer using primers to detect IS901, IS900, and IS6110, specific for Mycobacterium avium subsp. avium, Mycobacterium avium subsp. paratuberculosis, and Mycobacterium tuberculosis complex, including Mycobacterium bovis, respectively. The last 2 agents cause pathologies very similar to avian tuberculosis in deer. The 3 deer were strongly positive by both ELISAs, slightly positive by the IFN-γ test, and 1 of 2 was positive by the CCT test. As with domestic ruminants, ELISA could detect deer in an advanced stage of infection, with large numbers of mycobacteria.
Serum Glycated Albumin as a Glycemic Control Marker in Diabetic Cats
Authors: Mori Akihiro , Lee Peter , Mizutani Hisashi , Takahashi Tomoya , Azakami Daigo , Mizukoshi Mina , Fukuta Hiroko , Sakusabe Noriko , Sakusabe Arihito , Kiyosawa Yoshio , Arai Toshiro , Sako Toshinori
Published: May 2011

Measurements of glycated proteins such as serum fructosamine, glycated hemoglobin, and glycated albumin (GA) are increasingly used to complement serum glucose concentration for better management of diabetes mellitus. For example, the degree of glycemic control in diabetic cats can be determined by evaluating fructosamine concentration. Unfortunately, fructosamine tests are currently not performed in Japan, and as such, the measurement of GA may serve as a replacement test. The objectives of the current study were 2-fold. First, serum GA and fructosamine level were evaluated for positive correlation in cats as a preliminary gauge on whether serum GA use is applicable. Second, a GA percentage reference range was determined from healthy control cats for possible future diagnostic use. A positive correlation was determined for fructosamine and GA in both normal and diabetic cats. Moreover, the serum GA percentage reference interval based on control cats was determined to be 7.5–13.9% (95% nonparametric interfractile interval). Interestingly, no significant difference in serum GA percentages was observed between samples from diabetic cats with excellent glycemic control and control cats. However, good, fair, and poor glycemic control diabetic cats resulted in a significant increase in serum GA percentages in comparison to control cats. Therefore, these results indicate that serum GA may be a useful glycemic control indicator that could substitute for fructosamine to monitor glycemic control in diabetic cats.
Osteomyelitis and Discospondylitis due to Scedosporium Apiospermum in a Dog
Authors: Hugnet Christophe , Marrou Beatrice , Dally Claire , Guillot Jacques
Published: May 2011

A 6-year-old, 30-kg, female German Shepherd Dog, living in a leishmaniasis enzootic area, was presented with a severe rear limb motor disorder and a medical history of acute onset of fever. Routine hematology indicated neutrophilia. Spinal survey radiographs were consistent with osteomyelitis and discospondylitis. Because of the poor clinical prognosis and the painful nature of the lesions, the dog was euthanized at the owners' request. At necropsy, T13-L1 vertebrae had large areas of necrosis within the vertebral bodies. Histopathological findings were consistent with chronic, severe, fungal osteomyelitis and discospondylitis. Polymerase chain reaction identified Scedosporium apiospermum, a eutrophic filamentous fungus now recognized as an emerging agent of severe infections in immunosuppressed human patients.
Cryptococcus Gattii with Bimorphic Colony Types in a Dog in Western Oregon: Additional Evidence for Expansion of the Vancouver Island Outbreak
Authors: Byrnes Edmond J. , Bildfell Robert J. , Dearing Peggy L. , Valentine Beth A. , Heitman Joseph
Published: May 2011

Cryptococcus gattii was isolated from a 1.5-year-old dog with systemic cryptococcosis in Oregon. The dog had no link to Vancouver Island or British Columbia, Canada. Samples from a nasal swab and from a granulomatous mass within the cranial cavity were pooled for culture. Colonies on Sabouraud dextrose agar were mucoid and exhibited bimorphic morphology, melanin-pigmented and unpigmented. Pigmented colonies were encapsulated budding spherical yeast, whereas unpigmented colonies were of unencapsulated ovoid budding yeast. In addition to defective melanin production, the unpigmented colony type exhibited defective mating. Genetic analysis by high-resolution multilocus sequence typing revealed that the 2 isolates are genetically identical at 8 unlinked loci tested and that the 2 isolates are both the VGIIa Vancouver Island major genotype. Findings are consistent with expansion of the Vancouver Island outbreak onto the mainland Pacific Northwest region of the United States.
Detection of Bovine Viral Diarrhea Virus from Three Water Buffalo Fetuses (Bubalus Bubalis) in Southern Italy
Authors: Martucciello Alessandra , De Mia Gian Mario , Giammarioli Monica , De Donato Immacolata , Iovane Giuseppe , Galiero Giorgio
Published: May 2011

Bovine viral diarrhea virus (BVDV) is an important pathogen that primarily infects ruminants, leading to several clinical problems including abortion. BVDV-specific antibodies were reported in a wide range of hosts within domestic and wildlife animal populations, and serological studies also indicated BVDV infection in buffaloes. The purpose of this study was to analyze the presence of BVDV in 2 water buffalo ( Bubalus bubalis) herds with a history of abortion. Virus isolation from aborted fetuses and from maternal buffy coat and the molecular characterization of the isolates confirmed the presence of BVDV in these animals. The sequence analysis based on the 5′ UTR and Npro coding regions of the Pestivirus genome revealed that the isolates belong to subgenotype 1b of BVDV. The findings of this study also suggest a possible role of BVDV in causing congenital infection in water buffalo. Its presence in fetal tissues as well as in maternal blood raises questions about the possible development of clinical disease or its influence in abortions in water buffalo.
Metastatic Malignant Melanoma in an Alpaca (Vicugna Pacos)
Authors: Mollat Wendy Harless , Gailbreath Katherine L. , Orbell Geoffrey M.
Published: May 2011

A case of malignant melanoma in a 7-year-old, intact, black, male Huacaya alpaca with a history of a chronic, nonhealing wound involving the left external nostril, weight loss, and inappetence is described. Malignant melanoma was diagnosed by histology of punch biopsy specimens from a mass on the maxilla associated with the nonhealing wound and from a mass in the submandibular region. The alpaca was humanely euthanized 10 days after the diagnosis on the basis of the poor prognosis and rapid clinical deterioration. At postmortem examination, the alpaca had an ulcerated, multilobulated, black pigmented mass (8.0 cm × 6.0 cm × 4.0 cm) that infiltrated the left rostral maxilla extending into the marrow space and into the left nasal cavity. Numerous, discrete, coalescing masses were present in the subcutaneous tissue of the submandibular area, peritracheal connective tissue, pericardium, and diaphragmatic parietal pleura and were disseminated throughout the pulmonary parenchyma. The masses were diffusely black on cut surface and exuded black pigment. Histologically, all masses were composed of spindloid to polygonal cells with indistinct cell borders and moderate amounts of cytoplasm that contained abundant fine, black granules (melanin), confirming metastasis of a primary mucocutaneous melanoma. To the authors' knowledge, this is the first report of a malignant melanoma involving bone in a New World camelid.
Disseminated Bovine Viral Diarrhea Virus in a Persistently Infected Alpaca (Vicugna Pacos) Cria
Authors: Byers Stacey R. , Snekvik Kevin R. , Righter Daniel J. , Evermann James F. , Bradway Daniel S. , Parish Steven M. , Barrington George M.
Published: May 2011

Bovine viral diarrhea virus (BVDV) is an emerging infectious pathogen of concern to the alpaca industry. A 4-month-old, intact, male alpaca cria was diagnosed as persistently infected with BVDV on the basis of repeated positive antemortem polymerase chain reaction (PCR) and virus isolation (VI) assays and negative serologic titers to BVDV. Immunohistochemistry, real-time reverse transcription PCR, and VI performed on tissues collected at necropsy demonstrated disseminated BVDV-1b infection. Virus was detected in multiple tissues, including parotid salivary gland, testes, prostate, kidneys, skin, and gastrointestinal tract. Demonstration of BVDV in previously unreported tissues suggests additional potential routes of BVDV transmission in alpacas.
Sarcomatoid Carcinoma in the Lung of an Egyptian Fruit Bat (Rousettus Aegyptiacus)
Authors: McLelland David J. , Dutton Christopher J. , Barker Ian K.
Published: May 2011

A sarcomatoid carcinoma was diagnosed in the lung of a 10-year-old captive Egyptian fruit bat ( Rousettus aegyptiacus). Both carcinomatous and sarcomatous cytologic phenotypes were identified histologically. Cells of both types stained positive for pancytokeratin and S-100. Stromal cells stained positively for muscle actin. No staining for vimentin was noted in either neoplastic or normal internal control tissues. To the authors' knowledge, this is the first report of a pulmonary sarcomatoid carcinoma in a bat, and only the third report of sarcomatoid carcinoma outside the human literature.
Systemic Conidiobolus Incongruus Infection and Hypertrophic Osteopathy in a White-Tailed Deer (Odocoileus Virginianus)
Authors: Madson Darin M. , Loynachan Alan T. , Kariyawasam Subhashinie , Opriessnig Tanja
Published: May 2011

Postmortem examination of a free-range white-tailed deer ( Odocoileus virginianus) revealed severe emaciation, bilateral firm proliferation of the metatarsal diaphyses, and a large intrathoracic mass associated with the accessory lung lobe. Smaller masses were evident in the abomasum, duodenum, omentum, and the capsular surface of the liver. Microscopically, the masses were similar and were diagnosed as eosinophilic granulomas with intralesional fungal hyphae characteristic of Zygomycetes spp. Fungal hyphae were identified as Conidiobolus incongruus by 18S ribosomal RNA sequencing on fresh lung tissue. Furthermore, the proliferative lesions of the metatarsal bones along with the intrathoracic mass were compatible with hypertrophic osteopathy.
Osteogenic Intraocular Melanoma in an Alpaca (Vicugna Pacos)
Authors: Hill Fraser I. , Hughes Scott M.
Published: May 2011

A 10-year-old, female, pregnant alpaca ( Vicugna pacos) presented with a discolored, swollen right eye. The eye did not respond to treatment and was enucleated. Sectioning of the fixed eye revealed an ocular mass replacing the aqueous and vitreous humors, with multiple regions of bone formation. Histopathological, immunohistochemical, and transmission electron microscopy confirmed the tumor was an intraocular melanoma with bone formation. Osteogenic melanoma is a rare variant of melanoma not reported in the eye before.