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© IWA Publishing 2014 Journal of Water Reuse and Desalination





The use of treated wastewater and fertigation in
greenhouse pepper crop as affecting growth and
fruit quality
Eleni Dagianta, Dimitrios Goumas, Thrassyvoulos Manios
and Nikos Tzortzakis

The performance and suitability of tertiary treated wastewater (TW) and/or fertigation (F) in pepper
plants were studied over a 4-month period in greenhouse conditions. Four treatments were used
consisting of (1) water, (2) water þ F, (3) TW, and (4) TW þ F. The F and/or TW application increased
plant height and plant biomass compared with the control plants (irrigated with water) while no
differences were observed in the number of leaves produced. Plants grown with TW þ F were thicker
than control plants. The addition of F increased fruit number in both water and TW, which resulted in
increased plant yield and fruit marketability for the water application but decreased mean fruit
weight for TW application. The application of TW increased fruit total soluble solids but decreased
fruit firmness whereas adding F, these changes were normalized. The F and/or TW application
reduced fruit total phenolics, fruit acidity, and fruit length but not fruit diameter. No differences were
observed in fruit dry matter content, fruit color/lightness (a, b, and L value). Bacteria (total coliform

Eleni Dagianta
Dimitrios Goumas
Thrassyvoulos Manios
Nikos Tzortzakis
Department of Organic Greenhouse Crops and
School of Agricultural Technology,
Technological Educational Institute of Crete,
Nikos Tzortzakis (corresponding author)
Department of Agricultural Sciences,
Biotechnology and Food Science,
Cyprus University of Technology,
3603 Limassol,

and Escherichia coli) units on the fruits did not differ among the treatments. The results indicate that
wastewater may act as an alternative means of irrigation if following strict safety aspects while the
fertigation acted beneficially.
Key words

| fertigation, fruit quality, pepper, wastewater, yield

In recent years, the use of marginal-quality water for crop irri-

following primary, secondary, and tertiary (chlorotic, UV,

gation has gained importance in water-scarce regions (Qadir

O3, etc.) treatment aiming at the maximization of crop

et al. ) as the demand for water is continuously increasing

yields, not only quantitative, but also qualitative, without

in arid and semi-arid countries. One of the major types of

environmental risks (Manios et al. ; Petousi et al. ).

marginal-quality water is wastewater, which has been

Disinfection of wastewater is vital in addressing the potential

reused in agriculture for centuries; however, the amount of

health risks of urban water reuse and appropriate methods for

wastewater recycled has greatly increased in the last decade

disinfection as an essential purification step for safe urban

(Bhogal et al. ; Kalavrouziotis et al. ). Types of waste-

reuse have been studied (Bischoff et al. ). The above

water used for recycling include primary treated wastewater

methods of wastewater application will need to be

(PTW, Zabalaga et al. ), secondary treated wastewater

accompanied by restrictions that ensure the protection of

(STW, Pedrero & Alarcón ), and tertiary treated waste-

public health and the protection of air, aqueous, and soil

water (TTW, Pollice et al. ). Several scientific studies

sources. The use of wastewater affects the chemical properties

have researched this area in order to determine the reuse of

of soil in the first 30 cm below ground, as well as the consti-

processed wastewater for the irrigation needs of crops

tution of plants (Kiziloglu et al. ).

doi: 10.2166/wrd.2014.048


E. Dagianta et al.


Wastewater for pepper cultivation

Journal of Water Reuse and Desalination





In general, treated wastewater (TW) reuse has several

present study examined the impacts of mineral fertigation

advantages which include: (1) crop productivity improve-

combined with different water quality (TTW) in soil, in

ment in water-constrained systems (Raschid-Sally et al.

plant growth, and yield and fruit quality related parameters,

); (2) reduced amount of fresh water used for irrigation;

in greenhouse pepper production.

(3) reduced discharge of nutrients into surface waters; and
(4) a decrease in the cost of wastewater treatment by eliminating the need for nutrient removal (Rosenqvist et al.


). Furthermore, irrigation with municipal wastewater is
considered an environmentally friendly wastewater disposal

Plant material and tertiary treated wastewater source

practice that helps to minimize pollution of the ecosystem
which is subjected to contamination by direct disposal of

Pepper (Capsicum annuum L. cv. Oregon) plants were

wastewater into surface or groundwater (Mohammad &

grown under natural light from September to January (in

Mazahreh ). In addition, wastewater is a valuable

2011). PTW was obtained from the sewage treatment unit

source for plant nutrients (exchangeable Na, K, Ca, Mg,

of Heraklion (180,000 p.e.), Crete, Greece. TW was obtained

and P) and organic matter needed for maintaining fertility

by treating the effluent of packed bed filter (Advantex-AX20,

and productivity of arid soils. However, the reuse of waste-

Orenco; used for STW) using a sand filter and a chlorination

water for irrigation may potentially create environmental

process. The physicochemical characteristics of the TTW

problems if not properly treated and managed (Bahri & Bris-

used in the experiment were measured and presented in

saud ), such as nutrient accumulation. Consequently,

previous studies (Petousi et al. ). In detail, the physico-

management of irrigation with wastewater should consider

chemical characteristics of TTW and of tap water (in

the nutrient content in relation to the specific crop require-

parentheses) sources used in the experiment were: pH 7.5

ments and the concentrations of plant nutrients in the soil,

(7.8); electrical conductivity – EC (mS/cm) 1.1 (0.7);

and other soil fertility parameters. Thus, wastewater irriga-

chemical oxygen demand – COD (mg L1) 24 (16); total

tion in lettuce had a high influence on soil parameters:

soluble solids – TSS (mg L1) 9.1 (2.6); total nitrogen –

organic matter, N, P, Ca, Al, Fe, Pb, and Zn as well as patho-

TN (mg L1) 20.0 (4.7); total phosphorus – TP (mg L1)

genic microorganisms (Manas et al. ). Several studies

8.0 (2.0); boron – B (μg L1) 251.2 (16.4); magnesium –

examined the effects of wastewater used in agriculture for

Mg (mg L1) 51.2 (19.1); calcium – Ca (mg L1) 123.0

irrigation needs as well as the impact of fertilizers in the

(60.5); potassium – K (mg L1) 28.5 (not detected); and

crop-soil system, such as alfalfa, radish, and tomato (Albul-

zinc – Zn (μg L1) 7.0 (not detected).

basher et al. ; Manios et al. ; Najafi et al. ;
Aiello et al. ; Petousi et al. ), cucumber (Manios

Experimental design

et al. ; Pilatakis et al. ), eggplant (Najafi et al.
; Cirelli et al. ), citrus (Morgan et al. ), orna-

The experiment was carried out in an unheated plastic

mental (Lubello et al. ), cauliflower, and red cabbage

greenhouse with a north–south orientation at the Techno-

(Kiziloglu et al. ).

logical Educational Institute of Crete (TEI of Crete),

At the market interface, only produce that corresponds

Greece. Seedlings were produced in plastic seedling trays

to the expectations of the consumer can survive. Fruit qual-

filled with expanded clay and were acquired from a local

ity encompasses many aspects, and includes not only flavor,

agriculture nursery. Fertigation (F) and/or TW were used

color, nutritional aspects, and firmness, but also shelf life,

to create four treatments (three replications/treatment;

processing attributes, and resistance to pathogens (Brum-

three plants/replication) which were (1) water, (2) water þ

mell & Harpster ). Moreover, an increased interest in

F, (3) TW, and (4) TW þ F.

vegetables has been created by the fact that their consump-

Seedlings were transplanted in single pots (filled with

tion has been correlated with human health and the

substrate; 9 L capacity pot) and arranged in a single row

reduced risk of some types of cancer (Gerster ). The

with a completed randomized design for the replicates/


E. Dagianta et al.


Wastewater for pepper cultivation

Journal of Water Reuse and Desalination






treatment. Rows were 1.0 m apart and plants were separated

were expressed as the mean (%) of Brix. Sub-samples of

by 0.4 m. Drip irrigation emitters (one emitter/pot) were

homogenized fruit tissue were used to determine the pH

placed and irrigation took place twice (1 min/time) daily,

and EC of fruit juice using a standard pH/EC meter

via timer, by means of pressure pumps. Fertigation (EC:

(Orion 920A; Scientific Support, Hayward, CA, USA). Titra-

2.5–3.0 mS/cm; 200 mL/plant twice a week) with commer-

table acidity (TA) was determined by potentiometric

cial fertilizers took place manually. The drainage solution

titration, using fruit samples (5 g) diluted in 100 mL distilled

was collected in trays in each pot and was available for

water and titrated with 0.1 N NaOH, using phenolphthalein

plant water needs through capillary suction. Plants were

as pH indicator, and monitored up to 8.2 end point with a

treated according to a twin lateral pruning system (two lat-

pH meter. The reported values were expressed in terms of

eral stems growing vertically) on string.

citric acid percentage. Total phenolics was determined on
blended fruit tissue (5 g) extracts following repeated (four-


fold) addition of 2.5 mL of 50% (v/v) methanol, as reported
previously (Tzortzakis et al. ). Results were expressed in

The nutritional status of soil was observed. Thus, we

terms of gallic acid equivalents (GAE; Sigma Aldrich,

measured K and Na content (by a flame photometer), P

Athens, Greece) per 100 g fresh weight of tissue.






organic matter content and organic carbon, pH, and EC.

Fruits (n ¼ 23) were assessed for bacteria (total coliform
and Escherichia coli (E. coli)) units on the fruits as well as in

Beginning the second week after transplanting, we

the fruit, by employing ChromoCult® Coliform Agar (Merck

studied the impact of fertigation and/or water quality on

KGaA), which is a selective and differential chromogenic

plant growth/development, yield and fruit quality in pep-

culture medium. Harvested fruits were placed in sterile plas-

pers. Every second week the plant height, main stem

tic bags (one fruit per bag), which were then filled with

diameter, leaf number produced, flower and fruit number,

225 mL sterile Butterfield’s phosphate buffered water

and the leaf fluoresces (chlorophyll fluorometer, ΟS-30p,

(42.5 g/L KH2PO4, pH 7.2, Merck), and were gently

Opti-Sciences, UK) were measured. With the completion

shaken to rinse off most of the bacteria present. For isolation

of the experiment, plant biomass (fresh weight and the %

of coliform cultures, 1 mL of the solution was added to

of dry matter) and plant yield were determined.

0.45 μm membrane in vacuum under sterile contrition.

Fruit fresh weight, dry matter content (%), and fruit size

Inoculated membrane was transferred into a Petri dish con-

(length and diameter) were measured for each harvested

taining ChromoCult® Coliform Agar, and incubated at 35 C

fruit. Fruit marketability was observed by employing a 1–4

for 24–48 h. Pink colonies resulting from salmon-galactoside


scale (1: extra quality; 2: good quality; 3: medium quality

cleavage by β-D-galactosidase were classified as TCC,

(i.e., small size, and decolorization); and 4: not marketable

whereas dark blue colonies resulting from salmon-galacto-

quality (i.e., malformation, wounds, and infection)). Fruit

side and X-glucuronide cleavage by β-D-galactosidase and

color measurements were taken around the fruit equator

β-D-glucuronidase were classified as presumptive E. coli

(two measurements per fruit) with a Minolta Chroma

colonies (Byamukama et al. ). Similarly, soil samples

Meter CR300. Data are expressed in L × a × b units. Fruit

(n ¼ 5) were assessed for bacteria (total coliform and

firmness was measured at one point on the shoulder of the

E. coli) units.

fruit, for each treatment, using a penetrometer FT 011 (TR
Scientific Instruments, Forli, Italy). The amount of force

Statistical analysis

(in Newtons – N) required to break the radial pericarp
(i.e., surface) of each pepper was recorded at ambient (22–

24 C) temperature.

Data were tested for normality and then subjected to analysis of variance (ANOVA). Significant differences between

TSS concentration was determined on the fruit juice for
each treatment with a digital refractometer 300017 (Sper

Scientific Ltd, Scottsdale, AZ, USA) at 20 C and results

mean values were determined using Duncan’s Multiple
Range test (P < 0.05) following one-way ANOVA. Significant differences on percentage values (% dry weight) were


E. Dagianta et al.


Wastewater for pepper cultivation

Journal of Water Reuse and Desalination





logarithmatically transformed prior to analysis. Statistical

tap water (control) and this is probably due to the nutrient

analyses were performed using SPSS (SPSS Inc., Chicago,

lack of plants irrigated only with water. Plants irrigated

IL) and graphs produced using Prism v.2.0 (Graph Pad

with TW or water þ F produced similar stem thickness high-

Inc., San Diego, CA).

lighting the nutritive value that TW provided to the crop
following 105 days plant growth (Figure 2(a)). No differences








treatments (data not presented).
The addition of F increased fruit number produced in

Soil properties

both water and TW treatments and differed significantly
with the fruit number obtained in plants irrigated only

The nutritional status of soil used in the present study

with water (Table 1 and Figure 2(b)). However, plants irri-

were 0.83% organic matter (0.48% organic C), C/N of

gated with water formed fruits 15 days earlier compared

21.7, pH of 6.94, and EC 0.71 mS/cm as well as total N

with other treatments (Figure 2(b)), which is of interest as

content of 0.14% and 21.73 mg P L1, 5.55 mg K L1 and

fruit earliness is directly related with increased cost benefits

0.32 mg Na L1. The soil was of a white color, a calcareous

of the producers. Fruit number produced followed a similar

clay loam with a CEC of 2.9.

pattern as the number of the produced flowers (data not presented), implying that there was no flower absorption that

Effect on plant growth

was directly related to the applied treatments.

Examining the different F and/or TW application, plants
grown in soil enriched with F and/or irrigated with TW

Effect on plant yield

were taller with significant changes observed after 45 days
(average 40 cm) compared with the control plants (irrigated

Plants irrigated with water þ F increased yield (averaged

with tap water) while no differences were observed in leaf

327.61 g/plant) compared with plants irrigated with water

number produced (Figures 1(a) and 1(b)). The F-enrichment

or TW ± F (Table 1). Fruit fresh weight decreased in plants

in both water and TW application affected positively plant

irrigated with TW þ F compared with TW or water ± F, indi-

height and was in agreement with previous studies in

cating that the increased nutrition (due to the combination

cucumber and tomato crops when irrigated with TW and/

of TW and F, as both are nutrient sources) affected nega-

or fertigation (Manios et al. ). Plants grown with

tively the fruit fresh weight. No differences were obtained

TW þ F produced thicker stems than plants grown with

in fruit dry matter content among treatments (Table 1).

Figure 1


Effects of fertigation (F) and/or TW into soil on plant height and leaf number produced of greenhouse pepper crop. Values represent mean (±SE) of measurements made on six
plants per treatment.

E. Dagianta et al.



Figure 2


Wastewater for pepper cultivation

Journal of Water Reuse and Desalination





Effects of fertigation (F) and/or TW into soil on stem thickness and fruit number produced of greenhouse pepper crop. Values represent mean (±SE) of measurements made on
six plants per treatment.

Table 1


Effects of fertigation (F) and/or TW into soil on yield (g/plant), upper biomass fresh fruit (g/plant) and upper biomass dry matter (%) in greenhouse pepper crop

Yield (g/plant)

Biomass fwt (g/plant)

Biomass dry content (%)

Fruit no

Fruit fwt (g)

Fruit dry matter (%)


221.75 ba

57.86 ba

47.12 aa

2.12 bb

100.66 ab

8.55 ab

Water þ F

327.61 a

68.61 a

41.71 b

2.88 a

121.14 a

7.63 a


248.87 b

73.11 a

47.44 a

2.52 ab

104.60 a

5.71 a

TW þ F

235.45 b

79.31 a

37.74 c

3.63 a

67.72 b

6.99 a


Values (n ¼ 6) in columns followed by the same letter are not significantly different. P < 0.05.


Values (n ¼ 23) in columns followed by the same letter are not significantly different. P < 0.05.

The increased yield obtained in plants irrigated with water

The F and/or TW application increased plant biomass

þ F is due to the increased fruit number (more fruits)

(fresh weight; fwt) compared with the control plants (irri-

rather than differences in fruit fresh weight. When munici-

gated with water) while TW þ F reduced biomass dry

pal-TW was used in ornamental plants in nurseries, the

matter content (Table 1).

nutrient content of the tertiary effluent was able to maintain
as good plant growth as fertigated water for most of the

Effect on fruit quality

tested species (Lubello et al. ), which is in accordance
with the present study. Lettuce treated with wastewater

Fruit size fluctuated among treatments and the effects were

increased yield compared with control as well as increased

mainly in fruit length rather than fruit diameter (i.e., thicker

N, P, Pb, and Al content in plant tissues (Manas et al.

fruits) (Table 2). In detail, the greatest fruit length was observed

). Wastewater irrigation treatments also increased the

in the case of plants irrigated with water while plants irrigated

yield as well as N, P, K, Ca, Mg, Na, Fe, Mn, Zn, Cu, Pb,

with TW had a reduced (up to 34%) fruit length. Interestingly,

Ni, and Cd contents of cauliflower and red cabbage plants

the addition of fertilizers (TW þ F) alleviated the reduction in

(Kiziloglu et al. ). Additionally, a part of the increased

fruit length. Fruit firmness reduced when plants irrigated with

yield in tomato and eggplant irrigated with TW can be

TW were compared with the control treatment (water) while

related to better soil moisture and increased available nitro-

the addition of F resulted in fruit firmness being maintained

gen in the root zone (Najafi et al. ; Cirelli et al. ),

(see Table 2). Fruit firmness is an important quality attribute

which is in line with the present study with the increased

and is directly related to enhancing the storability potential

nutrient concentration, i.e., N, of TW compared with tap

and to inducing greater resistance to decay and mechanical


damage (Barret & Gonzalez ).

E. Dagianta et al.


Table 2



Wastewater for pepper cultivation

Journal of Water Reuse and Desalination





Effects of fertigation (F) and/or TW into soil on fruit size (length and diameter in mm), fruit firmness (N) and fruit color (L, a, and b values), fruit marketability on 1–4 scale (1: extra
quality; 2: good quality; 3: medium quality; and 4: not marketable quality) in greenhouse pepper crop

Fruit size (mm)


Fruit color



Fruit firmness





7.31 a

6.77 a

4.40 a

33.69 b 

10.81 a

14.37 a

2.00 b

Water þ F

5.75 b

6.31 a

4.40 a

34.16 b 

10.57 a

13.62 a

2.55 a


4.84 c

6.53 a

3.16 b

37.02 a 

10.62 a

14.22 a

2.21 ab

TW þ F

5.43 b

5.82 a

4.36 a

34.44 b 

9.55 a

13.21 a

2.36 ab

Values (n ¼ 23) in columns followed by the same letter are not significantly different. P < 0.05.

Table 3



Effects of fertigation (F) and/or TW into soil on TSS (TSS: Brix), titratable acidity (TA: (% citric acid)), sweetness (TSS/TA), pH, EC (mS/cm), and total phenols (gallic acid equivalent:
GAE/ 100 g fwt) in greenhouse pepper crop






Total phenols


3.26 b

4.68 a

0.79 c

5.30 a

1.78 a

296.91 a

Water þ F

3.20 b

3.09 b

1.13 b

5.30 a

1.48 a

184.57 c


3.76 a

3.10 b

1.28 a

5.11 b

1.62 a

263.15 b

TW þ F

3.53 b

3.52 b

1.01 b

5.38 a

2.01 a

186.61 c

Values (n ¼ 23) in columns followed by the same letter are not significantly different. P < 0.05.

Fruit marketability was maintained as good quality

agreement with previous studies in tomato and eggplant

(marked as 2 out of the four values) for the water application

(Cirelli et al. ). In detail, the average total coliform

but was enhanced when fertigation was applied and this is

units were 5.3 CFU/100 g fruit fwt while fertigation

probably due to the better nutrition that plants achieved.

enhanced the presence of bacteria. The average number of

No differences were observed when TW was used in combi-

E. coli was approximately 0.36 CFU/100 g fruit fwt and it

nation with fertigation (Table 2). Fruit lightness (L)

was only present in the case of the application of fertilizer

increased when TW was used while the addition of fertilizer

in soil (water þ F and TW þ F). When selected fruits were

reduced L value (Table 2). Fruit green color (Chroma a

examined for the presence of bacteria inside the fruit, bac-

and b), fluctuated among treatments but did not differ

teria units were not detected (data not presented),


implying that bacteria did not move through plant tissue.

The application of TW increased fruit TAA and fruit
sweetness (TSS/TA ratio) whereas upon the addition of F,

Table 4


Effects of fertigation (F) and/or TW into bacteria (total coliform and E. coli) units
(CFU/100 g fruit fwt) on the fruits as well as in the soil (CFU/100 g soil) in green-

these changes were normalized. The F and/or TW appli-

house pepper crop

cation reduced fruit total phenolics and fruit acidity
(Table 3). Pepper fruits were less acid in plants irrigated


with TW but no differences were observed in the pepper





juice EC. In previous studies, irrigation with TW did not






affect tomato fruit pH, increased their size up to 2 cm in diam-


3.81 a

0.00 a

eter, and weight up to 78.7 g. Additionally, a decrease of 1.5%

Water þ F

4.48 a

0.56 a

420 b

40 a

in the TSS, 0.59 kg in firmness, and 5.1% in weight loss of


5.82 a

0.00 a

5352 a

51 a

tomato fruit were recorded (Al-Lahham et al. ).

TW þ F

7.26 a

0.95 a

8005 a

50 a

Bacteria (total coliform and E. coli) units on the fruits
did not differ among the treatments (Table 4) being in

93 c

66 a

Values (n ¼ 23) in columns followed by the same letter are not significantly different. P <

E. Dagianta et al.



Wastewater for pepper cultivation

However, this fact needs to be examined more precisely and
in greater detail in the future, before final statements. It is
worthwhile mentioning that the bacteria load into the soil
was approximately 103 times more than the one in fruits
(which is in agreement with previous studies; Manios et al.
) for the equivalent treatments (see Table 4) while the
wastewater application in tomato crops resulted in increased
microbial contamination (E. coli and fecal Streptococci) on
the soil surface (Aiello et al. ). In previous studies, the
washing solution of tomato fruits grown with wastewater
was analyzed for fecal coliforms. It appeared that the fruit
skins were free of viable fecal coliforms 24 h after the wastewater application. Subsurface drainage analyses did not show
any alarming levels of constituents irrespective of the source
of the water: wastewater or freshwater (Albulbasher et al.
The use of TW in agriculture may act as an alternative
method for crop irrigation considering the great shortage
of water in the last years. Moreover, wastewater contains
nutritive value (mainly N and P) that should be considered
as additives in crop needs while the microbial load that
wastewater may contain might act as a negative parameter
for plant growth and fruit production. The use of TW for
vegetable production, which are considered as edible fresh
products, needs further exploration. Indeed, the negligible
microbial contamination of tomato fruit and washing solution suggested that the TW can be used as a valid
alternative for irrigation of tomatoes. However, the TW
can be used as an alternative for irrigation practice in vegetables, i.e., tomatoes, eaten after cooking but not for
those eaten raw provided that the effluent quality is continuously monitored to avoid contamination (Al-Lahham et al.
). In conclusion, the results indicate that wastewater
may be used as an additional water resource and act as an
alternative means of irrigation for pepper cultivation in
water-scarce Mediterranean environments, following strict
safety aspects while the fertigation acted beneficially.

This work was funded by the Greek National Research
program XM EOX: EL0031.

Journal of Water Reuse and Desalination





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First received 27 October 2013; accepted in revised form 9 December 2013. Available online 2 January 2014