Olive mill wastewater

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OMWW’s fractions separation, phenolic compounds identification and stabilization  

  • Productions of valuable fractions from OMWWs coming from six cultivars, Coratina, organic Coratina, Cellina, Koroneiki, Asprolia and Lianolia, using the multiple steps filtration system present in the laboratory of ISPA-CNR of Bari, Italy;
  •  The performance of separation plant was: MF fraction 29%, UF fraction 14.5 % and NF fraction 10 %, all respect the OMWW row material used; Characterization of the fractions recovered from multiple steps filtration system (Microfiltrate MF, Ultrafiltrate UF, Nanofiltrate NF) by HPLC-DAD analysis;
  • Characterization of the VW using HPLC-DAD analysis and HPLC-MS. Compounds identified were: Oleuropein, decarboxymethyloleuropeinaglycon (3,4-DHPEA-EDA), verbascoside, isoberbascoside, ligstroside, acyclodihydroelenolate, elenolic acid, oxidized verbascoside form , β-hydroxyverbascoside diastereoisomer (Cardinali et al. J. Agric. Food Chem. 2012, 60, 1822−1829), caffeyol ester of secologanoside and 6’-p-coumaroyl secologanoside, as well as, tyrosol, hydroxytyrosol, caffeic acid and coumaric acid.
  • All the fractions of Vegetative water that have been threated using the pilot scale extraction system were analyzed for their dry weight and was added in each fraction equal amount of the dry weight of fiber, in order to have the same ratio between fiber and dry weight. The fiber that has been used are maltodextrins, and acacia gum in the ratio 50:50 between polyphenols and coating material.
  • An innovative process using carbon-based nanomaterials and zero-valent iron has been developed for the treatment of OMW and the reduction of the initial polluting load. The OMW treatment with graphene led to a 60% reduction of COD and up to 90% of total phenols. The application of multi-walled carbon nanotubes showed a 65% reduction of COD and 60% of total phenols, whereas treatment with nano-Fe0/Fe2O3 caused a 55% reduction of COD and 70% of total phenols. All the processes are promising for the recovery or degradation of the phenolic content of OMWW.

Antimicrobial activity

  • The effect of OMWW obtained from Italian and Greek cultivars, and some of their pure biophenols compounds, on the A.flavus growth and aflatoxin production was investigated. A total inhibition of A. flavus growth in the samples amended with OMWW Lianolia cv, at a maximum dose (15%) was recorded. Furthermore, at the doses tested, a change in the mycelia morphology of the fungi cultures grown on Lianolia and Koroneiki OMWW was observed.
  • Tyrosol, Verbascoside and Caffeic acid showed a significant increase in mycelium dry weight, while the reduction (P<0.05) of AFB1 content ranging from 80 to 100%.
  • OMWW may be a promising natural source of bioactive compounds useful in food safety control.

Biological assay

  • Protocols for the evaluation of the antioxidant activity of the UF fractions coming from the six OMWWs cultivars, by using Low Density Lipoprotein (LDL) assay;
  • All the cultivars analyzed showed a high potential of LDL inhibition.
  • Also Verbascoside and caffeic acid were investigated for their potential as antioxidant in the inhibition effect on LDL oxidation
  • A photo-protective role, on skin cells,  of some polyphenols usually present in OMWW, such as verbascoside, isoverbascoside and tyrosol, was found. The polyphenols were investigated for their effect on HEKa (Human Epidermal Keratinocytes adult) cell cultures challenged from UVA-rays (365 nm). The effect of UVA-rays on HEKa cultures induces a decrease of cell viability, caused by cell necrosis.
  • The simultaneous treatment with each polyphenol caused a decrease of ROS production, corroborating the antioxidant role, by ROS quenching, in the skin cells, keratinocytes.
  • Treatment with polyphenols after the UVA- irradiation, exerted a protective, anti-cancer action, by ROS-mediated apoptosis in UVA-damaged cells. The increasing of ROS levels, acting as signal molecules, was associated with the pro-apoptotic effect on keratinocytes, mainly evident in UVA-damaged surviving cells treated with the polyphenols after the UVA-irradiation.
  • The Ultra-filtered OMWW coming from the milling of three Italian cultivars  (Coratina, organic Coratina and Cellina) and three Greek cultivars (Koroneiki, Lianolia, Asprolia) of olives, have exerted similar protective effect on human keratinocytes. 
  • Effects on "anti-aging" activities (stimulation of antioxidant defenses of the cells of the dermis). The preparations tested showed activity at concentrations between 0.01 and 0.05% comparable to those of control consists of Vitamin E 100 micromolar.
  • Effects on anti-inflammatory capacity. The preparations obtained from the vegetation water at the same concentrations used for “anti-aging” activities are able to inhibit the formation of nitric oxide (NO) induced by bacterial infections.
  • Effect of detoxification. The formulations are able to slow the damage induced by UV rays stimulating the detoxifying activity of the proteasome Cell