In Vitro Host-Cell Susceptibility to Usutu Virus

We investigated the susceptibility to Usutu virus (Flavivirus) of 13 permanent cell lines, 3 primary cell cultures, and chicken embryos. Vero, PK-15, and goose embryo fibroblast cells developed cytopathic effects; however, viral multiplication was detected in all mammalian cell types by immunohistochemical tests. Chicken embryo fibroblast cells and chicken embryos were resistant

Age-related presence of selected viral and bacterial pathogens in paraffin-embedded lung samples of dogs with pneumonia

The aim of this retrospective study was to detect selected pathogens in pneumonic lung tissue of dogs of different age groups by immunohistochemistry (IHC), in situ hybridisation (ISH) or polymerase chain reaction (PCR) in order to get information about their involvement in pneumonia formation. In archived formalin-fixed and paraffin wax-embedded lung samples from 68 cases with the clinical and histologic diagnosis of pneumonia the histological pattern of pneumonia was re-evaluated and the samples were further investigated for the following infectious agents: canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine respiratory coronavirus (CRCoV), Bordetella (B.) bronchiseptica, Pasteurella (P.) multocida, Mycoplasma spp., and Pneumocystis spp. In 47.1% of the samples at least one of the featured respiratory pathogens was detected. In 31.3% of these positive samples more than one pathogen could be found. The correct detection of CDV had been achieved in ten out of eleven positive cases (90.9%) upon initial investigation, but the presence of bacterial pathogens, like B. bronchiseptica (10 cases) and P. multocida (17 cases) had been missed in all but one case. While CDV and CRCoV infections were exclusively found in dogs younger than one year, the vast majority of infections with P. multocida and B. bronchiseptica were both common either in dogs younger than 4 months or older than one year. Thus, this retrospective approach yielded valuable data on the presence, absence and prevalence of certain respiratory pathogens in dogs with pneumonia

Coinfection with Entamoeba polecki and Brachyspira hyodysenteriae in a pig with severe diarrhea

Enteric disease in pigs is usually of multifactorial etiology, including infectious and non-infectious factors. In many cases of endemic diarrhea in weaner-to-finisher pigs, the combination of 2 or more microorganisms leads to aggravation of intestinal lesions and, consequently, clinical signs. We autopsied a 4-mo-old fattening pig with diarrhea and diagnosed severe fibrinonecrotizing typhlocolitis. Numerous spiral-shaped bacteria and amoeba-like PAS-positive protozoa were observed in the cecal and colonic mucosa and submucosa. Brachyspira hyodysenteriae was detected by PCR from colonic content. By in situ hybridization, large numbers of Entamoeba polecki were found within the lamina propria and submucosa; moderate numbers of Blastocystis sp. and scattered trichomonads were present in intestinal content. In addition, Entamoeba polecki, Balantidium spp., Blastocystis sp., and Trichomonas sp. were also detected by PCR.info:eu-repo/semantics/acceptedVersio

Identification of a putatively novel trichomonad species in the intestine of a common quail (Coturnix coturnix)

AbstractA common quail (Coturnix coturnix) from a private keeping died unexpectedly and showed a moderate lymphocytic infiltration of the colonic mucosa associated with numerous protozoa-like objects at the pathological examination. These organisms were further identified using chromogenic in situ hybridization (ISH) and gene sequencing. ISH was performed on paraffin embedded tissue sections and produced a positive signal using a probe specific for the 18S ribosomal RNA (rRNA) gene of the order Trichomonadida, but remained negative with probes specific for the 18S rRNA gene of the common bird parasites Histomonas meleagridis, Tetratrichomonas gallinarum or Trichomonas gallinae. The trichomonads were found on the mucosal surface, inside the crypts and also immigrating into the lamina propria mucosae. DNA was extracted from the paraffin embedded tissue and the entire 18S rRNA gene, ITS-1 region, 5.8S rRNA gene, ITS-2 region and a part of the 28S rRNA gene were sequenced using primer walking. The acquired sequence showed 95% homology with Tritrichomonas foetus, a trichomonad never described in birds. A phylogenetic analysis of a part of the 18S rRNA gene or of the ITS-1, 5.8S and ITS-2 region clearly placed this nucleotide sequence within the family of Tritrichomonadidae. Therefore, the authors propose the detection of a putative new Tritrichomonas sp. in the intestine of a common quail

Lineage 1 and 2 Strains of Encephalitic West Nile Virus, Central Europe

An encephalitic lineage 2 strain of WNV is observed for the first time outside Africa

Immunohistochemical characterization of type II pneumocyte proliferation after PRRSV (Type I) challenge

The study aimed to histologically and immunohistochemically characterize lung lesions after a challenge with a recently isolated PRRSV field strain in growing pigs 10 and 21 days post infection (DPI). In the first phase of the study lung lesions were evaluated microscopically on routine haematoxylin and eosin stained slides. The evaluation was performed as a blinded analysis and the lesions were scored based on the following criteria: (1) pneumocyte hypertrophy and hyperplasia, (2) septal mononuclear infiltration, (3) intraalveolar necrotic debris, (4) intraalveolar inflammatory cell accumulation and (5) perivascular inflammatory cell accumulation. For further characterization of the lung lesions, immunohistochemical stainings were performed using anti-cytokeratin, anti-Ki67, anti-TTF-1 (Thyroid Transcription Factor-1), anti-myeloid receptor (MAC387), and anti-PRRSV antibodies to identify alveolar epithelial cells, proliferating cells, type II pneumocytes, macrophages, and PRRSV antigen, respectively. The evaluation of the immunohistochemical stainings revealed that humanized anti TTF-1 antibodies can successfully identify type II pneumocytes in porcine lung tissue. Marked proliferation of these cells was confirmed by a significant (p<0.05) increase of TTF-1 positive cells in acute cases compared to the lungs of control pigs. Cytokeratin labeling marked the type I, and type II pneumocytes as well as bronchial epithelial cells, however this staining was not suitable for cell counting purposes. When the routine histological scores were compared to the number of immunohistochemically positive cells, Ki67 cell counts were found to show positive correlation (p<0.05) with the overall severity of the lesions

Mystery of fatal ‘staggering disease’ unravelled: novel rustrela virus causes severe meningoencephalomyelitis in domestic cats

‘Staggering disease’ is a neurological disease entity considered a threat to European domestic cats (Felis catus) for almost five decades. However, its aetiology has remained obscure. Rustrela virus (RusV), a relative of rubella virus, has recently been shown to be associated with encephalitis in a broad range of mammalian hosts. Here, we report the detection of RusV RNA and antigen by metagenomic sequencing, RT-qPCR, in-situ hybridization and immunohistochemistry in brain tissues of 27 out of 29 cats with non-suppurative meningoencephalomyelitis and clinical signs compatible with’staggering disease’ from Sweden, Austria, and Germany, but not in non-affected control cats. Screening of possible reservoir hosts in Sweden revealed RusV infection in wood mice (Apodemus sylvaticus). Our work indicates that RusV is the long-sought cause of feline ‘staggering disease’. Given its reported broad host spectrum and considerable geographic range, RusV may be the aetiological agent of neuropathologies in further mammals, possibly even including humans

Emergence of porcine epidemic diarrhea virus in southern Germany

Background Over the last years, porcine epidemic diarrhea virus (PEDV) has caused devastating enteric diseases in the US and several countries in Asia, while outbreaks in Europe have only been reported sporadically since the 1980s. At present, only insufficient information is available on currently circulating PEDV strains in Europe and their impact on the European swine industry. In this case report, we present epidemic outbreaks of porcine epidemic diarrhea in three farms in South-Western Germany. Case presentation Epidemic outbreaks of diarrhea affecting pigs of all age groups were reported in three farms, one fattening farm and two piglet producing farms, in South-Western Germany between May and November 2014. In the fattening farm yellowish, watery diarrhea without evidence of mucus or blood was associated with a massive reduction of feed consumption. Severity of clinical signs and mortality in young suckling pigs varied significantly between the two affected sow farms. While mortality in suckling piglets reached almost 70 % in one sow herd, no increase in suckling piglet mortality was observed in the second sow farm. In all three cases, PEDV was confirmed in feces and small intestines by RT-qPCR. Phylogenetic analyses based on full-length PEDV genomes revealed high identity among strains from all three herds. Moreover, the German strains showed very high nucleotide identity (99.4 %) with a variant of PEDV (OH851) that was isolated in the United States in January 2014. This strain with insertions and deletions in the S-gene (so called INDEL strains) was reported to show lower virulence. Slightly lower identities were found with other strains from the US and Asia. Conclusion Phylogenetic information on the distribution of PEDV strains in Europe is severely lacking. In this case report we demonstrate that acute outbreaks of PEDV occurred in southern Germany in 2014. Current strains were clearly different from isolates found in the 1980s and were closely related to a PEDV variant found in the US in 2014. Moreover, the present case report indicates that variant strains of PEDV, containing insertions and deletions in the S gene, which were reported to be of lower virulence, might be able to cause high mortality in suckling piglets