Occurrence and Molecular Typing of Giardia psittaci in Parakeets in Germany-A Case Study.

A grey-hooded parakeet (Psilopsiagon aymara) and two budgerigars (Melopsittacus undulatus) from different owners presented with decreased activity, vomitus, and diarrhea. A microscopic examination of feces showed trophozoites of the protozoan flagellate Giardia. A commercial immunochromatographic dipstick test for Giardia sp. antigens confirmed the infection. These findings were assured by PCR of the small subunit ribosomal RNA (SSU rRNA) gene and coproantigen ELISA. Sequencing of PCR products of the SSU rRNA (292 bp) and ß-giardin genes (511 bp) identified Giardia psittaci as the species involved. Therefore, our results show that a GSA 65-based coproantigen ELISA, which was established for diagnosis of Giardia duodenalis is applicable for the detection of G. psittaci. A treatment with ronidazole was started. Additionally, fecal examination and dissection of the dead birds revealed coinfection with the fungal pathogen Macrorhabdus ornithogaster. One budgerigar survived and repeatedly tested negative after treatment with ronidazole. The described cases indicate that a single infection with G. psittaci has a good prognosis, whereas the prognosis is poor when coinfections occur, especially with M. ornithogaster.

Reporte de caso- Presentación y tipificación molecular de Giardia psittaci en periquitos en Alemania: Un estudio de caso. Un periquito catita aimará (Psilopsiagon aymara) y dos periquitos australianos (Melopsittacus undulatus) de diferentes propietarios presentaron actividad disminuida, vómito y diarrea. El examen microscópico de las heces mostró trofozoitos del protozoo flagelado Giardia. Una prueba de tira reactiva inmunocromatográfica comercial para antígenos de Giardia sp. confirmó la infección. Estos resultados fueron confirmados por PCR para el gene de ARN de la subunidad pequeña ribosomal (SSU rRNA) y por ELISA de coproantígeno. La secuenciación de los productos de PCR del ARNr de SSU (292 pb) y los genes de ß-giardina (511 pb) identificaron a Giardia psittaci como la especie involucrada. Por lo tanto, estos resultados muestran que el método de ELISA de coproantígeno basado en GSA 65, que se estableció para el diagnóstico de Giardia duodenalis, es aplicable para la detección de G. psittaci. Se inició un tratamiento con ronidazol. Además, el examen fecal y la disección de las aves muertas revelaron coinfección con el patógeno fúngico Macrorhabdus ornithogaster. Un periquito australiano sobrevivió y dio negativo repetidamente después del tratamiento con ronidazol. Los casos descritos indican que la infección única con G. psittaci tiene un buen pronóstico, mientras que el pronóstico es malo cuando ocurren coinfecciones, especialmente con M. ornithogaster. Abbreviations: GSA = Giardia-specific antigen; OD = optical density; rRNA = ribosomal ribonucleic acid; SSU = small subunit.

Neurotrophic effects of GM1 ganglioside, NGF, and FGF2 on canine dorsal root ganglia neurons in vitro.

Dogs share many chronic morbidities with humans and thus represent a powerful model for translational research. In comparison to rodents, the canine ganglioside metabolism more closely resembles the human one. Gangliosides are components of the cell plasma membrane playing a role in neuronal development, intercellular communication and cellular differentiation. The present in vitro study aimed to characterize structural and functional changes induced by GM1 ganglioside (GM1) in canine dorsal root ganglia (DRG) neurons and interactions of GM1 with nerve growth factor (NGF) and fibroblast growth factor (FGF2) using immunofluorescence for several cellular proteins including neurofilaments, synaptophysin, and cleaved caspase 3, transmission electron microscopy, and electrophysiology. GM1 supplementation resulted in increased neurite outgrowth and neuronal survival. This was also observed in DRG neurons challenged with hypoxia mimicking neurodegenerative conditions due to disruptions of energy homeostasis. Immunofluorescence indicated an impact of GM1 on neurofilament phosphorylation, axonal transport, and synaptogenesis. An increased number of multivesicular bodies in GM1 treated neurons suggested metabolic changes. Electrophysiological changes induced by GM1 indicated an increased neuronal excitability. Summarized, GM1 has neurotrophic and neuroprotective effects on canine DRG neurons and induces functional changes. However, further studies are needed to clarify the therapeutic value of gangliosides in neurodegenerative diseases.

Canine dorsal root ganglia satellite glial cells represent an exceptional cell population with astrocytic and oligodendrocytic properties.

Dogs can be used as a translational animal model to close the gap between basic discoveries in rodents and clinical trials in humans. The present study compared the species-specific properties of satellite glial cells (SGCs) of canine and murine dorsal root ganglia (DRG) in situ and in vitro using light microscopy, electron microscopy, and immunostainings. The in situ expression of CNPase, GFAP, and glutamine synthetase (GS) has also been investigated in simian SGCs. In situ, most canine SGCs (>80%) expressed the neural progenitor cell markers nestin and Sox2. CNPase and GFAP were found in most canine and simian but not murine SGCs. GS was detected in 94% of simian and 71% of murine SGCs, whereas only 44% of canine SGCs expressed GS. In vitro, most canine (>84%) and murine (>96%) SGCs expressed CNPase, whereas GFAP expression was differentially affected by culture conditions and varied between 10% and 40%. However, GFAP expression was induced by bone morphogenetic protein 4 in SGCs of both species. Interestingly, canine SGCs also stimulated neurite formation of DRG neurons. These findings indicate that SGCs represent an exceptional, intermediate glial cell population with phenotypical characteristics of oligodendrocytes and astrocytes and might possess intrinsic regenerative capabilities in vivo.

Skeletal muscle hypoplasia represents the only significant lesion in peripheral organs of ruminants infected with Schmallenberg virus during gestation.

Schmallenberg virus (SBV), an arbovirus within the family Bunyaviridae, represents a ruminant pathogen that has caused epidemic abortion and birth of malformed or stillborn animals in many European countries since August 2011. Histological and immunohistochemical analysis of peripheral tissues of SBV-infected animals, including lymphoid tissues, endocrine organs and tissues of the gastrointestinal, urogenital and respiratory system, were analyzed in order to elucidate the occurrence of SBV-associated changes and the presence of viral antigens and RNA. Twenty calves and 12 lambs as well as age-matched controls were included in this study. Significant muscular hypoplasia with fatty replacement was noted in affected calves and lambs. In addition, hepatocellular degeneration with lymphohistiocytic inflammation, interstitial fibrosis and biliary hyperplasia was detected in calves. All animals lacked SBV-positive cells in the peripheral organs. These observations resemble those found in Akabane virus- and Cache Valley virus-infected animals and support the occurrence of few residual lesions in peripheral organs following SBV infection.

Lack of schmallenberg virus in ruminant brain tissues archived from 1961 to 2010 in Germany.

Schmallenberg virus (SBV) is an orthobunyavirus of the family Bunyaviridae that is associated with stillbirth and malformations in ruminants. The infection has been identified in many European countries since August 2011. The present study investigated retrospectively the occurrence of SBV infection in ruminants using immunohistochemistry and in-situ hybridization in brain tissues archived between 1961 and 2010 (112 cattle, 57 sheep, 16 goats and 27 wild ruminants). Eighty-five animals with inflammatory brain lesions and 47 animals with malformations were included. Due to the lack of SBV protein and RNA detection, SBV appears to have been introduced recently into Northern parts of Europe from tropical or subtropical regions.

Theiler's murine encephalomyelitis virus induced phenotype switch of microglia in vitro.

The present in vitro study aimed to define the involvement of astrocytes and microglia in the initial inflammatory response of Theiler's murine encephalomyelitis (TME), a virus-induced mouse model of multiple sclerosis, and whether intralesional microglia exert pro- (M1) or anti-inflammatory (M2) effects following TME virus (TMEV) infection. Therefore astrocytes and microglia were purified from neonatal murine brains and inoculated either with TMEV or mock-solution. Gene expression of IL-1, IL-2, IL-10, IL-12, TNF, TNF receptors (TNFR1, TNFR2), TGFß1, IFNγ and transcription factors NF-κB (p50, p65) and AP-1 (c-jun, c-fos) were quantified using RT-qPCR at 6, 48, and 240h post infection (hpi). In addition, IL-1, IL-10, IL-12, TNF and TGFß1 mRNA transcripts were investigated at 168 hpi in TMEV- and mock-infected SJL/J mice. Overall in vitro astrocytes showed a significant higher amount of viral RNA compared to microglia. In addition, TMEV-infected astrocytes showed higher numbers of IL-1, IL-12 and TNF transcripts at 48 hpi. In microglia high IL-10 and low IL-12 mRNA levels were detected at 48 hpi, while the opposite was the case at 240 hpi. In addition, TNF mRNA was increased in microglia at 240 hpi. In addition, the observed up-regulation of IL-1, IL-12 and IL-10 in the early phase of TME in vivo substantiates the relevance of these cytokines during the disease induction. Summarized data indicate that TMEV infection of microglia induces a switch from the anti-inflammatory (M2) during the early phase to the pro-inflammatory (M1) phenotype in the later phase of the infection. The simultaneous expression of TNF and its receptors by both cell types might generate autocrine feedback loops possibly associated with pro-inflammatory actions of astrocytes via TNFR1.

Perosomus elumbis, cerebral aplasia, and spina bifida in an aborted thoroughbred foal.

Perosomus elumbis represents a rare congenital anomaly characterized by aplasia of the lumbosacral spinal cord and vertebrae. This anomaly is often associated with arthrogryposis and malformations of the urogenital and intestinal tract. This report describes the first case of perosomus elumbis in an aborted Thoroughbred foal associated with cerebral aplasia with meningocele, cranioschisis, spina bifida, a fused urogenital and intestinal tracts lined by a cutaneous mucosa without uterine glands, atresia ani, and arthrogryposis of the hind legs. Immunohistochemistry detected no abnormalities in the GFAP, S-100, vimentin, NeuN, doublecortin, and neurofilament expression pattern in the remnants of the brainstem. Pathogenic bacteria or equine herpesviruses one and four were not found using a microbiological investigation or polymerase chain reaction, respectively. The observed malformations may be induced by a severe developmental disturbance during the formation of the neural tube with secondary distortion of the surrounding mesoderm. Although the aetiology of this dysorganogenesis remains undetermined, inherited chromosomal mutations, an intake of different xenobiotics, and/or a disturbed metabolism of the mare may have caused the defect.

Two new cases of polysomy 13 in canine prostate cancer.

Besides man, the dog is the only known mammalian species that spontaneously develops carcinomas of the prostate with considerable frequency. For this reason, the dog is considered to be the only useful animal model for spontaneously occurring prostate malignancies in man. Cytogenetic investigations of human prostate cancers have revealed the frequent occurrence of trisomies 7, 8, and 17. Chromosome analyses of canine prostate carcinomas are rare. In this report we present 2 cases of canine prostate cancer showing a clonal polysomy 13 along with complex karyotype changes. Along with a previous report demonstrating polysomy 13 as the only karyotype deviation in a canine prostate cancer the present report supports the hypothesis that in canine prostate cancer, polysomy 13 is a recurrent cytogenetic aberration linked to the development of the disease. As human chromosomes (HSA) 8q and 4q and the canine chromosome (CFA) 13 share high homology, these results suggest that a conserved area on these chromosomes is involved in tumorigenesis in both species.

In vitro characterization and preferential infection by canine distemper virus of glial precursors with Schwann cell characteristics from adult canine brain.

AIMS: Canine distemper virus (CDV)-induced demyelinating leukoencephalomyelitis is a naturally occurring model for multiple sclerosis. The aim of this study was to establish primary glial cell cultures from adult canine brain for the analysis of CDV spread and cell tropism. METHODS: Cultures were inoculated with the CDV-R252 and a CDV-Onderstepoort strain expressing the green fluorescent protein (CDV-OndeGFP). CDV antigen expression was studied using cell type-specific antibodies at different days post infection. Glial cells expressing p75(NTR) were purified using antibody-based techniques and characterized with regard to antigen expression and proliferation. RESULTS: Three weeks after seeding, cultures contained spindle-shaped cells expressing p75(NTR), oligodendrocytic cells, astrocytes, microglia and fibroblasts. Both CDV strains induced a mild to moderate cytopathic effect that consisted of single necrotic and few syncytial giant cells, but displayed in part a differential cell tropism. Whereas CDV-OndeGFP expression in microglia and astrocytes did not exceed 1% and 50%, respectively, CDV-R252 infected 100% and 80% of both cell types, respectively. The cells most early infected by both CDV strains expressed p75(NTR) and may correlate to cells previously identified as aldynoglia. Treatment of p75(NTR+) cells with Schwann cell mitogens and serum deprivation increased proliferation and A2B5 expression, respectively, indicating common properties compared with Schwann cells and oligodendrocyte precursors. CONCLUSIONS: Infection of adult canine astrocytes and microglia revealed CDV strain-specific cell tropism. Moreover, this is the first identification of a glial cell type with Schwann cell-like properties in adult canine brain and, more importantly, these cells displayed a high susceptibility to CDV infection.

Unusual type of reactive astrocytes in the feline central nervous system.

An unusual type of hypertrophic astrocytes termed plump polygonal astrocytes (PPA) has been observed in the feline central nervous system which was characterized in a first preliminary study of 17 cats. These cells presented an oval to polygonal shape, measured about 20 microm in diameter, and displayed short, barely detectable processes. The condensed, hyperchromatic, eccentric nucleus was surrounded by an abundant, homogenous, eosinophilic cytoplasm. These GFAP-and S-100-positive and vimentin-negative cells were predominantly found in brains showing status spongiosus and less frequently in association with inflammation and in brains lacking histological lesions. They were mainly detected in white matter areas of the hindbrain. In addition, these cells were also observed in the dentate hilar region adjacent to degenerated neurons and a small amount of PPA were positive for caspase-3. It remains to be determined if PPA represent a specific type of reactive astrocytes and whether they are characteristic for a specific cause or response in the feline brain.

[Familial occurrence of a congenital defect of the keratinisation of the skin in German Angus calves]. / Familiäres Auftreten einer angeborenen Verhornungsstörung der Haut bei zwei Kälbern der Rasse Deutsche Angus.

Congenital dysfunction of the keratinisation of the epithelium was diagnosed in two female German Angus calves born on the same farm. The relationship coefficient between the two affected Angus calves was 34.38%. The clinical findings were similar to ichthyosis congenita as the alterations of the skin were present at birth and the levels of zinc in the blood were not decreased. However, parakeratosis could not be completely excluded as skin alterations were partly parakeratotic. On account of the close relationship between the two affected calves a genetic cause is likely for the present cases.

A case of interface perianal dermatitis in a dog: is this an unusual manifestation of lupus erythematosus?

Discoid lupus erythematosus (DLE) is a well-known autoimmune disorder described in dogs and humans. In dogs, DLE is considered the second most common immune-mediated dermatitis and is usually localized to the nasal planum. DLE does not evolve to generalized disease, however lesions may spread to the bridge of the nose and less commonly may extend to periocular region, pinnae, distal limbs, and mucocutaneous junctions (lips, oral cavity, and genital region). A 4-year-old male Bavarian Mountain Scenthound developed a chronic, erosive, cutaneous lesion located exclusively in the perianal region without facial skin involvement. Clinical signs included erythema, depigmentation, severe alopecia, crusting, and ulceration. Histologically, the hallmarks of the changes were an interface dermatitis consisting of plasma cells, lymphocytes, neutrophils, and macrophages, hydropic degeneration of basal cells, few apoptotic cells in the basal layer, pigmentary incontinence, and a focal thickening of the basement membrane, which was characterized by linear deposition of IgG. Despite the unusual localization the lesion was diagnosed as DLE based on the characteristic histologic and immunohistologic features. Following diagnosis, corticosteroid therapy resulted in a complete resolution of perianal lesions.