Anna Kübber-Heiss

Prevalence of feline coronavirus types I and II in cats with histopathologically verified feline infectious peritonitis.

Abstract Feline coronaviruses (FCoV) vary widely in virulence causing a spectrum of clinical manifestations reaching from subclinical course to fatal feline infectious peritonitis (FIP). Independent of virulence variations they are separated into two different types, type I, the original FCoV, and type II, which is closely related to canine coronavirus (CCV). The prevalence of FCoV types in Austrian cat populations without FIP has been surveyed recently indicating that type I infections predominate. The distribution of FCoV types in cats, which had succumbed to FIP, however, was fairly unknown. PCR assays have been developed amplifying parts of the spike protein gene. Type-specific primer pairs were designed, generating PCR products of different sizes. A total of 94 organ pools of cats with histopathologically verified FIP was tested. A clear differentiation was achieved in 74 cats, 86% of them were type I positive, 7% type II positive, and 7% were positive for both types. These findings demonstrate that in FIP cases FCoV type I predominates, too, nonetheless, in 14% of the cases FCoV type II was detected, suggesting its causative involvement in cases of FIP.

Characterization of methicillin-resistant Staphylococcus spp. carrying the mecC gene, isolated from wildlife

OBJECTIVES A recently identified mecA homologue, mecC, in methicillin-resistant Staphylococcus aureus (MRSA) has been isolated from humans and different animal hosts. The aim of this study was to determine antimicrobial resistance and provide molecular characterization of MRSA and methicillin-resistant non-Staphylococcus aureus staphylococci (MRnSA) isolated from wildlife that carried the gene mecC. METHODS Five S. aureus and one coagulase-negative Staphylococcus isolate displaying phenotypic oxacillin resistance, but not recognized with conventional PCR for mecA, were further characterized by a polyphasic approach. The presence of mecC in all isolates was determined using specific PCR. PCR targeting Panton-Valentine leucocidin (PVL) genes of MRSA was performed. MRSA isolates were genotyped by spa typing and multilocus sequence typing. All isolates were genotyped by staphylococcal cassette chromosome mec (SCCmec) typing. 16S rDNA sequence analysis for MRnSA identification was performed. Antimicrobial susceptibility testing was performed for all isolates. RESULTS All five MRSA isolates contained the mecC gene, were PVL negative, carried SCCmec type XI and belonged to ST130 (where ST stands for sequence type), with spa types t843, t10513 or t3256, or to ST2620, with spa type t4335. The MRnSA isolate, most closely related to Staphylococcus stepanovicii, carried mecA and blaZ genes related to SCCmec XI. MRSA isolates exhibited resistance to the β-lactams only. CONCLUSIONS The MRSA isolates described in this study represent the first detection of mecC-positive MRSA in a European otter (Lutra lutra) and a European brown hare (Lepus europaeus). The MRnSA isolate represents the first isolation of MRnSA from a Eurasian lynx (Lynx lynx).

Fox on the run--molecular surveillance of fox blood and tissue for the occurrence of tick-borne pathogens in Austria.

BackgroundThe red fox (Vulpes vulpes) is a widespread species, harbouring many pathogens relevant for humans and pets. Indeed, Anaplasma spp., Ehrlichia canis and Rickettsia spp. among the bacteria and Hepatozoon canis as well as Babesia sp. among the parasites have been the focus of several studies.FindingsIn a cohort of 36 foxes shot on one day in the north-eastern part of Austria, Babesia microti-like pathogens were found in 50%, while H. canis was detected in 58.3% of the samples. The spleen was more useful for detection of H. canis, whereas B. microti-like parasites were more frequently found in the blood. Bacteria could not be confirmed in any of the cases to demonstrate the occurrence of such tick-borne pathogens using PCR and sequencing on blood and spleen samples.ConclusionsThe occurrence of B. microti-like and H. canis parasites raised many questions, because these infections have never been found autochthonously in dogs. Furthermore in the case of H. canis the main vector tick, Rhipicephalus sanguineus, is absent in the sampling area, leaving space for further hypotheses for transmission such as vertical transmission, transmission via ingestion of prey animals or other vector ticks. Further studies are needed to evaluate the risks for pets in this area. PCRs delivered differing results with the different tissues, suggesting the use of both spleen and blood to obtain an integral result.

A golden jackal (Canis aureus) from Austria bearing Hepatozoon canis--import due to immigration into a non-endemic area?

The protozoan Hepatozoon canis, which is transmitted via ingestion of infected ticks by canine hosts, is not endemic to mid-latitude regions in Europe. Its distribution is supposed to be linked to the occurrence of its primary tick vector Rhipicephalus sanguineus. A young male golden jackal (Canis aureus) found as road kill close to Vienna, Austria, was infected by this pathogen. Cloning and sequencing of the PCR product revealed 6 different haplotypes of H. canis. Based on the sequences, no clear relationship to the origin of infection could be traced. This is the first report of H. canis for Austria, and wild canines such as the currently found jackal may provide a source of natural spread of this parasite into non-endemic areas. This natural immigration of wild animals represents a way of pathogen introduction, which has to be considered in disease prevention in addition to human-made introduction due to animal import and export.

Phylogenetic analysis of the L and HN gene of ophidian paramyxoviruses

Summary.  Two reptilian paramyxoviruses, isolated from a neotropical rattlesnake (neotropical virus, NTV, ATCC VR-1408) and a bush viper (bush viper virus, BVV, ATCC VR-1409), respectively, were analysed to determine their taxonomic position among other reptilian paramyxoviruses investigated previously by Ahne et al. [7]. A 679 bp long region of the hemagglutinin-neuraminidase (HN) gene and a 627 bp long region of the large (L) gene were reverse transcribed, amplified by polymerase chain reaction (PCR), and sequenced. The deduced amino acid sequences were compared to mammalian paramyxoviruses belonging to the genera Respirovirus and Rubulavirus. The deduced amino acid sequences revealed 58.9 to 62% homology for the partial L protein and 41% to 47.1% homology for the partial HN protein. For phylogenetic analyses, a 518 bp L gene and a 352 bp HN gene fragment were used, both generating similar trees consisting of two distinct main groups, and some intermediate isolates. BVV clustered within group “b” while NTV clustered together with the intermediate ophidian paramyxovirus isolate Crot2-OH90.

IDIOPATHIC ACUTE ONSET MYELOPATHY IN CHEETAH (ACINONYX JUBATUS) CUBS

Abstract Numerous cases of ataxia, hind limb paresis, and paralysis have occurred in cheetah (Acinonyx jubatus) cubs over the past 10 yr within the European Endangered Species Program population, including 12 in mainland Europe, two in the British Isles, one in Namibia, and one in Dubai. The condition is the most important medical factor limiting European cheetah population growth. Eight cubs at the Salzburg Zoo, Austria, were affected. They demonstrated upper motor neuron lesions when alive and bilateral, symmetrical myelin degeneration of the spinal cord on necropsy. Ballooning of myelin sheaths surrounded mostly preserved axons, and no spheroids, characteristic of acute axonal degeneration, were found. Myelin loss markedly exceeded axonal degeneration. The syndromes etiology is unclear, although viral, bacterial, parasitic, genetic, nutritional–metabolic, toxic, and physical causes have been considered.

Detection of Cryptosporidium spp., Entamoeba spp. and Monocercomonas spp. in the Gastrointestinal Tract of Snakes by In-situ Hybridization

This report describes the development of a diagnostic method for protozoal infections of the gastrointestinal tract of captive snakes, based on chromogenic in-situ hybridization with probes designed for the detection of 18S rRNA genes from Cryptosporidium spp., Entamoeba spp., Entamoeba invadens and Monocercomonas spp. The specificity of the probes was confirmed with the help of parasitic cultures and gene sequence analysis. The probes gave clear positive signals. Of 182 snakes examined, seven were positive with the Cryptosporidium probe, 13 with the Entamoeba probe (of which nine were also positive with the E. invadens probe), and 34 with the Monocercomonas probe.

Characterization of ESBL- and AmpC-Producing and Fluoroquinolone-Resistant Enterobacteriaceae Isolated from Mouflons ( Ovis orientalis musimon ) in Austria and Germany

The aim of this study was to investigate the presence of β-lactamase producing or fluoroquinolone-resistant members of the family Enterobacteriaceae in European mouflons (Ovis orientalis musimon). The mouflon samples originated from nasal and perineal swabs and/or organ samples in cases of a suspected infection. Only one of the 32 mouflons was tested positive for the presence of Enterobacteriaceae that displayed either an ESBL/AmpC phenotype or were resistant to ciprofloxacin. The positively tested swab originated from a sample of the jejunal mucosa of a four-year old female mouflon. Two different colony morphotypes were identified as Escherichia coli and Klebsiella pneumoniae. These isolates were phenotypically and genotypically characterized in detail by a polyphasic approach. Both isolates were multi-drug resistant. The E. coli isolate belonged to the phylogenetic group B1 and sequence type (ST) 744 and harboured the β-lactamase genes blaCTX-M-15 and blaOXA-1. The K. pneumoniae, identified as ST11, harboured the β-lactamase genes blaSHV-11, blaOXA-1, and blaDHA-1 as well as the plasmid-mediated quinolone resistance (PMQR) gene qnrB55. The present study demonstrates that wild animals can acquire human-derived resistance determinants and such findings may indicate environmental pollution with resistance determinants from other sources.

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

A 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.

Neosporosis in a captive Parma wallaby (Macropus parma).

Infection with Neospora caninum has been diagnosed in a variety of animal species; however, reports in marsupials are rare. A captive Parma wallaby (Macropus parma) died suddenly and was subjected to necropsy examination. The main finding was necrotizing myocarditis associated with protozoan parasites. The protozoa were identified as N. caninum by use of immunohistochemistry and partial gene sequence analysis. Neospora and Toxoplasma should be considered a possible cause of disease in captive marsupials. Further work is required to determine whether marsupials are an accidental or terminal host of this protozoan in order to better understand the host-parasite relationship.