The effects of a water extract of cotton bracts (CBE) on guinea pig isolated trachealis smooth muscle was studied. The ability of CBE to evoke tension responses, to modify tissue reactivity to excitatory and inhibitory agents, and to modify electric field stimulation-induced neurogenic responses was evaluated. CBE caused contraction in low concentrations, which were not mediated by histamine H1-, muscarinic, or 5-hydroxytryptamine (5-HT) receptors, and caused relaxation in high concentrations. In the presence of CBE, the maximum contractile response to 5-HT and the sensitivity to KCl were reduced. The maximum relaxation responses and sensitivities to adenosine and ATP were increased by CBE. In contrast, contractile responses to histamine and methacholine and relaxation responses to isoproterenol were unaffected. Neurogenic cholinergic excitatory responses and neurogenic adrenergic responses did not appear to be affected by CBE. However, the relaxant effect of nonadrenergic inhibitory nerve stimulation was increased in the presence of CBE. These findings differ from previous ones observed in our laboratory for dog isolated trachealis smooth muscle preparations, which indicates that the effects of CBE in isolated airways is species-dependent. Virtually all of the contractile activity of CBE was dialyzable (molecular weight less than 14,000) and retained in ashed samples. Inorganic constituents may contribute to the spasmogenic activity of CBE.