Disseminated Mycobacterium genavense infection in a chinchilla (Chinchilla lanigera).

A 1-year-old neutered male chinchilla (Chinchilla lanigera) was presented with emaciation and a 1-month history of progressive weight loss. The animal was bright and responsive on clinical examination, but had poor body condition. Serum biochemical analysis revealed elevated alanine amino transferase and alkaline phosphatase. Ultrasound examination was unremarkable. Thoracic radiography showed changes consistent with bullous emphysema and severe pneumonia. Antibiotic therapy was initiated, but the chinchilla died 6 weeks later. Necropsy examination revealed granulomatous lesions in the lungs and liver. Numerous acid-fast bacilli were present in the cytoplasm of macrophages. Sequencing of genetic material isolated from fixed tissue classified the pathogen as Mycobacterium genavense.

Hemoprotozoa of domestic animals in France: prevalence and molecular characterization.

Very limited information is available on epizootiology of haematozoan infections in French domestic animals. In an attempt to address this issue, prevalence of piroplasmida was studied in carnivores and ruminants, whereas prevalence of Hepatozoon spp. was only investigated in carnivores. In total, 383 animals were included in the survey (namely 116 cats, 108 dogs, 91 sheep and 68 cows). Parasite diagnosis was carried out using molecular methods such as PCR and sequencing of the 18S rRNA gene. In addition, ruminant samples were analyzed with the reverse line blotting technique (RLB). Results of RLB and PCR plus sequencing were in total agreement. In carnivores, haematozoan prevalence was close to 1%. Two cats were infected by H. canis (1.7% prevalence) and one of them was co-infected by Cytauxzoon sp. (0.8%). This represents the first finding of both pathogens in French cats. One dog was infected by H. canis (0.9%) and another by Babesia canis vogeli (0.9%). In ruminants, haematozoan prevalence (piroplasmida) was significantly higher than in carnivores (4.8% in sheep and 8.8% in cow). Theileria ovis was found in 1 sheep, Theileria sp. in 2 sheep, Theileria buffeli in 5 cows and B. major in 1 cow. Evidence presented in this contribution indicates that haematic protozoa are not widely distributed in domestic mammal populations of France.

Development and evaluation of a quantitative PCR assay for detection of Hepatozoon sp.

With the aim to improve current molecular diagnostic techniques of Hepatozoon sp. in carnivore mammals, we developed a quantitative PCR (qPCR) assay with SYBR Green I((R)). The method, consisting of amplification of a 235bp fragment of the 18S rRNA gene, is able to detect at least 0.1fg of parasite DNA. Reproducible quantitative results were obtained over a range of 0.1ng-0.1fg of Hepatozoon sp. DNA. To assess the performance of the qPCR assay, DNA samples from dogs (140) and cats (50) were tested with either standard PCR or qPCR. Positive samples were always confirmed by partial sequencing of the 18S rRNA gene. Quantitative PCR was 15.8% more sensitive than standard PCR to detect H. canis in dogs. In cats, no infections were detected by standard PCR, compared to two positives by qPCR (which were infected by H. canis as shown by sequencing).

A novel polyomavirus (goose hemorrhagic polyomavirus) is the agent of hemorrhagic nephritis enteritis of geese.

We have identified the etiological agent of hemorrhagic nephritis enteritis of geese (HNEG), a fatal disease of European geese. HNEG has been recognized in almost all goose breeding areas, with an epizootic pattern, and up to now, the infectious agent has remained unknown. In order to identify the causative agent, infected tissues from HNEG-affected geese were inoculated to 1-day-old goslings, which then developed clinical signs typical of HNEG. Tissue homogenates from these birds were subjected to Freon extraction followed by sucrose density gradient ultracentrifugation. The resulting main band was examined by electron microscopy and consisted of spherical, naked, papovavirus-like particles approximately 45 nm in diameter. The virus was isolated and propagated in goose kidney cell primary culture. Tissue- or culture-purified virus allowed the experimental reproduction of the disease in goslings. Random PCR amplification of viral nucleic acid produced a 1,175-bp fragment which was shown to be associated with field samples collected from geese affected by HNEG on commercial farms in France. Sequence analysis of the PCR product revealed a unique open reading frame, showing 63 to 72% amino acid similarity with the major capsid protein (VP1) of several polyomaviruses. Finally, based on phylogenetic analysis, we conclude that the causative agent of HNEG is closely related to but clearly distinct from other polyomaviruses; we thus have named this newly identified virus Goose hemorrhagic polyomavirus.