Reduction in vegetative growth of the water mold Saprolegnia parasitica (Coker) by humic substance of different qualities

Humic substances (HS) account for 50-80% of the dissolved organic matter in non-eutrophicated freshwater ecosystems. HS are not inert, but are taken up by and may interact with aquatic organisms. However, at present no information is available on the interaction of HS with fungi, for instance, the fish-pathogenous species Saprolegnia parasitica. To fill this gap, we tested effects of HS on S. parasitica growth in-vitro using 25-500 mg L-1 carbon of HS on GY-agar. We investigated 20 HS including natural organic matter (NOM) samples, two lignite derived HS, and one synthetic HS. The aim was (1) to find out, if there are inhibiting effects and (2) if potential effects can be explained by humic matter properties by structure activity relationships. The growth of S. parasitica was related to the growth on HS-free agar controls. Characterization of HS and NOMs included elemental analysis, high-pressure size exclusion chromatography (HPSEC), UV/VIS, FTIR-, and EPR-spectroscopy in order to obtain information on elemental and structural composition including various metals, molecular weights of the HS fraction, aromaticity, free organic radicals, and functional groups. NOMs with high moieties of high-molecular carbohydrates supported the growth of S. parasitica, all other HS and NOMs reduced it. However, no inhibition of the development of the sporangia and primary zoospores was found. Therefore, the impact of the HS on S. parasitica has to be classified as fungistatic, rather than fungicidal. Synthetic and lignite-derived HS were among the most efficient HS sources. Growth inhibition was correlated (p < 0.05) with the molecular weights of the HS-fraction, sUVa, COOH groups, C and H. Our results suggest that especially HS with higher molecular weights and aromaticity which contain a high number of organic radicals are the most efficient in reducing fungal growth. Furthermore, highly functionalized HS seem to be important for the observed effect. The development of internal oxidative stress could be a mechanism explaining the observed growth inhibition of S. parasitica.