Nathalie Cordonnier

European Surveillance for Pantropic Canine Coronavirus

ABSTRACT Highly virulent pantropic canine coronavirus (CCoV) strains belonging to subtype IIa were recently identified in dogs. To assess the distribution of such strains in Europe, tissue samples were collected from 354 dogs that had died after displaying systemic disease in France (n = 92), Hungary (n = 75), Italy (n = 69), Greece (n = 87), The Netherlands (n = 27), Belgium (n = 4), and Bulgaria (n = 1). A total of 124 animals tested positive for CCoV, with 33 of them displaying the virus in extraintestinal tissues. Twenty-four CCoV strains (19.35% of the CCoV-positive dogs) detected in internal organs were characterized as subtype IIa and consequently assumed to be pantropic CCoVs. Sequence and phylogenetic analyses of the 5′ end of the spike protein gene showed that pantropic CCoV strains are closely related to each other, with the exception of two divergent French viruses that clustered with enteric strains.

Encephalomyocarditis virus mortality in semi‐wild bonobos (Pan panicus)

Background  Fatal myocarditis from encephalomyocarditis virus (EMCV) infection has previously been identified in sporadic and epidemic forms in many species of captive non‐human primates probably including one bonobo (Pan paniscus).

Evidence of transplacental transmission of bluetongue virus serotype 8 in goats

During the incursion of bluetongue virus (BTV) serotype 8 in Europe, an increase in the number of abortions in ruminants was observed. Transplacental transmission of BTV-8 in cattle and sheep, with subsequent foetal infection, is a feature of this specific bluetongue serotype. In this study, BTV-8 ability to cross the placental barrier at the beginning of the second third of pregnancy and at the end of pregnancy was investigated in goats in two separate experiments. In the first experiment, nine goats were experimentally infected with BTV-8 at 61 days of pregnancy. Foetuses were collected 21 dpi. BTV-8 was evidenced by real time RT-PCR and by viral isolation using blood from the umbilical cord and the spleens of 3 out of the 13 foetuses. All dams were viraemic (viral isolation) at the moment of sampling of the foetuses. Significant macroscopic or histological lesions could not be observed in foetuses or in their infected dams (notably at the placenta level). In the second experiment, 10 goats were infected with BTV-8 at 135 days of pregnancy. Kids were born by caesarean section at the programmed day of birth (15 dpi). BTV-8 could not be detected by rt-RT-PCR in blood or spleen samples from the kids. This study showed for the first time that BTV-8 transplacental transmission can occur in goats that have been infected at 61 days of pregnancy, with infectious virus recovered from the caprine foetuses. The observed transmission rate was quite high (33%) at this stage of pregnancy. However, it was not possible to demonstrate the existence of BTV-8 transplacental transmission when infection occurred at the end of the goat pregnancy.

IS-98-ST1 West Nile Virus Derived from an Infectious cDNA Clone Retains Neuroinvasiveness and Neurovirulence Properties of the Original Virus

Infectious clones of West Nile virus (WNV) have previously been generated and used to decipher the role of viral proteins in WNV virulence. The majority of molecular clones obtained to date have been derived from North American, Australian, or African isolates. Here, we describe the construction of an infectious cDNA clone of a Mediterranean WNV strain, IS-98-ST1. We characterized the biological properties of the recovered recombinant virus in cell culture and in mice. The growth kinetics of recombinant and parental WNV were similar in Vero cells. Moreover, the phenotype of recombinant and parental WNV was indistinguishable as regards viremia, viral load in the brain, and mortality in susceptible and resistant mice. Finally, the pathobiology of the infectious clone was examined in embryonated chicken eggs. The capacity of different WNV strains to replicate in embryonated chicken eggs closely paralleled their ability to replicate in mice, suggesting that inoculation of embryonated chicken eggs could provide a practical in vivo model for the study of WNV pathogenesis. In conclusion, the IS-98-ST1 infectious clone will allow assessment of the impact of selected mutations and novel genomic changes appearing in emerging European strains pathogenicity and endemic or epidemic potential. This will be invaluable in the context of an increasing number of outbreaks and enhanced severity of infections in the Mediterranean basin and Eastern Europe.

Encephalomyocarditis Virus 2A Protein Is Required for Viral Pathogenesis and Inhibition of Apoptosis

ABSTRACT The encephalomyocarditis virus (EMCV), a Picornaviridae virus, has a wide host spectrum and can cause various diseases. EMCV virulence factors, however, are as yet ill defined. Here, we demonstrate that the EMCV 2A protein is essential for the pathogenesis of EMCV. Infection of mice with the B279/95 strain of EMCV resulted in acute fatal disease, while the clone C9, derived by serial in vitro passage of the B279/95 strain, was avirulent. C9 harbored a large deletion in the gene encoding the 2A protein. This deletion was incorporated into the cDNA of a pathogenic EMCV1.26 strain. The new virus, EMCV1.26Δ2A, was capable of replicating in vitro, albeit more slowly than EMCV1.26. Only mice inoculated with EMCV1.26 triggered death within a few days. Mice infected with EMCV1.26Δ2A did not exhibit clinical signs, and histopathological analyses showed no damage in the central nervous system, unlike EMCV1.26-infected mice. In vitro, EMCV1.26Δ2A presented a defect in viral particle release correlating with prolonged cell viability. Unlike EMCV1.26, which induced cytopathic cell death, EMCV1.26Δ2A induced apoptosis via caspase 3 activation. This strongly suggests that the 2A protein is required for inhibition of apoptosis during EMCV infection. All together, our data indicate that the EMCV 2A protein is important for the virus in counteracting host defenses, since Δ2A viruses were no longer pathogenic and were unable to inhibit apoptosis in vitro.

Skin fragility syndrome in a cat with multicentric follicular lymphoma

An 11-year-old, spayed female domestic shorthair cat was presented for a right flank wound. On clinical examination, a single non-painful skin tear lesion with irregular edges was detected. During the examination, star-shaped cigarette paper-like skin lesions appeared spontaneously. An abdominal mass was also palpated. Feline skin fragility syndrome (FSFS) was suspected and a multicentric lymphoma was diagnosed by fine needle aspiration. The cat’s condition declined and it died spontaneously. Post-mortem examination confirmed the diagnosis of lymphoma. Neoplastic lymphocytes were not observed in the skin. Histological analysis of the skin was consistent with the morphological aspects of FSFS. A possible direct link between the two conditions remains a matter of speculation, but this case report provides the first description of FSFS associated with multicentric follicular lymphoma. Thus, multicentric follicular lymphoma should be considered as a differential diagnosis in cats presenting with FSFS.

Parotid Salivary Duct Sialocele Associated with Glandular Duct Stenosis in a Cat

Feline parotid salivary duct sialocele is an uncommon disorder that has been previously reported in association with traumatic rupture of the duct in only two cats. Both cases were successfully treated by proximal duct ligation. We describe the successful surgical treatment of a parotid duct sialocele, secondary to spontaneous salivary duct stenosis, in an adult domestic shorthair cat. The cat was referred for assessment of a recurrent fluid-filled swelling on the left side of the face. Cytology of the aspirated fluid was consistent with serous saliva. The anatomical localisation of the lesion and the nature of the fluid were indicative of parotid gland/duct involvement. Retrograde sialography by parotid duct cannulation was unsuccessful because the left parotid duct opening was stenosed and obstructed by scar tissue. Surgical exploration revealed a parotid salivary duct sialocele, which was completely removed along with the parotid gland without complications.

Assessment of Aspergillus fumigatus burden in lungs of intratracheally-challenged turkeys (Meleagris gallopavo) by quantitative PCR, galactomannan enzyme immunoassay, and quantitative culture

Aspergillus fumigatus remains a major respiratory pathogen in birds and treatment is still difficult. We challenged different groups of few-day-old turkeys via intratracheal aerosolisation with increasing concentrations (10(5) up to 10(8)) of conidia using a MicroSprayer(®) device. The fungal burden was assessed by real-time PCR, galactomannan dosage, CFU counting and histopathological evaluation in order to provide a comparison of these results within each inoculum groups. Significant mortality, occurring in the first 96h after inoculation, was only observed at the highest inoculum dose. Culture counts, GM index and qPCR results on the one hand and inoculum size on the other hand appeared to be clearly correlated. The mean fungal burden detected by qPCR was 1.3log10 units higher than the mean values obtained by CFU measurement. The new model and the markers will be used to evaluate the efficacy of antifungal treatments that could be used in poultry farms.

Immunohistochemical characterization of a hepatic neuroendocrine carcinoma in a cat

A hepatic mass was identified in a 5-year-old, female mixed-breed cat that died spontaneously after a clinical history of progressive emaciation, ptyalism, and persistent coryza. At necropsy, a 7-cm-diameter, yellow-brown, firm, multilobulated tumor was identified in the liver. Microscopically, the mass consisted of neoplastic cells arranged in small, closely packed nests within a thin fibrovascular stroma. These cells were of medium sized and polygonal, with fine argyrophilic cytoplasmic granules. Nuclei were predominantly round with finely stippled chromatin and indistinct nucleoli. Mitotic figures were numerous. Immunohistochemically, most of the neoplastic cells were immunoreactive for chromogranin A, neuron-specific enolase (NSE), and cytokeratin AE1/AE3 and weakly labeled for synaptophysin. The tumor was negative for glial fibrillary acidic protein (GFAP), vimentin, and cytokeratins 5, 6, 8, and 17. Vascular emboli and intrahepatic micrometastasis were also identified with chromogranin A. All these features were consistent with a hepatic neuroendocrine carcinoma and emphasized the importance of using a panel of antibodies to diagnose such rare tumors.

Generalized verrucosis associated with canine papillomavirus 9 infection in a dog

In dogs, different types of canine papillomavirus (CPV) have been associated with distinct skin lesions, including exophytic or endophytic warts, pigmented plaques, hyperkeratotic to horny lesions and squamous cell carcinoma. Previously, CPV1 has been associated with extensive oral and cutaneous papillomas in a shar-pei dog. This letter describes a new clinical presentation of a severe and widespread skin papillomavirus infection and the third identification of CPV9. A 7-year-old intact female Staffordshire bull terrier was presented for generalized alopecia and ‘warts’, along with a history of recurrent demodicosis from 9 months of age. Previous therapies included benzoyl peroxide shampoo (Paxcutol ; Virbac, Carros, France) and topical amitraz rinses (Ectodex ; MSD Animal Health, Angers, France) once weekly, with periodic improvements but never total recovery. Physical examination revealed mild depression and submandibular, prescapular and popliteal lymphadenomegaly. Dermatological examination showed generalized alopecia and a mix of multiple exophytic warts and slightly raised plaques over the entire body. Most of these lesions were darkly pigmented, but some were clearly hypopigmented. Lesions were widespread on the ventrum and coalescing on the distal part of the limbs to form a shell-like appearance with no visible uninvolved skin (Figure 1). Pruritus was not observed. Numerous Demodex canis mites were found in deep skin scrapings. Fungal culture was negative for dermatophytes. Multiple skin biopsies were collected for histopathological examination and virus examination. Histopathological findings included multiple heavily keratinized finger-like projections of thickened squamous epithelium (Figure 2). A highly developed granular layer with numerous and large keratohyalin granules, hyperpigmentation, keratinocytes with pale cytoplasm, oval hypochromic nuclei and marginated chromatin (koilocytes) were present. Some Demodex mites were also found. A diagnosis of papillomavirus infection and demodicosis was made. DNA from the samples was extracted using a DNeasy extraction kit (Qiagen, Hombrechtikon, Switzerland) according to the manufacturer’s instructions. Polymerase chain reaction, performed as published previously, identified CPV9. Haematology, CD4+ to CD8+ T-lymphocyte ratio and biochemistry profile, radiographs and ultrasound examinations were unremarkable.