Agriculture, Ecosystems and Environment 192 (2014) 105–114

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Agriculture, Ecosystems and Environment

journal homepage: www.elsevier.com/locate/agee

Impact assessment of the reuse of two discrete treated wastewaters

for the irrigation of tomato crop on the soil geochemical properties,
fruit safety and crop productivity
Anastasis Christou a,∗ , Grivas Maratheftis a , Elena Eliadou b , Costas Michael b,c ,
Evroula Hapeshi b,c , Despo Fatta-Kassinos b,c
a
Agricultural Research Institute, P.O. Box 22016, 1516 Nicosia, Cyprus
b
Department of Civil and Environmental Engineering, University of Cyprus, P.O. Box 20537, 1678 Nicosia, Cyprus
c
NIREAS-International Water Research Center, University of Cyprus, P.O. Box 20537, 1678 Nicosia, Cyprus

a r t i c l e i n f o a b s t r a c t

Article history: Advanced tertiary treatment and disinfection technologies have enabled the production of wastewater
Received 18 November 2013 (WW) with quality complying with the established criteria for reuse in agriculture. This study assessed
Received in revised form 9 April 2014 the impacts of tomato crop irrigation with two qualitatively distinct treated WW effluents, as compared to
Accepted 11 April 2014
control tubewell water (TW) irrigation, on the soil geochemical properties, tomato fruit safety and crop
Available online 5 May 2014
productivity. The treated effluents reused for irrigation were produced in two Municipal Wastewater
Treatment Plants (MWTPs) utilizing two discrete tertiary treatment and disinfection technologies, i.e.
Keywords:
Slow Sand Filtration and chlorination (MWTP I), and Membrane Bioreactor and UV radiation (MWTP
Electrical conductivity
Escherichia coli
II), respectively. The impacts on soil pH, electrical conductivity, total organic C, Cl− , NO3 − and heavy
Heavy metal content metal (Zn, Mn, Ni, Cu, Co) content were evaluated. In addition, the heavy metal content in tomato fruits
Microbial assay and leaves, as well as the microbial load in fruit flesh and peel was determined. Crop productivity was
Tomato fruit measured by the mean fruit weight and maximum diameter, and by the number of fruits per harvest.
Transfer factor Irrigation with either WW did not significantly affect the soil pH, organic C and heavy metal content, as
Wastewater reuse well as crop productivity, in comparison to control TW irrigation. Furthermore, the heavy metal content
of tomato fruits and leaves in all irrigation treatments was found to be below the maximum permissible
levels set for fruit safety and the critical tissue concentration for phytotoxicity, respectively. Moreover,
no microbiological contamination (total coliform, fecal coliform, Escherichia coli, Salmonella spp., Listeria
spp.) of tomato fruits was found from any irrigation treatment. Overall, results obtained with regard to
the parameters examined strongly suggest that advanced tertiary treated effluent of good quality might
be safely reused, in terms of both environmental sustainability and public health safety, for vegetable
irrigation, concurrently promoting water use efficiency in dry areas.
© 2014 Elsevier B.V. All rights reserved.

1. Introduction
Quality fresh water for agriculture is becoming an increasingly
scarce resource due to climate change effects and escalating com-
petition from other water use sectors (Mesa-Jurado et al., 2012;
Milano et al., 2012). Thus, wastewater (WW) reuse for irrigation
represents an advantageous alternative for the mitigation of the
ever increasing irrigation water scarcity and demand. Treated
WW represents a potentially valuable, nutrient-rich and reliable
∗ Corresponding author. Tel.: +357 22403113; fax: +357 22316770.
E-mail address: anastasis.christou@ari.gov.cy (A. Christou).
source of water for the agricultural sector, available all year round
(Hamilton et al., 2007). Therefore, WW reuse, mainly for irrigation,
becomes increasingly important as an indispensible component of
all integrated water recourses management schemes in arid and
semi-arid areas around the world (Bixio et al., 2006; Angelakis and
Durham, 2008). Although WW reuse for irrigation has gained an
acceptance as an economic alternate that could substitute nutri-
ent needs and water requirement of crop plants (Khurana and
Singh, 2012), WW may contain undesirable chemical constituents
and pathogens that pose negative environmental and health
impacts (Muchuweti et al., 2006; Bernstein, 2011; Fatta-Kassinos
et al., 2011). It is becoming widely accepted that WW reuse for
irrigation, accompanied with the use of sewage sludge, constitute

http://dx.doi.org/10.1016/j.agee.2014.04.007
0167-8809/© 2014 Elsevier B.V. All rights reserved.
106 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114

Table 1
Physico-chemical and microbiological analysis of the two MWTP’s treated flows and the tubewell water abstracted from a borehole used for the irrigation of the tomato crop.

Parameter MWTP I WW MWTP II WW Control TW Irrigation water quality guidelines1

pH 8.19 ± 0.03 8.31 ± 0.04 8.45 ± 0.06 6.50–8.40

EC (mS cm−1 ) 1.71 ± 0.04 1.56 ± 0.03 3.13 ± 0.06 0.70–3.00b />3.00c
BOD5 (mg O2 L−1 ) 5.00 ± 0.45 3.58 ± 0.21 0.98 ± 0.03
COD (mg O2 L−1 ) 36.67 ± 3.49 30.52 ± 1.23 7.09 ± 0.58
SS (mg L−1 ) 4.38 ± 0.40 3.00 ± 0.45 0.53 ± 0.12 <50.00a
Total N (mg L−1 ) 7.38 ± 1.49 6.07 ± 1.19 0.61 ± 0.17
Total P (mg L−1 ) 2.29 ± 0.14 0.70 ± 0.27 0.18 ± 0.02
Cl− (mg L−1 ) 261.05 ± 3.73 317.70 ± 18.54 315.65 ± 23.06
Zn (mg L−1 ) 0.04 ± 0.01 0.04 ± 0.01 0.11 ± 0.01 2.00
Mn (mg L−1 ) 0.03 ± 0.01 0.02 ± 0.01 0.01 ± 0.00 0.20
Ni (mg L−1 ) 0.04 ± 0.03 0.01 ± 0.00 b.l.q.2 0.20
Cu (mg L−1 ) 0.03 ± 0.01 0.01 ± 0.00 0.01 ± 0.00 0.20
Co (mg L−1 ) 0.03 ± 0.00 0.01 ± 0.00 0.02 ± 0.00 0.05
E. coli (CFU 100 mL−1 ) ND3 ND ND
Helminth eggs (egg L−1 ) ND ND ND

Data are the mean values ± standard errors (SE) of 5 samples taken during the tomato crop growing season.
1
As set by the FAO’s water quality for agriculture report (Ayers and Westcot, 1985).
2
Below limit of quantification.
3
Not detected (< 1CFU 100 mL−1 or < 1 egg L−1 ).
a
No restrictions.
b
Slight to moterate restrictions.
c
Severe restrictions for irrigation use.

the main causes of soil contamination with heavy metals and other
pollutants (Li et al., 2009; Xu et al., 2010). Moreover, WW reuse
may cause additional negative impacts, such as soil salinization
(Klay et al., 2010), with salinity being recognized as a major fac-
tor reducing crop productivity worldwide (Krasensky and Jonak,
2012), and agricultural produce contamination with microbial and
other anthropogenic pathogens (Sacks and Bernstein, 2011; Cirelli
et al., 2012), if mismanagement and improper practices are taking
place.
Several studies have recently documented an emerging risk
concerning heavy metal accumulation in the topsoil of WW irri-
gated sites, worldwide (Mapanda et al., 2005; Li et al., 2009;
Xu et al., 2010). Repeated WW applications may result in heavy
metal accumulation in cultivated top-soil to toxic concentrations
for plant growth (Megateli et al., 2009), while subsequent entry
into the food-chain through various food crops and fodders repre-
sents the main pathway of human exposure to soil contamination
(Rajaganapathy et al., 2011). Crop plants irrigated with treated
WW have been found to absorb and accumulate excess heavy
metals in the edible parts beyond maximum permissible lim-
its (MPLs) (EC, 2001; WHO/FAO, 2007), set for guidance of their
safety (Muchuweti et al., 2006; Khan et al., 2008; Singh et al.,
2010a). Importantly, high levels of heavy metals in foodstuff evoke
concern of potential chronic negative health impacts, in both
children and adults (Szkup-Jablonska et al., 2012; Wang et al.,
2012). Thus information about heavy metal concentrations, both
in cultivated soils and agricultural produce, is very important for
assessing the risks to public health. The presence of bacteria, such
as Escherichia coli, and other human health related pathogens
in WW irrigated crops’ edible parts is also a potential concern
(Petterson et al., 2001; Palese et al., 2009; Cirelli et al., 2012;
Forslund et al., 2012). In this regard, comprehensive guidelines
and criteria have been established in order to safeguard envi-
ronmental sustainability and public health safety as a result of
WW irrigation (WHO, 2006; Brissaud, 2008; U.S. Environmental
Protection Agency, 2012). In addition, advanced WW treatment
technologies have been developed, enabling contaminants and
pathogens removal from treated WW (Fatta-Kassinos et al., 2010;
Kalbar et al., 2012).
A holistic approach was employed in this study with the aim
to assess the impacts of the reuse of effluents produced from two
different WW treatment systems for the irrigation of a tomato
crop, as compared to control tubewell water (TW) irrigation, on
soil geochemical properties, fruit safety and crop productivity. The
first treatment system includes Slow Sand Filtration (SSF) and chlo-
rination, while the second one Membrane Bioreactor (MBR) and
ultraviolet disinfection (UV). The two selected discrete technolo-
gies for the tertiary treatment and disinfection of WW are utilized
worldwide, since they efficiently produce WW with a quality com-
plying with the established criteria for WW reuse in agriculture,
while simultaneously being cost effective (Meneses et al., 2010).
2. Materials and methods
2.1. Experimental design and treatments
Tomato seedlings were transplanted in a field dominated by
sandy clay loam soil at the experimental station of the Agricul-
tural Research Institute in Nicosia, Cyprus, in April 2012. Tomato
plants were subjected to 3 treatments, based on the water source
used for their irrigation. More precisely, treatments involved the
WW irrigation of tomato plants with the tertiary treated effluent of
MWTP I (MWTP I WW) and MWTP II (MWTP II WW), as well as con-
trol tubewell water irrigation (Control TW) with water abstracted
from a nearby borehole within the experimental station. Tertiary
treatment and disinfection in MWTP I is accomplished through SSF
and chlorination process, whereas, in MWTP II through MBR and
UV treatment, respectively. It is worth noting that these tertiary
treatment and disinfection technologies are used in all WWTPs in
Cyprus to produce 22 million cubic meters (MCM) of treated WW,
from which nearly 65% (∼14 MCM) is reused for the irrigation of for-
age crops, citrus, olives and vegetables, as regulated by the Cyprus
guideline for WW reuse. The chemical and microbial load of the
three water sources used for the irrigation of tomato plants is pre-
sented in Table 1. The climate in the experimental region during the
growing season is characterized by high temperatures (day light
30–40 ◦ C; night 25–30 ◦ C), and low relative humidity (less than
20%), having as a result high evapotranspiration rates. A completely
randomised block design was applied, while each treatment was
independently run in five replicates; each replicate consisted of 20
individual tomato plants. As a result 300 tomato plants were used
in this experiment. Tomato plants were drip irrigated based upon
direct measurements of soil moisture status (15 centibars) by the
use of tensiometers. The tomato growing season lasted 150 d, and
eight harvests took place.
 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114
2.2. Sample collection and preparation
Water samples were regularly taken in pyrex storage bot-
tles from the end of the drip irrigation pipeline and immediately
transferred to laboratory and stored at 4 ◦ C, till analysis. The
physico-chemical analysis of the three water sources concerned the
pH, electrical conductivity (EC), BOD5 , COD, SS, total N and total P,
Cl− and heavy metal content, while the microbial analysis included
the E. coli and Helminth eggs content. Soil samples (5 replicates)
representing each treatment replication were collected with an
auger sampler from the 0–25 cm topsoil layer and the 26–50 cm soil
layer as composite samples, consisting of 20 subsamples (each plant
represented a supsample). Soil sampling was undertaken in prede-
fined intervals during the growing period (day 0, 10, 50, 100, 150).
All soil samples were allowed to dry laid in a glasshouse, ground,
and sieved (<10 mesh). From the sieved samples, pH, EC, Cl− and
NO3 − anions content, total organic C, as well as heavy metal (Mn,
Co, Cu, Zn, Ni) content, were determined. Tomato leaves and fruits
randomly collected from all 20 tomato plants in each replication
during the last harvest (d 150) constituted the composite samples of
each treatment. Tomato leaves where washed with ultrapure Milli-
Q H2 O (Milli-Q Integral Water Purification System; EMD Millipore
Co., Billerica, USA) in order to remove any soil particles, blotted
dried with tissue paper, weighed to obtain fresh mass and dried in
a preheated oven at 80 ◦ C for 72 h, until constant weight. Half of
the tomato fruits were immediately placed into polyethylene ster-
ile bags and transferred to the laboratory for the microbiological
assays, while the rest were treated as leaves. Dry tomato leaves
and fruit samples were weighed again to obtain dry mass, ground,
sieved (30 mesh) and the content of heavy metals (Mn, Co, Cu, Zn,
Ni) was estimated.
The selection of heavy metals for analyses in both soil, and
tomato leaves and fruits was based upon their concentrations quan-
tified in the three water sources used for tomato crop irrigation
(Table 1), since various other heavy metals (Cd, Pb, Hg, Cr) were
below the limit of quantification in all water sources utilized.
2.3. Analytical methods and measurements
2.3.1. Tubewell water and wastewater
The pH and EC of the three water resources used for the irrigation
of tomato crop were estimated using a pH meter (Thermo Scientific
Orion 4-star benchtop pH/ISE meter; Cole-Parmer Co., IL, USA) and
an electrical conductivity meter (Hanna HI4321 benchtop electrical
conductivity meter; Hanna Instruments Inc., MI, USA), respectively.
Chloride (Cl− ) anions were determined according to the Mohr’s
titration method (APHA, 1998), while NO3 − anions were deter-
mined with the reflectometric method with test strips, using the
RQflex plus 10 reflectometer (Merck KGaA, Darmstadt, Germany).
In addition, COD was determined by the APHA 5220B method, using
the Merck® Spectroquant Kits (WTW Photolab S6; Merck KGaA,
Darmstadt, Germany), while the BOD5 by the APHA 5210A method,
using the Lovibond OxiDirec apparatus (Tintometer GmbH, Dort-
mund, Germany) (APHA, 1998). The analysis of total suspended
solids (TSS) was based on the filtration of a well-mixed subsam-
ple through a weighed standard glass-fiber filter (0.45 ␮m) (APHA,
1998). Total N was determined via persulfate digestion, followed by
the colorimetric measurement of nitrate (APHA 4500-N C method),
whereas total P was also determined with the colorimetric method
after its conversion to orthophosphates via digestion (APHA 4500-
P B method) (APHA, 1998). The Escherichia coli and Helminths eggs
were determined as described in APHA’s standard method for the
examination of water and WW. Finally, for the heavy metal content
quantification, 45 mL samples were mixed with 5 mL concentrated
(69–71%) HNO3 and mineralised using the MARS Xpress Microwave
Digestion System (CEM Corp., Matthews, USA) following the U.S.
107
Environmental Protection Agency (USEPA) Method 3015 A (U.S.
Environmental Protection Agency, 1998). The heavy metal content
in digested water samples was determined by a Thermo Scientific
SOLLAR/AA Flame Atomic Absorption Spectrophotometer (AAS)
(Thermo Fisher Scientific Inc., Waltham, USA).
2.4. Soil samples
Soil pH was determined using a pH meter (Thermo Scientific,
Orion 4 Star) in a 1:2 suspension in ultrapure Milli-Q H2 O, as
described by Datta et al. (1997). Soil EC was measured in the
saturated soil-paste extract, using the Hanna HI4321 benchtop
conductivity meter. Chloride and nitrate anions were determined
in saturated soil paste extract according to the Mohr’s titration
method and the reflectometric method with test strips, respec-
tively, as previously described. Total organic C was measured
using the EURO EA Elemental Analyser (EuroVector S.P.A., Milan,
Italy). For heavy metal determination, 0.5 g of grounded sieved soil
samples were mineralised into solution using the MARS Xpress
Microwave Digestion System (CEM Corp., Matthews, USA) with a
mixture of concentrated (69–71%) HNO3 and concentrated (37%)
HCl (9:3 mL) in a total digestion procedure equivalent to the USEPA
Method 3051 A (U.S. Environmental Protection Agency, 1998). Sam-
ples were then allowed to cool, filtered (Whatman 42 Filter Paper)
into polypropylene centrifuge tubes and diluted to 50 mL with
ultrapure Milli-Q H2 O. A blank digest was run in the same way
in all digestion events. The soil heavy metal content was deter-
mined using an AAS. A Certified European sandy soil Reference
material (ERM-CRM) (Code ERM-CC018 trace elements in contami-
nated sandy soil) purchased from BAM Federal Institute for Material
Research and Testing (Berlin, Germany) was used to evaluate ana-
lytical accuracy and precision regarding soil heavy metal content
determination (elemental recoveries ranged from 92% to 102%).
2.5. Tomato fruits and leaves
Dried ground tomato fruit and leaves samples (0.4 g) were dis-
solved in a mixture of concentrated (69–71%) HNO3 and H2 O
(4:4 mL) in a microwave PTFE vessels. Samples decomposition was
accelerated by microwave digestion in the MARS Xpress Microwave
Digestion System, in a program consisting of an 8 min ramp to
175 ◦ C, a 15 min step at 175 ◦ C and then a ventilated cooling
period (Carbonell et al., 2011). Solution samples were transferred to
polypropylene centrifuge tubes and diluted with ultrapure Milli-Q
H2 O to 25 mL. A blank digest was run also, as previously described.
A centrifugation was followed in order to obtain clear supernatant
(7000 × g, 2 min). Metal concentrations in tomato fruits and leaves
digests were determined using an AAS. The soil to tomato fruit and
leaves heavy metal transfer factor (TF) was calculated as the ratio of
the concentration of metal to the fruit or leaves respectively (on dry
weight basis), to its respective total content in the sampled topsoil
layer (0–25 cm), as proposed by Cui et al. (2004).
2.5.1. Microbial contamination of fruits and crop productivity
Tomato fruits microbiological examination concerned the aero-
bic plate count (APC) (Hydrophobic Grid Membrane Filter Method,
AOAC official method 986.32), as well as the quantification of total
coliform (TC) and fecal coliform (FC) (Hydrophobic Grid Mem-
brane Filter Method, AOAC official method 983.25) in both fruit
flesh and peel (AOAC, 2005). The presence of Escherichia coli in
fruit homogenate was assessed following the AOAC official method
2004.03 (AOAC, 2005). In addition, the presence of Salmonella spp.
and Listeria spp. in the tomato fruit homogenate was evaluated
using the AOAC approved kits of Neogen Corporation (Neogen Cor-
poration Inc, Lansing, USA), as per the manufacturer instructions.
Finally, the commercial yield of each harvest, hand collected at an
108 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114
advanced full red stage of maturity, was measured both by mean
fruit weight and by number of fruits per treatment. Furthermore,
the fruit size was determined by measuring the diameter at the
maximum circumference of the fruit using a caliper, as proposed
by Al-Lahham et al. (2003).
2.6. Statistical analysis
Statistical analysis was carried out using the software package
SPSS v21.0 (SPSS Inc., Chicago, USA) and the comparison of averages
of each treatment was based on the analysis of variance (One-Way
ANOVA) according to Tukey’s range test at significance level 5%
(P < 0.05). Data are means ± standard error (SE) of five replications.
3. Results and discussion
3.1. Irrigation water quality
The average values of physico-chemical and microbial proper-
ties of the two effluents and the control TW met irrigation water
quality standards according to FAO’s guidelines (Ayers and Westcot,
1985), with the sole exception of the high EC recorded in TW
(3.13 mS cm−1 ), as shown in Table 1. The EC of TW was slightly
above the threshold of 3 mS cm−1 defined as the water salinity
beyond which severe restrictions for crop irrigation may occur.
Tubewell water’s high salinity may be attributed to the deteriora-
tion of ground water quality, due to the sustained ground water
pumping for irrigation and the consequent saltwater upcoming
(Reilly and Goodman, 1987; Werner et al., 2009). Thus, WW reuse
for irrigation purposes in the experimental site may contribute to
the amelioration of ground water over-abstraction and to its qual-
itative restoration. In general, the three irrigation water sources
were alkaline with pH values ranging from 8.19 to 8.45. Control
TW had significantly lower values of BOD5 and COD, whereas Zn
was quantified in values greater than those of the two treated efflu-
ents. The EC, BOD5 , COD, SS, total P and total N, and heavy metal
content in MWTP I WW were higher compared to those of MWTP
II WW. In addition, both Escherichia coli and Helminths eggs were
not detected in all the water sources used for irrigation. Moreover,
both WWs reused for irrigation were fully in line with both the
WHO (WHO, 2006) and the USEPA (U.S. Environmental Protection
Agency, 2012) guidelines for WW reuse in agriculture.
3.2. Effects of the irrigation treatments on the soil geochemical
properties and heavy metal content
A time-scale assessment was employed for evaluating soil pH,
EC, Cl− and NO3 − content, in response to WW or TW irrigation.
Interestingly, none of the water sources significantly impacted
the soil pH, since the pH values quantified did not differ among
irrigation treatments, as well as from values found at the imposition
of treatments (day 0) (Fig. 1A). Notable variations were registered
regarding the soil EC, NO3 − and Cl− content. More specifically, con-
trol TW irrigation resulted in the excessive accumulation of soluble
salts in soil, as evidenced by the marked increase of soil EC in both
soil profile layers sampled, as compared to day 0. Irrigation with
both treated effluents also enhanced soil salinity, albeit lower than
TW irrigation. The impact of the irrigation water source on the soil
EC was readily apparent within 50 d of irrigation, reaching its high-
est level 100 d after the initiation of the experiment. The highest EC
values were recorded in the 0–25 cm topsoil (12.81 mS cm−1 ) and
the 26–50 cm soil layer (9.64 mS cm−1 ) irrigated with TW for 100 d.
Soil EC in both soil layers sampled, irrigated for 100 d with either
treated flows ranged from 3.68 to 6.03 mS cm−1 . A notable reduc-
tion in soil EC occurred during the last sampling (d 150), due to
salt leaching as a result of heavy precipitation. Nonetheless, soil
irrigated with TW appeared to have higher EC levels compared
to WW irrigated ones, even after salt leaching (Fig. 1B). The soil
Cl− content followed similar to the soil EC pattern also, reach-
ing its maximum variance 100 d after the initiation of tomato
crop cultivation (Fig. 1C). On the other hand, the NO3 − content
was found to be higher in the soil irrigated with MWTP I WW
in all samplings after the initiation of the experiment, whereas
no differences were registered among the MWTP I WW and the
control TW irrigated soil layers sampled (Fig. 1D). Similar find-
ings, demonstrating no significant impacts of WW irrigation on the
soil pH, have been reported by Mohammad Rusan et al. (2007),
whereas results corroborating the progressive soil EC elevation
due to the short-term WW irrigation recorded in our study, have
been reported also (Al-Lahham et al., 2007; Morugan-Coronado
et al., 2011). The elevated EC and Cl− content in TW irrigated soil
can be attributed to the high EC of the control TW as a result of
ground water quality deterioration in the experimental site, prob-
ably due to over-abstraction (Reilly and Goodman, 1987; Werner
et al., 2009). The observed soil alkalinization is expected to impact
tomato plant’s growth and productivity, since tomato yield starts to
decline at soil EC of 2.50 mS cm−1 , while a 10% decrease in yield per
unit increase in salinity beyond this threshold has been indicated
(Maas and Hoffman, 1977). Reported salinity effects on tomato
plant growth and productivity, either due to saline soil or irrigation
water (Tuna et al., 2007; Martinez-Rodriguez et al., 2008; Rady,
2012; Gong et al., 2013), further corroborate the expected negative
impacts of all irrigation treatments on tomato crop, in the current
study.
The sandy clay loam soil in the experimental site is poor in
organic matter, as evident by the low average values of total organic
C found (0.57–0.78%). Although no statistically significant differ-
ences on soil total organic C among all irrigation treatments have
been found, an increasing trend in the topsoil of WW irrigated
soil was observed, possibly due to slight accumulation of organic
matter in the soil (Table 2). Taking into consideration the poor fer-
tility and the alkaline nature of this soil, the minor increase in total
organic C registered could have beneficial effects in soil fertility,
especially if WW application is being practiced for a prolonged
period.
Importantly, none of the three water sources used impacted soil
heavy metal content significantly, since the concentrations quan-
tified were similar to those quantified at day 0 (Table 2). Soil Zn
content ranged from 63.11 to 78.77 mg kg−1 , whereas Mn from
451.99 to 526.05 mg kg−1 , Ni from 27.37 to 30.16 mg kg−1 , Cu from
43.06 to 47.10 mg kg−1 and Co from 14.50 to 16.16 mg kg−1 . The
heavy metal concentrations quantified were below the MPLs set
by the EU Directive 86/278/EEC (EEC, 1986) (last updated in 2002),
which are 300, 75 and 140 mg kg−1 for Zn, Ni and Cu respectively.
Importantly, taking into consideration the heavy metals’ concen-
tration in the WW reused for irrigation (maximum concentration
0.04 mg L−1 ) and the yearly irrigation water demands of the tomato
crop under the climatic conditions in Cyprus (∼6540 m3 ha−1 )
(Metochis, 1997), as well as the bulk density of the soil, no more
than a couple of mg of a single heavy metal can be added to a
25 cm soil profile layer of the experimental site, after a continuous
long-term (e.g. 10 years) WW irrigation.
3.3. Effects of the irrigation treatments on the tomato fruit and
leaves heavy metal content
As shown in Table 3, fruit heavy metal content was significantly
affected by the water source used for irrigation. The heavy metal
content in fruits harvested from plants irrigated with WW were
less than or equal to that in fruits harvested from the control
TW irrigation treatment. More precisely, higher Zn, Mn and Cu
concentrations were found in tomato fruits harvested from plants
 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114 109

Fig. 1. Soil pH (A), electrical conductivity (B), Cl− (C) and NO3 − content (D) in the topsoil (0–25 cm) and the 26–50 cm soil profile layer of the tomato crop, as affected by
the irrigation with the two discrete treated wastewaters and tubewell water. Data are means ± standard error (SE) of five replications. Treatments with different letters are
significantly different according to Tukey’s range test at significance level 5% (P < 0.05).

irrigated with control TW. However, higher concentrations of plants irrigated with the treated effluent of MWTP II was lower,
these heavy metals were recorded in fruits harvested from plants compared to fruits harvested from plants irrigated with the other
irrigated with MWTP I WW than those irrigated with MWTP II WW. two water sources, while no changes were registered regarding
Furthermore, the Ni concentration in tomato fruits harvested from the fruit’s Co concentration. The quantified Zn and Cu content were
110 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114

Table 2
Effect of treated wastewater effluents from two discrete technologies (MWTP I and MWTP II) and tubewell water irrigation on soil total organic C and heavy metal content
at 2 soil profiles layers (0–25 and 26–50 cm) of the tomato crop.

Treatment Soil profile Total organic C Zn (mg kg−1 ) Mn (mg kg−1 ) Ni (mg kg−1 ) Cu (mg kg−1 ) Co (mg kg−1 )
(source of irrigation) layer (cm) (%)

Control TW 0–25 0.67 ± 0.06 ab 78.77 ± 5.12 a 526.05 ± 20.73 a 28.84 ± 1.53 a 43.28 ± 3.35 a 14.98 ± 0.60 a
26–50 0.61 ± 0.07 ab 68.17 ± 2.26 ab 497.57 ± 8.08 a 27.37 ± 1.73 a 43.06 ± 2.82 a 14.50 ± 0.53 a

MWTP I WW 0–25 0.78 ± 0.09 a 67.43 ± 3.17 ab 519.89 ± 11.20 a 30.16 ± 1.66 a 45.62 ± 2.81 a 15.90 ± 0.82 a
26–50 0.60 ± 0.07 b 63.11 ± 3.14 b 451.99 ± 36.14 a 29.32 ± 1.67 a 47.10 ± 1.96 a 15.82 ± 0.99 a

MWTP II WW 0–25 0.77 ± 0.08 a 68.77 ± 2.09 ab 521.28 ± 19.80 a 28.62 ± 1.27 a 44.58 ± 2.74 a 15.26 ± 0.39 a
26–50 0.57 ± 0.14 ab 68.37 ± 3.29 ab 463.89 ± 16.39 a 29.79 ± 2.16 a 46.80 ± 2.61 a 15.68 ± 0.89 a

Concentration at the initiation 0–25 0.67 ± 0.09 ab 67.61 ± 2.51 ab 502.43 ± 21.01 a 29.60 ± 1.61 a 43.34 ± 2.91 a 16.16 ± 0.68 a
of the experiment 26–50 0.62 ± 0.10 ab 66.46 ± 3.11 ab 508.48 ± 19.01 a 29.05 ± 1.99 a 43.78 ± 2.13 a 14.99 ± 1.68 a

MPL1 (EU standards, 1986)2 300 75 140

Data are means ± SE of five replications.

1
Maximum Permissible Level.
2
Last updated in 2002. Treatment values with different letters among each parameter examined are significantly different according to Tukey’s range test at significance
level 5% (P < 0.05).

Table 3
Effect of treated wastewater effluent from two discrete technologies (MWTP I and MWTP II) and tubewell water irrigation on tomato fruit’s heavy metal content (on dry
weight basis).

Element Irrigation water source MPL (WHO/FAO, 2007)

Control TW MWTP I WW MWTP II WW

Zn (mg kg−1 ) 47.77 ± 1.30 a 43.41 ± 0.40 b 37.86 ± 0.09c 60

Mn (mg kg−1 ) 21.76 ± 0.56 a 17.60 ± 0.17 b 14.45 ± 0.08c –
Ni (mg kg−1 ) 1.36 ± 0.06a 1.39 ± 0.13a 1.00 ± 0.07b –
Cu (mg kg−1 ) 21.11 ± 0.37 a 19.64 ± 0.22 b 14.26 ± 0.10 c 40
Co (mg kg−1 ) 2.14 ± 0.15a 1.81 ± 0.11a 2.13 ± 0.06a –

MC (%) 94.12 94.51 94.62

Data are means ± SE of five replications; MC, Moisture content; MPL, Maximum Permissible Level; Treatment values with different letters among each heavy metal examined
are significantly different according to Tukey’s range test at significance level 5% (P < 0.05).

lower than the MPLs (60 and 40 mg kg−1 , respectively) set by the
WHO/FAO Codex Committee on Food Hygiene (WHO/FAO, 2007).
The effects of short- and long-term WW irrigation on vegeta-
bles’ fruits heavy metal content have been extensively studied.
Short-term tomato crop irrigation with tertiary filtered municipal
WW with lower heavy metal content than the WW used in our
study, revealed no significant difference in fruit heavy metals con-
centrations, as compared to conventional water irrigation, while
concentrations did not exceed toxic values (Lonigro et al., 2007).
In contrast, Al-Lahham et al. (2007) found that the short-term
tomato crop irrigation with WW having several folds higher heavy
metal content than the ones used in the current study (45, 12,
300 and 5 times higher Cu, Mn, Zn, and Ni content, respec-
tively), resulted in increased concentrations of heavy metals in
tomato fruits as compared to potable water irrigated tomato fruits,
albeit below the Jordanian standard limits. Furthermore, stud-
ies in India, aiming at assessing the long-term WW irrigation
impacts on tomato and other vegetable fruit’s heavy metal con-
tent, as compared to conventional water irrigation, revealed similar
Cu and Zn concentrations in tomato fruits, while Ni concentra-
tion was several folds higher (∼13), exceeding the nationals safe
limits (Singh et al., 2010a, 2010b). However, it should be stated
that WW irrigation was practiced for a prolonged period (20
years) and had 2–3 times higher Cu, Zn and Ni concentration, as
compared to the MWTP I WW and MWTP II WW used in this
study.
Variations between irrigation treatments were verified
regarding tomato leaves’ heavy metal content, as well. However, no
definite trend in the leave’s heavy metal content among irrigation
treatments could be registered. The heavy metals concentrations in
tomato leaves were in the order of Mn > Zn > Cu > Co > Ni (Table 4).
Interestingly, Khan et al. (2013) reported similar Zn, Cu and Mn,
and higher Ni concentrations in tomato leaves irrigated with WW
of higher heavy metal content (10–25 times higher), in Pakistan.
The Mn, Zn and Ni quantified concentrations in tomato leaves
were below the critical limits of phytotoxicity for these metals, as
defined by Macnicol and Beckett (1985).
3.4. Heavy metal transfer from soil to tomato fruits and leaves
The calculation of the TF accurately reflects the differences
among heavy metals’ phytoavailablity and translocation to the
above-ground tomato plant parts, as affected by the water source
reused for irrigation. Results revealed that the variations of TF for
Zn, Ni, Cu and Co between leaves and fruits were less pronounced
as compared to the TF calculated for Mn (Table 5). This suggests
that Mn is selectively accumulated in tomato leaves, rather than
fruits, since the TF calculated, as well as the Mn content in leaves,
were consistently higher compared to the respective values for
fruits. The TF values calculated for Zn were similar to the ones
reported by Singh et al. (2010b) in tomato fruits harvested from
WW irrigated sites in India, whereas those for Cu and Ni were
significantly lower. The low TF values calculated in the present
study may be attributed to the alkaline soil of the experimental site
(pH values ranging from 8.36 to 9.02), which in turn facilitates the
adsorption of heavy metals by soil particles and the precipitation
of metal hydroxides and carbonates, or the formation of insoluble
organic complexes (Smith et al., 1996). These results, along with
the measured heavy metal content in fruits being below MPLs,
suggest that a variety of vegetable crops could be safely drip
irrigated with the WW used in this study. As the tomato crop is
characterized by its long growing period, high irrigation water
 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114 111

Table 4
Effect of treated wastewater effluent from two discrete technologies (MWTP I and MWTP II) and tubewell water irrigation on tomato leaves’ heavy metal content (on dry
weight basis).

Element Irrigation water source Critical tissue concentration

of phytotoxicity1

Control TW MWTP I WW MWTP II WW

−1
Zn (mg kg ) 55.47 ± 0.91 a 28.91 ± 0.25 c 33.71 ± 0.60 b 350
Mn (mg kg−1 ) 76.73 ± 1.28 b 56.15 ± 0.79c 96.58 ± 1.01 a 500
Ni (mg kg−1 ) 1.76 ± 0.14a 1.34 ± 0.15b 1.47 ± 0.04b 1030
Cu (mg kg−1 ) 17.21 ± 0.14 b 18.90 ± 0.19 a 17.87 ± 0.15b
Co (mg kg−1 ) 1.20 ± 0.16b 1.71 ± 0.05a 1.77 ± 0.12a

Data are means ± SE of five replications.

1
According to Macnicol and Beckett (1985). Treatment values with different letters among each heavy metal examined are significantly different according to Tukey’s
range test at significance level 5% (P < 0.05).

Table 5
Transfer factor (TF) of heavy metals from the topsoil (0–25 cm) of the tomato crop field to the fruits and leaves of tomato plants irrigated either with treated wastewater
effluent from two discrete technologies (MWTP I and MWTP II) or tubewell water (on dry weight basis).

Irrigation water source Element

Zn Mn Ni Cu Co

Fruits
Control TW 0.598 ± 0.038 a 0.042 ± 0.002 a 0.047 ± 0.005 a 0.525 ± 0.043 a 0.135 ± 0.012 a
MWTP I WW 0.649 ± 0.030 a 0.034 ± 0.001 a 0.046 ± 0.007 a 0.437 ± 0.026 a 0.118 ± 0.013 a
MWTP II WW 0.552 ± 0.018 b 0.028 ± 0.001 a 0.035 ± 0.003 a 0.322 ± 0.020 b 0.140 ± 0.004 a
Leaves
Control TW 0.703 ± 0.056 a 0.148 ± 0.008 b 0.062 ± 0.002 a 0.407 ± 0.032 a 0.081 ± 0.014 a
MWTP I WW 0.432 ± 0.019 b 0.108 ± 0.003 c 0.048 ± 0.006 a 0.421 ± 0.026 a 0.108 ± 0.007 a
MWTP II WW 0.491 ± 0.010 b 0.186 ± 0.009 a 0.053 ± 0.004 a 0.406 ± 0.023 a 0.115 ± 0.008 a

Data are means ± SE of five replications; Treatment values with different letters in fruits or leaves among each heavy metal examined are significantly different according to
Tukey’s range test at significance level 5% (P < 0.05).

needs and biomass production, as compared to other vegetables
(e.g. pepper, eggplant, beans, melons, etc.).
3.5. Tomato fruits’ microbial quality
The level of the microbial contamination with APC, TC and FC on
tomato fruit peel and flesh, as well as the presence of Escherichia
coli, Salmonella spp. and Listeria spp. in the fruit homogenate, are
presented in Table 6. The assay of APC revealed that the aerobic,
mesophilic micro-organisms were predominantly found in fruit
peel, rather than fruit flesh. The irrigation treatment did not affect
the APC levels in tomato flesh (22 CFU g−1 ), whereas the APC levels
in tomato fruit peel were higher in fruits collected from plants irri-
gated with MWTP I WW (810.23 CFU g−1 ) compared to ones from
plants irrigated with MWTP II WW (472.36 CFU g−1 ) and the control
TW (278.45 CFU g−1 ). The APC levels in tomato fruit peel and flesh in
our experiment could be attributed to the external environmental
contamination, as suggested by Frank (2001). It is noteworthy that
the TC and FC in fruit flesh and peel, as well as the E. coli, Salmonella
spp. and Listeria spp. in fruit homogenate, were not detected in
any fruits, strengthening the safety of the application of WW for
the drip irrigation of tomato crop. Results highlighting that WW
drip irrigation (both surface and subsurface) of tomato crop is safe
with regard to microbial contamination have been also recently
reported (Aiello et al., 2007; Cirelli et al., 2012). Moreover, by
using molecular approaches, Forslund et al. (2012) demonstrated
that both sprinkler, and surface and subsurface irrigation with ter-
tiary treated WW is safe for the production of tomatoes. At the
same time, these authors stressed the importance of the external
environment, typically wildlife, as a source of fecal contamination.
In addition, Bernstein et al. (2008) reported no bacterial popula-
tions in an agricultural greenhouse hydroponic production system
irrigated with reclaimed WW. In contrast to the above findings, Al-
Lahham et al. (2003) reported considerable coliform contamination

Table 6
Effects of the tomato crop irrigation with two discrete treated wastewater effluents (MWTP I WW and MWTP II WW) and tubewell water, on the tomato fruit flesh and peel,
and tomato homogenate, microbiological load.

Microbial & pathogen test Tomato fruit part tested Irrigation water source

Control TW MWTP I WW MWTP II WW

APC (CFU g−1 ) Flesh 22.00 ± 17.43 22.00 ± 12.41 22.00 ± 15.62
Peel 278.45 ± 20.32 810.23 ± 174.13 472.36 ± 55.08
TC (CFU g−1 ) Flesh ND1 ND ND
Peel ND ND ND
FC (CFU g−1 ) Flesh ND ND ND
Peel ND ND ND
Escherichia coli (CFU g−1 ) Homogenate Absent2 Absent Absent
Salmonella spp. (CFU g−1 ) Homogenate Absent Absent Absent
Listeria spp. (CFU g−1 ) Homogenate Absent Absent Absent

Data are means ± SE of five replications. APC, aerobic plate count; TC, total coliform; FC, fecal coliform.
1
Not detected (<1 CFU g−1 ).
2
<1 CFU 25 g−1 .
112 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114

(A) 200

Control TW
160 MWTP I WW

Mean fruit weight (g)

MWTP II WW

120

80

40

(B) 100

80
Maximum diameter (mm)

60

40

20

(C) 500

400
Total number of fruits

300

200

100

Harvest

Fig. 2. Effect of the tomato crop irrigation with two discrete wastewaters and tubewell water on the fruit mean weight (A) and maximum diameter (B), and on the total
number of fruits (C) in each harvest. Data are means ± standard error (SE) of all fruits in each harvest.

of tomato fruit skin, as a result of WW irrigation, suggesting that compared to fresh water irrigation. On the other hand, Al-Lahham
tomatoes produced from WW irrigated crops should be eaten after et al. (2003) found that tomato fruit diameter and weight was sig-
cooking, rather than raw. nificantly higher in WW irrigated tomato plants, as compared to
those in plants irrigated with potable water.

3.6. Effects of the irrigation treatment on tomato fruits’ quality

and crop’s productivity 4. Conclusions

WW irrigation did not significantly affect mean fruit weight and
maximum fruit diameter, as compared to control TW irrigation. As
shown in Fig. 2, fruit weight ranged from 67.66 to 142.89 g, while
fruit maximum diameter from 45.94 to 68.05 mm. In addition, the
total number of fruits in each harvest was not affected by the
irrigation treatment, reaching its highest values in the 5th harvest.
Similarly, Aiello et al. (2007) and Cirelli et al. (2012) reported that
tomato mean fruit weight was not affected by WW irrigation, as
The effects of tomato crop irrigation with treated WW effluent
produced by two discrete technological processes, as compared to
groundwater irrigation, on soil geochemical properties, fruit safety
and crop productivity, were evaluated. Irrigation using either efflu-
ent did not significantly affect soil pH, organic C or heavy metal
content. Also, no impacts of WW irrigation on crop productivity, as
compared to control irrigation, were registered. Furthermore, the
heavy metal content of tomato fruits and leaves in all irrigation
 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114
treatments was below the MPLs set for fruit safety and the critical
tissue concentration for phytotoxicity. Moreover, no microbiolog-
ical contamination (TC, FC, E. coli, Salmonella spp., Listeria spp.) of
tomato fruits was found, in any treatment. Higher Zn, Mn, Ni and Cu
content in tomato fruits, as well as the elevated APC concentration
in tomato fruit peels were measured in samples harvested from
plants irrigated with the MWTP I WW compared to those irrigated
with MWTP II WW. In addition, MWTP I WW irrigation resulted
in higher NO3 − accumulation in soil, as compared to MWTP II WW
irrigation. Overall, results lend support to the increasing use of WW
for the drip irrigation of tomato crop, since no negative impacts on
soil geochemical parameters or on fruit heavy metal bioaccumu-
lation and microbial contamination, were registered. Thus, WW of
the quality used here, which complied with the established criteria,
can be reused for vegetable crops irrigation in water scarce areas
helping to improve water use efficiency, however regular monitor-
ing will be required.
Acknowledgements
The authors wish to thank all the personnel of the Cyprus Agri-
cultural Research Institute involved in the study for their excellent
technical assistance. This study was undertaken in collaboration
with Nireas – International Water Research Center of the University
of Cyprus and supported by Cyprus Agricultural Research Institute
Internal Grant to Anastasis Christou.
References
Aiello, R., Cirelli, G.L., Consoli, S., 2007. Effects of reclaimed wastewater irrigation
on soil and tomato fruits: a case study in Sicily (Italy). Agric. Water Manage. 93,
65–72.
Al-Lahham, O., El Assi, N.M., Fayyad, M., 2003. Impact of treated wastewater
irrigation on quality attributes and contamination of tomato fruit. Agric. Water
Manage. 61, 51–62.
Al-Lahham, O., El Assi, N.M., Fayyad, M., 2007. Translocation of heavy metals to
tomato (Solanum lycopersicom L.) fruit irrigated with treated wastewater. Sci.
Hortic. 113, 250–254.
Angelakis, A.N., Durham, B., 2008. Water recycling and reuse in EUREAU countries:
trends and challenges. Desalination 218, 3–12.
AOAC, 2005. Official methods of analysis of the Association of Official Analytical
Chemists, 18th ed. AOAC Press, Washington DC.
APHA, 1998. Standard method for the examination of water and wastewater, 20th
ed. APHA, AWWA and WEF, Washington, DC.
Ayers, R.S., Westcot, D.W. (Eds.), 1985. Water Quality for Irrigation. FAO, Rome.
Bernstein, N., 2011. Potential for contamination of crops by microbial human
pathogens introduced into the soil by irrigation with treated effluent. Israel J.
Plant Sci. 59, 115–123.
Bernstein, N., Guetsky, R., Friedman, H., Bar-Tal, A., Rot, I., 2008. Monitoring bacte-
rial populations in an agricultural greenhouse production system irrigated with
reclaimed wastewater. J. Hortic. Sci. Biotechnol. 83, 821–827.
Bixio, D., Thoeye, C., De Koning, J., Joksimovic, D., Savic, D., Wintgens, T., Melin, T.,
2006. Wastewater reuse in Europe. Desalination 187, 89–101.
Brissaud, F., 2008. Criteria for water recycling and reuse in the Mediterranean
countries. Desalination 218, 24–33.
Carbonell, G., Imperial, R.M.d., Torrijos, M., Delgado, M., Rodriguez, J.A., 2011. Effects
of municipal solid waste compost and mineral fertilizer amendments on soil
properties and heavy metals distribution in maize plants (Zea mays L.). Chemo-
sphere 85, 1614–1623.
Cirelli, G.L., Consoli, S., Licciardello, F., Aiello, R., Giuffrida, F., Leonardi, C., 2012.
Treated municipal wastewater reuse in vegetable production. Agric. Water Man-
age. 104, 163–170.
Cui, Y.-J., Zhu, Y.-G., Zhai, R.-H., Chen, D.-Y., Huang, Y.-Z., Qiu, Y., Liang, J.-Z., 2004.
Transfer of metals from soil to vegetables in an area near a smelter in Nanning,
China. Environ. Int. 30, 785–791.
Datta, S.P., Subba Rao, A., Ganeshamurthy, A.N., 1997. Effect of electrolytes coupled
with variable stirring on soil pH. Indian Soc. Soil Sci. 45, 185–187.
EC, 2001. Commission Regulation (EC) 466/2001. Setting maximum levels for cer-
tain contaminants in foodstuffs. Official Journal of the European Communities.,
pp. 77.
EEC, 1986. Council Directive of 12 June 1986 on the protection of
the environment, and in particular of the soil, when sewage
sludge is used in agriculture (86/278/EEC), http://eur-lex.europa.eu/
legal-content/EN/TXT/PDF/?uri=CELEX:31986L0278&from=EN (accessed
25.06.13).
Fatta-Kassinos, D., Bester, K., Kümmerer, K., Hapeshi, E., Malato, S., Mantzavinos, D.,
Rizzo, L., Xekoukoulotakis, N.P., 2010. Removal of xenobiotic compounds from
113
water and wastewater byadvanced oxidation processes. In: Fatta-Kassinos, D.,
Bester, K., Kümmerer, K. (Eds.), Xenobiotics in the Urban Water Cycle. Springer,
Netherlands, pp. 387–412.
Fatta-Kassinos, D., Kalavrouziotis, I.K., Koukoulakis, P.H., Vasquez, M.I., 2011. The
risks associated with wastewater reuse and xenobiotics in the agroecological
environment. Sci. Total Environ. 409, 3555–3563.
Forslund, A., Ensink, J.H.J., Markussen, B., Battilani, A., Psarras, G., Gola, S., Sandei,
L., Fletcher, T., Dalsgaard, A., 2012. Escherichia coli contamination and health
aspects of soil and tomatoes (Solanum lycopersicum L.) subsurface drip irrigated
with on-site treated domestic wastewater. Water Res. 46, 5917–5934.
Frank, J.F., 2001. Microbial attachment to food and food contact surfaces. Adv. Food
Nutr. Res. 43, 319–370.
Gong, B., Wen, D., VandenLangenberg, K., Wei, M., Yang, F., Shi, Q., Wang, X., 2013.
Comparative effects of NaCl and NaHCO3 stress on photosynthetic parameters,
nutrient metabolism, and the antioxidant system in tomato leaves. Sci. Hortic.
157, 1–12.
Hamilton, A.J., Stagnitti, F., Xiong, X., Kreidl, S.L., Benke, K.K., Maher, P., 2007. Waste-
water irrigation: the state of play. Vadose Zone J. 6, 823–840.
Kalbar, P.P., Karmakar, S., Asolekar, S.R., 2012. Technology assessment for waste-
water treatment using multiple-attribute decision-making. Technol. Soc. 34,
295–302.
Khan, A., Javid, S., Muhmood, A., Mjeed, T., Niaz, A., Majeed, A., 2013. Heavy metal
status of soil and vegetables grown on peri-urban area of Lahore district. Soil
Environ. 32, 49–54.
Khan, S., Cao, Q., Zheng, Y.M., Huang, Y.Z., Zhu, Y.G., 2008. Health risks of heavy metals
in contaminated soils and food crops irrigated with wastewater in Beijing, China.
Environ. Pollut. 152, 686–692.
Khurana, M.P.S., Singh, P., 2012. Waste water use in crop production: a review.
Resour. Environ. 2, 116–131.
Klay, S., Charef, A., Ayed, L., Houman, B., Rezgui, F., 2010. Effect of irrigation with
treated wastewater on geochemical properties (saltiness, C, N and heavy metals)
of isohumic soils (Zaouit Sousse perimeter, Oriental Tunisia). Desalination 253,
180–187.
Krasensky, J., Jonak, C., 2012. Drought, salt, and temperature stress-induced
metabolic rearrangements and regulatory networks. J. Exp. Bot. 63,
1593–1608.
Li, P., Wang, X., Allinson, G., Li, X., Xiong, X., 2009. Risk assessment of heavy metals
in soil previously irrigated with industrial wastewater in Shenyang, China. J.
Hazard. Mater. 161, 516–521.
Lonigro, A., Rubino, P., Brandonisio, O., Spinelli, R., Pollice, A., Laera, G., 2007. Veg-
etable crop irrigation with tertiary filtered municipal wastewater. Plant Biosyst.
141, 275–281.
Maas, E.V., Hoffman, G.J., 1977. Crop salt tolerance – current assessment. J. Irrig.
Drain. Div. ASCE 103, 115–134.
Macnicol, R.D., Beckett, P.H.T., 1985. Critical tissue concentrations of potentially toxic
elements. Plant Soil 85, 107–129.
Mapanda, F., Mangwayana, E.N., Nyamangara, J., Giller, K.E., 2005. The effect
of long-term irrigation using wastewater on heavy metal contents of
soils under vegetables in Harare, Zimbabwe. Agric. Ecosyst. Environ. 107,
151–165.
Martinez-Rodriguez, M.M., Estañ, M.T., Moyano, E., Garcia-Abellan, J.O., Flores, F.B.,
Campos, J.F., Al-Azzawi, M.J., Flowers, T.J., Bolarín, M.C., 2008. The effectiveness
of grafting to improve salt tolerance in tomato when an ‘excluder’ genotype is
used as scion. Environ. Exp. Bot. 63, 392–401.
Megateli, S., Semsari, S., Couderchet, M., 2009. Toxicity and removal of heavy
metals (cadmium, copper, and zinc) by Lemna gibba. Ecotoxicol. Environ. Saf.
72, 1774–1780.
Meneses, M., Pasqualino, J.C., Castells, F., 2010. Environmental assessment of urban
wastewater reuse: Treatment alternatives and applications. Chemosphere 81,
266–272.
Mesa-Jurado, M.A., Martin-Ortega, J., Ruto, E., Berbel, J., 2012. The economic value of
guaranteed water supply for irrigation under scarcity conditions. Agric. Water
Manage. 113, 10–18.
Metochis, C., 1997. Assessment of irrigation water needs of main crops of Cyprus.
Cyprus Agricultural Research Institute Series. Ministry of Agriculture, Natural
Resources and Environment, Cyprus, Nicosia.
Milano, M., Ruelland, D., Fernandez, S., Dezetter, A., Fabre, J., Servat, E., 2012. Facing
climatic and anthropogenic changes in the Mediterranean basin: what will be
the medium-term impact on water stress? C. R. Geosci. 344, 432–440.
Mohammad Rusan, M.J., Hinnawi, S., Rousan, L., 2007. Long term effect of wastewater
irrigation of forage crops on soil and plant quality parameters. Desalination 215,
143–152.
Morugan-Coronado, A., Garcia-Orenes, F., Mataix-Solera, J., Arcenegui, V., Mataix-
Beneyto, J., 2011. Short-term effects of treated wastewater irrigation on
Mediterranean calcareous soil. Soil Till. Res. 112, 18–26.
Muchuweti, M., Birkett, J.W., Chinyanga, E., Zvauya, R., Scrimshaw, M.D., Lester, J.N.,
2006. Heavy metal content of vegetables irrigated with mixtures of wastewater
and sewage sludge in Zimbabwe: Implications for human health. Agric. Ecosyst.
Environ. 112, 41–48.
Palese, A.M., Pasquale, V., Celano, G., Figliuolo, G., Masi, S., Xiloyannis, C., 2009.
Irrigation of olive groves in Southern Italy with treated municipal wastewater:
effects on microbiological quality of soil and fruits. Agric. Ecosyst. Environ. 129,
43–51.
Petterson, S.R., Ashbolt, N.J., Sharma, A., 2001. Microbial risks from wastewater
irrigation of salad crops: a screening-level risk assessment. Water Environ. Res.
73, 667–672.
114 A. Christou et al. / Agriculture, Ecosystems and Environment 192 (2014) 105–114
Rady, M.M., 2012. A novel organo-mineral fertilizer can mitigate salinity stress
effects for tomato production on reclaimed saline soil. S. Afr. J. Bot. 81, 8–14.
Rajaganapathy, V., Xavier, F., Sreekumar, D., Mandal, P.K., 2011. Heavy metal contam-
ination in soil, water and fodder and their presence in livestock and products: a
review. J. Environ. Sci. Technol. 4, 234–249.
Reilly, T.E., Goodman, A.S., 1987. Analysis of saltwater upconing beneath a pumping
well. J. Hydrol. 89, 169–204.
Sacks, M., Bernstein, N., 2011. Utilization of reclaimed wastewater for irrigation of
field-grown melons by surface and subsurface drip irrigation. Israel J. Plant Sci.
59, 159–169.
Singh, A., Sharma, R.K., Agrawal, M., Marshall, F.M., 2010a. Health risk assessment
of heavy metals via dietary intake of foodstuffs from the wastewater irrigated
site of a dry tropical area of India. Food Chem. Toxicol. 48, 611–619.
Singh, A., Sharma, R.K., Agrawal, M., Marshall, F.M., 2010b. Risk assessment of heavy
metal toxicity through contaminated vegetables from waste water irrigated area
of Varanasi, India. Trop. Ecol. 51, 375–387.
Smith, C.J., Hopmans, P., Cook, F.J., 1996. Accumulation of Cr, Pb, Cu, Ni, Zn and Cd in
soil following irrigation with treated urban effluent in Australia. Environ. Pollut.
94, 317–323.
Szkup-Jablonska, M., Karakiewicz, B., Grochans, E., Jurczak, A., Nowak-Starz, G., Rot-
ter, I., Prokopowicz, A., 2012. Effects of blood lead and cadmium levels on the
functioning of children with behaviour disorders in the family environment.
Ann. Agric. Environ. Med. 19, 241–246.
Tuna, A.L., Kaya, C., Ashraf, M., Altunlu, H., Yokas, I., Yagmur, B., 2007. The
effects of calcium sulphate on growth, membrane stability and nutrient
uptake of tomato plants grown under salt stress. Environ. Exp. Bot. 59,
173–178.
U.S. Environmental Protection Agency, 1998. Microwave Assisted Acid Digestion
of Sediments, Sludges, Soils, and Oils. Office of Solid Waste and Emergency
Response. U.S. Government Printing Office, Washington, DC.
U.S. Environmental Protection Agency, 2012. Guidelines for Water Reuse. United
States Environmental Protection Agency.
Wang, Y., Qiao, M., Liu, Y., Zhu, Y., 2012. Health risk assessment of heavy metals in
soils and vegetables from wastewater irrigated area, Beijing-Tianjin city cluster,
China. J. Environ. Sci. 24, 690–698.
Werner, A.D., Jakovovic, D., Simmons, C.T., 2009. Experimental observations of salt-
water up-coning. J. Hydrol. 373, 230–241.
WHO, 2006. WHO Guidelines for the Safe use of Wastewater, Excreta and Greywa-
ter, Vol. 2. Wastewater use in agriculture. World Health Organisation, Geneva,
Switzerland.
WHO/FAO, 2007. Joint FAO/WHO food standard programme codex alimentarius
commission. 13th Session. Report of the thirty eight session of the codex com-
mittee on food hygiene. United States of America, Houston, ALINORM 07/30/13.
Xu, J., Wu, L., Chang, A.C., Zhang, Y., 2010. Impact of long-term reclaimed wastewater
irrigation on agricultural soils: a preliminary assessment. J. Hazard. Mater. 183,
780–786.