Termite alates and accompanying soldiers were collected during a 5-year period from diverse habitats on the Delmarva Peninsula, including inland hardwood sites (Newark, Delaware, U.S.A.; Galena, Maryland, U.S.A.), and a pine scrub beach (Lewes, Delaware, U.S.A.). Alates from 34 colonies were identified to species based on taxonomic keys. Pronotal width was measured for 1,447 accompanying soldiers from 33 of the colonies and compared to similar studies in Florida, U.S.A. Mitochondrial DNA 16S sequencing was conducted on soldiers from 31 of the colonies. Reticulitermes flavipes and R. virginicus were identified from pine scrub, whereas only R. flavipes was collected from the hardwood sites. DNA sequences showed three lineages of R. virginicus, two of which matched that from specimens previously reported to be a new species (R. malletei nomen nudum). Soldier pronotal width ranges at the 95% confidence level were: R. flavipes, 0.84–1.04 mm; R. virginicus, 0.63–0.83 mm. Statistical analyses indicated that species identification could be based on a sample of 5 soldiers with a confidence level of >95%. The observed pronotal width range for R. virginicus overlapped with that reported for R. hageni Banks in Florida, U.S.A.
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