ARTICLE IN PRESS

International Biodeterioration & Biodegradation 61 (2008) 127–134

www.elsevier.com/locate/ibiod

Review

Bioremediation of olive mill wastewater

Christopher J. McNamaraa,b,c,, Christos C. Anastasiouc,1,
Vincent O’Flahertyd, Ralph Mitchellb
a
Exposure, Epidemiology and Risk Program, Department of Environmental Health, Harvard School of Public Health, Boston, MA 02215, USA
b
Laboratory of Applied Microbiology, Harvard School of Engineering and Applied Sciences, Cambridge, MA 02138, USA
c
Cyprus International Institute for the Environment and Public Health in association with Harvard School of Public Health, Nicosia, Cyprus
d
Microbial Ecology Laboratory, Department of Microbiology and Environmental Change Institute, National University of Ireland, Galway, Ireland
Received 22 October 2007; received in revised form 9 November 2007; accepted 9 November 2007
Available online 21 December 2007

Abstract

The vast majority of olive oil production (498%) occurs in the Mediterranean region, utilizing a tremendous volume of water
(10–30 million m3) in an area of the world in which water resources are limited. Treatment and reuse of olive mill wastewater (OMWW)
presents significant challenges both due to the nature of olive oil production (seasonal and small scale) and due to the characteristics of
the wastewater (high chemical oxygen demand (COD), high phenolic content, and dark color). A number of different microorganisms
(Archaea, Bacteria and fungi) and processes (aerobic or anaerobic bioreactors, composting) have been tested to treat OMWW. Aerobic
bacteria have been tested primarily as an approach for removal of phytotoxic compounds from OMWW, although some studies have
also focused on reduction of COD. Fungi on the other hand, have proven effective at reducing COD and toxicity. Anaerobic consortia
can effectively reduce COD, but are sensitive to phenolics in OMWW. Biological processes provide some of the most viable options for
the treatment of OMWW. Effective application of these techniques, yielding significant reductions in COD, phenolics, and color, will
allow safe and economical disposal of OMWW.
r 2007 Elsevier Ltd. All rights reserved.

Keywords: Olive mill wastewater; Bioremediation; Phytotoxicity; Phenolics; COD

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
1.1. Olive oil production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
1.2. OMWW characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
1.3. Olive oil production and the environment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
1.4. OMWW treatment methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
2. Microbial treatment of OMWW . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
2.1. Aerobic microorganisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
2.1.1. Aerobic bacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
2.1.2. Fungi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
2.1.3. Combined bacterial–fungal systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
2.2. Anaerobic microorganisms. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
2.3. Combined aerobic–anaerobic systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131

Corresponding author. Exposure, Epidemiology and Risk Program, Department of Environmental Health, Harvard School of Public Health, Boston,
MA 02215, USA. Tel.: +1 617 384 8749.
E-mail address: cmcnamar@hsph.harvard.edu (C.J. McNamara).
1
Current address: Department of Civil Engineering, Frederick University Cyprus, Nicosia, Cyprus.

0964-8305/$ - see front matter r 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.ibiod.2007.11.003
 ARTICLE IN PRESS
128 C.J. McNamara et al. / International Biodeterioration & Biodegradation 61 (2008) 127–134

3. Bioreactors used for direct OMWW treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131

4. Composting . . . . . . . . . . ............... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
5. Conclusions . . . . . . . . . . ............... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
Acknowledgments . . . . . . ............... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
References . . . . . . . . . . . ............... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132

1. Introduction number of small olive mills scattered throughout the olive

oil-producing countries.
1.1. Olive oil production
1.2. OMWW characteristics
Mediterranean countries produce more than 98% of the
world’s olive oil, which is estimated at over 2.5 million OMWW has a very high organic load (chemical oxygen
metric tons per year. About 75% is produced in the demand (COD) typically ranges from 50 to 150 g l 1, or
European Union (EU). The largest olive oil producers are about two orders of magnitude higher than municipal
Spain, with 36% of the global production, Italy, which wastewater). OMWW also contains high levels of phyto-
produces 24%, and Greece, which manufactures 17% of toxic and microbially inhibitory compounds, such as
the world’s total. The next largest producer is Portugal, phenolics and long-chain fatty acids. The main organic
with production one order of magnitude lower than the constituents of OMWW are sugars and polyphenols.
three leading countries, followed by France, Cyprus and Sugars reported in OMWW include fructose, mannose,
Croatia (FAOSTAT, 2007). Over the last decade, olive oil glucose, saccharose, sucrose and pentose. Phenolic com-
production has increased about 40% worldwide and pounds that are present in olive stones and pulp tend to be
Europe has witnessed a 45% increase in production. more soluble in the water phase than oil, resulting in
Spanish olive oil production, which represents about half concentrations ranging from 0.5 to 25 g l 1 in OMWW.
of European production, has shown an increase of almost More than 30 different phenolic compounds have been
80% during that time (FAOSTAT, 2007). identified in OMWW and the types and concentrations of
Multiple methods are used in the production of olive oil, phenolics reported in OMWW vary tremendously. Wide
resulting in different waste products. Olive presses, the ranges in all of these characteristics have been reported and
technology traditionally used for the extraction of olive oil, the composition of OMWW varies substantially not only
have been replaced in modern olive oil plants by decanter by region, but also by the type of process used, daily usage
centrifuges that separate the oil from the solids. Currently, conditions, and local operational procedures (Niaounakis
two-phase and three-phase centrifugation systems are most and Halvadakis, 2006).
commonly used (Niaounakis and Halvadakis, 2006). The The presence of phytotoxic phenolics generally prohibits
environmental impact of olive oil production is consider- the use of untreated OMWW for irrigation purposes in
able, mainly due to the utilization of large amounts of agricultural production (e.g., El Hadrami et al., 2004;
water and the production of large amounts of either Mekki et al., 2006). The complex composition of OMWW
wastewater or sludge. Pressure and three-phase centrifuga- coupled with the seasonal nature of olive production and
tion systems produce substantially more olive mill waste- the wide geographical dispersion of mills poses consider-
water (OMWW) than two-phase centrifugation, able technical and economic barriers for efficient effluent
which significantly reduces liquid waste yet produces treatment and disposal (Marrara et al., 2002).
large amounts of semi-solid or slurry waste commonly
referred to as two-phase pomace or alperujo in Spanish 1.3. Olive oil production and the environment
(Alburquerque et al., 2004). In the past 10 years, the
two-phase system has become dominant in Spain and An estimated 10–30 million m3 of OMWW is generated
in Croatia. However, the two-phase-system has not every year from the production of olive oil (Niaounakis
penetrated significantly into the other olive oil-producing and Halvadakis, 2006). As an example of the scale of the
countries, mainly due to difficulty in handling the environmental impact of OMWW, it should be noted that
sludge. Most countries continue to make use of the three- 10 million m3 year 1 of liquid effluent from three-phase
phase centrifugation system (Kotronarou and Mendez, systems corresponds to an equivalent load of the waste-
2003). water generated from about 20 million people. Further-
Treatment of OMWW is complicated by the seasonal more, the fact that most olive oil is produced in countries
and geographically diffuse nature of olive oil production. that are deficient in water and energy resources makes the
Olive production varies significantly from year to year need for effective treatment and reuse of OMWW critical.
(trees typically bear the most fruit every second year). The dark coloration, high biological and chemical
Furthermore, olives are collected and oil is extracted in the oxygen demand (BOD/COD), and the presence of phyto-
winter months (primarily from early November to late toxic substances prohibit direct discharge of wastewater
February). Finally, the oil is typically extracted in a large produced in the processing of olives into fresh and coastal
 ARTICLE IN PRESS
C.J. McNamara et al. / International Biodeterioration & Biodegradation 61 (2008) 127–134 129

waters or onto land. Discharge of OMWW directly onto et al., 1996; Ehaliotis et al., 1999; Piperidou et al., 2000),
soil may impact soil physical and chemical properties such Pseudomonas putida and Ralstonia sp. (Di Gioia et al.,
as porosity and pH (Niaounakis and Halvadakis, 2006). 2001a, b), and various bacterial consortia (Ranalli, 1992;
However, the main obstacle to direct use of OMWW for Borja et al., 1995a; Zouari and Ellouz, 1996a; Benitez et al.,
irrigation is the high concentration of phenolic compounds, 1997).
which are phytotoxic and can inhibit plant seed germina- Aerobic bacteria have been tested primarily as an
tion. OMWW also has significant impacts when discharged approach for removal of phytotoxic compounds (i.e.,
directly into surface waters (Niaounakis and Halvadakis, monoaromatic or simple phenolics) from OMWW,
2006). The high concentration of darkly colored poly- although some studies have also focused on reduction of
phenols in OMWW can discolor streams and rivers. In COD. Ramos-Cormenzana et al. (1996) reported a 50%
addition, the high concentration of reduced sugars can reduction in the phenolic content of OMWW by B. pumilis.
stimulate microbial respiration, lowering dissolved oxygen Likewise, A. vinelandii has been shown to effectively reduce
concentrations, while the high phosphorus content can lead the phytotoxicity of OMWW (Constantinos et al., 1999;
to eutrophication. Piperidou et al., 2000). Zouari and Ellouz (1996a) reported
a 50% reduction in COD and almost complete removal of
1.4. OMWW treatment methods phytotoxicity using an enrichment culture of organisms
isolated from industrial and municipal wastes and agricul-
OMWW treatment processes that employ physical, tural soil. Generally, the aerobic bacteria that have been
chemical, biological and combined technologies have been tested appear to have a minimal effect on the more complex
tested, and reviewed by others (e.g., Niaounakis and polyphenolics responsible for the dark coloration of
Halvadakis, 2006; Rozzi and Malpei, 1996). In addition OMWW.
to being technically feasible, OMWW treatment processes The effectiveness of aerobic bacteria in reducing the
must be efficient, allow for easy and economical operation phytotoxicity of OMWW varies greatly. Aerobic bacteria
in small-scale farm settings, and consider the seasonality appear to be very effective against some phenolic
and the spatial distribution of olive oil production. compounds and relatively ineffective against others. For
Common disposal practices for OMWW from three- example, B. pumilus was able to completely degrade
phase systems in Europe include direct discharge into soils protocatechuic acid and caffeic acid, but had much less
or streams and use of evaporation ponds or lagoons affect on tyrosol (Ramos-Cormenzana et al., 1996). On the
(Kotronarou and Mendez, 2003). These methods do not other hand, Knupp et al. (1996) reported complete
alleviate the high COD or toxicity of OMWW. For transformation of tyrosol to 4-hydroxyphenylacetic acid
example, the use of evaporation ponds reduces the volume by Arthrobacter sp.
of waste without treating the pollutants in the wastewater. Several studies of bioremediation of OMWW in Greece
Additionally, the use of evaporation ponds contributes to have focused on A. vinelandii. Papadelli et al. (1996)
odor problems as well as leading to the loss of water from isolated a strain of A. vinelandii from soil treated with
areas that are mostly deficient in this resource. OMWW. Eventually, 490% removal of phytotoxic
A variety of biological processes (e.g., aerobic or compounds from OMWW was achieved using this strain
anaerobic bioreactors, composting) and microorganisms (Ehaliotis et al., 1999; Piperidou et al. 2000). Di Gioia et al.
have been tested to treat OMWW to remove the dark (2001a, b) used two bacteria, Ralstonia sp. and P. putida, to
coloration, reduce the organic load and remove phytotoxic treat Italian OMWW. Cultured together, these bacteria
compounds. In this review, we describe the microorganisms were able to degrade many phenolic compounds found in
and methods used to directly treat OMWW. We have not OMWW.
included the small, but growing body of literature A number of studies have utilized bacterial consortia for
describing the abiotic use of microbially derived enzymes bioremediation of OMWW. The consortia have come from
for treatment of OMWW. These studies have, with some activated sludge (Borja et al., 1995a; Benitez et al., 1997),
degree of success, employed peroxidases and laccases commercial communities (Ranalli, 1992), soil, and waste-
produced by white rot fungi (e.g., see Sayadi and Ellouz, water (Zouari and Ellouz, 1996a). Bioremediation of
1995; Perez et al., 1998; D’Annibale et al., 2000). OMWW using aerobic consortia has been quite successful
in these studies, achieving significant reductions in COD
2. Microbial treatment of OMWW (up to 80%) and the concentration of phytotoxic com-
pounds, and complete removal of some simple phenolics.
2.1. Aerobic microorganisms

2.1.1. Aerobic bacteria 2.1.2. Fungi

A number of different bacteria have been tested in Fungal remediation of OMWW has been studied using
aerobic processes to treat OMWW, including Bacillus three types of organisms: white rot fungi (including the
pumilus (Ramos-Cormenzana et al., 1996), Arthrobacter sp. edible mushrooms Lentinula and Pleurotus), Aspergillus
(Knupp et al., 1996), Azotobacter vinelandii (Papadelli sp., and several different yeasts. In addition to reduction of
 ARTICLE IN PRESS
130 C.J. McNamara et al. / International Biodeterioration & Biodegradation 61 (2008) 127–134

COD and removal of simple phenolics, fungi are also C. tropicalis. Reduction of COD (55–63%) and phenolics
effective at reducing coloration of OMWW. (44–52%) was similar for all three fungi. Garcia Garcia
A variety of white rot fungi have been used for et al. (2000) compared two white rot fungi (Phanerochaete
remediation of OMWW, including Coriolus versicolor and chrysosporium and G. candidum) and two species of
Funalia trogii (Yesilada et al., 1995; Yesilada et al., 1998), Aspergillus (A. niger and A. terreus). No removal of
Geotrichum candidum (Assas et al., 2000), Lentinula phenols was observed for G. candidum. For the other
(Lentinus) edodes (Vinciguerra et al., 1995; Vinciguerra organisms, the efficiency of the removal of total phenols
et al., 1997; D’Annibale et al., 1998) and Phanerochaete sp. relative to the total organic load consumed occurred in the
(Sayadi and Ellouz, 1992; Ben Hamman et al., 1999). order P. chrysosporium4A. niger4A. terreus.
A number of studies have examined the ability of the edible
white rot fungus Pleurotus to degrade OMWW (Sanjust
2.1.3. Combined bacterial–fungal systems
et al., 1991; Tomati et al., 1991; Kissi et al., 2001; Tsioulpas
De Felice et al. (1997) used a combination of bacteria
et al., 2002). The white rot fungi appear quite effective,
and yeast to degrade OMWW. The yeast, Yarrowia
achieving removal rates as high as 88% for COD, 100% for
lipolytica, reduced the COD of OMWW by 80% in 24 h,
phenolics, and 81% for coloration (Table 1).
leaving aromatic compounds undegraded. Effluent from
Use of Aspergillus for bioremediation of OMWW has
the yeast fermentation was then treated with P. putida,
not been as common as the white rot fungi. Hamdi and co-
producing further reduction of COD and removal of
workers, in a number of studies, characterized the ability of
aromatics in 12–16 h.
Aspergillus niger to degrade OMWW (Hamdi et al.,
1991a, b; Hamdi and Ellouz, 1992a, b, 1993). Reduction
of COD and phenol concentrations of greater than 60% 2.2. Anaerobic microorganisms
were achieved using A. niger. Similar reductions in COD
and phenolics were found using Aspergillus terreus Anaerobic bioremediation of OMWW has employed,
(Martinez Nieto et al., 1993). almost exclusively, uncharacterized microbial consortia
Yeasts used in the bioremediation of OMWW include derived from municipal and other waste facilities (Rozzi
Trichosporon cutaneum (Chtourou et al., 2004), Candida et al., 1989; Martin et al., 1991; Borja et al., 1996; Dalis et
tropicalis (Ettayebi et al., 2003) and Saccharomyces sp. al., 1996; Fiestas Ros de Ursinos and Borja-Padilla, 1996;
(Gharsallah, 1993). Like the other fungi, these yeasts were Zouari and Ellouz, 1996b; Marques et al., 1997; Marques,
effective at reduction of COD and removal of mono- and 2001). A significant advantage of anaerobic processes over
polyphenols. aerobic may be the generation of methane, which could
Two studies have compared the effectiveness of different potentially be used in remediation or as an energy
types of fungi for the bioremediation of OMWW. Fadil source for other processes (Fiestas Ros de Ursinos and
et al. (2003) compared Geotrichum sp., Aspergillus sp. and Borja-Padilla, 1996).

Table 1
Effectiveness of white rot fungi for bioremediation of OMWW

Fungus Maximum % removal Reference

COD Total phenolics Color

Coriolus versiclor 63 90 65 Yesilada et al. (1998)

Geotrichum candidum 62 70 Assas et al. (2000)

Funalia trogii 40 77 38 Yesilada et al. (1995)

70 93 81 Yesilada et al. (1998)
Lentinula (Lentinus) edodes 75 66 45 Vinciguerra et al. (1995)
88 89 75 D’Annibale et al. (1998)
Phanerochaete chyrsosporium 80 74 Sayadi and Ellouz (1992)
64 70 67 Kissi et al. (2001)
Phanerochaete flavido-alba 90 70 Ben Hamman et al. (1999)
Pleurotus sp. 76 Tsioulpas et al. (2002)
Pleurotus eryngii 490 Sanjust et al. (1991)
Pleurotus floridae 490 Sanjust et al. (1991)

Pleurotus ostreatus 490 Sanjust et al. (1991)

100 Tomati et al. (1991)

Pleurotus sajor-caju 490 Sanjust et al. (1991)

COD—chemical oxygen demand.

 ARTICLE IN PRESS
C.J. McNamara et al. / International Biodeterioration & Biodegradation 61 (2008) 127–134 131

Reductions in COD from 70% to 89% have been OMWW pretreated by G. candidum, A. terreus and
reported for anaerobic processes (Borja et al., 1996; A. chroococcum were enhanced 2.5-, 4.2- and 4.0-fold,
Marques et al., 1997; Marques, 2001). In addition to a respectively. One interesting factor of note from this study
substantial reduction of COD, Dalis et al. (1996) reported is that while the white rot fungi appear to the most effective
large reductions (475%) in the concentrations of both microorganisms in aerobic treatment processes (i.e., super-
toxic phenols and volatile fatty acids using a two stage ior to Aspergillus and bacteria), they are the least effective
anaerobic reactor with an inoculant obtained from a organisms for pretreating OMWW for anaerobic digestion.
domestic wastewater facility. In contrast, other studies
have reported that the build up of recalcitrant phenolics 3. Bioreactors used for direct OMWW treatment
(e.g., condensed tannins, Zouari and Ellouz, 1996b) as well
as the presence of long-chain fatty acids (Hwu and Bioreactor technology, which could be applied to remove
Lettinga, 1997) in anaerobic reactors inhibited microbial organic pollutants (measured as COD) from OMWW, is
activity. based on either aerobic or anaerobic bioreactor configura-
The presence of compounds toxic to methanogens in tions. Aerobic bioreactor configurations, such as activated
OMWW appears to be a significant problem for anaerobic sludge or sequencing batch reactors are energy intensive,
digestion of OMWW. One approach to the problem has require high nutrient inputs to support treatment, are
been to dilute the OMWW to reduce the concentration of complex to maintain and operate, produce large quantities
phenolics and fatty acids. However, this results in the need of waste biomass and have high capital construction costs
to treat a substantially larger volume of wastewater. (Gray, 1999). These are particularly significant drawbacks
A second approach, discussed below, has been the use of for treatment of relatively small-scale OMWW streams.
aerobic pretreatment of OMWW to remove compounds A variety of anaerobic bioreactor types have been
that are toxic to methanogenic consortia. tested for the treatment of OMWW. There are several
potential advantages with this approach including the
2.3. Combined aerobic–anaerobic systems production of a useable biogas fuel (typical yields of
7–8 m3 m 3 OMWW), low waste biomass generation
Fungi have been used effectively in the pretreatment of (0.15–0.25 kg kg 1 COD removed) and the generation of
OMWW prior to anaerobic digestion. Hamdi (1991) found an excellent soil conditioner and fertilizer (Lettinga, 1995;
that pretreatment of OMWW with A. niger more than Niaounakis and Halvadakis, 2006). A number of basic
doubled methane production in subsequent anaerobic bioreactor designs have been employed, including sus-
digestion. In other studies, anaerobic digestion after pended biomass systems (e.g., continuously stirred tank
pretreatment with A. niger removed over 60% of COD reactors (CSTR) and retained biomass systems), the upflow
and resulted in high methane yields (Hamdi et al., 1992; anaerobic sludge blanket (UASB), the anaerobic baffled
Hamdi and Ellouz, 1993). Similarly, studies using reactor (ABR), upflow and downflow anaerobic filters and
A. terreus have demonstrated that aerobic pretreatment the fluidized bed. More recently, the development of
greatly reduced the concentration of phenolics and granular activated carbon-based bioreactor systems has
significantly increased methane production (up to 23%, been reported (Bertin et al., 2006).
Borja et al., 1995b, c). Martinez-Garcia et al. (2007) used In order to overcome the poor availability of nitrogen in
the yeast C. tropicalis to aerobically pretreat OMWW prior OMWW and to stabilize bioreactor pH, concentrated
to anaerobic digestion. The combined system resulted in a ammonia is often added to the OMWW. A number of
93% reduction in COD and degradation of 54% of the authors have also successfully anaerobically co-digested the
phenolic content of the OMWW. OMWW with pig manures (Angelidaki and Ahring, 1997;
Two different white rot fungi have been used in the Angelidaki et al., 2002) and dairy effluents (Gavalla et al.,
pretreatment of OMWW prior to anaerobic digestion, with 1996), an approach which is potentially attractive as low-
quite different results. P. chrysosporium reduced the COD cost nutrients are provided for digestion, inhibitory
of OMWW, but apparently had little effect on polyphe- compounds present in the OMWW are diluted and the
nolics, which remained in the effluent and inhibited digester can be operated on a year-round basis (Gavalla et
subsequent methane production (Gharsallah et al., 1999). al., 1996; Angelidaki et al., 2002).
However, pretreatment with G. candidum reduced the The initial trials on direct anaerobic biotreatment of
COD, phenolic and volatile fatty acid content of OMWW OMWW were carried out using CSTR and anaerobic
and increased substrate uptake during anaerobic digestion contact bioreactors. However, a number of features,
(Martin et al., 1993). including the requirement for mechanical mixing and the
Borja et al. (1998) compared anaerobic digestion of need for prolonged (10–25 day) hydraulic retention times
OMWW pretreated by two different fungi and a bacterium: (HRTs) make these systems unattractive for full-scale
G. candidum, A. terreus and Azotobacter chroococcum. applications. A number of trials have also been carried out
These organisms decreased the phenolic concentration and with ‘‘second generation’’ bioreactors. For example,
toxicity of OMWW by 59%, 87% and 79%, respectively. stable performance has been reported in laboratory-scale
Subsequently, the kinetics of anaerobic digestion of UASB bioreactors at organic loading rates (OLRs) of
 ARTICLE IN PRESS
132 C.J. McNamara et al. / International Biodeterioration & Biodegradation 61 (2008) 127–134

5–18 kg m 3 day 1 (Boari and Mancini, 1990; Zouari and 5. Conclusions

Ellouz, 1996b; Ubay and Öztürk, 1997), with COD
removal efficiency of approximately 75% at HRTs of 1–2 Generation of OMWW in the Mediterranean region has
days. Khabbaz et al. (2004) reported stable and efficient a significant environmental impact. The seasonal nature of
COD removal in an ABR bioreactor system, but at a olive oil production, the geographic dispersion of mills and
maximum OLR of only 2.5 kg m 3 day 1. The use of an economic limitations for cost effective treatment all present
anaerobic filter, which combines a UASB and an anaerobic significant challenges in designing treatment options for
filter for treatment of diluted OMWW was reported by OMWW. Overall, the incorporation of biological processes
Borja-Padilla et al. (1996) for treatment of wash waters provides some of the most viable options for the treatment
derived from the purification of virgin olive oil with of OMWW. Effective bioremediation resulting in signifi-
encouraging results (COD removal efficiencies of 489% at cant reductions in COD, phenolics and color allows safe
an OLR of 17.9 kg m 3 day 1 and a HRT of 4.8 h). and economical disposal of OMWW onto land or into
A number of multi-phase anaerobic digester configura- surface waters. As an added benefit, bioremediation may
tions have also been employed. For example, the aqua- produce valuable products. Bioremediated OMWW is an
tecOLIVIA process is a two-phase anaerobic treatment excellent fertilizer (Cereti et al., 2004; Mekki et al., 2006)
approach, consisting of an initial acidification step (HRT and can serve as a substrate for nitrogen-fixing bacteria or
2.5 days) and a second methanogenic stage using a moving polymer production (Balis et al., 1996).
bed reactor (HRT 8 days), with hollow cylindrical supports
as a carrier. A semi-pilot process was installed in Crete in
1999 and achieved COD removal efficiencies of 92–97% at Acknowledgments
imposed OLRs (Niaounakis and Halvadakis, 2006). A
second pilot configuration involving an upflow-sludge bed We thank K. Bearce Lee and B. Ross for assistance in
and an anaerobic filter operated in series was described by collecting references and compiling the bibliography. This
Dalis et al. (1996), and achieved a combined 91% COD work was supported by the Harvard School of Public
removal efficiency at an OLR of 11 kg m 3 day 1. The use Health Cyprus Initiative for the Environment and Public
of two stage systems is potentially attractive as a degree of Health.
phase separation can be achieved. Saturation of long-chain
fatty acids and acidification would predominate in the first
References
phase, allowing efficient methanogenesis of the acidified
stream in the second phase. Alburquerque, J.A., Gonzalvez, J., Garcia, D., Cegarra, J., 2004.
Agrochemical characterisation of ‘‘alperujo,’’ a solid by-product of
the two-phase centrifugation method for olive oil extraction.
4. Composting Bioresource Technology 91, 195–200.
Angelidaki, I., Ahring, B.K., 1997. Codigestion of olive oil mill
wastewaters together with manure, household waste or sewage sludge.
In addition to the use of bioreactors, composting has Biodegradation 8, 221–226.
been used to treat OMWW. The most common methodol- Angelidaki, I., Ahring, B.K., Deng, H., Schmidt, J.E., 2002. Anaerobic
ogy employed has been the Rutgers strategy (Finstein et al., digestion of olive oil mill effluents with swine manure in UASB
1985). A number of different bulking agents have been used reactors. Water Science and Technology 45, 213–218.
in compost with OMWW, including solid olive and olive Assas, N., Marouani, L., Hamdi, M., 2000. Scale down and optimization
of olive mill wastewaters decolorization by Geotrichum candidum.
tree waste (Papadimitriou et al., 1997; Vlyssides et al., Bioprocess Engineering 22, 503–507.
1996, 1999; Filippi et al., 2002; Plaza et al., 2005), wheat Balis, C., Chatzipavlidis, J., Flouri, F., 1996. Olive mill waste as a
straw (Tomati et al., 1995; Galli et al., 1997), maize straw substitute for nitrogen fixation. International Biodeterioration &
(Parades et al., 1996; Parades et al., 2000) and cotton waste Biodegradation 38, 169–178.
Ben Hamman, O., De la Rubia, T., Martinez, J., 1999. Decolorization of
(Parades et al., 2000; Parades et al., 2002). Composting
olive oil mill wastewaters by Phanerochaete flavido-alba. Environ-
typically results in complete removal of phytotoxicity mental Toxicology and Chemistry 18, 2410–2415.
within a few weeks (e.g., Parades et al., 1996). Benitez, J., Beltran-Heredia, J., Torregrosa, J., Acero, J.L., Cercas, V.,
Parades et al. (2001) compared different composts for 1997. Aerobic degradation of olive mill wastewaters. Applied
treatment of OMWW using cotton waste or maize straw as Microbiology and Biotechnology 47, 185–188.
bulking agents and poultry manure, municipal waste, or Bertin, L., Colao, M.C., Ruzzi, M., Marchetti, L., Fava, F., 2006.
Performances and microbial features of an aerobic packed-bed biofilm
industrial waste from orange juice extraction as nutrient reactor developed to post-treat an olive mill effluent from an anaerobic
sources. At the end of composting, the germination index GAC reactor. Microbial Cell Factories 5, 5–16.
of all treatments was over 50%, indicating reduction of Boari, G., Mancini, I.M., 1990. Combined treatments of urban and olive
phytotoxicity. The wide range of bulking agents and other mill effluents in Apulia, Italy. Water Science and Technology 22,
wastes that can be successfully composted with OMWW 235–240.
Borja, R., Alba, J., Banks, C.J., 1995a. Activated sludge treatment of wash
suggest that this is a versatile technology that could be waters derived from the purification of virgin olive oil in a new
utilized and adapted to local conditions in olive producing manufacturing process. Journal of Chemical Technology and Biotech-
countries. nology 64, 25–30.
 ARTICLE IN PRESS
C.J. McNamara et al. / International Biodeterioration & Biodegradation 61 (2008) 127–134 133

Borja, R., Alba, J., Garrido, S.E., Martinez, L., Garcia, M.P., Incerti, C., Fadil, K., Chahlaoui, A., Ouahbi, A., Zaid, A., Borja, R., 2003. Aerobic
Ramos-Cormenzana, A., 1995b. Comparative study of anaerobic biodegradation and detoxification of wastewaters form the olive oil
digestion of olive mill wastewater (OMW) and OMW previously industry. International Biodeterioration & Degradation 51, 37–41.
fermented with Aspergillus terreus. Bioprocess Engineering 13, FAOSTAT: Food and Agricultural Organization of the United Nations
317–322. Statistics Division, 2007. ProdSTAT: crops: olive oil (19 April 2007)—
Borja, R., Alba, J., Garrido, S.E., Martinez, L., Garcia, M.P., based on 2005 data, online at: /http://faostat.fao.org/site/567/
Monteoliva, M., Ramos-Cormenzana, A., 1995c. Effect of aerobic default.aspxS.
pretreatment with Aspergillus terreus on the anaerobic digestion of Fiestas Ros de Ursinos, J.A., Borja-Padilla, R., 1996. Biomethanization.
olive-mill wastewater. Biotechnology and Applied Biochemistry 22, International Biodeterioration & Degradation 38, 145–153.
233–246. Filippi, C., Bedini, S., Levi-Minzi, R., Cardelli, R., Saviozzi, A., 2002.
Borja, R., Alba, J., Banks, C.J., 1996. Anaerobic digestion of wash waters Cocomposting of olive oil mill by-products: chemical and micro-
derived from the purification of virgin olive oil using a hybrid reactor biological evaluations. Compost Science and Utilization 10, 63–71.
combining a filter and a sludge blanket. Process Biochemistry 31, Finstein, M.S., Miller, F.C., MacGregor, S.T., Psarianos, K.M., 1985. The
219–224. Rutgers strategy for composting: process design and control. United
Borja, R., Alba, J., Mancha, A., Martin, A., Alonso, V., Sanchez, E., 1998. States Environmental Protection Agency Report EPA/600/S2-85/059,
Comparative effect of different aerobic pretreatments on the kinetics Washington, DC.
and macroenergetic parameters of anaerobic digestion of olive mill Galli, E., Pasetti, L., Fiorelli, F., Tomati, U., 1997. Olive-mill wastewater
wastewater in continuous mode. Bioprocess Engineering 18, 127–134. composting: microbiological aspects. Waste Management and Re-
Borja-Padilla, R., Alba-Mendoza, J., Banks, C.J., 1996. Anaerobic search 15, 323–330.
digestion of wash waters derived from the purification of virgin olive Garcia Garcia, I., Jimenez Pena, P.R., Bonilla Vaceslada, J.L., Martin
oil in a new manufacturing process. Process Biochemistry 31, 25–30. Martin, A., Martin Santos, M.A., Ramos Gomez, E., 2000. Removal
Cereti, C.F., Rossini, F., Federici, F., Quaratino, D., Vassilev, N., Fenice, of phenol compounds from olive mill wastewater using Phanerochate
M., 2004. Reuse of microbially treated olive mill wastewater as chrysosporium, Aspergillus niger, Aspergillus terreus and Geotrichum
fertilizer for wheat (Triticum durum Desf.). Bioresource Technology 91, candidum. Process Biochemistry 35, 751–758.
135–140. Gavalla, H.N., Skiadas, I.V., Lyberatos, G., 1996. Anaerobic codigestion
Chtourou, M., Ammar, E., Nasri, M., Medhioub, K., 2004. Isolation of a of agricultural industries’ wastewaters. Water Science and Technology
yeast, Trichosporon cutaneum, able to use low molecular weight 34, 67–75.
phenolic compounds: application to olive mill waste water treatment. Gharsallah, N., 1993. Production of single cell protein from olive mill
Journal of Chemical Technology and Biotechnology 79, 869–878. wastewater by yeast. Environmental Technology 14, 391–395.
Constantinos, E., Papadopoulou, K., Kotsou, M., Mari, I., Constantinos, Gharsallah, N., Labat, M., Aloui, F., Sayadi, S., 1999. The Effect of
B., 1999. Adaptation and population dynamics of Azotobacter Phanerochaete chrysosporium pretreatment of olive mill waste waters
vinelandii during aerobic biological treatment of olive-mill wastewater. on anaerobic digestion. Resources, Conservation and Recycling 27,
FEMS Microbiology Ecology 30, 301–311. 187–192.
D’Annibale, A., Crestini, C., Vinciguerra, V., Giovannozzi Sermanni, G., Gray, N.F., 1999. Water Technology: an Introduction for Scientists and
1998. The biodegradation of recalcitrant effluents from an olive mill by Engineers. Arnold Publishers, London.
a white-rot fungus. Journal of Biotechnology 61, 209–218. Hamdi, M., 1991. Effects of agitation and pretreatment on the batch
D’Annibale, A., Rita Stazi, S., Vinciguerra, V., Giovannozzi Sermanni, anaerobic digestion of olive mill wastewater. Bioresource Technology
G., 2000. Oxirane immobilized Lentinula edodes laccase: stability and 36, 173–178.
phenolics removal efficiency in olive mill wastewater. Journal of Hamdi, M., Ellouz, R., 1992a. Bubble column fermentation of olive mill
Biotechnology 77, 265–273. wastewaters by Aspergillus niger. Journal of Chemical Technology and
Dalis, D., Anagnostidis, K., Lopez, A., Letsiou, I., Hartmann, L., 1996. Biotechnology 54, 331–335.
Anaerobic digestion of total raw olive-oil wastewater in a two-stage Hamdi, M., Ellouz, R., 1992b. Use of Aspergillus niger to improve
pilot-plant (up-flow and fixed-bed bioreactors). Bioresource Technol- filtration of olive mill waste-waters. Journal of Chemical Technology
ogy 57, 237–243. and Biotechnology 53, 195–200.
De Felice, B., Pontecorvo, G., Carfagna, M., 1997. Degradation of waste Hamdi, M., Ellouz, R., 1993. Treatment of detoxified olive mill waste-
waters from olive oil mills by Yarrowia lipolytica ATCC 20255 and waters by anaerobic filter and aerobic fluidized bed process. Environ-
Pseudomonas putida. Acta Biotechnologica 17, 231–239. mental Technology 14, 183–188.
Di Gioia, D., Bertin, L., Fava, F., Marchetti, L., 2001a. Biodegradation of Hamdi, M., BouHamed, H., Ellouz, R., 1991a. Optimization of the
hydroxylated and methoxylated benzoic, phenylacetic and phenylpro- fermentation of olive mill waste-waters by Aspergillus niger. Applied
penoic acids present in olive mill wastewaters by two bacterial strains. Microbiology and Biotechnology 36, 285–288.
Research Microbiology 152, 83–93. Hamdi, M., Khadir, A., Garcia, J., 1991b. The use of Aspergillus niger for
Di Gioia, D., Fava, F., Bertin, L., Marchetti, L., 2001b. Biodegradation of the bioconversion of olive mill waste-waters. Applied Microbiology
synthetic and naturally occurring mixtures of mono-cyclic aromatic and Biotechnology 34, 828–831.
compounds present in olive mill wastewaters by two aerobic bacteria. Hamdi, M., Garcia, J.L., Ellouz, R., 1992. Integrated biological process
Applied Microbiology and Biotechnology 55, 619–626. for olive mill wastewater treatment. Bioprocess Engineering 8, 79–84.
Ehaliotis, C., Papadopoulou, K., Kotsou, M., Mari, I., Balis, C., 1999. Hwu, C.-S., Lettinga, G., 1997. Acute toxicity of oleate to acetate-utilizing
Adaptation and population dynamics of Azotobacter vinelandii during methanogens in mesophilic and thermophilic anaerobic sludges.
aerobic biological treatment of olive-mill wastewater. FEMS Micro- Enzyme and Microbial Technology 21, 297–301.
biology Ecology 30, 301–311. Khabbaz, M.S., Vossoughi, M., Shakeri, M., 2004. Performance of an
El Hadrami, A., Belaqziz, M., El Hassni, M., Hanifi, S., Abbad, A., anaerobic baffled reactor for olive mill wastewater treatment. In:
Capasso, R., Gianfreda, L., El Hadrami, I., 2004. Physico-chemical Proceedings of the 39th Central Canadian Symposium On Water
characterization and effects of olive oil mill wastewaters fertirrigation Quality Research, Burlington, Ontario, Canada.
on the growth of some Mediterranean crops. Journal of Agronomy 3, Kissi, M., Mountadar, M., Assobhei, O., Gargiulo, E., Palmieri, G.,
247–254. Giardina, P., Sannia, G., 2001. Roles of two white-rot Basidiomycete
Ettayebi, K., Errachidi, F., Jamai, L., Tahri-Jouti, M.A., Sendide, K., fungi in decolorisation and detoxification of olive mill wastewater.
Ettayebi, M., 2003. Biodegradation of polyphenols with immobilized Applied Microbiology and Biotechnology 57, 221–226.
Candida tropicalis under metabolic induction. FEMS Microbiology Knupp, G., Rucker, G., Ramos-Cormenzana, A., Garrido Hoyos, S.,
Letters 223, 215–219. Neugebauer, M., Ossenkop, T., 1996. Problems of identifying phenolic
 ARTICLE IN PRESS
134 C.J. McNamara et al. / International Biodeterioration & Biodegradation 61 (2008) 127–134

compounds during the microbial degradation of olive mill wastewater. induced by Azotobacter vinelandii. Journal of Agriculture and Food
International Biodeterioration & Biodegradation 38, 277–282. Chemistry 48, 1941–1948.
Kotronarou, N., Mendez, M., 2003. IMPEL Olive Oil Project. European Plaza, C., Senesi, N., Brunetti, G., Mondelli, D., 2005. Cocomposting of
Union network for the implementation and enforcement of environ- sludge from olive oil mill wastewater mixed with tree cuttings.
mental law, Rome, Italy. Compost Science and Utilization 13, 217–236.
Lettinga, G., 1995. Anaerobic digestion and wastewater treatment Ramos-Cormenzana, A., Juarez-Jimenez, B., Garcia-Pareja, M.P., 1996.
systems. Antonie van Leeuwenhoek 67, 3–28. Antimicrobial activity of olive mill waste-waters (alpechin) and
Marques, I.P., 2001. Anaerobic digestion treatment of olive mill waste- biotransformed olive oil mill wastewater. International Biodeteriora-
water for effluent re-use in irrigation. Desalination 137, 233–239. tion & Biodegradation 38, 283–290.
Marques, I.P., Teixeira, A., Rodrigues, L., Martins Dias, S., Novais, J.M., Ranalli, A., 1992. Microbiological treatment of oil mill waste waters.
1997. Anaerobic co-treatment of olive mill and piggery effluents. Grasas y Aceites 43, 16–19.
Environmental Technology 18, 265–274. Rozzi, A., Malpei, F., 1996. Treatment and disposal of olive mill effluents.
Marrara, G., Tamburino, V., Zimbone, S.M., 2002. Storage and land International Biodeterioration & Biodegradation 38, 135–144.
application of olive oil mill wastewater: experiences in Calabria. Paper Rozzi, A., Passino, R., Limoni, M., 1989. Anaerobic treatment of olive
no. 026136. American Society of Agricultural and Biological Engineers mill effluents in polyurethane foam bed reactors. Process Biochemistry
Annual Meeting, St. Joseph, MI. 24, 68–74.
Martin, A., Borja, R., Garcia, I., Fiestas, J.A., 1991. Kinetics of methane Sanjust, E., Pompei, R., Rescigno, A., Rinaldi, A., Ballero, M., 1991.
production form olive mill wastewater. Process Biochemistry 26, Olive milling wastewater as a medium for growth of four Pleurotus
101–107. species. Applied Biochemistry and Biotechnology 31, 223–235.
Martin, A., Borja, R., Chica, A., 1993. Kinetic study of an anaerobic Sayadi, S., Ellouz, R., 1992. Decolourization of olive mill waste-waters by
fluidized bed system used for the purification of fermented olive mill the white-rot fungus Phanerochaete chrysosporium: involvement of the
wastewater. Journal of Chemical Technology and Biotechnology 56, lignin-degrading system. Applied Microbiology and Biotechnology 37,
155–162. 813–817.
Martinez-Garcia, G., Johnson, A.C., Bachmann, R.T., Williams, C.J., Sayadi, S., Ellouz, R., 1995. Roles of lignin peroxidase and manganese
Burgoyne, A., Edyvean, R.G.J., 2007. Two-stage biological treatment peroxidase from Phanerochaete chrysosporium in the decolorization of
of olive mill wastewater with whey as co-substrate. International olive mill wastewaters. Applied and Environmental Microbiology 61,
Biodeterioration & Biodegradation 59, 273–282. 1098–1103.
Martinez Nieto, L., Garrido Hoyos, S.E., Camacho Rubio, F., Garcia Tomati, U., Galli, E., Di Lena, G., Buffone, R., 1991. Induction of laccase
Pareja, M.P., Ramos Cormenzana, A., 1993. The biological purifica- in Pleurotus ostreatus mycelium grown in olive oil waste waters.
tion of waste products from olive oil extraction. Bioresource Agrochimica 35, 275–279.
Technology 43, 215–219. Tomati, U., Galli, E., Paseti, L., Volterra, E., 1995. Bioremediation of
Mekki, A., Dhouib, A., Aloui, F., Sayadi, S., 2006. Olive wastewater as an olive-mill wastewaters by composting. Waste Management and
ecological fertilizer. Agronomy for Sustainable Development 26, Research 13, 509–518.
61–67. Tsioulpas, A., Dimou, D., Iconomou, D., Aggelis, G., 2002. Phenolic
Niaounakis, M., Halvadakis, C.P., 2006. Olive processing waste manage- removal in olive oil mill wastewater by strains of Pleurotus spp. in
ment: literature review and patent survey, second ed. Elsevier, respect to their phenol oxidase (laccase) activity. Bioresource
Amsterdam. Technology 84, 251–257.
Papadelli, M., Roussis, A., Papadopoulou, K., Venieraki, A., Chatzipav- Ubay, G., Öztürk, I., 1997. Anaerobic treatment of olive mill effluents.
lidis, I., Katinakis, P., Balis, K., 1996. Biochemical and molecular Water Science and Technology 36, 287–294.
characterization of an Azotobacter vinelandii strain with respect to its Vinciguerra, V., D’Annibale, A., Delle Monache, G., Giovannozzi Sermanni,
ability to grow and fix nitrogen in olive mill wastewater. International G., 1995. Correlated effects during the bioconversion of waste olive
Biodeterioration & Biodegradation 38, 179–181. waters by Lentinus edodes. Bioresource Technology 51, 221–226.
Papadimitriou, E.K., Chatjipavlidis, I., Balis, C., 1997. Application of Vinciguerra, V., D’Annibale, A., Gacs-Baitz, E., Delle Monache, G., 1997.
composting to olive mill wastewater treatment. Environmental Biotransformatiuon of tyrosol by whole-cell and cell-free preparation
Technology 18, 101–107. of Lentinus edodes. Journal of Molecular Catalysis B: Enzymatic 3,
Parades, C., Cegarra, J., Senchez Monedero, M.A., Galli, E., Fiorelli, F., 213–220.
1996. Composting of fresh and pond-stored olive-mill wastewater by Vlyssides, A.G., Bouranis, D.L., Loizidou, M., Karvouni, G., 1996. Study
the Rutgers system. In: de Bertoldi, M., Sequi, P., Lemmes, B., Papi, T. of a demonstration plant for the co-composting of olive-oil-processing
(Eds.), The Science of Composting 2. Blackie Academic and wastewater and solid residue. Bioresource Technology 56, 187–193.
Professional, Glasgow, UK, pp. 1266–1270. Vlyssides, A.G., Loizidou, M., Zorpas, A.A., 1999. Characteristics of solid
Parades, C., Roig, A., Bernal, M.P., Sanchez-Monedero, M.A., Cegarra, residues from olive oil processing as bulking agents for co-composting
J., 2000. Evolution of organic matter and nitrogen during co- with industrial wastewaters. Journal of Environmental Science and
composting of olive mill wastewater with solid organic wastes. Biology Health A 34, 737–748.
and Fertility of Soils 32, 222–227. Yesilada, O., Fiskin, K., Yesilada, E., 1995. The use of white rot fungus
Parades, C., Bernal, M.P., Roig, A., Cegarra, J., 2001. Effects of olive mill Funalia Trogii (Malatya) for the decolourization and phenol removal
wastewater addition in composting of agroindustrial and urban wastes. from olive mill wastewater. Environmental Technology 16, 95–100.
Biodegradation 12, 225–234. Yesilada, O., Sik, S., Sam, M., 1998. Biodegradation of olive oil mill
Parades, C., Bernal, M.P., Cegarra, J., Roig, A., 2002. Bio-degradation of wastewater by Coriolus versicolor and Funalia trogii: effects of
olive mill wastewater sludge by its co-composting with agricultural agitation, initial COD concentration, inoculum size and immobiliza-
wastes. Bioresource Technology 85, 1–8. tion. World Journal of Microbiology & Biotechnology 14, 37–42.
Perez, J., de la Rubia, T., Ben Hamman, O., Martinez, J., 1998. Zouari, N., Ellouz, R., 1996a. Microbial consortia for the aerobic
Phanerochaete flavido-alba laccase induction and modification of degradation of aromatic compounds in olive oil mill effluent. Journal
manganese peroxidase isoenzyme pattern in decolorized olive oil mill of Industrial Microbiology 16, 155–162.
wastewaters. Applied and Environmental Microbiology 64, Zouari, N., Ellouz, R., 1996b. Toxic effect of coloured olive compounds
2726–2729. on the anaerobic digestion of olive oil mill effluent in UASB-like
Piperidou, C., Chaidou, C., Stalikas, C., Soulti, K., Pilidis, G., Balis, C., reactors. Journal of Chemical Technology and Biotechnology 66,
2000. Bioremediation of olive oil mill wastewater: chemical alterations 414–420.