Employing adult Hymenolepis diminuta SMP and exogenous pyridine nucleotide-generating systems, reduced pyridine nucleotide-dependent net 32P incorporation into ATP was examined. NADH supported rotenone-sensitive 32P incorporation and this rate increased markedly with fumarate addition, in keeping with an active fumarate reductase. Interestingly, corresponding evaluations with NADPH did not result in detectable phosphorylation in the absence or presence of fumarate. However, with NAD addition, but without NAD generation, active NADPH-dependent phosphorylation occurred, thereby demonstrating mitochondrial transhydrogenase involvement, and 32P incorporation increased significantly with fumarate addition. More importantly, in the presence of rotenone and both NADPH and NAD generation, significant net 32P incorporation was noted, but was undetectable in the presence of DCCD or protonophores (e.g., niclosamide). Without NAD generation, minimal phosphorylation occurred. These data demonstrate that with ongoing NADPH and NAD generation, the H. diminuta, proton-translocating, mitochondrial transhydrogenase can serve as an additional anaerobic phosphorylation site. A model is presented.