Microbial diversity in the predatory mite Metaseiulus occidentalis (Acari: Phytoseiidae) and its prey, Tetranychus urticae (Acari: Tetranychidae(

Microorganisms associated with the predatory mite Metaseiulus (=Typhlodromus or Galendromus) occidentalis (Nesbitt) and its prey, the two-spotted spider mite Tetranychus urticae (Koch), were assessed using a high-fidelity polymerase chain reaction (PCR) protocol and primers designed to identify Eubacteria, Archaeabacteria, iridoviruses, Helicosporidia, Cytophaga-like microorganisms, Wolbachia and its bacteriophage WO, fungi and yeast-like organisms. Sequences from four bacterial species related to Wolbachia ((alpha)-Proteobacteria), Cardinium, Bacteroidetes, and Enterobacter ((gamma)-Proteobacteria) were obtained from M. occidentalis, and three sequences related to Wolbachia, Rickettsia, and Caulobacter ((alpha)-Proteobacteria) were obtained from T. urticae. No nucleotide differences were detected between the 16S rRNA, wspA or wspB Wolbachia sequences obtained from M. occidentalis and T. urticae, which suggest that horizontal transfer of Wolbachia could have occurred. Southern blot analyses of genomic DNA from both M. occidentalis and T. urticae using wspA probes were negative, indicating that this Wolbachia sequence is not integrated into the nuclear genome of either species. Two of the T. urticae colonies tested contained the WO bacteriophage, but none of the six M. occidentalis populations were infected. New M. occidentalisspecific forward and reverse 16S rRNA primers based on the Wolbachia, Cardinium, Bacteroidetes, and Enterobacter sequences obtained were designed and used to amplify PCR products from each of two laboratory and four field-collected samples of M. occidentalis females and eggs, indicating that these infections are widespread. Likewise, species-specific primers for T. urticae were designed for the Wolbachia, Rickettsia, and Caulobacter sequences obtained and used to evaluate T. urticae from strawberries, wine grapes, hops, almonds, and cherries from California, Washington, and Florida; all were positive for Wolbachia and Caulobacter but two of the six were negative for Rickettsia. None of the M. occidentalis colonies tested were positive for the microsporidium Oligosporidium occidentalis, which previously had been associated with a pathogenic condition in some of our laboratory colonies. The Gainesville colonies of M. occidentalis and T. urticae were negative for iridovirus, Archaeabacteria, fungi, Helicosporidia, and yeast-like organisms. So far, Wolbachia is the only symbiont that is shared by this predator and its prey.