Developmental physiology of cestodes: characterization of putative crowding factors in Hymenolepis diminuta

It was shown previously that worm-conditioned saline (WCS) prepared from crowded 10-day-old H. diminuta inhibited the incorporation of 3H-thymidine into DNA in the anterior regions of uncrowded worms and that the inhibition was partially accounted for by succinate and acetate excreted by the worms. The present study describes further characterization of the active components of WCS. An ultrafiltrate was fully as potent as untreated WCS, indicating that all detectable inhibitory components were less than about 500 daltons in molecular mass. Inhibitory factors in WCS were stable to heat (80 C for 30 min), cold (4 C for 48 hr), drying and reconstitution, alkaline pH (11 to 12 for 3 hr), and ethanolic extraction. Active compounds were probably not lipoidal in nature. Although the acidic ethanol extract of WCS was inhibitory, no activity was observed in fractions of WCS that contained basic, acidic and neutral amino acids. Amino compounds in the WCS were further investigated. Twenty-four amino acids were identified, 3 of which (phosphoserine, 1-methylhistidine, and 3-methylhistidine) have not been reported previously for H. diminuta. On a molar basis, alanine accounted for 40-50% of the amino acids released. The amino sugar, D-glucosaminic acid, was found in the WCS and also has not been heretofore reported from H. diminuta or any other cestode. In concentrations comparable to those in the WCS, D-glucosaminic acid inhibited incorporation of 3H-thymidine into the DNA of the tapeworms by 25-35%, suggesting that D-glucosaminic acid may be one of the crowding factors.