Spontaneous phasic contractions recorded from isolated body strips of Fasciola hepatica were increased in frequency and amplitude by GYIRFamide, an FMRFamide-related peptide (FaRP). Superfusion with guanosine 5'-O-(2-thiodiphosphate) (100 µM, n = 5) reduced the effects of GYIRFamide on both frequency (by 82%) and amplitude (by 75%). The adenylate cyclase inhibitor MDL-12330A (25 µM) increased spontaneous activity. MDL-12330A completely inhibited the frequency response to GYIRFamide and reduced the amplitude response by 66% as measured relative to this elevated basal activity (n = 6). Inhibition of phospholipase C (PLC) with neomycin sulfate (1 mM) had no direct effect on activity but reduced the frequency response to GYIRFamide by 64% and the amplitude increase by 95% (n = 9). The protein kinase C (PKC) inhibitor chelerythrine chloride (10 µM) also reduced frequency and amplitude responses by 98 and 99%, respectively, without affecting basal contractility (n = 5). Phorbol 12-myristate 13-acetate, an activator of PKC, increased contraction frequency and amplitude (n = 6). It was concluded that GYIRFamide stimulates mechanical activity in F. hepatica through a G protein, via a PLC- and PKC-dependent second messenger pathway.