As myxozoan actinospores are stimulated by fish epidermal mucus to attach to their hosts via extrusion of filaments from specialized organelles, the polar capsules, mucus components were tested for discharge triggering activity on Myxobolus cerebralis actinospores. Using various methodological approaches, a selective exclusion of candidate substances based on experimental outcome is provided and the physiochemical traits of the putative agents are explored to create a basis for the isolation of the host recognition chemostimuli. Activity was detected in compounds that can be characterized as small molecular, amphiphilic to slightly hydrophobic organic substances. They were separable by chromatographic methods using reversed phase C18 supports. An active fraction was isolated by solid phase extraction comprising at least nine UV-detectable constituents as shown by thin-layer chromatography. By means of biochemical fractionation and analysis of host fish mucus, non-volatile inorganic electrolytes, all volatiles, free L-amino acids, glycoproteins, bound and free hexoses, sialic acids, glycans, proteins, urea, amines and inositols were shown not to trigger polar filament discharge. The results contribute to the identification of the attachment host cues and enable a more focused laboratory activation of myxozoan actinospores.