Isolation and propagation of the animal rotaviruses in MA-104 cells--30 years of practical experience.

A total of 136 rotavirus positive samples from diarrhoeic animals of different species were submitted for isolation and cultural propagation of rotavirus on MA-104 cells. The samples were collected from animals with diarrhoea, between 1980 and 2010, originating from herds or farms located in several parts of Germany. Rotaviruses of species A were isolated from 102 faecal samples in cultures of MA-104 cells under the following conditions: pre-treatment of virus with trypsin, incorporation of trypsin into culture medium, use of roller cultures, and centrifugation of the samples on the cells. The cell culture adapted viruses produced a cytopathic effect, accompanied by the release of cells from the glass surface of the cultivation vessels. After 10 passages the virus isolates yielded titres between 10(5.5) and 10(7.5)ml(-1) TCID50. Isolation and serial propagation of the virus in MA-104 cells was confirmed by immunofluorescence assay, transmission electron microscopy, and polyacrylamide-gel electrophoresis of viral dsRNA. Eight (5.9%) of the electrophoretic profiles were characteristic of species B or D rotaviruses, which were not replicated in MA-104 cells.

Two outbreaks of virulent systemic feline calicivirus infection in cats in Germany.

Over the last years, several outbreaks of virulent systemic feline calicivirus (VS-FCV) infection have been described in the USA and several European countries. The paper describes two outbreaks of VS-FCV infection in cats in Germany. Data concerning clinical, laboratory, and histopathological features ofVS-FCV infection were collected from two outbreaks affecting 55 and 4 cats, respectively. Presence of feline calicivirus was confirmed by PCR followed by sequencing of the PCR-products. Clinical signs were variable, including severe upper respiratory tract infection, dyspnoea, oral and footpad ulceration, facial oedema, enteritis, pneumonia, bleeding disorder, high fever, and icterus. Both outbreaks were characterized by a high mortality rate.The present report describes the first documented outbreaks of VS-FCV infection in cats in Germany. Clinical and histopathological features are comparable to outbreaks described in the USA and Europe. However, phylogenetic analysis of the virus genome suggests that virus strains involved in these outbreaks were different from each other and from virulent strains isolated before, confirming the known genetic variability of FCV.

Isolation and molecular characterisation of equine rotaviruses from Germany.

A total of 26 rotavirus positive faecal samples of diarrhoeal foals, and 8 equine rotavirus isolates were examined. Viral RNA patterns were generated, G typing was performed by PCR, and a P[12]-specific DNA probe was developed for P typing. Furthermore, five equine rotavirus isolates were sequenced in the genomic regions coding for VP7 and part of VP4. Rotaviruses of genotype G3 P[12] were found in 22 faecal samples and G14 P[12] type could be found in 4 faecal samples. These findings confirm that in Germany G3 P[12] is the predominating type of equine rotaviruses.