bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic Characterization of Canine Rotavirus Isolated from a Puppy in Korea and Experimental Reproduction of Disease
Authors: Kang Bo Kyu , Song Dae Sub , Jung Kwon Il , Lee Chul Seung , Park Sung Jun , Oh Jin Sik , An Dong Jun , Yang Jeong Sun , Moon Hyoung Joon , Lee Sang Sun , Yoon Young Dhuk , Park Bong Kyun
Published: May 2011

Canine rotavirus was isolated from feces of a Korean Jindo dog with mild diarrhea, and the isolate was genetically characterized. Rotaviral antigen was detected in the feces using a commercial rotavirus antigen detection kit and cytopathic effects were observed in a cell line inoculated with the feces. The virus isolate (GC/KS05) was identified as subtype G3P[3] using reverse transcription polymerase chain reaction (RT-PCR). The strain displayed 98% and 90% identity with the VP7 genes of a canine rotavirus isolate (RV52/96) from Italy and the simian rotavirus strain (RRV) respectively. However, the GC/KS05 isolate exhibited only 83% and 82% identity, respectively, with the G3 serotype canine strains, RV198/95 and K9. Phylogenetic analysis of the VP7 and VP4 genes of GC/KS05 strain led to the classification of VP7 in a different cluster than other canine rotavirus VP7 genes, and VP4 within the cluster of canine rotavirus VP4 genes. The Korean isolate was thus more closely related to the RV52/96 isolate than the other isolates for which sequence data is available. Detailed analysis of the VP7 region revealed 6 amino acid variations between the new isolate and RV52/96. After 5 passages in cell culture, the GC/KS05 strain remained pathogenic for young pups, in which inoculation resulted in diarrhea and virus shedding in the feces.
Brain Lesions in Pigs Affected with Postweaning Multisystemic Wasting Syndrome
Authors: Corrěa André Mendes Ribeiro , Zlotowski Priscila , de Barcellos David Emilio Santos Neves , da Cruz Cláudio Estěvão Farias , Driemeier David
Published: May 2011

Anti-porcine circovirus type 2 (anti-PCV2) immunostaining was associated with cerebellar lymphohistiocytic vasculitis combined with hemorrhages (50 pigs) or with lymphohistiocytic meningitis (23 pigs) in pigs naturally affected with postweaning multisystemic wasting syndrome (PMWS). The animals originated from 12 farms in Rio Grande do Sul, Brazil. In total, 456 unthrifty 3- to 5-month-old postweaning pigs confirmed as PMWS cases were necropsied. Although most findings mimicked those extensively reported in PMWS-affected pigs, there were distinctive brain lesions that included multiple hemorrhages in the cerebellar leptomeninges associated with lymphohistiocytic vasculitis and fibrinoid degeneration in vessels of the cerebellum and periventricular areas (69 pigs). These vascular lesions were also seen in conjunction with lymphohistiocytic meningitis (38 additional pigs). PCV2 antigen was immunohistochemically demonstrated in the cytoplasm and nuclei from intralesional perivascular macrophages and endothelial-like cells in brain tissues. Together these findings suggest that these lesions were caused by PCV2.
Accidental Intra-Auricular Artery Injection of Ceftiofur in Two Beef Steers
Authors: Bolin David C.
Published: May 2011

Two beef steers accidentally injected into a branch of the auricular artery with an oil-based formulation of ceftiofur died within 5 minutes of injection. Notable pathologic findings included distention and obstruction of cerebral and cerebellar arteries by a whitish tan material and hemorrhages within meningeal spaces, the choroid plexus, cerebrum, and cerebellum. Lipid material was identified within cerebral blood vessels in frozen sections stained with oil red O. This report describes an unusual case of brain ischemia in beef cattle.
Serological Prevalence of Canine Respiratory Coronavirus in Southern Italy and Epidemiological Relationship with Canine Enteric Coronavirus
Authors: Priestnall Simon Lawrence , Pratelli Annamaria , Brownlie Joe , Erles Kerstin
Published: May 2011

Canine respiratory coronavirus (CRCoV) has been detected in dogs suffering from respiratory disease and is thought to be involved in canine infectious respiratory disease (CIRD) complex. Canine enteric coronavirus (CECoV) is a widespread pathogen of dogs, responsible for mild to severe diarrhea in pups. The purpose of this study was to establish the seroprevalence of CRCoV in Italy and its relationship to CECoV type II seroprevalence. The age and year of sample collection from seropositive dogs was also assessed. Of adult domestic dogs, 23.3% had antibodies to CRCoV, compared with 86.1% with antibodies to CECoV. Amongst a population of kenneled pups, 4.0% had antibodies to CRCoV, and 97.0% had antibodies to CECoV.
Clostridium Perfringens type A Toxin Production in 3 Commonly Used Culture Media
Authors: Fernandez-Miyakawa Mariano E. , Marcellino Romanella , Uzal Francisco A.
Published: May 2011

In vitro toxin production is an important tool not only for diagnostic purposes but also for the study of pathogenesis of Clostridium perfringens infections. The present study was carried out to compare the level of toxin production by several strains of C. perfringens type A, isolated from the intestine of animals, when cultured in 3 different conventional culture media. Six strains of C. perfringens type A isolated from the small intestine of healthy sheep were cultured in commercial cooked meat medium (CMM), brain heart infusion (BHI), and tryptone glucose yeast (TGY). Intravenous lethality in mice and phospholipase C (PLC) activity were measured in filtered culture supernatants. Lethality of culture supernatants was highest for all isolates when grown in BHI, followed by CMM. No supernatants from any isolates grown in TGY produced lethality in mice. Phospholipase C activity was highest when the isolates were grown in BHI and CMM and significantly lower when grown in TGY.
Unilateral Orchitis and Epididymitis Caused by Salmonella Enterica Subspecies Diarizonae Infection in a Ram
Authors: del Carmen Ferreras Ma , Muñoz María , Pérez Valentín , Benavides Julio , García-Pariente Carlos , Fuertes Miguel , Adúriz Gorka , García-Marín Juan Francisco
Published: May 2011

A case of unilateral suppurative epididymo-orchitis associated with Salmonella enterica subsp. diarizonae serovar 61:k:1,5,(7) infection is described in a 2-year-old ram. Gross lesions were characterized by severe enlargement of the scrotal contents, fibrous adhesions between testicular layers, coexistence of epididymal abscesses and foci of fibrinous exudate, and testicular atrophy. Microscopically, testicular and epididymal microabscesses and diffuse inflammatory infiltrates with abundant macrophages containing short Gram-negative rods were observed. Superimposed on the chronic lesions were fibrin deposits with clusters of neutrophils, as well as walled-off granulation tissue. Bacterial colonies were also identified in thrombosed spermatic cord vessels, scrotal lymph nodes, lung, and liver. S. enterica subsp. diarizonae serovar 61:k:1,5,(7) was isolated from the affected testis. To the authors' knowledge, this is the first report of infection of the testis and epididymis by Salmonella in rams. This organism must be taken into account in the differential diagnosis of ovine genital infections.
Muscular Pseudohypertrophy (Steatosis) in a Bovine Fetus
Authors: Langohr Ingeborg M. , Stevenson Gregory W. , Valentine Beth A.
Published: May 2011

Muscular pseudohypertrophy was diagnosed in the cervical musculature of a full-term crossbred Simmental fetus delivered by fetotomy. Only head and cervical regions were submitted for pathologic examination; the rest of the fetal body was reportedly normal. The neck musculature of the fetus was markedly deformed by 23 cm and 18 cm in diameter, firm, spherical masses that consisted of enlarged and pale left splenius and right serratus ventralis cervicis muscle, respectively, covered by intact skin. Additionally, lipomatous masses were present within the cervical vertebral canal, compressing the spinal cord. Microscopically, the prominent muscular enlargement was due to massive adipose and fibrous connective tissue replacement of atrophic muscle. Focal myelodysplasia and astrocytosis affecting the grey matter was detected in the mid-cervical region of the spinal cord, accompanied by degeneration in the ascending and descending tracts of the remaining cord segments. Abnormal spinal cord development as a result of severe spinal cord compression by the lipomatous masses within the spinal canal leading to replacement of muscle by fat and fibrous tissue was considered to be the cause of the muscular malformation in this fetus.
Kernicterus in a Neonatal Foal
Authors: Loynachan Alan T. , Williams N. M. , Freestone J. F.
Published: May 2011

A 5-day-old Thoroughbred foal was submitted to the necropsy service at the University of Kentucky Livestock Disease Diagnostic Center. The foal had a clinical history of seizure activity and severe icterus. A complete blood count and serum chemistry analysis indicated that the foal was anemic (hematocrit, 16%), hyperbilirubinemic (45 mg/dl), and hypoglycemic. At necropsy, all tissues were discolored various shades of yellow. Microscopically, there was degeneration and necrosis of cerebral neurons and cerebellar Purkinje cells; severe hepatocellular degeneration and necrosis; and deposition of amorphous golden-yellow material in the cerebellar granular cell layer, pulmonary alveoli, renal tubular epithelium, splenic trabecula, and the lamina propria of the small and large intestine. The golden-yellow material in the brain, lung, spleen, and small intestine was identified as bilirubin by histochemistry. Based on the macroscopic and microscopic findings, a diagnosis of kernicterus (bilirubin encephalopathy) was made. This report describes a rare case of equine neonatal kernicterus.
Fatal Ricin Toxicosis in a Puppy Confirmed by Liquid Chromatography/Mass Spectrometry when Using Ricinine as a Marker
Authors: Mouser Pamela , Filigenzi Michael S. , Puschner Birgit , Johnson Valerie , Miller Margaret A. , Hooser Stephen B.
Published: May 2011

Ricin, a lectin from the castor bean plant ( Ricinis communis), is considered one of the most potent plant toxins. Ingestion of masticated seeds results in high morbidity, with vomiting and watery to hemorrhagic diarrhea. The prognosis varies with the number of seeds ingested, the degree of mastication, individual susceptibility, and the delay in treatment. Low mortality restricts assessment of histologic lesions, and the literature on toxicologic analysis for ricin is limited. This report describes a fatal case of castor bean ingestion in a 12-week-old Mastiff puppy, with confirmation of ricin exposure through detection of the biomarker ricinine by liquid chromatography/mass spectrometry (LC/MS). Despite supportive therapy, the puppy died several hours after presentation for acute vomiting, diarrhea, and lethargy. At necropsy, a segment of jejunum and mesenteric lymph nodes were congested. When the owner reported the presence of castor beans in the dog's feces, selected formalin-fixed and unfixed tissues were submitted for diagnostic evaluation. Histopathologic findings included superficial necrotizing enteritis of the jejunum and occasional, random foci of coagulative necrosis in the liver. The alkaloid ricinine was detected in gastric content by using a newly developed LC/MS method. This confirmation of exposure is important in the diagnosis of ricin toxicosis, because ingestion of castor beans is not always fatal, histologic lesions are nonspecific, and the degree of mastication can influence the effective dose of ricin.
Failure to Detect Bovine Viral Diarrhea Virus in Necropsied Farm-Raised White-Tailed Deer (Odocoileus Virginianus) in Pennsylvania
Authors: Brooks Jason W. , Key Douglas W. , Hattel Arthur L. , Hovingh Ernest P. , Peterson Ryan , Shaw Daniel P. , Fisher Jenny S.
Published: May 2011

Between January 1 and December 31, 2005 gross and histologic examinations were performed on carcasses of 61 farm-raised white-tailed deer originating from Pennsylvania. Single-tube real-time reverse transcription polymerase chain reaction (real-time RT-PCR) for the detection of bovine viral diarrhea virus type 1 (BVDV-1) and type 2 (BVDV-2) was performed on each animal. Virus isolation was performed on tissue samples from 25 of 61 animals. Immunohistochemical (IHC) staining of ear-notch skin to identify BVDV antigen was performed on each animal. All tissues samples tested negative for both BVDV-1 and BVDV-2 by real-time RT-PCR, virus isolation, and IHC. Gross or histopathologic lesions suggestive of BVDV infection were not detected. Results of this study suggest that BVD is not a common cause of mortality in farm-raised white-tailed deer in Pennsylvania.
Superoxide Dismutase Expression and Oxidative Damage in a Case of Myopathy in Brown Pelicans (Pelecanus Occidentalis)
Authors: Giri Dipak K. , Miller Debra L. , Thompson Larry J. , Mailler Lesley , Styer Eloise , Baldwin Charles
Published: May 2011

Four brown pelicans ( Pelecanus occidentalis) housed at a rehabilitation facility were found dead after a 3-day history of muscle weakness and after being fed for about 2 weeks from a recent shipment of fish. The birds had pale streaking of the skeletal and heart muscles. Microscopically, the skeletal muscle, and to a lesser extent the cardiac muscle, had severe myocyte degeneration and necrosis characterized by microvacuolation with loss of cross-striations, condensation of cytoplasm, fragmentation, mineralization, and inflammatory cell infiltrates consisting of multinucleated cells, macrophages, and few heterophils. The findings were consistent with myopathy, and a nutritional myopathy caused by eating rancid fish was suspected. Immunohistochemical staining revealed abundant immunoreactive copper zinc superoxide dismutase and manganese superoxide dismutase either as diffuse homogeneous precipitates or granular aggregates in the cytoplasm of affected cells. Immunoreactivity was directly related to degree of cellular damage as estimated by light microscopic examination. We suggest that the lack of protection, despite upregulation of superoxide dismutase, is most likely attributable to supersaturation of oxidants beyond the capacity of superoxide dismutases to scavenge.
Mortality in Pennsylvania Captive Elk (Cervus Elaphus): 1998–2006
Authors: Hattel Arthur L. , Shaw Daniel P. , Fisher Jenny S. , Brooks Jason W. , Love Brenda C. , Drake Thomas R. , Wagner Donald C.
Published: May 2011

The necropsy records of 65 elk ranging from 1 day to 13 1/2 years of age were reviewed to determine the primary cause of death of each animal. The animal carcasses were submitted from 22 separate Pennsylvania elk farms over an approximately 8-year period. The most common causes of mortality in animals in which the cause of death was determined were gastrointestinal parasitism (21 cases), pneumonia (7 cases), enterocolitis (5 cases), malnutrition (5 cases), and rumenitis/acidosis (5 cases). The cause of death was undetermined in 4 of the 65 elk. Nematode organisms (20 cases) were the primary parasites identified in cases of mortality owing to gastrointestinal parasitism. Arcanobacterium pyogenes (3 cases), Escherichia coli (3 cases), and Streptococcus sp. (2 cases) were the most commonly isolated bacteria from the lung in the cases of pneumonia. Fungal organisms, consistent with Aspergillus sp. were present histologically within the pulmonary lesions in 5 cases. Bacterial agents associated with enterocolitis included Clostridium perfringens (2 cases), E. coli (1 case), Salmonella Newport (1 case) and Salmonella Thompson (1 case). The majority (56.3%) of the death loss in elk of known ages occurred in animals 1 year of age or less, with 71.4% of the gastrointestinal parasitism, 60.0% of the malnutrition, 60.0% of the enterocolitis, and 57.1% of the pneumonia cases occurring in animals within this age range. In conclusion, gastrointestinal parasitism, pneumonia, malnutrition, enterocolitis and rumenitis acidosis were considered the most common causes of death in captive elk in this study.
Effect of Time and Temperature on PrPCWD Immunoreactivity as Evidenced by Western Blot
Authors: Triantis Joni , Dennis Michelle M. , Salman Mo D. , Gould Daniel H.
Published: May 2011

The protease-resistant infectious prion protein, PrPres, that causes transmissible spongiform encephalopathies, is remarkably resistant to conventional physical and chemical sterilization methods, including heat. It was hypothesized that thermal-dependent PrPres degradation has been underestimated, and the effect of prolonged incubation at 37°C, 55°C, and 80°C on PrPres detection was examined using brain homogenates from chronic wasting disease-affected elk and mule deer (PrPCWD). Immunoblotting demonstrated progressive loss of PrPCWD immunoreactivity with time in all incubated samples as temperature increased, and PrPCWD was virtually undetectable after 90 days of incubation at 55°C and 80°C. These results indicate that decontamination methods and tissue disposal systems maintaining elevated temperatures for long periods of time could interfere with immunodetection, and the reliability of assays for PrPres detection could be compromised when applied to tissues exposed to heat with time. Although these results may suggest that such prolonged heat treatment could destroy prions, the observed loss of immunoreactivity does not necessarily correlate with a concurrent loss of infectivity. Bioassay is needed to determine if samples that have been incubated under these conditions retain infectivity.
Tympany, Acidosis, and Mural Emphysema of the Stomach in Calves: Report of Cases and Experimental Induction
Authors: Panciera Roger J. , Boileau Melanie J. , Step Douglas L.
Published: May 2011

Necropsy of 4 neonatal calves with a peracute syndrome of abdominal distention, diarrhea, dehydration, shock, and death revealed abomasal tympany, marked edema, hemorrhage, and emphysema of ruminal and abomasal walls and histopathologic lesions characteristic of forestomach acidosis. The presence of acid damage and the production of substantial quantities of gas strongly suggested that the pathogenesis of the syndrome involved exuberant fermentation of intragastric substrate. This supposition led to attempts to experimentally induce the syndrome by dosing neonatal calves with an alimentary tract flora, followed by milk replacer supplemented with excessive fermentable carbohydrate (D-glucose and cornstarch). Two of the 5 calves thus treated developed a syndrome very similar to that observed in naturally affected calves. Literature suggests involvement of several Clostridial species, Sarcina spp., and possibly other microbes or a combination thereof to be involved in very similar syndromes. Therefore it is suggested that this complex of lesions and clinical signs is not dependent on a single etiologic agent. Rather, it is proposed that the natural occurrence of disease requires a quantity of highly fermentable substrate (starch, glucose, lactose, etc.) and a bacterial flora that is capable of rapidly fermenting that particular substrate with a resultant production of gas and acid.
A Universal Test to Determine the Integrity of RNA, and its Suitability for Reverse Transcription, in Animal Tissue Laboratory Specimens
Authors: Oakey H. Jane
Published: May 2011

Degradation of RNA in diagnostic specimens can cause false-negative test results and potential misdiagnosis when tests rely on the detection of specific RNA sequence. Current molecular methods of checking RNA integrity tend to be host species or group specific, necessitating libraries of primers and reaction conditions. The objective here was to develop a universal (multi-species) quality assurance tool for determining the integrity of RNA in animal tissues submitted to a laboratory for analyses. Ribosomal RNA (16S rRNA) transcribed from the mitochondrial 16S rDNA was used as template material for reverse transcription to cDNA and was amplified using polymerase chain reaction (PCR). As mitochondrial DNA has a high level of conservation, the primers used were shown to reverse transcribe and amplify RNA from every animal species tested. Deliberate degradation of rRNA template through temperature abuse of samples resulted in no reverse transcription and amplification. Samples spiked with viruses showed that single-stranded viral RNA and rRNA in the same sample degraded at similar rates, hence reverse transcription and PCR amplification of 16S rRNA could be used as a test of sample integrity and suitability for analysis that required the sample's RNA, including viral RNA. This test will be an invaluable quality assurance tool for determination of RNA integrity from tissue samples, thus avoiding erroneous test results that might occur if degraded target RNA is used unknowingly as template material for reverse transcription and subsequent PCR amplification.
Development of a Nested Polymerase Chain Reaction Method to Detect Oncogenic Marek's Disease Virus from Feather Tips
Authors: Murata Shiro , Chang Kyung-Soo , Lee Sung-Il , Konnai Satoru , Onuma Misao , Ohashi Kazuhiko
Published: May 2011

For the easy survey of Marek's disease virus (MDV), feather tip–derived DNA from MDV-infected chickens can be used because feather tips are easy to collect and feather follicle epithelium is known to be the only site of productive replication of cell-free MDV. To develop a diagnostic method to differentiate highly virulent strains of MDV from the attenuated MDV vaccine strain, CVI988, which is widely used, nested polymerase chain reaction (PCR) was performed to detect a segment of the meq gene in feather tip samples of chickens experimentally infected with MDV. In chickens infected with Md5, a strain of oncogenic MDV, the meq gene was consistently detected, whereas the L- meq gene, in which a 180–base pair (180-bp) sequence is inserted into the meq gene, was detected in CVI988-infected chickens. Moreover, the meq gene was mainly detected even in chickens co-infected with both Md5 and CVI988. These results suggest that this method is appropriate for the surveillance of the highly virulent MDV infection in the field.
Molecular Assays for Detection of Falcon Adenovirus
Authors: Schrenzel Mark , Snook Eric , Gagneux Pascal
Published: May 2011

Falcon adenovirus is a newly recognized member of the family Aviadenoviridae and includes 2 closely related strains that are pathogenic to several species of falcons. Peregrine falcons appear to be one of the primary reservoirs, but recent outbreaks suggest that other carrier species probably exist. To allow screening of captive birds for virus shedding and investigations of disease outbreaks, conventional and realtime, quantitative polymerase chain reaction (PCR) assays and an in situ hybridization technique were developed. The diagnostic protocols were used on tissue and fecal samples from 7 species or subspecies of falcons infected with adenovirus as well as adenoviruses from other birds and mammals. The assays were specific for falcon adenovirus and detected both strains of virus in fecal samples from living animals or frozen and formalin-fixed, paraffinized tissues. Together with established serologic tests for falcon adenovirus, these molecular assays are valuable tools for management and conservation of falcons in captivity and the wild.
Evaluation of the Sensitivity of Reverse-Transcription Polymerase Chain Reaction to Detect Porcine Reproductive and Respiratory Syndrome Virus on Individual and Pooled Samples from Boars
Authors: Rovira Albert , Clement Travis , Christopher-Hennings Jane , Thompson Bob , Engle Mark , Reicks Darwin , Muñoz-Zanzi Claudia
Published: May 2011

Boar studs are continuously monitored for the presence of porcine reproductive and respiratory syndrome virus (PRRSV) by testing different biological samples by reverse-transcription polymerase chain reaction (RT-PCR). In most cases, samples are run in pools, even though the impact of pooling on the sensitivity of RT-PCR is unknown. The objective of this study was to evaluate the feasibility of using PCR on pooled samples through the estimation of the sensitivity of RT-PCR on different biological samples run individually, in pools of 3 and in pools of 5. Twenty-nine boars were inoculated with a low virulent PRRSV isolate. Serum, blood swab, and semen samples were obtained from each boar every 2 to 3 days for 2 weeks. Each sample was tested by RT-PCR undiluted or diluted 1:3 and 1:5 with negative samples. Eleven of the 29 boars did not appear to get infected from the inoculum, as evidenced by no seroconversion 15 days after inoculation. Data from the other 18 boars showed that serum was the best sample to detect PRRSV during acute infection, with the blood swab sample performing almost as well. Semen samples failed to detect PRRSV infection in most of the cases. Pooling samples at pool sizes of 3 and 5 resulted in a decrease in the sensitivity of RT-PCR. Sensitivity was reduced by 6% and 8%, respectively, when serum or blood swab samples were run in pools of 5. The impact of pooling on the sensitivity of PCR was higher in samples taken during the beginning of the viremic period.
Transmission of Bovine Viral Diarrhea Virus to Adult Goats from Persistently Infected Cattle
Authors: Broaddus Charles C. , Holyoak G. Reed , Dawson Lionel , Step D. L. , Funk Rebecca A. , Kapil Sanjay
Published: May 2011

The transmission of bovine viral diarrhea virus (BVDV) from persistently infected (PI) heifers to adult seronegative goats was examined in this study. Ten seronegative adult goats were exposed to 4 PI heifers. None of the goats developed any clinical signs but all goats seroconverted by 42 days after exposure to the PI cattle. Results indicate that goats are susceptible to BVDV infection when housed with PI cattle.
Detection of the Disease-Associated Isoform of the Prion Protein in Formalin-Fixed Tissues by Western Blot
Authors: Nicholson Eric M. , Kunkle Robert A. , Hamir Amir N. , Lebepe-Mazur Semakaleng , Orcutt Dennis
Published: May 2011

Clinical signs of prion disease are not specific and include a variety of differential diagnoses. Serological tests and nucleic acid-based detection methods are not applicable to prion-disease-agent detection because of the unusual nature of the infectious agent. Prion-disease diagnosis is primarily conducted by means of immunodetection of the infectious agent, typically by at least 2 distinct procedures with immunohisto-chemistry and Western blot being the most informative. These approaches differ in the need for formalin-fixed and frozen or fresh tissue respectively. This work describes a method for the detection of the disease-associated isoform of the prion protein by Western blot using formalin-fixed tissues. The approach requires only minimal modification of existing Western-blot procedures and could readily be incorporated into existing detection schemes for confirmatory purposes when fresh or frozen tissues are unavailable.
Abomasal and Uterine Adenocarcinomas with Ovarian Metastasis in a Captive elk (Cervus Elaphus Nelsoni)
Authors: Duncan Colleen , Powers Jenny , Davis Tracy , Spraker Terry
Published: May 2011

A captive, 20-year-old female elk ( Cervus elaphus nelsoni) euthanized due to progressive lameness and weight loss was presented to Colorado State University Veterinary Diagnostic Laboratory for postmortem examination. Within the uterus there was a poorly demarcated, multilobulated mass measuring 10 cm in diameter. Histologically the tumor was an adenocarcinoma. Histologic examination of the ovaries revealed unilateral metastasis. A focal, 1-cm diameter adenocarcinoma was identified within the abomasum; this tumor was histologically distinct from the neoplasm found in the uterus and ovary. Although this elk had a history of experimental reproductive treatments, including leuprolide, gonadotropin-releasing hormone vaccine, and Brucella abortus vaccination, it was most likely that both tumors represent spontaneous, independent neoplastic transformations and were unrelated.
Sarcocystis sp.-Associated Meningoencephalitis in a Bald Eagle (Haliaeetus Leucocephalus)
Authors: Olson Erik J. , Wünschmann Arno , Dubey J. P.
Published: May 2011

Protozoal meningoencephalitis is uncommon in raptors. An adult female bald eagle ( Haliaeetus leucocephalus) was euthanized after several months of treatment for progressive neurologic signs. The predominant histologic lesion was lymphoplasmacytic and histiocytic meningoencephalitis involving the cerebrum and cerebellum. There was a marked segmental loss of granular cells and Purkinje cells, as well as segmental atrophy of the molecular layer in the cerebellum. Protozoal merozoites and schizonts were observed in the gray matter of the cerebellum. Ultrastructurally, the merozoites were classified as a species of Sarcocystis due to the lack of rhoptries. Immunohistochemistry of the agent revealed a positive reaction for Sarcocystis neurona, while sections were negative for Toxoplasma gondii and Neospora caninum. Sarcocystis sp. infection should be considered as a differential diagnosis in bald eagles with chronic neurologic disease.
Sinonasal Plasmacytoma in a Cat
Authors: Schöniger Sandra , Bridger Nicole , Allenspach Karin , Mantis Panagiotis , Rest Joan , Steeves Elizabeth
Published: May 2011

A 13-year-old female spayed Domestic Shorthair cat presented with a history of right-sided mucopurulent nasal discharge for 18 months. Computed tomography revealed a mass within the right nasal cavity and the right frontal sinus. The animal was euthanized, and a postmortem examination was performed. On macroscopic examination, the right nasal cavity and the right frontal sinus were partially occluded by a soft whitish mass. Microscopically, the mass was composed of well-differentiated plasma cells that were immunopositive for immunoglobulin G and lambda light chains. These findings were consistent with a mature-type sinonasal plasmacytoma. In addition, there was right-sided mucopurulent rhinitis and sinusitis caused by a Pasteurella infection, which probably developed secondary to the sinonasal plasmacytoma. To the authors’ knowledge, this is the first report of a sinonasal plasmacytoma in a cat. The present communication shows that feline sinonasal plasmacytomas should be included in the differential diagnosis for tumors located in the upper respiratory tract of cats.
Antemortem Diagnosis and Characterization of Nasal Intranuclear Coccidiosis in Sulawesi Tortoises (Indotestudo Forsteni)
Authors: Innis Charles J. , Garner Michael M. , Johnson April J. , Wellehan James F. X. , Tabaka Chris , Marschang Rachel E. , Nordhausen Robert W. , Jacobson Elliott R.
Published: May 2011

Intranuclear coccidia and Mycoplasma spp. were identified from the nasal cavity of 5 Sulawesi tortoises (Indotestudo forsteni) affected by chronic rhinosinusitis and oronasal fistulae. This study provides the first antemortem diagnosis of intranuclear coccidiosis in tortoises and the first cytomorphologic descriptions of this disease. Histopathologic and ultrastructural morphology of the intranuclear coccidia were identical to those previously described in tortoises. Nucleic acid sequence data of a 1715 base-pair fragment of the 18S small subunit rRNA gene identify this coccidian as a novel species.
Experimental Infection of Crickets (Gryllus Bimaculatus) with an Invertebrate Iridovirus Isolated from a High-Casqued Chameleon (Chamaeleo Hoehnelii)
Authors: Weinmann Nadine , Papp Tibor , de Matos Antonio Pedro Alves , Teifke Jens P. , Marschang Rachel E.
Published: May 2011

Invertebrate iridoviruses (IIV) have been a regular problem for insect breeders. They have also recently been isolated from various lizard species. An iridovirus isolated from several tissues of a high-casqued chameleon (Chamaeleo hoehnelii) was identified as an IIV on the basis of electron microscopy (EM), sequencing of a portion of the major capsid protein (MCP) gene, and restriction endonuclease analysis of viral DNA. The pathogenicity of this isolate for crickets of the species Gryllus bimaculatus was tested by using 3 experimental infection studies. The mortality rates in the infected crickets ranged between 20% and 35%. The fat bodies of the crickets were examined on cell culture, with a nested PCR targeting the MCP gene, histologically, with in situ hybridization and by EM. Nested PCR was the most sensitive method for detecting IIV in the fat-body samples. Virus was re-isolated from several fat-body samples. In some fat bodies of infected crickets, massive arrays of viruses could be detected by EM. These findings support the hypothesis that IIV from insects are able to infect reptiles.
Value of Skin Punch Biopsies for the Diagnosis of Acute Classical Swine Fever
Authors: Kaden Volker , Lange Elke , Faust Anja , Teifke Jens P.
Published: May 2011

The objective of this study was to determine if skin punch biopsies are appropriate for the diagnosis of classical swine fever. For this purpose, 6 wild boars and 2 domestic pigs were experimentally infected with the highly virulent classical swine fever virus (CSFV) Koslov and 5 domestic pigs with a CSFV field isolate (genotype 2.3 Uelzen) derived from wild boar. Skin biopsy specimens were tested using virus isolation, real-time reverse transcription polymerase chain reaction (rtRT-PCR), and fluorescent antibody test (FAT) on cryosections. Whereas CSFV Koslov was first detected at 4 days postinfection (DPI) by rtRT-PCR and virus isolation, FAT failed to detect CSFV antigen until 9 DPI. In domestic pigs infected with CSFV 2.3 Uelzen, viral RNA and CSFV were detected at 7 DPI. FAT was negative until 11 DPI. CSFV antigen was detected in endothelial cells of the vascular plexus in the upper dermis as shown by confocal laser-scanning microscopy and double labeling with von Willebrand factor. At 18 DPI, CSFV antigen was present diffusely in capillaries and spindle shaped cells of the dermis, multifocally within keratinocytes of the epidermis and in numerous cells of the inner and outer root sheath epithelium, hair bulb, and intravascular leukocytes. The rtRT-PCR proved to be the test with the highest sensitivity followed by virus isolation and FAT. Taken together, this study demonstrates that skin is easy to sample antemortem and is also suitable as postmortem tissue, and suggests that rtRT-PCR of skin should be included for CSF diagnosis in the acute period of disease.
Hepatic Lipidosis and other Test Findings in Two Captive Adult Porcupines (Erethizon Dorsatum) Dying from a “Sudden Death Syndrome”
Authors: Barigye Robert , Schamber Ev , Newell Teresa K. , Dyer Neil W.
Published: May 2011

Routine postmortem examination and histologic evaluation of tissue sections demonstrated hepatic lipidosis (HL) in 2 adult captive porcupines with a history of sudden death. The male porcupine had a markedly enlarged pale liver that microscopically showed large unilocular vacuoles within hepatocellular cytoplasm. The periparturient female had similar but less marked hepatic lesions and an incidental pulmonary mycosis. These findings suggest HL as an important differential of spontaneous death in captive porcupines. It is hypothesized that in addition to the widely documented causes, HL in captive porcupines may be specifically associated with nutritional imbalances caused by the feeding of unsuitable commercial diets. The possible association of the condition with dietary and other factors in captive porcupines needs to be thoroughly investigated.
Bovine Spongiform Encephalopathy in Sweden: An H-Type Variant
Authors: Gavier-Widén Dolores , Nöremark Maria , Langeveld Jan P. M. , Stack Mick , Biacabe Anne-Gaëlle , Vulin Johann , Chaplin Melanie , Richt Jürgen A. , Jacobs Jorg , Acín Cristina , Monleón Eva , Renström Lena , Klingeborn Berndt , Baron Thierry G. M.
Published: May 2011

Bovine spongiform encephalopathy (BSE) had never been detected in Sweden until 2006, when the active surveillance identified a case in a 12-year-old cow. The case was an unusual form, because several molecular features of the protease-resistant prion protein (PrPres) were different from classical BSE. The differences included higher susceptibility for proteinase K, higher molecular weight of the PrPres bands, affinity to the N-terminus-specific antibodies 12B2 and P4, and peculiar banding pattern with antibody SAF84 showing an additional band at the 14 kDa position. The molecular characteristics were in accordance to previous descriptions of H-type BSE. This report shows that a range of Western blot techniques and antibodies can be applied to confirm H-type BSE and further describes that the ratio of the amounts of PrPres#1 and PrPres#2, after deglycosylation, depends on the antibody used during processing. Immunohistochemistry on sections of medulla at the level of the obex applying antibodies with epitopes covering a broad range of the PrP sequence showed accumulation of disease-specific PrP (PrP d ) in the gray matter. Fine punctate deposition in the neuropil was the most predominant type and was more severe in BSE target nuclei. The types of PrP d deposition are described in comparison with classical BSE. PrP-gene sequencing showed 6 copy octarepeat alleles and no abnormalities. It is postulated that the disease had a spontaneous origin, rather than having had been acquired in the BSE epidemic.
Comparison of Serologic Techniques for the Detection of Antibodies against Feline Coronaviruses
Authors: Pratelli Annamaria
Published: May 2011

The seroprevalence of feline Coronavirus (FCoV) antibodies was studied in cats in southern Italy. One hundred twenty sera collected from cats belonging to catteries or community shelters and to households were tested for FCoV type I and II antibodies. The virus neutralization (VN) was performed and compared with indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). Ninety-six sera tested positive for FCoV antibodies by VN and ELISA. Interestingly, ELISA revealed 2 more positive sera than did the VN test and 3 more positive sera than did the IFAT. All results were confirmed by Western blotting. ELISA proved to be more sensitive and detected a seroprevalence of about 82%. Considering the cross-reactivity of FCoV type I and type II, ELISA was able to detect antibodies against both serotypes, allowing the use of the assay as a reference test for sera screening. The high prevalence of antibodies observed indicates that FCoVs are common in southern Italian cat populations.
Comparison of Bacterial Culture, Histopathology, and Immunohistochemistry for the Diagnosis of Johne's Disease in Culled Dairy Cows
Authors: Martinson Shannon A. , Hanna Paul E. , Ikede Basil O. , Lewis Jeff P. , Miller Lisa M. , Keefe Greg P. , McKenna Shawn L. B.
Published: May 2011

Paired samples of formalin-fixed, paraffin-embedded ileum and lymph node from 204 culled dairy cows were investigated for evidence of infection by Mycobacterium avium subsp. paratuberculosis. Of the samples, 151 were from animals that were tissue-culture positive for M. avium subsp. paratuberculosis, and 53 were from animals that were tissue and fecal culture negative. From the culture-positive animals, M. avium subsp. paratuberculosis was isolated from 78 samples of ileum and from 107 samples of lymph node. Ziehl-Neelsen acid-fast and immunoperoxidase stained slides were examined for 15 minutes each. Acid-fast organisms were identified in 7 of 78 (8.97%) and 6 of 106 (5.61%) culture-positive ileum and lymph node samples, respectively. Immunohistochemical (IHC) analysis of the same tissues identified infection in the ileum of 9 of 78 (11.54%) and in the lymph node of 5 of 106 (4.67%) culture-positive tissues. All tissues from culture-negative animals tested negative when using acid-fast and IHC staining. The sensitivity of these 2 tests in detecting M. avium subsp. paratuberculosis in culled dairy cows was not significantly different, and the tests exhibited substantial to almost perfect agreement. Both tests were much less sensitive than bacterial culture, detecting less than 6% of tissues positive compared with culture.
Urine Sampling for Real-Time Polymerase Chain Reaction-Based Diagnosis of Canine Leishmaniasis
Authors: Manna Laura , Reale Stefano , Picillo Esther , Vitale Fabrizio , Gravino Angelo Elio
Published: May 2011

A real-time polymerase chain reaction (PCR) assay was used for quantifying Leishmania infantum DNA in urine samples from naturally infected dogs. Forty-one infected dogs were divided into 3 groups: 22 dogs showing only cutaneous signs (group 1), 12 dogs showing hematuria (group 2), and 7 dogs affected by severe nephropathy (group 3). Groups 2 and 3 dogs showed altered laboratory parameters related to an impairment of renal function. The real-time PCR analysis showed higher levels of Leishmania DNA in the lymph node aspirates from all groups of infected dogs versus those measured in their blood or urine. Interestingly, urine samples from dogs belonging to groups 2 and 3 contained a higher Leishmania DNA load than that detected in their blood. This finding suggests that a real-time PCR analysis of urine from infected dogs could be a useful and noninvasive tool for monitoring the severity of leishmaniasis.
Vitamin E and Selenium Concentrations in Month-Old Beef Calves
Authors: Maas John , Hoar Bruce R. , Myers Daniel M. , Tindall Justin , Puschner Birgit
Published: May 2011

Three groups of beef cow and calf pairs were studied to determine plasma vitamin E and blood selenium (Se) concentrations of calves at 1 month old. Group 1 was managed on irrigated pasture and calves received no Se/vitamin E injections at birth. Group 2 was managed on irrigated pasture, and the calves were injected with Se/vitamin E at birth. Group 3 was managed on dry foothill grasslands, and these cows were supplemented with 56.3 mg vitamin E and 3 mg Se daily, and the calves received a Se/vitamin E injection at birth. The plasma concentration of vitamin E in group 1 and 2 cows (9.5 ± 1.24 and 8.43 ± 1.0 μg/ml. respectively) was significantly higher than that of the group 3 cows (2.28 ± 0.42 μg/ml; P < 0.05). The blood Se concentrations in group 3 cows (169 ± 37 ng/ml) were significantly higher than those in group 1 and 2 cows (36.4 ± 15.9 and 31.1 ± 12.5 ng/ml, respectively; P < 0.05). Calf Se was highly correlated to cow Se ( r = 0.965), and calf vitamin E was moderately correlated to cow vitamin E ( r = 0.605). Calf vitamin E concentrations were consistently lower than cow vitamin E concentrations, and many values would be considered deficient.
Diagnosis of Anatoxin-a Poisoning in Dogs from North America
Authors: Puschner Birgit , Hoff Brent , Tor Elizabeth R.
Published: May 2011

Anatoxin-a, a toxin produced by several genera of blue-green algae, is considered a potent neurotoxin. Ingestion of water contaminated with the toxin results in acute neurological signs and often death. This report describes fatal cases of anatoxin-a ingestion in 6 dogs, with confirmation of anatoxin-a exposure by liquid chromatography/tandem mass spectrometry (LC-MS/MS/MS). In 1 outbreak, 3 dogs developed seizures and died within an hour after swimming in a river in California, while the other outbreak involved 3 dogs that died within 1 hour after swimming in a pond in Ontario. Anatoxin-a poisoning is rarely reported in dogs as a cause of acute neurological signs and death. However, increased occurrences of blue-green algae blooms in North America make this neurotoxin an important consideration in the diagnosis of sudden death associated with environmental water exposure. This brief communication reports on the isolation and detection of anatoxin-a from environmental water sources and the stomach contents of North American dogs dying of acute neurotoxicosis. This demonstrates the first documented cases of anatoxin-a poisoning in dogs in North America and the importance of LC-MS/MS/MS in identifying neurotoxins responsible for sudden death in cases of suspected blue-green algae toxicosis; especially those cases showing no gross or histological lesions.
Streptococcus Dysgalactiae Cellulitis and Toxic Shock-Like Syndrome in a Brown Swiss Cow
Authors: Chénier Sonia , Leclère Mathilde , Messier Serge , Fecteau Gilles
Published: May 2011

An adult Brown Swiss cow was presented to the Large Animal Hospital of the Centre Hospitalier Universitaire Vétérinaire de l'Université de Montréal due to a postpartum downer cow syndrome. The animal had severe and generalized swelling of all 4 limbs and was in shock, as demonstrated by hypotension, dehydration, hypothermia, altered mental status, and abnormal blood parameters. It died rapidly, and necropsy revealed a generalized, severe cellulitis caused by Streptococcus dysgalactiae subsp. dysgalactiae, bronchopneumonia, and lesions of disseminated intravascular coagulation (DIC) in the kidneys. The portal of entry of the bacteria in the subcutaneous tissue was not found, as there was no history of skin trauma or mastitis. The pure growth of large quantities of an invasive Streptococcus species, associated with hypotension, coagulopathy, and renal failure, are supportive of streptococcal toxic shock-like syndrome in a bovine.
Clotting Profiles in Newborn Maltese Kids during the First Week of Life
Authors: Piccione Giuseppe , Bertolucci Cristiano , Giannetto Claudia , Giudice Elisabetta
Published: May 2011

The neonatal period is probably the only time when a higher incidence of spontaneous thromboembolic complications may occur in the otherwise normal, healthy individual. This study was designed to determine the postnatal development of the kid coagulation system. Ten clinically healthy and full-term-born Maltese kid goats (5 males and 5 females) were used. In each kid, during the first week after birth, the prothrombin time (FT), the activated partial thromboplastin time (aPTT), the thrombin time (TT), and fibrinogen were assessed. Analysis of variance showed a highly significant effect of age on PT, TT, and fibrinogen. Our results of this study indicate that the clotting mechanism in kids is influenced by growth. This investigation contributes to the knowledge of clotting adaptations in kids during the first 7 days of life and provides useful information for the diagnosis and treatment of some neonatal diseases.
Fatal Pulmonary Hemorrhage Associated with RTX Toxin-Producing Actinobacillus Equuli Subspecies Haemolyticus Infection in an Adult Horse
Authors: Pusterla Nicola , Jones Megan E. B. , Mohr F. Charles , Higgins Jamie K. , Mapes Samantha , Jang Spencer S. , Samitz Eileen M. , Byrne Barbara A.
Published: May 2011

A case of fatal pulmonary hemorrhage in a 6–year-old American Paint mare with a 2–week history of intermittent coughing, fever, and epistaxis is described. Significant macroscopic abnormalities at postmortem examination were restricted to the respiratory system, and microscopically, severe pulmonary hemorrhage with suppurative bronchopneumonia was found. Actinobacillus equuli subsp. haemolyticus was cultured from a transtracheal wash performed antemortem as well as from the lungs at necropsy. The presence of airway-associated hemorrhage in conjunction with bacterial bronchopneumonia suggested endothelial damage caused by a locally elaborated bacterial toxin, possibly produced by the A. equuli strain isolated from the lungs. The objective of this report was to indirectly document the presence of hemolysin repeat in structural toxin (RTX) in the lungs of the reported mare. A real-time polymerase chain reaction (PCR) assay targeting the recently described aqx gene of A. equuli subsp. haemolyticus was established and validated. Transcriptional activity of the aqx gene was used as a surrogate method to document toxin production. Real-time PCR analysis of the transtracheal fluid and lung tissue of the affected mare confirmed the presence and the transcriptional activity of the aqx gene at the genomic (gDNA) and complementary DNA (cDNA) levels, respectively. The presence of pneumonia associated with hemorrhagic pulmonary fluid and the culture of large numbers of hemolytic A. equuli should prompt the clinician to consider endothelial damage caused by bacterial toxins.