Concentrations of dissolved organic carbon (DOC) and total phenols (TP), and the phytotoxicity to cress (Lepidium sativum L.) were determined for three molecular-sized fractions of olive mill wastewater (OMW), <1000, 1000-5000, and >5000 Da, before and after incubation with Pleurotus ostreatus. The <1000-Da fraction contained 82% of the total DOC and 48% of the TP, and was the most phytotoxic. Ethyl acetate separation of aqueous and solvent fractions showed that the aqueous fraction contained 93% of the total DOC, 83% of the TP, and was most phytotoxic, indicating low importance of monomeric phenols. Incubation of whole OMW and of the separate size fractions with P. ostreatus mycelia reduced TP by factors of 4.3-5.3, but exerted diverse impact on phytotoxicity; overall, P. ostreatus efficacy in organic load removal and OMW detoxification was limited. Additional size fractionation of the incubated fractions revealed that most residual phytotoxicity was associated with low-molecular weight (MW) compounds originated from the <1000 Da fraction and not with low-MW byproducts from the degradation of higher-MW fractions and that polymerized metabolites were nonphytotoxic. Total phenols should not be used as sole indicators of the successful remediation of OMW.
Controlled land spreading of untreated olive mill wastewater (OMW) has been widely practiced as a means of its disposal. However, potential phytotoxic effects are critical for the selection of sites and crop types and for proper synchronization between land application and cropping. This study traced the fate of dissolved organic carbon (DOC), total phenols (TP), electrical conductivity, pH, microbial counts, and phytotoxicity to cress ( L.) after soil application at doses equivalent to 80, 160, and 320 m ha. Vertisol (fine-clayey) and Loess (sandy loam) soils were treated and incubated at 12 or 25°C and at moisture contents maintained at 70% of field water capacity or gradually reduced from 70 to 20% without compensation. Temperature, rather than moisture content, had a major effect on removal rates of DOC and TP. The maximum combined effect of warm temperature and higher moisture content resulted in removal rates greater than those under cooler, drier conditions by factors of up to 1.8 and 4.1 for DOC and TP, respectively. Favorable biodegradation conditions were indicated by increased numbers of total soil microorganisms and fungi by factors of up to 26 and 5, respectively. A whole-soil bioassay was developed to assess the dynamics of residual soil phytotoxicity after OMW application. Phytotoxicity measurement in soil extract generally showed stronger inhibition or stimulation activity than measurement in whole soil, depending on soil type and OMW dose. The newly developed bioassay seems to be useful for the refinement of general recommendations regarding permitted OMW application doses.
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