BackgroundThe feline endogenous retrovirus RD114 is contained in the genome of cats. The virus may contaminate live canine vaccines based on cultured feline cells. The in vivo infectivity, acute and subacute pathogenicity, and viral proliferation of the RD114 virus were evaluated by experimental infection of dogs.MethodsNine specific pathogen free dogs were divided into three groups, with each group consisting of one female and two male dogs. Dogs were subcutaneously inoculated in the neck with either 1 ml RD114 stock virus (group A), inactivated RD114 virus suspension (group B), or cell culture medium (group C) as a negative control. To assess blood cell counts and biochemical properties, blood samples from each group were collected 5 days before inoculation, just prior to inoculation, and 1, 3, 7 and 10 days post-inoculation.ResultDuring the experimental period of 51 days, none of the dogs inoculated with RD114 virus showed any clinical signs, significant increases in rectal temperature or abnormal blood biochemical characteristics including C-reactive protein when compared with the negative controls. We were not able to re-isolate the RD114 virus from buffy coat cells of group A dogs. Additionally, we could not detect RD114 provirus in the genomic DNA isolated from peripheral blood leukocytes, lymph node, spleen and sternal bone marrow cells.ConclusionsSigns of RD114 virus proliferation were not found after subcutaneous infection of dogs. Although the potential risk caused by infection with RD114 virus in dogs could not be assessed in this study, we suspect that RD114 virus has little or no virulence in dogs.

It is known that certain feline cell lines, such as the Crandell-Rees feline kidney cell, produce an RD-114-like virus. As a feline endogenous retrovirus, RD114 virus, exists in the genome of all cats, it can be assumed that contamination with the virus in feline and canine live vaccines manufactured by culturing cells of feline origin occurs. To detect an infectious RD114 virus in vitro, a LacZ marker rescue assay has recently been established. In feline and canine live vaccines approved in Japan, feline cell lines are widely used to produce vaccines, especially those containing canine parvovirus components. The LacZ marker rescue assay detects infectious viral particles, but the real-time reverse-transcription-PCR detects both infectious and defective viruses. The canine live vaccines manufactured in cells of feline origin showed positive results for the env gene by the real-time reverse-transcription-PCR, including all of the 8 vaccines produced in feline cell lines that were negative in the LacZ marker rescue assay. In conclusion, the present investigation suggests that the newly developed method has the advantages of shorter time requirements and can be applied as a valuable screening method to detect RD114 viral RNA in vaccines.

ABSTRACT. Stray dogs are a public health risk factor when canine rabies is endemic. The Rabies Prevention Law has introduced measures to control stray dogs, but many dogs are still captured in Japan. In order to estimate the immune status of stray dogs for the purposes of risk management, we conducted a serological survey at the Hyogo Prefecture Animal Well-being Center. Only 27.7% of dogs brought into the Center (n=166) had protective immune status. This result suggests that there is the potential for reintroduction of canine rabies into stray dogs, leading to endemic rabies and its transmission to humans. Continued removal of stray dogs, education on rabies prevention and vaccination of dogs therefore remain important public health issues.

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