Detection of Chronic bee paralysis virus ( CBPV ) genome 1 and its replicative RNA form in various hosts and possible 2 ways of spread . 3 4

22 Detection of Chronic bee paralysis virus (CBPV) is reported for the first time in two 23 species of ants (Camponotus vagus and Formica rufa) and in Varroa destructor. A 24 quantitative real-time PCR (qPCR) method was used to detect and quantify CBPV in 25 infected bees, ants and mites. A minus-strand-specific RT-PCR was used to assess viral 26 replication. These results suggest a new way by which the infection may be spread and 27 other sites of viral persistence in the close apiary environment. 33 34 A large variety of viruses multiply in the honey bee Apis mellifera L. Knowledge of the spreading mechanism of honey bee pathogens within the hive and 36 apiary is essential to our understanding of bee disease dynamics. Among the viruses 37 infecting honey bees, Chronic bee paralysis virus (CBPV) is the causal agent of chronic 38 paralysis known to induce significant losses in honey bee colonies (Ball & Bailey, 1997). 39 The pathology is characterized by clusters of trembling, flightless, crawling bees and by 40 individual bees, sometimes hairless, standing at the hive entrance (Bailey et al., 1983). A 41 correlation between chronic paralysis and high viral loads of CBPV was demonstrated 42 particularly in symptomatic bees (Blanchard et al., 2007a). Moderate viral loads were also 43 demonstrated in colonies without symptoms (Blanchard et al., 2007a). To date, CBPV 44 has been detected only in A. mellifera (Allen & Ball, 1996) and the presence of this virus 45 has been observed on every continent (Bailey, 1967). However, several honey bee 46 viruses have been identified in the ectoparasitic mite Varroa destructor (Ball & Allen, 47 1988). Acute bee paralysis virus (ABPV) and deformed wing virus (DWV) have been 48 identified as major factors contributing to the mortality of bees in colonies infested by V. 52 Tentcheva et al., 2004), the presence of mites in CBPV symptomatic hives was observed, 53 suggesting its potential role in the viral dissemination. Like the mite V. destructor, which is 54 recognised as the main vector of some honeybee viruses, other organisms could also 55 play this vector role in the honeybee. In fact, during our field observations, we noted the 56 presence of ants close to apiaries, carrying dead bees around hives. Two species of ants 57 were observed: Camponotus vagus, a very common ant with a worldwide distribution 58 and Formica rufa, present in many coniferous …