Previous inventories of the diversity of lepidopteran symbionts have been limited to Eubacteria. We conducted a microbial survey of Calyptra thalictri Borkhausen (Lepidoptera: Noctuidae) by using polymerase chain reaction (PCR) primers for 16S rRNA sequences for Eubacteria, and primers for Archaea, fungi including yeast-like organisms, Microsporidia, and Wolbachia. Heads and abdomens of adult males of this fruit-piercing and blood-feeding moth were assayed separately. High-fidelity PCR and subsequent DNA analyses indicated that at least five microorganisms belonging to the α-, β-, and γ-Proteobacteria were present. Two eubacterial sequences, related to a Klebsiella sp. and a Sinorhizobium sp., were detected in the abdomens of all nine individuals sampled, and three additional sequences, two related to species in the genus Alcaligenes and one related to a Rhizobium sp., were found in some of the abdominal samples, suggesting all five could be associated with abdominal structures. No Archaea, fungi including yeast-like organisms, Microsporidia, or Wolbachia were detected. These results document the first microbial associates in a fruit-piercing and blood-feeding moth.