The fluted kidneyshell Ptychobranchus subtentum (Say, 1825) is a candidate for listing under the Endangered Species Act by the US Fish and Wildlife Service. Fecundity, fish hosts, and selected population demographics were determined during 2005–2006 for the fluted kidneyshell in the upper Clinch River, Hancock County, Tennessee. Females were fertilized in Aug. within a 5 d period and contained viable glochidia about 4 wk later. As the embryos began to develop, the marsupium gradually changed color from white to dark brown. Glochidia were contained within conglutinates that resemble Simuliidae pupae likely to attract benthic insectivorous fish and were held over winter and released in May. Fecundity was positively related to mussel length (r²  =  0.81) and ranged from 43,000 to 500,000 glochidia. Eight species of darters (Etheostoma spp. and Percina spp.) were infested with glochidia in the laboratory to examine potential hosts and host suitability. Juveniles transformed on bluebreast darters E. camurum and dusky darters P. sciera and previously reported hosts: rainbow darters E. caeruleum and fantail darters E. flabellare. In addition, fantail darters and rainbow darters were infested with glochidia from two river systems. The median time of glochidial metamorphosis did not differ significantly between the two mussel populations. The observed ratio of adult females to males (1.9∶1) in the Clinch River differed significantly from 1∶1. Based upon thin-sections, individuals live to at least 26 y and females become sexually mature at age five.