Identification of snake arenaviruses in live boas and pythons in a zoo in Germany

Objective: Recent studies have described the detection and characterisation of new, snake specific arenaviruses in boas and pythons with inclusion body disease (IBD). The objective of this study was to detect arenaviral RNA in live snakes and to determine if these were associated with IBD in all cases. Samples for arenavirus detection in live animals were compared. Detected viruses were compared in order to understand their genetic variability. Materials and methods: Esophageal swabs and whole blood was collected from a total of 28 boas and pythons. Samples were tested for arenaviral RNA by RT-PCR. Blood smears from all animals were examined for the presence of inclusion bodies. Internal tissues from animals that died or were euthanized during the study were examined for inclusions and via RT-PCR for arenaviral RNA. All PCR products were sequenced and the genomic sequences phylogenetically analysed. Results: Nine live animals were found to be arenavirus-positive. Two additional snakes tested positive following necropsy. Five new arenaviruses were detected and identified. The detected viruses were named Boa Arenavirus Deutschland (Boa Av DE) numbers 1-4 and one virus detected in a python (Morelia viridis) was named Python Av DE1. Results from sequence analyses revealed considerable similarities to a portion of the glycoprotein genes of recently identified boid snake arenaviruses. Conclusions: Both oral swabs and whole blood can be used for the detection of arenaviruses in snakes. In most cases, but not in all, the presence of arenaviral RNA correlated with the presence of inclusions in the tissues of infected animals. There was evidence that some animals may be able to clear arenavirus infection without development of IBD. This is the first detection of arenaviruses in live snakes. Clinical relevance: The detection of arenaviruses in live snakes is of importance for both disease detection and prevention and for use in quarantine situations. The findings in this study support the theory that arenaviruses are the cause of IBD, but indicate that in some cases it may be possible for animals to clear arenavirus infections without developing IBD.