Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets

AbstractA novel coronavirus, designated as ferret enteric coronavirus (FECV), was identified in feces of domestic ferrets clinically diagnosed with epizootic catarrhal enteritis (ECE). Initially, partial sequences of the polymerase, spike, membrane protein, and nucleocapsid genes were generated using coronavirus consensus PCR assays. Subsequently, the complete sequences of the nucleocapsid gene and the last two open reading frames at the 3′ terminus of the FECV genome were obtained. Phylogenetic analyses based on predicted partial amino acid sequences of the polymerase, spike, and membrane proteins, and full sequence of the nucleocapsid protein showed that FECV is genetically most closely related to group 1 coronaviruses. FECV is more similar to feline coronavirus, porcine transmissible gastroenteritis virus, and canine coronavirus than to porcine epidemic diarrhea virus and human coronavirus 229E. Molecular data presented in this study provide the first genetic evidence for a new coronavirus associated with clinical cases of ECE

Mutations in Exons 8 and 11 of c-kit Gene in Canine Subcutaneous Mast Cell Tumors and Their Association with Cell Proliferation

The prognostic significance of internal tandem duplication (ITD) mutations in exons 8 and 11 of c-kit has been well-described for canine cutaneous mast cell tumors (MCTs), but c-kit mutations have rarely been reported in subcutaneous MCTs. The objective of this study was to identify the prevalence of ITD mutations in exons 8 and 11 of c-kit in canine subcutaneous MCTs and to investigate its association with histologic grade, KIT pattern, and proliferation markers. ITD mutations in exons 8 and 11 of c-kit, mitotic count, Ki67 index, AgNOR number, Ki67xAgNOR score, KIT pattern, and histologic grade (two-tier system) were retrospectively recorded for 216 dogs with subcutaneous MCTs. ITD mutations in exons 8 and 11 of c-kit were detected in 23 (10.6%) and 12 (5.56%) subcutaneous MCTs, respectively. Exon 11 mutations were significantly associated with Kiupel high grade (p < 0.001) and increased mitotic count (p < 0.001) compared to subcutaneous MCTs with no mutations in exons 8 or 11 (p = 0.002) or subcutaneous MCTs with a mutation in exon 8 (p = 0.001). There was no significant association of either c-kit mutation with KIT patterns or proliferation activity. This study identified a higher prevalence of ITD mutations in exons 8 and 11 of c-kit in subcutaneous MCTs than previously reported. Like their cutaneous counterpart, subcutaneous MCTs with exon 11 mutations were more likely to be histologically high grade and have a higher mitotic count, whereas such associations were not observed in subcutaneous MCTs with exon 8 mutations

Patterns of Lymphocytic Infiltrates Can Differentiate Feline Hepatic Lymphoma from Lymphocytic Portal Hepatitis

Hepatic lymphoma is poorly characterized in cats and differentiating between inflammation and lymphomas is often difficult. The diagnosis of hepatic lymphoma in humans relies on recognition of specific patterns of lymphocytic infiltrates and clonality testing of antigen receptors. Herein, we defined similar patterns of lymphocytic infiltrates in hepatic biopsies of cats and correlated them with clonality to determine which patterns are predictive of lymphoma. A retrospective study was performed on surgical biopsies from 44 cats. The immunophenotype was characterized using CD3 and CD20 on all 44 samples. All 44 samples were tested using PCR for T-cell receptor gamma-gene rearrangements. PCR for immunoglobulin heavy chain gene rearrangements was performed on 24 of these cats. Four patterns of lymphocytic infiltrates were characterized: (1) tightly periportal, (2) periportal and centrilobular, (3) nodular, and (4) periportal with sinusoidal extension. Other histomorphologic features (fibrosis, biliary hyperplasia, bile ductopenia, bile duct targeting, hepatic hematopoiesis, lipogranulomas, lymphonodular aggregates, other inflammatory cells) were also evaluated. The sensitivity and specificity of the lymphocytic patterns to diagnose lymphomas were determined using Bayesian Hui–Walter analysis (BLCM) against clonality results. Lymphocytic patterns 2, 3, and 4 accurately diagnosed hepatic lymphomas with a sensitivity and specificity of 82% (CI 95%: 0.65, 0.96) and 77% (CI 95%: 0.54, 1.00), respectively. None of the other microscopic features evaluated were predictive of a lymphoma or inflammation. Our study identified specific patterns of lymphocytic infiltration that differentiate feline hepatic lymphoma from inflammation while other histologic features were not associated with an accurate diagnosis

"Big Data" im Physikunterricht mit Handy und Tablet-PC

Moderne Smartphones und Tablet-PCs haben eine Reihe von fest eingebauten Sensoren, die zur intuitiven Bedienbarkeit der Geräte erheblich beitragen. Es gibt inzwischen eine Vielzahl von Beiträgen, die zeigen, wie die Sensordaten über unterschiedliche Programme ("Apps") auch für den Physikunterricht genutzt werden können.Hier wird ein System vorgestellt, mit dem die Sensoren eines Android-Gerätes direkt ausgelesen werden. Durch hohe Abtastrate stehen verhältnismäßig viele Rohdaten zur Verfügung, die dann auf verschiedene Fragestellungen hin ausgewertet werden können. Durch die Möglichkeit, die Daten auf einen Server zu laden stehen diese prinzipiell auch allen Lernenden zu Verfügung.Die Verfügbarkeit und Nutzung eines populären Gerätes, die Qualität der Sensoren und die einfache lokale oder dezentrale Speichermöglichkeit der Daten ermöglichen so die exemplarische Thematisierung eines Auswertungs- und Interpretationsprozesses für (relativ) große Datenmengen

Atypical Mitotic Figures Are Prognostically Meaningful for Canine Cutaneous Mast Cell Tumors

Cell division through mitosis (microscopically visible as mitotic figures, MFs) is a highly regulated process. However, neoplastic cells may exhibit errors in chromosome segregation (microscopically visible as atypical mitotic figures, AMFs) resulting in aberrant chromosome structures. AMFs have been shown to be of prognostic relevance for some neoplasms in humans but not in animals. In this study, the prognostic relevance of AMFs was evaluated for canine cutaneous mast cell tumors (ccMCT). Histological examination was conducted by one pathologist in whole slide images of 96 cases of ccMCT with a known survival time. Tumor-related death occurred in 11/18 high-grade and 2/78 low-grade cases (2011 two-tier system). The area under the curve (AUC) was 0.859 for the AMF count and 0.880 for the AMF to MF ratio with regard to tumor-related mortality. In comparison, the AUC for the mitotic count was 0.885. Based on our data, a prognostically meaningful threshold of ≥3 per 2.37 mm2 for the AMF count (sensitivity: 76.9%, specificity: 98.8%) and >7.5% for the AMF:MF ratio (sensitivity: 76.9%, specificity: 100%) is suggested. While the mitotic count of ≥ 6 resulted in six false positive cases, these could be eliminated when combined with the AMF to MF ratio. In conclusion, the results of this study suggests that AMF enumeration is a prognostically valuable test, particularly due to its high specificity with regard to tumor-related mortality. Additional validation and reproducibility studies are needed to further evaluate AMFs as a prognostic criterion for ccMCT and other tumor types

Gastrinoma and Zollinger-Ellison Syndrome in Canids: A Literature Review and a Case in a Mexican Gray Wolf

Gastrinoma, an infrequent diagnosis in middle-aged dogs, occurs with nonspecific gastrointestinal morbidity. Laboratory tests can yield a presumptive diagnosis, but definitive diagnosis depends on histopathology and immunohistochemistry. We describe a malignant pancreatic gastrinoma with lymph node metastases and corresponding Zollinger–Ellison syndrome in a Mexican gray wolf (Canis lupus baileyi) and review this endocrine neoplasm in domestic dogs. A 12-y-old, captive, male Mexican gray wolf developed inappetence and weight loss. Abdominal ultrasonography revealed a thickened duodenum and peritoneal effusion. Two duodenal perforations were noted on exploratory celiotomy and were repaired. Persisting clinical signs led to a second celiotomy that revealed a mesenteric mass, which was diagnosed histologically as a neuroendocrine carcinoma. During the following 16 mo, the wolf received a combination of H2-receptor antagonists, proton-pump inhibitors, gastroprotectants, and anti-emetics, but had recurrent episodes of anorexia, nausea, acid reflux, and remained underweight. Worsening clinical signs and weakness prompted euthanasia. The antemortem serum gastrin concentration of 414 ng/L (reference interval: 10–40 ng/L) corroborated hypergastrinemia. Autopsy revealed a mass expanding the right pancreatic limb; 3 parapancreatic mesenteric masses; duodenal ulcers; focal duodenal perforation with septic fibrinosuppurative peritonitis; chronic-active ulcerative esophagitis; and poor body condition. The pancreatic mass was diagnosed histologically as a neuroendocrine carcinoma and the parapancreatic masses as lymph node metastases. Immunohistochemistry of the pancreatic mass was positive for gastrin and negative for glucagon, insulin, pancreatic polypeptide, serotonin, somatostatin, and vasoactive intestinal peptide

"Big Data" im Physikunterricht mit Handy und Tablet-PC

Moderne Smartphones und Tablet-PCs haben eine Reihe von fest eingebauten Sensoren, die zur intuitiven Bedienbarkeit der Geräte erheblich beitragen. Es gibt inzwischen eine Vielzahl von Beiträgen, die zeigen, wie die Sensordaten über unterschiedliche Programme ("Apps") auch für den Physikunterricht genutzt werden können.Hier wird ein System vorgestellt, mit dem die Sensoren eines Android-Gerätes direkt ausgelesen werden. Durch hohe Abtastrate stehen verhältnismäßig viele Rohdaten zur Verfügung, die dann auf verschiedene Fragestellungen hin ausgewertet werden können. Durch die Möglichkeit, die Daten auf einen Server zu laden stehen diese prinzipiell auch allen Lernenden zu Verfügung.Die Verfügbarkeit und Nutzung eines populären Gerätes, die Qualität der Sensoren und die einfache lokale oder dezentrale Speichermöglichkeit der Daten ermöglichen so die exemplarische Thematisierung eines Auswertungs- und Interpretationsprozesses für (relativ) große Datenmengen

Prevalence of canid herpesvirus-1 infection in stillborn and dead neonatal puppies in Denmark

BACKGROUND: Canid herpesvirus-1 (CaHV-1) infection in puppies less than three weeks of age is often reported to be associated with a lethal generalized necrotizing inflammation and since the discovery of the virus in 1965 several reports of neonatal infections have been published. However, the significance of CaHV-1 for peri- and neonatal mortality in puppies remains unclear. Therefore, we examined stillborn and dead neonatal puppies in Denmark to determine the prevalence of infection and further to correlate infection levels with necropsy findings to assess the possible significance of the infection. RESULTS: From a cross-sectional study of 57 dead puppies, 22.8% (n = 13) were confirmed positive for CaHV-1 by real-time polymerase chain reaction (PCR) of tissue pools of lung/liver and/or spleen/kidney. Specimens from PCR positive cases were further investigated by histology and in situ hybridization (ISH). High levels of CaHV-1 DNA were present in only one case in which lesions and ISH staining consistent with CaHV-1 infection were found as well. CaHV-1 concentrations in the other cases were low and a range of lesions not consistent with CaHV-1 were found. Similar, ISH staining was mostly negative in these except for one case with a few positive cells. CONCLUSION: CaHV-1 infection in stillborn and dead neonatal puppies in Denmark seems to be common, but the direct significance for puppy mortality remains unclear as only one of 13 PCR positive puppies (7.7%) had pathognomonic lesions. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13028-014-0092-9) contains supplementary material, which is available to authorized users

Auricular cutaneous T-cell lymphoma, subtype mycosis fungoides and otitis externa/media in a domestic rat (Rattus norvegicus forma domestica)

An adult female, entire domestic rat (Rattus norvegicus forma domestica) presented with swollen auricles and incoordination. The rat was diagnosed with bilateral otitis externa. Radiographs of the skull revealed bilateral otitis externa/media. Bacterial culture of a swap from the external ear canals identified large numbers of Pasteurella pneumotropica as well as fewer Streptococcus merionis, Bacteroides fragilis and Fusobacterium spp. Microscopic examination of a deep skin scraping and a core needle biopsy of the pinnae diagnosed lymphoma. The animal was euthanased after being treated unsuccessfully symptomatically for 10 days. Postmortem examination revealed a cutaneous T-cell lymphoma, mycosis fungoides, associated with bilateral otitis externa/media and metastases into several lymph nodes, thymus and lungs. Cutaneous T-cell lymphoma has not been reported in pet rats to date. Lymphoma should be considered as a differential diagnosis in rats presented with swelling of the pinnae