Three North Carolina populations of Belonolairnus longicaudatus differed significantly from three Georgia populations in stylet measurements, the c ratio, the distance of the excretory pore from the anterior end for both sexes; the a ratio for females only; and the total body length, tail length, and spicule length for males only. The Georgia nematodes were stouter, and the females possessed sclerotized vaginal pieces. The distal portion of the spicules of North Carolina males had an indentation and hump lacking in those of the Georgia males. The haploid number of chromosomes was eight for males from all populations of B. longicaudatus and a North Carolina population of B. maritimus. Interpopulation matings of the Tarboro, N.C. and Tifton, Ga. populations indicated that the offspring produced were infertile. Morphological differences and reproductive isolation suggest that the North Carolina and the Georgia populations belong to different species.