Geographic distribution of Ovavesicula popilliae in the United States and sensitivity of visual diagnosis compared with qPCR detection.

The Japanese beetle (Popillia japonica) is one of the most destructive invasive pests in North America, causing significant economic impact to many fruit crops, turfgrass and the nursery industry. A microsporidian pathogen of Japanese beetle, Ovavesicula popilliae, discovered in 1985, proliferates in the Malpighian tubules of larvae and adults, disrupting waste-removal, mineral filtering, and fluid balance in heavily infected individuals. Most infected larvae do not survive from fall to spring, and egg production by infected females is reduced by 50%. Ovavesicula popilliae is promising as a classical biological control agent for Japanese beetle, but outside of surveys completed in Connecticut and Michigan little is known about its geographic distribution in North America. The objective of this research is to obtain a better understanding of the distribution of O. popilliae in North America. Japanese beetles were collected at 59 locations in a total of 19 different states in the USA for pathogen analysis. Overall, the proportion of Japanese beetle adults infected by O. popilliae was much greater in Michigan, Ohio, Tennessee and four states in the northeastern USA compared with sites located west of the Mississippi River (18.6 ± 13.3% and 0.6 ± 1.2%, respectively). Nucleotide sequences of the gene encoding a small subunit of nuclear ribosomal RNA (ssrDNA), obtained from GenBank for O. popilliae was used to develop a highly specific qPCR test for O. popilliae DNA. A subsample of 110 individual Japanese beetles were visually diagnosed first, then analyzed via qPCR. Visual diagnosis and qPCR detection agreed for 80.9% of the beetles tested. The qPCR assay is more sensitive than visual diagnosis (56 visually positive, 73 qPCR positive), is highly specific for O. popilliae, and will be useful for detecting the pathogen in large batches of beetles, or in beetle frass.