Aquacult Int

DOI 10.1007/s10499-014-9821-3

Effect of water flow rates on growth of Cyprinus carpio

var. koi (Cyprinus carpio L., 1758) and spinach plant
in aquaponic system

Tanveer Hussain A. K. Verma V. K. Tiwari Chandra Prakash

G. Rathore A. P. Shete Neelam Saharan

Received: 22 January 2014 / Accepted: 11 August 2014

Springer International Publishing Switzerland 2014

Abstract The experiment was aimed at standardization of water flow rate in aquaponic
system in order to correlate nutrient removal and water quality with growth of Cyprinus
carpio var. koi (koi carp fingerlings) and Beta vulgaris var. bengalensis (spinach). Dif-
ferent flow rates, i.e., 3.2, 1.5, and 1.0 l min-1, were assigned as treatments T1, T2, and
T3, respectively, with spinach plants (28 plant m-2), whereas S1 and S2 were the treat-
ments having flow rates of 1.5 and 1.0 l min-1, respectively, without plants. Control
(C) was set at flow rates of 3.2 l min-1 without plants. Treatment T2 (1.5 l min-1) showed
highest weight gain of koi carp fingerlings and also height gain of spinach plants as
compared to other treatments. There was no significant difference in length gain, per-
centage weight gain, specific growth rate, feed conversion ratio, feed efficiency ratio, and
protein efficiency ratio as compared to other treatments and control. All the treatments, T1,
T2, and T3, effectively remove nitrate (7778 %), phosphate (47.0455.06 %), and
potassium (22.8529.16 %) from fish effluent tanks. These results suggest that flow rates
3.2, 1.5, and 1.0 l min-1 were effective under aquaponic system. Of which, 1.5 l min-1
can be suggested as optimum water flow rate for the growth of spinach and koi carp in
aquaponic system as percentage weight gain in fish, percentage height gain, and yield of
plants were higher compared to flow rates 3.2 and 1.0 l min-1.

Keywords Aquaponic system
Koi carp
Spinach
Flow rate
Nutrient dynamics

Nutrient removal

Introduction

Aquaculture has emerged as a major food producing sector, and it is now a major global
industry with total annual production exceeding 63.6 million tons (FAO 2012). Nowadays,
scarcity of water has become a major problem in many of the countries because of the

T. Hussain
A. K. Verma (&)
V. K. Tiwari
C. Prakash
G. Rathore
A. P. Shete
N. Saharan

Central Institute of Fisheries Education, Panch Marg, Yari Road, Versova, Andheri (W),
Mumbai 400 061, India
e-mail: akverma45@yahoo.com

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increased human population. Water is a prerequisite for successful aquaculture operations.

Aquaculture practices also generate lots of wastewater, which is one of the major causes of
environmental pollution. Aquaponics is one of the solutions to both these problems.
Wastewater from the aquaculture system, which is nutrient-rich water, can be used for
growing of plants without addition of any additional chemical nutrients to be added to the
system. Aquaponics is an integration of recirculating aquaculture systems with soil-less
production of plants (Rakocy et al. 2006). Recirculating systems are designed in such a
manner so as to raise large quantities of fish in relatively small volumes of water and then
reusing the wastewater after treating the water for the removal of toxic waste products.
Aquaponic systems require substantially less water quality monitoring than separate
hydroponic or recirculating aquaculture systems. Although the practices of fish farming
and hydroponics have been traced to ancient times, the combination of these two is quite
new. Research in aquaponics has began in the 1970s, and the integration of aquaculture and
the hydroponic cultivation of plants has been examined repeatedly over the past three
decades with a wide variety of system designs, protocols, plant, and aquatic animal species
(Rakocy and Hargreaves 1993). Integrated hydroponics and aquaculture was not much
successful until the 1980s; however, innovations since the 1980s have transformed aqua-
ponics technology into a feasible system of food production (Diver 2006). McMurtry et al.
(1993, 1997b) created the first known closed-loop aquaponic system (called an aqua-
vegeculture system) in 1986 that used tilapia culture effluent into sand-planted tomato
beds. Integration of these techniques can possibly reduce wastes and associated environ-
mental impacts, with additional crop at the same time (Naegel 1977; Quillere et al. 1995;
Rakocy et al. 2006). Recirculating hydroponic systems integrate fish production and plant
production, and have been proposed for the control of waste nutrients accumulation from
fish culture in a way that consumes less water and produces additional saleable crops
(Ghaly et al. 2005). Hydroponic plants have been widely used in wastewater treatment
systems because they efficiently absorb dissolved compounds in wastewater as nutrients
for plant growth (Rakocy et al. 1989; Dushenkov et al. 1995).
Vegetables are good candidate plants for use in recirculating hydroponic systems as
they grow rapidly in response to the high levels of nutrients in aquaculture water. Different
species of vegetables such as ice-lettuce, tomatoes, leaf lettuce, and basil have been suc-
cessfully grown in aquaculture wastewater (Naegel 1977; Quillere et al. 1995; McMurtry
et al. 1997b; Rakocy et al. 2006). However, the goal was to culture a vegetable that will
generate the highest level of income per unit area per unit time. With this criterion,
culinary herbs are the excellent choice as they grow rapidly and fetch high market prices.
The income from herbs such as basil, cilantro, chives, portulaca, parsley, and mint is higher
than that from fruiting crops such as tomatoes, cucumbers, egg plant, and okra (Rakocy
et al. 2006).
Spinach, Beta vulgaris var. bengalensis (Chenopodiaceae), is an annual herb with deep
taproot and shallow secondary root (Nonnecke 1989). Commonly called spinach, the crop
is mainly a winter vegetable crop that survives low temperatures (Nonnecke 1989). The
crop can, however, be successfully grown under partial shade in summer provided there is
sufficient moisture at the root zone (Klein 2007). Spinach is considered to be a popular
salad ingredient rich in iron content and a good source of folic acid. Spinach can be grown
hydroponically in greenhouse production (Rosik-Dulewska and Grabda 2002). Aquarium
fishes are rapidly gaining importance not only because of their aesthetic value but also due
to their immense commercial value in the export trade all over the world. The ornamental
fish production is slowly gaining momentum in India, and efforts have been made to
increase the countrys share in global market. The world trade of ornamental fish is

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estimated to tune of US $ 4.5 billion and striding further, with annual growth rate of about
10 % (Ayyappan et al. 2011). Among ornamental fishes, Koi carp, Cyprinus carpio var.
koi of the cyprinidae family, is a popular fish. Koi carp are hardy, so they make an
excellent aquarium species as well as laboratory species. They are easy to culture and are
also available in several varieties. Koi carp of Asian origin is an important ornamental
species and cultured throughout the world (Hickling et al. 2007). It is a fast-growing fish
species best suited to farming in ponds and lakes (Hashem et al. 1997). Due to ever
growing demand of koi carp, there is an urgent need to intensify its culture. Like most
carps, koi carp excretes large amount of waste because it does not have a stomach and only
has an intestinal tract, and thus cannot digest excess of proteins unlike most tropical fishes.
According to Al-Hafedh (1999), growth rate of fish increases with increase in the level of
dietary protein till the optimum level is reached. Fish mortality occurs due to excessive
accumulation of wastes to toxic levels. Thus, high stocking density of koi carp culture
necessitates high water exchange, discharge of effluent water leads to pollution of the water
bodies. Moreover, koi culture system requires a continuous supply of freshwater source
that is generally not available in most of the water scarce areas so aquaponic systems are
well suited to these conditions.
In this study, hydroponic barrels planted with spinach B. vulgaris var. bengalensis were
integrated with a recirculating aquaculture tank of koi carp. The main objectives of the
study were to investigate the effect of flow rate on the growth performance of the koi carp
fingerlings and spinach and to analyze the effect of flow rate on water quality and nutrient
removal in the aquaponic system.

Materials and methods

Design of experimental aquaponic system

The aquaponic recirculating system was designed and set up at the wet laboratory of
aquaculture division, CIFE, Mumbai. The system consisted of 18 individual, identical
aquaponic units, allowing replication of experimental treatments. Each aquaponic re-
circulating system consisted of a fish tank, 2 hydroponics tanks filled with gravel, a
submersible pump with pipe arrangement, and ball valves to regulate the water flow
rate (Fig. 1). Pipelines (15 mm) made of polyvinyl chloride with ball valve (15 mm)
were installed to recirculate water between the fish culture tank and hydroponic tanks.
Rectangular tanks of 180 l (81.2 9 57 9 38.8 cm) capacity were selected for fishes.
Water circulation in the aquaponic system was done by using submersible water pump
(Sobo WP1650 1,500 l h-1 at 1.5 m head). Two halves of a HDPE barrel each having size
of 0.51 m2 and a depth of 29.4 cm were used for hydroponics tank. Hydroponic tanks were
filled with the gravel of size ranging from 5 to 15 mm. Water from the fish tank to the
hydroponics was regulated by 15-mm ball valves provided at both sides of hydroponic tank
and water from the hydroponic tank returned by gravity force to the fish tank with help of
15-mm PVC drain pipe.
For experimental purpose, water was filled up to 120 l. The aquaponic system operated
constantly with known density of fishes, continuous supply of water (24 h day-1) pumped
from the fish tank to the hydroponic barrels via submersible pump, and the flow rate of
water is regulated by ball valve. Thus, the constant filtration of culture water is maintained.

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Effluent
Clean water Ball valve (water flow regulator)

Hydroponic barrel

Drainage pipe

Aeration

Fish tank

Submersible Pump

Fig. 1 Schematic representation of the aquaponic system

Fish and spinach plantlets

Koi carp fingerlings were procured from commercial breeders of Kurla, Mumbai, and
acclimatized for 30 days in a 1,000-l capacity tank. Fingerlings of koi carp were stocked in
each tank at 1.4 kg m-3 stocking density. The size of the fingerlings was measured before
stocking. The average size and weight of koi carp fingerling at the time of stocking were
6.08 0.04 cm and 5.96 0.05 g, respectively. The fingerlings were provided with
artificial pelleted feed with 32 % protein, 7.07 % moisture, 8.31 % ether extract, and fed at
the rate 2 % of body weight twice in a day.
Seed of spinach (B. vulgaris var. bengalensis) was sown in nursery tray with coconut
husk medium, adjacent to the experimental setup, and allowed to grow for 15 days before
transplanting into the experimental aquaponic systems. Spinach plants were transplanted
from the trays to hydroponics tanks at a stocking density of 28 no m-2. The size of the
plants at the time of stocking was 6.83 0.09 cm.

Experiment methodology

Optimization of flow rate was studied by conducting experiments with different flow rates
in an aquaponic system for the period of 45 days. Before starting of the experiment, all
tanks were properly cleaned and disinfected with KMnO4 solution. The groundwater used
for the experiment was stored in the reservoir and provided with aeration for 2 days prior
to start of the experiment. Chemical composition of groundwater used for the experiment
was presented in the (Table 1). The experimental design consisted of 3 treatments, each
having 3 replicates. Different flow rates, i.e., 3.2, 1.5, and 1.0 l min-1, were assigned for
T1, T2, and T3 treatments, respectively. S1 and S2 were the treatments with water flow

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rates 1.5 l min-1 and 1.0 l min-1, respectively, without spinach plants to compare nutrient
accumulation (nitrate, potassium, and phosphate). Control (C) was set with water flow rate
3.2 l min-1 without plants in hydroponic tanks with 3 replicates. Initially, the system was
operated for 2 weeks with few fishes to enhance ammonia level for the growth of nitrifying
bacteria. After that, spinach plantlets were transplanted in the hydroponic component of the
system.

Sampling

Sampling of fishes was carried out at 15 days interval for the assessment of growth (length
and weight). Plant growth was monitored once in 15 days by measuring the plant height.

Water sample analysis

Water quality parameters were analyzed during the experimental period with an interval of
10 days using the standard methods outlined in APHA (2005). Sampling was carried out
between 8.30 am and 9.30 am on each sampling date. Samples were refrigerated at 4 C in
labeled polythene bottles for chemical analysis. The water temperature was measured by
the thermometer, and pH was measured by using universal pH indicator for all the
experimental tanks. Dissolved oxygen, free CO2, hardness, alkalinity, ammonia, nitrite,
and nitrate were analyzed by standard methods outlined in APHA (2005). Sodium,
potassium, and calcium in the water samples were estimated by flame atomic emission
spectrometry (FAES) using a flame photometer (Elico CL 378, India).
Magnesium, iron, and zinc in the water samples were analyzed by digesting water
samples using Supra Pure concentrated acids (Merck) in a microwave-based digestion
system (Microwave 3000, Anton Parr, USA). Digested samples were diluted subjected to
the analysis of the three elements (Mg, Fe, and Zn) by atomic absorption spectropho-
tometer (Analyst 800, PerkinElmer, USA) using flame atomization.

Statistical analysis

The obtained data were analyzed using SPSS version 16 in which one-way ANOVA and
Duncans multiple range test were performed at a significance level of (P \ 0.05) at 95 %
confidence limit to know the significant difference between the treatments and control
means for different parameters.

Results and discussion

Fish growth parameters

This study was set up to compare three different flow rates suitable for the growth of
spinach and koi carp under aquaponic system. Fish growth, spinach growth, nutrient
removal, and water quality parameters were used as indicators of suitability. The mean
weight of koi carp did not vary significantly (P [ 0.05) on 15 and 30th days (Table 2). The
body weight of koi carp at the time of harvest varied significantly (P \ 0.05) among the
treatments and control with slightly higher value observed in T2 (8.65 0.01) followed by
S1 (8.63 0.02), T3 (8.61 0.02), C (8.60 0.02), S2 (8.59 0.01), and T1

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Table 1 Chemical composition

Water quality parameters
of groundwater used for the
experiment
Salinity (ppt) 1.61 0.00
pH 7.38 0.01
DO (mg l-1) 6.36 0.09
Free CO2 (mg l-1) 5.23 0.15
Hardness (mg l-1) 418.33 2.88
Alkalinity (mg l-1) 239 0.58
Ammonia (mg l-1)
NitriteN (mg l-1)
NitrateN (mg l-1)
Phosphate (mg l-1)
Potassium (mg l-1) 18.63 0.07
Calcium (mg l-1) 127.83 0.44
Magnesium (mg l-1) 52.0 0.58
Sodium (mg l-1) 323.17 1.69
Iron (mg l-1) 0.08 0.01
Zinc (mg l-1) 0.12 0.00

(8.58 0.01) (Table 2). The different flow rates did not induce any significant effect on
the survival of koi carp among the treatments. All the treatments and control showed
100 % survival (Table 2). The percentage of weight gain at the end of 45-day experiment
was not significantly different (P [ 0.05) among all treatments and control. The highest
percentage of weight gain was observed in T2 (45.24 0.46 %) followed by S1
(44.94 0.42 %), T3 (44.47 0.45 %), C (44.12 0.88 %) and S2 (43.85 0.41 %),
and T1 (43.89 0.89 %) (Table 2). Specific growth rate (SGR) (% per day) did not show
any significant difference (P [ 0.05) among all the treatments and control and followed
same trend as that of weight gain showing highest value in T2 (0.83 0.01) and lowest in
T1 (0.79 0.01) (Table 2). Treatments and control did not show any significant difference
in length gain at the end of 45-day experiment (P [ 0.05) (Table 1).
The feed conversion ratio (FCR) did not show any significant difference (P [ 0.05)
among all the treatments and control. The FCR values varied between 2.28 and 2.34
(Table 1). The feed efficiency ratio (FER) did not show any significant difference
(P [ 0.05) among all the treatments and varied between very narrow range from 0.43 to
0.42 (Table 2). The protein efficiency ratio (PER) values varied within narrow range of
1.331.36, which was not significantly different in all the treatments and control (Table 2).
Total amount of feed delivered was (153.74 0.12 g) in all treatments and control, and
the FCR values obtained in this study support the findings of Korkmaz and Cakirogullari
(2011); reported FCR value for koi carp was 2.21 fed with 34 % crude protein.

Plant growth parameters

Plant growth is another important aspect in the aquaponic system. In our study, spinach
was grown in gravel medium in an aquaponic system and supplied with fish wastewater in
three different flow rates. As Lennard and Leonard (2006) reported 90.9 % removal of
nitrate using gravel bed as media for lettuce production, so the gravel media were used for
experiment. In the present study, height of plant showed significant difference between

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Table 2 Growth performance of koi carp under different flow rates in treatments and control
Parameter Treatments

C T1 T2 T3 S1 S2

Flow rate (l min-1) 3.2 (without plant) 3.2 (with plant) 1.5 (with plant) 1.0 (with plant) 1.5 (without plant) 1.0 (without plant)
Fish growth parameters
Initial weight (g) 5.97a 0.02 5.95a 0.01 5.96a 0.02 5.97a 0.06 5.96a 0.02 5.96a 0.02
a a a a a
Initial length (cm) 6.03 0.13 6.13 0.12 6.10 0.12 6.06 0.12 6.13 0.09 6.03a 0.09
a a a a a
15-day weight (g) 6.85 0.02 6.83 0.09 6.84 0.03 6.82 0.05 6.80 0.02 6.78a 0.02
a a a a a
15-day length (cm) 6.70 0.20 6.56 0.12 6.66 0.09 6.50 0.06 6.66 0.03 6.73a 0.09
ab a ab ab abc
30-day weight (g) 7.73 0.02 7.64 0.03 7.72 0.03 7.66 0.03 7.68 0.01 7.65a 0.02
a a a a a
30-day length (cm) 7.30 0.15 7.20 0.12 7.23 0.09 7.33 0.12 7.26 0.09 7.20a 0.12
ab e b ab ab
Final Weight (g) 8.60 0.02 8.58 0.01 8.65 0.13 8.61 0.02 8.63 0.02 8.59a 0.01
a a a a a
Final Length (g) 7.70 0.06 7.76 0.09 7.83 0.03 7.73 0.09 7.80 0.06 7.76a 0.12
a a a a a
Percentage weight gain 44.12 0.88 43.89 0.89 45.25 0.46 44.47 0.45 44.95 0.42 43.85a 0.41
a a a a a
SGR 0.81 0.01 0.80 0.01 0.83 0.01 0.82 0.01 0.82 0.01 0.81a 0.01
a a a a a
FCR 2.34 0.04 2.32 0.05 2.28 0.02 2.31 0.02 2.28 0.02 2.34a 0.02
a a a a a
FER 0.42 0.01 0.42 0.01 0.43 0.00 0.43 0.00 0.43 0.00 0.42a 0.01
a a a a a
PER 1.33 0.01 1.34 0.01 1.36 0.01 1.34 0.01 1.36 0.01 1.33a 0.01
a a a a a
Survival rate (%) 100 100 100 100 100 100a
Mean values with same superscript did not show any significant difference (P [ 0.05)

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treatments. On 15th and 30th day of sampling, average height of spinach plant showed
significant difference (P \ 0.05); T2 (1.5 l min-1) shows highest plant height of
10.86 0.06 cm and 18.30 0.12 cm on 15th and 30th day, respectively. Similarly, at
the end experiment, better height was obtained in the flow rate of 1.5 l min-1 as compared
to other treatments, whereas no significant difference between treatments was obtained in
terms of yield (Table 3). Spinach plants grown in aquaponic tanks were healthy, indicating
that there were no major mineral deficiencies or toxicities caused by the wastewater. The
results of the present study were comparable with the findings of Endut et al. (2009) and
reported that 1.6 l min-1 showed best plant growth as well as fish growth under an
aquaponic system using water spinach and African cat fish.

Correlation between water volume, fish production, and plant production

The mean initial biomass of fish at the start of the experiment did not vary significantly
(P [ 0.05), and at the end of 45 days, there was no significant (P [ 0.05) difference found
among the treatments (Table 4) in terms of fish biomass, spinach yield, and this clearly
indicate that aquaponic treatments with flow rates 3.2, 1.5, and 1.0 l min-1 were found
effective producing 1.2 kg of spinach with stocking density 142 g of koi carps, and the
observation from the present study states that 142 g of koi carp biomass with water volume
of 120 l fulfills the nutrient requirement of spinach plant, at constant water flow rates (3.2,
1.5, and 1.0 l min-1). Thus, plants assimilated the available nutrients from the culture
water and maintained the water quality parameters congenial for the growth of koi carp.

Water quality parameters and nutrient dynamics

The water temperature during the study period varied within a range of 2527.6 C, with
no marked variation between the treatments and control at any given time of sampling. The
mean value of salinity of water did not show any significant difference (P [ 0.05) among
the treatments and control (Table 5). In general, pH varied within a narrow range of
7.387.29, and no marked variation was observed among the treatments and control
(Table 5). In the present study, water renovation per day was approximately 4 % due to the
loss of water through evaporation and transpiration by plant. Similarly, Lewis et al. (1978)
reported that average daily water renovation in a gravel/sand bed system is around 6.6 %,
and also, McMurtry et al. (1997a) suggested daily water exchange rates were between 1.2
and 4.7 %.

Table 3 Growth performance of spinach in different treatments

Parameters Treatments

T1 T2 T3

Initial height (cm) 6.80 0.06a 6.86 0.03a 6.83 0.03a

a b
15-day height 10.30 0.10 10.86 0.06 10.70b 0.11
a b
30-day height 17.60 0.12 18.30 0.12 18.10b 0.17
a b
Final height (cm) 23.86 0.03 24.26 0.09 24.00a 0.10
a a
Percentage height gain 251.04 3.41 253.40 0.65 251.24a 2.47
a a
Yield (kg) 1.25 14.10 1.28 8.21 1.26a 15.31

Mean values with same superscript did not show any significant difference (P [ 0.05)

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Table 4 Mean weight of biomass of koi carp, C. carpio var. koi, and spinach at the end of 45-day rearing
Parameters Aquaponic treatments

T1 T2 T3

Flow rate (l min-1) 3.2 1.5 1.0

Initial biomass of koi carp (0 day) 142.72a 0.35 142.48a 0.35 142.14a 0.64
Final biomass of koi carp (45 day) 206.32a 0.21 206.80a 0.42 206.40a 0.14
a a
Yield of spinach (kg) 1.25 14.10 1.28 8.21 1.26a 15.31

Mean values with same superscript did not show any significant difference (P [ 0.05)

The mean dissolved oxygen content in all treatments and control varied significantly
(P \ 0.05). The level of dissolved oxygen was highest in C (6.47 0.07 mg l-1), fol-
lowed by T1 (6.46 0.08 mg l-1), whereas lowest dissolved oxygen concentration was
observed in T3 (6.12 0.14 mg l-1) (Table 5). Maintenance of dissolved oxygen is
important for fish health and aerobes of biofilter. Dissolved oxygen levels should be
maintained above 5 mg l-1 for optimum growth of warm water fish (Masser et al. 1999).
Similarly, activity of nitrifying bacteria to convert harmful ammonia to less harmful nitrate
is also dependent on dissolved oxygen. Nitrifying bacteria are known to become inefficient
at DO levels below 2 mg l-1 (Masser et al. 1999). In the present experiment, DO levels in
all treatments and control were found to be in favorable ranges (Fig. 2).
The mean free CO2 was significantly different (P \ 0.05) in all the treatments and
control. The highest value of CO2 was observed in T3 (5.04 1.07 mg l-1) and S2
(5.00 0.22 mg l-1), whereas lowest was observed in T1 (4.27 0.19 mg l-1). The total
alkalinity values did not show any significant difference (P [ 0.05) among the treatments
and control (Table 5). Similarly, hardness of water among all treatments and control did
not show any significant difference (P [ 0.05) (Table 5).
The suggested value of ammonia in a recirculating system should be \1.00 mg l-1
(Van Rijn and Rivera 1990; Nijhof and Bovendeur 1990). A main waste product of the
protein catabolism in fish is ammonia (Wood 1993). Ammonia toxicity depends on the
water parameters including pH and temperature (Randall and Tsui 2002). Chronic expo-
sure to ammonia reduces growth and survival of the fish at early development stages
(Brinkman 2009). Gomuka et al. (2011) suggested critical level of the unionized ammonia
nitrogen for Leuciscus idus larvae (cyprinidae) was 0.21 mg l-1. (Wood 2004) reported
low concentration of ammonia in the water stimulates the growth of fish. Nowosad et al.
(2013) reported that the biomass of the reared fish is the only factor that can influence the
ammonia concentration in water, under intensive rearing of Tench tench in controlled
condition. In the present study, the observed values of ammonia were in the range of
0.030.15 mg l-1. NH4N concentration was highest in control as well as treatment
without plants S1 and S2 as compared to treatments T1, T2, and T3 (Fig. 3), which
indicates the uptake efficiency of the spinach plants. Vaillant et al. (2004) reported that
ammonium (NH4?) is one of the major sources of inorganic nitrogen taken up by the roots
of higher plants. During the experiment, nitriteN showed significant difference between
treatments and control. The mean nitratenitrogen concentration varied significantly
(P \ 0.05) between treatments and control. The highest nitratenitrogen level was detected
in control (C) (14.65 1.79) followed by treatments S2 (12.38 1.89), S1
(12.22 1.89), T1 (3.48 0.39), T2 (2.78 0.30), and T3 (2.57 0.26) (Table 6). This
shows greater assimilation of nitrates by spinach plants. NitrateN is relatively harmless

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Table 5 Mean of physicochemical parameters and nutrients in fish tank after the experimental period of 45 days for different treatments
Parameters Chemical composition of raw C T1 T2 T3 S1 S2
water at the beginning of
the experiment

Flow rate (l min-1) 3.2 3.2 1.5 1.0 1.5 1.0

Temperature (C) 25 26.30a 0.22 26.31a 0.24 26.25a 0.23 26.30a 0.21 26.23a 0.22 26.26a 0.25
Salinity (ppt) 1.61 0.00 1.60a 0.08 1.61a 0.08 1.61a 0.08 1.61a 0.08 1.60a 0.08 1.60a 0.08
a a a a a a
pH 7.38 0.01 7.34 0.01 7.33 0.01 7.33 0.01 7.34 0.01 7.34 0.01 7.34a 0.01
-1 b ab ab a ab
DO (mg l ) 6.36 0.09 6.47 0.07 6.46 0.08 6.22 0.11 6.12 0.14 6.24 0.12 6.18ab 0.11
-1 a a a ab b ab
Free CO2 (mg l ) 5.23 0.15 4.23 0.18 4.27 0.19 4.78 0.94 5.04 1.07 4.87 0.26 5.00b 0.22
-1 a a a a a
Hardness (mg l ) 418.33 2.88 344.02 1.49 348.93 2.59 349.27 2.58 349.93 2.43 349.87 2.67 350.53a 2.52
-1 a a a a a a
Alkalinity (mg l ) 239 0.58 236.20 3.49 237.01 3.02 237.67 2.93 238.20 2.98 238.20 3.00 238.10a 2.90

Mean values with same superscript did not show any significant difference (P [ 0.05)
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Fig. 2 Variation in dissolved oxygen concentration throughout the experimental period in treatments

Fig. 3 Variation in NH4? concentration throughout the experimental period in treatments

and is also the preferred form of nitrogen for growing higher plants (Rakocy et al. 2006).
NO3N concentrations were detected in the favorable ranges in all treatments and control
for koi carp culture (Fig. 4). NitrateN is not generally of great concern to cultivable
organisms, and it has also been proved that aquatic species can tolerate extremely high
concentrations ([25 mg l-1) of nitrateN (Ebeling et al. 1993). Poxton and Allouse (1982)
recommended that NO3N concentrations should not exceed 50 mg l-1 in waters used for
the culture of fish and shellfish. In the present study, mean phosphate levels were lower in
the treatments T1, T2, and T3, because of the phosphate utilization by spinach plant as
compared to control and S1, S2 (Table 6; Fig. 5). Food residues and fecal matter are the
major sources of phosphorus in aquaculture effluent.
The mean value of potassium concentration varied significantly (P \ 0.05) between
treatments. The potassium level was higher in treatments without plant groups as compared

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Table 6 Mean of physicochemical parameters and nutrients in fish tank after the experimental period of 45 days for different treatments
Parameters Chemical composition of raw C T1 T2 T3 S1 S2
water at the beginning
of the experiment

Flow rate (l min-1) 3.2 3.2 1.5 1.0 1.5 1.0

Ammonia (mg l-1) 0.12b 0.01 0.06a 0.01 0.04a 0.01 0.04a 0.11 0.14bc 0.01 0.15c 0.01
Nitrite-N (mg l-1) 0.09b 0.00 0.04a 0.01 0.05a 0.01 0.05a 0.09 0.08b 0.00 0.08b 0.01
-1 a a a b
Nitrate-N (mg l ) 14.65b 1.79 3.48 0.39 2.57 0.26 2.78 0.30 12.22 1.89 12.38b 1.89
-1 a a a ab
Phosphate (mg l ) 2.08b 1.08 0.53 0.07 0.46 0.07 0.49 0.08 0.98 0.15 1.04ab 0.16
-1 c b a b c
Potassium (mg l ) 18.63 0.07 19.36 0.22 15.62 0.43 14.25 0.70 15.59 0.43 19.38 0.86 19.68c 0.28
-1 b a a a b
Calcium (mg l ) 127.83 0.44 129.70 1.24 127.30 1.51 127.32 1.30 127.2 1.47 130.32 1.03 130.63b 1.05
-1 b a a a b
Magnesium (mg l ) 52.0 0.58 51.75 1.08 46.40 0.93 45.59 1.17 45.89 1.28 52.01 1.41 52.61b 1.37
-1 b a a a b
Sodium (mg l ) 323.17 1.69 328.41 3.91 321.45 3.63 321.01 4.72 319.13 5.47 329.30 3.74 334.63c 2.72
-1 b a a a b
Iron (mg l ) 0.08 0.01 0.06 0.00 0.03 0.00 0.03 0.00 0.03 0.00 0.06 0.00 0.05b 0.00
-1 c a a a b
Zinc (mg l ) 0.12 0.00 0.13 0.00 0.07 0.05 0.07 0.01 0.07 0.00 0.10 0.01 0.12c 0.01
Mean values with same superscript did not show any significant difference (P [ 0.05)
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Fig. 4 Variation in NO3 concentration throughout the experimental period in treatments

Fig. 5 Variation in phosphorus concentration throughout the experimental period in treatments

to treatments with plants. The highest level of potassium was observed in S2

(19.68 0.28), whereas lowest level was observed in T2 (14.25 0.70) (Table 6).
Potassium levels were higher in the treatments without plants. Uptake of potassium in
plants was affected by the presence of higher Na? levels, which will interfere with the
uptake of K? and Ca?2 (Rakocy et al. 2006).
It was observed that mean value of calcium showed significant difference (P \ 0.05)
between treatments and control. The mean value of magnesium showed significant dif-
ference (P \ 0.05) between treatments and control. The higher concentration of calcium
and magnesium, due to water used for experiment, was hard. The mean value of sodium
varied significantly (P \ 0.05) between treatments and control (Table 6), and the

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Fig. 6 Percentage of nutrient removal after experimental period of 45 days in different treatments

concentration of sodium was high because the source of groundwater borewell was located
near the Versova coast; this was the possible reason for higher sodium levels in water. The
mean value of iron in treatments and control showed very negligible variation, and it was
ranged between 0.02 and 0.06 mg l-1. It was observed that the mean value of zinc on
whole showed significant difference (P \ 0.05) among treatments and control (Table 6). In
the present study, iron and zinc were not supplemented so this was the possible reason for
negligible concentration of Fe and Zn.

Percentage of nutrient removal at different flow rates

All the treatments T1 (3.2 l min-1), T2 (1.5 l min-1), and T3 (1 l min-1) effectively
removed nitrate in the range of 77.278.65 % (Fig. 6); this indicates that all the treatments
efficiently remove nitrate from the polluted aquaculture wastewater. Similarly, Ghaly et al.
(2005) examined the use of hydroponically grown barley for the removal of NO3N from
aquaculture wastewater and reported that NO3N reductions range from 54.7 to 91.0 %.
Lennard and Leonard (2006) reported 90.9 % removal of nitrate using gravel bed as media
for lettuce production.
Phosphate removal also showed no significant difference among the treatments, and
phosphate removed was in the range of 47.0455.06 % (Fig. 6). Similarly, Lin et al. (2002)
reported that construction of wetlands system receiving aquaculture effluent effectively
removed 3271 % phosphate. Ghaly et al. (2005) examined the use of hydroponically
grown barley for the removal of PO4P from aquaculture wastewater and reported
91.893.6 % PO4P removal. Clarkson and Lane (1991) evaluated the use of the nutrient
film technique for PO4P removal from aquarium wastewater and observed the reduction
of PO34P from 4.4 to 0.3 mg l-1 using barley in a period of 4 weeks.
Potassium concentration showed significant difference between the treatments (Fig. 6).
The percentage removal of potassium was studied at the end of the experiment, which
showed 22.8529.16 %, whereas Ghaly et al. (2005) examined the use of hydroponically
grown barley for the removal of potassium from aquaculture wastewater and reported
potassium reductions ranging from 99.6 to 99.8 %. Dontje and Clanton (1999) reported
2571 % potassium removal in recirculating aquaculture systems using cattails, reed
canary grass, and tomatoes grown in sand beds. Mant et al. (2003) achieved 24.9 %
potassium removal using Salix viminalis grown in gravel hydroponic system to treat pri-
mary settled sewage. In the present study, potassium was not supplemented from outside

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source to the aquaponic system. Groundwater used for the study contained 18.6 mg l-1 of
potassium.

Conclusion

From the study, it was concluded that all the flow rates were found to be effective in terms
of fish growth, plant growth, and nutrient removal, but flow rate of 1.5 l min-1 was
observed as more effective for better growth of spinach (height gain and yield) and koi carp
(percentage weight gain). Water quality and nutrient removal were found best in aquaponic
treatments as compared to the treatments without plants, spinach plants effectively utilized
the nutrients and maintained water quality, which is ideal for Koi carp production.
However, nutrient removal percentage of was found better in all treatments. This clearly
indicates aquaponic systems are effectively reducing the environmental pollution, utilizing
the biological waste that is generated from fish wastes for vegetable production, and are
organic, eco-friendly and sustainable.

Acknowledgments This work was supported and funded by Indian Council of Agricultural Research, New
Delhi, India. The authors are grateful to Dr W. S. Lakra, Director, Central Institute of Fisheries Education,
Mumbai, for providing all the necessary facilities for this research.

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