Toxicity evaluation of olive oil mill wastewater and its polar fraction using multiple whole-organism bioassays

Olive mill wastewater (OMW) as a by-product of olive oil extraction process has significant polluting properties mainly related to high organic load, increased COD/BOD ratio, high phenolic content and relatively acidic pH. Raw OMW from Slovenian Istria olive oil mill and its polar fraction were investigated in this study. Chemical characterization of OMW polar fraction identified tyrosol as the most abundant phenolic product, followed by catechol. Lethal and sub-lethal effects of OMW matrix and its polar fraction were tested using a battery of bioassays with model organisms: bacteria Vibrio fischeri , algae Chlorella vulgaris , water fleas Daphnia magna , zebrafish Danio rerio embryos, clover Trifolium repens and wheat Triticum aestivum . Raw OMW sample was the most toxic to V. fischeri (EC 50 = 0.24% of OMW sample final concentration), followed by D. magna (EC 50 = 1.43%), C. vulgaris (EC 50 = 5.20%), D. rerio (EC 50 = 7.05%), seeds T. repens (EC 50 = 8.68%) and T. aestivum (EC 50 = 11.58%). Similar toxicity trend was observed during exposure to OMW polar fraction, showing EC 50 values 2.75–4.11 times lower comparing to raw OMW. Tested samples induced also sub-acute effects to clover and wheat (decreased roots, sprouts elongation); and to zebrafish embryos (increased mortality, higher abnormality rate, decreased hatching and pigmentation formation rate). A comprehensive approach using a battery of bioassays, like those used in this study should be applied during ecotoxicity monitoring of untreated and treated OMW. • Raw OMW and polar fraction exhibited the highest toxicity on the photobacterium Vibrio fischeri . • Tyrosol was the most common phenolic compound in the tested OMW. • Multi-biomarker approach showed as a good indicator of OMW toxic potential.