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Aquaculture 418–419 (2014) 79–86

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Aquaculture
journal homepage: www.elsevier.com/locate/aqua-online

Effects of addition of maize starch on the yield, water quality and


formation of bioflocs in an integrated shrimp culture system
Liping Liu a,⁎, Zhenxiong Hu a, Xilin Dai a, Yoram Avnimelech b
a
Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources of the Shanghai Ocean University and Ministry of Education, Shanghai Ocean University, 999 Huchenghuan Road,
Shanghai 201306, China
b
Department of Civil and Environmental Eng., Technion, Israel Inst. of Technology, 32000 Haifa, Israel

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

Article history: Integrated Multi-Trophic Aquaculture (IMTA) is an ecological and economic farming strategy that minimizes
Received 6 April 2013 waste from culture systems, reduces the risk of diseases and provides additional income source. A trial was
Received in revised form 26 September 2013 conducted to evaluate the effects of adding maize starch on the yield, water quality, formation of bioflocs and
Accepted 3 October 2013
economic return in an IMTA system comprising white shrimp (Litopenaeus vannamei), spotted scat (Scatophagus
Available online 12 October 2013
argus) and water spinach (Ipomoea aquatic). Shrimp were randomly assigned to 12 cemented tanks (T1, T2, T3
Keywords:
and T4 with three replicates) and reared in monoculture (T1) without the addition of starch, or in polyculture
Litopenaeus vannamei (with spotted scat and water spinach) with (T3 and T4) or without (T2) the addition of starch. Shrimp in T4
Scatophagus argus were fed at 80% of the feeding amount of other treatments. The results showed that shrimp survival was higher
Integrated aquaculture in T3 and T4 (95.6% and 94.3%, respectively) than in T1 and T2 (51.1% and 56.5%, respectively) (P b 0.01). The
Biofloc technology shrimp yield was the highest in T3 (0.495 kg m−3), which also had the lowest feed conversion ratio (1.02) and
best economic performance. The levels of nitrite-N (NO2-N) were significantly lower in T3 and T4 at the end of
the experiment than in other treatments (P b 0.01). Similarly, total ammonia nitrogen (TAN) was lower and
total suspended solids (TSS) was higher in T3 and T4 than in T1 and T2 (P b 0.05). The results suggested that
the addition of starch into the IMTA system of shrimp, spotted scat and water spinach improved productivity,
profitability and water quality. The combination of the IMTA model and biofloc technology had a synergistic
effect on overall FCR and economic return, and was more effective at improving shrimp survival, production
and reducing nitrite-N and TAN than use of the IMTA model alone.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction The rationale for IMTA is the reuse of waste, i.e., the waste products
from one species are used to feed a second species (Chopin, 2006).
Shrimp are reared primarily by intensive culture in China, the leading Stocking species in an ideal polyculture pond occupy different niches
producer of shrimp in the world. The expansion of intensive shrimp and possess feeding habits that differ from, and are complementary to
culture has been associated with increasingly serious environmental each other. Some species are capable of utilizing uneaten and undigested
problems and the outbreak of diseases (Boyd and Clay, 1998; Burford feed (Tian et al., 2001; Tookwinas, 2003; Yi, 1998), whereas extractive
and Williams, 2001; Jackson et al., 2003; Zhang et al., 2012). To address species, such as shellfish and seaweeds, make use of suspensions, organic
some of these issues, culturists have begun to utilize a new approach, matter and nitrogenous and phosphorus nutrients (Stirling and Okumus,
termed Integrated Multi-Trophic Aquaculture (IMTA). IMTA is generally 1995; Taylor et al., 1992; Troell et al., 1999, 2006). Since shrimp produce
considered to represent a more sustainable alternative to traditional nitrogenous and phosphorus wastes and eutrophic waste water due to
culture models (monoculture), because it allows for more efficient poor utilization of food (Fast and Menasveta, 2000), in recent decades,
resource utilization, produces less waste, has an increased range of researchers have experimented with culturing shrimp with a number
outputs, and can supply markets more continuously (Diana et al., 2013; of alternative food sources, including tilapia (Wang et al., 1998; Yuan
Soto, 2009). Asian countries have for centuries been practicing IMTA et al., 2010), milkfish (Eldani and Primavera, 1981), mullet (James,
through “trial and error” and experimentation (Troell et al., 2009; Zhao 1996), bivalves (Hopkins et al., 1993; Hunt, 1991) and seaweed (Jones
et al., 2012). et al., 2002). In most instances, water quality and overall production
were improved relative to monoculture.
Biofloc technology (BFT), a new type of water treatment technology,
Abbreviations: IMTA, Integrated Multi-Trophic Aquaculture; TAN, total ammonia is often cited as an approach to reduce feed costs and environmental
nitrogen; FCR, feed conversion ratio; TSS, total suspended solids; Chl a, chlorophyll a;
BFT, biofloc technology; BCR, benefit cost ratio; SVI, sludge volume index.
problems associated with discharge of waste products (Avnimelech,
⁎ Corresponding author. Tel.: +86 2161900411; fax: +86 2161900405. 1999). Biofloc technology involves stimulating the process of inorganic
E-mail address: lpliu.sou@gmail.com (L. Liu). nitrogen assimilation by heterotrophic bacteria. These bacteria utilize

0044-8486/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.aquaculture.2013.10.005
80 L. Liu et al. / Aquaculture 418–419 (2014) 79–86

ammonium, in addition to the organic nitrogenous wastes, to synthesize After washing away residual ClO2, the tanks were filled to a depth
new cells by consumption of carbohydrates, simultaneously utilizing of 1 m and salinity was maintained at 1 ppt. Litopenaeus vannamei
organic matters, microorganisms and some suspended solids to form postlarvae (PL) from a commercial hatchery were reared in tanks
bioflocs (Hargreaves, 2006). This process reuses uneaten feed and for six weeks before being randomly assigned to the experimental
excrement and reduces the need for water exchange, thereby reducing tanks. The average initial weight was 3.11 g and shrimp were stocked
the risk of disease infection through water intake (Crab et al., 2009). into tanks at 80 PLs m−2 (Table 1). Juvenile spotted scats averagely
The C/N ratio of most of the feeds used in semi-intensive aquaculture weighted 31.83 g from a local commercial aquarium were cultured
ponds is around 10, whereas bacteria require about 20 units of carbon together temporarily for 3 d then added to the experimental tanks
per unit of nitrogen assimilated (Avnimelech, 1999). Therefore, if the (2 individual m−2) one week after shrimp stocking. Two floating beds
C/N ratio is increased by adding a carbohydrate source, such as tapioca (100 × 100 × 1 cm) made of polyethylene foam were placed in each
starch, in addition to the regular feed, the increased availability of carbon IMTA system tank, and young water spinach seedlings purchased from
allows the heterotrophic bacterial population to increase in abundance a local market were then planted in holes in the floating bed one
(Asaduzzaman et al., 2010a), resulting in bioflocs. Burford et al. (2004) week after shrimps were stocked (Table 2). There was no water
evaluated the consumption of bioflocs by shrimp using 15N labeling exchange throughout the entire experimental period and water loss
and noted that 18–29% of the protein consumed by shrimp on a daily due to evaporation was compensated during later period. The aerator
basis was derived from bioflocs. Hari et al. (2006) also noted that the was operated daily (24 h), except during feeding. Hyper-concentrated
addition of tapioca starch in shrimp culture systems improved water composite bacillus (HaiFeng™, Dalian Aotai) (10 g tank−1) was
quality, shrimp growth and feed utilization efficiency. added to all tanks weekly from week 3 to improve water quality and
Spotted scat (Scatophagus argus), like tilapia, are benthic animals supplement the bacillus community.
that are cultured because of their ease of breeding, tolerance to wide Shrimp in all tanks were fed three times daily (8:30, 13:30 and
range of environmental conditions, adaptation to both fresh and 18:30) at a rate of 3–10% of body weight per day using commercial
brackish water environment and resistance to diseases. The species is pellet (41% crude protein for feed #2 with a diameter of 2 mm,
also ideal for use in polyculture systems because of its mild behavior Guangdong Haid Group Co. Ltd). Feeding tray was used for feed
and broad diet (Barry and Fast, 1992). Similarly, water spinach (Ipomoea adjustment according to feed consumption, water quality and weather,
aquatic) has been widely used in water purification research because of as described by Martinez-Cordova et al. (1998) with slight modification.
its high productivity, low cost and ease of planting on floating beds Locally purchased maize starch (Nutrient content: carbohydrate 85%,
(Chen et al., 2010; Fu et al., 2011) and had remarkable effects (Li et al., water 13.5%, lipid 0.1%, protein 1.2%, fibre 0.1%, ash 0.1%) was used as
2007; Xin et al., 2010; Yang et al., 2012). carbohydrate source for manipulating the C/N ratio at 15. The pre-
The use of IMTA in combination with biofloc technology has the weighed maize starch was mixed in a pot with tank water and uniformly
potential to improve shrimp culture efficiency and water quality, distributed over the tank surface directly after feed application. Spotted
though currently there is no evidence to support this. The research scats were not fed separately and total feed and starch addition in each
objective was to evaluate the effects of adding maize starch on the tank were recorded daily.
yield, water quality, formation of bioflocs and economic return in an
IMTA system of white shrimp, spotted scat and water spinach in 2.4. Measuring and monitoring
outdoor cement tanks.
2.4.1. Harvest of shrimp, spotted scat and water spinach and estimation of
2. Materials and methods production parameters
Shrimps and spotted scats were harvested after draining the
2.1. Experimental design tanks. Thirty shrimps from each tank were weighed to determine
mean weight. Net yield, survival rate, specific growth rate (SGR), feed
Four treatment groups were set up as follows: T1 consisted of three conversion ratio (FCR, the amount of feed needed per unit weight gain
tanks containing only shrimp (control), whereas T2, T3 and T4 were set of shrimps) and overall feed conversion rate (overall FCR, the amount
up to simulate the IMTA system approach and contained shrimp, of feed needed per unit weight gain of shrimps and spotted scats)
spotted scat and water spinach. The animals in T1, T2 and T3 were fed were calculated. At the end of the experiment, water spinach was
equal amounts of commercial feed pellets, while those in T4 were fed carefully removed from the floating beds, gently washed with tap
at 80% of the feeding amount of other treatments. Maize starch was water, blotted with absorbing paper, and weighed.
added to the tanks in T3 and T4, the amount of which was maintained
at 90% of the feeding amount to keep the C/N ratio at 15. Each treatment 2.4.2. Water quality analysis
group was triplicated and the design was completely randomized. Water samples were collected at 09:00 on a weekly basis from each
tank during the experiment using a horizontal water sampler. Water
2.2. Experimental site and infrastructure temperature, surface dissolved oxygen (portable DO meter: SevenGo
Pro-SG6), pH (Hanna HI98128 pH meter) and transparency (Secchi
The experiment was carried out at the Specific Fisheries Field of disc) were monitored in situ at dawn. Water quality parameters e.g. total
Shanghai Ocean University (SHOU), China in 12 rectangular outdoor ammonia nitrogen (TAN), nitrite nitrogen (NO2-N), nitrate nitrogen
cement tanks (5 × 3 × 1.2 m) for 49 d between 9 September 2012 and (NO3-N), total phosphorous (TP), total nitrogen (TN), chlorophyll a
27 October 2012. The tanks were supplied with low saline water pumped (Chl a) and total suspended solids (TSS) were analyzed following the
from a local nearby canal. Aeration was supplied by an air blower standard methods for water and wastewater analysis (APHA, 1998).
(HRE40L 1.1KW 220v, Roots blower) and delivered via a nanometer
micro-porous aeration disc to all experimental tanks. Feed trays were 2.4.3. Determination of biofloc parameters
applied in all tanks to monitor shrimp growth and consumption of feed. Water in tanks was sampled weekly. The biofloc volume was
determined using an Imhoff cone, recording the volume (FV-15) taken
2.3. Tank preparation and feed management in by the flocs in 1L of tank water after 15min sedimentation. The sludge
volume index (SVI) is defined as the volume in milliliters occupied
Prior to the start of the experiments, all tanks were completely by 1 g of TSS after settling. At the time of the last water sampling, an
drained and fully exposed to prevailing sunlight. In addition, the effort was made to collect biofloc samples by sorting the tank water
tanks were disinfected with 2 g m−3 chlorine dioxide (ClO2) for 24 h. undisturbed in a cylinder then decanting and separating the settled
L. Liu et al. / Aquaculture 418–419 (2014) 79–86 81

Table 1
Growth performance of shrimp cultured for 49 d and spotted scat cultured for 42 d in each treatment (T1: Single shrimp tank system with 100% of the feeding amount; T2: Shrimp-spotted
scat-water spinach tank system with 100% of the feeding amount; T3: Shrimp-spotted scat-water spinach tank system with 100% of the feeding amount and starch supplementation;
T4: Shrimp-spotted scat-water spinach tank system with 80% of the feeding amount and starch supplementation).

Performances T1 T2 T3 T4

Shrimp
Harvesting
Biomass (kg m−3) 0.292 ± 0.007Bb 0.304 ± 0.002Bb 0.495 ± 0.038Aa 0.423 ± 0.027ABa
Mean weight (g shrimp−1) 6.85 ± 0.15Aa 6.56 ± 0.07Aab 6.10 ± 0.16ABb 5.52 ± 0.22Bc
SGR (%) 1.61 ± 0.04Aa 1.52 ± 0.02ABa 1.37 ± 0.06ABab 1.17 ± 0.09 Bb
Survival (%) 51.1 ± 6.3Bb 56.5 ± 5.3Bb 95.6 ± 6.1Aa 94.3 ± 7.7Aa
Shrimp FCR 5.64 ± 0.25Aa 4.43 ± 0.15 Ab 1.02 ± 0.32 Bc 1.15 ± 0.27 Bc

Spotted scat
Harvesting
Biomass (kg m−3) 0.112 ± 0.009a 0.100 ± 0.003a 0.094 ± 0.009a
SGR (%) 1.34 ± 0.18a 1.07 ± 0.14a 0.98 ± 0.24a
Survival (%) 98.9 ± 1.10 a 97.8 ± 1.10a 96.7 ± 0.00a
Mean weight (g fish−1) 56.07 ± 4.04a 50.13 ± 2.99a 48.51 ± 4.78a

Combined
Total biomass (kg m−3) 0.292 ± 0.007Cc 0.416 ± 0.009Bb 0.595 ± 0.036Aa 0.517 ± 0.028ABa
Overall FCR 5.64 ± 0.22 Aa 2.37 ± 0.16Bb 0.88 ± 0.13Cc 0.98 ± 0.26Cc

Shrimps at average 3.11 g in weight were stocked at a density of 80 individual m−2, while spotted scat averagely 31.83 g were stocked at a density of 2 individual m−2.
Data represent means ± S.E. of three replicates. Mean values in the same row with different superscript letters were significantly different. Uppercase letters indicate the significant
difference at α = 0.01, while the lowercase letters show the significant difference at α = 0.05.

biofloc from the bottom of the cylinder. The samples were stored in a were compared using one-way analysis of variance (ANOVA), and LSD
refrigerator until biochemical composition analysis using standard test followed to determine which treatments differed significantly
methods (AOAC, 2005). from each other at α = 0.05 and α = 0.01. All statistical analyses were
performed using SPSS 17.0 for Windows.
2.4.4. Economic analysis
An economic analysis was conducted to evaluate the net return and 3. Results
benefit cost ratio in the different treatments. The following formulae
were used (Asaduzzaman et al., 2010b): 3.1. Growth performance

R ¼ I−ðFC þ VC þ Ii Þ; Treatment T3 had the highest shrimp yield (0.495 kg m−3) and
lowest feed conversion ratio (FCR: 1.02) at harvest, which were
Benefit cost ratio ðBCRÞ ¼ R=ðFC þ VC þ Ii Þ; significantly better than those in T1 and T2 (P b 0.01), but not
significantly different from those in T4 (Table 1). T1 tended to have
Where, R = net return, I = income from spotted scat, water spinach the lowest shrimp yield and highest FCR, but no significant difference
and shrimp sale, FC = fixed costs, VC = variable costs and Ii = interest was observed between T1 and T2. The shrimp survival was significantly
on inputs. higher in T3 and T4 (95.6% and 94.3%, respectively) than in T1 and T2
This analysis was based on farm-gate prices on October 24th 2012 (P b 0.01). However, specific growth rate (SGR) and mean weight at
for harvested shrimp and spotted scat. The market prices for cost harvest were higher in T1 than in other treatments, while SGR in T4
items in China were expressed in US dollars (US$ 1.0 = 6.2 RMB). The was significantly lower than in T2 (P b 0.05) and T1 (P b 0.01).
farm-gate price for harvested shrimp was fixed at US$ 3.87 kg−1 (7 g) Treatment T2 had the highest spotted scat yield at harvest, but there
and US$ 3.55 kg−1 (6 g), and for spotted scat (above 30 g) at US$ was no significant difference relative to T3 and T4. The fish SGR and
15.32 kg−1. Market prices for shrimp postlarvae (US$ 3.55 for 1000 mean weight were highest in T2, followed by T3, but there was no
pieces), water spinach (US$ 0.48 kg−1), maize starch (US$ 0.40 kg−1), significant difference among all IMTA treatments. The spotted scat
shrimp feed (US$ 1.41 kg−1) and electricity (US$ 0.08 kwh−1) were survival rates were above 96.7% in all IMTA treatments. The highest
applied to the analysis. The calculation for cost of working capital was combined biomass (0.595 kg m−3) and lowest overall FCR (0.88) of
based on an annual interest rate of 8%. harvested shrimp and spotted scat were obtained in T3, while combined
biomass and overall FCR were significantly worse in T1 than in other
2.5. Statistical analysis treatments (P b 0.01). T2 had the highest water spinach biomass at
harvest, which was significantly higher than T4 (P b 0.01), but not
Prior to the analysis, the assumptions of normal distribution and significantly different from that in T3 (Table 2).
homogeneity of variance were checked. Means between treatments
3.2. Water quality
Table 2
Production performance of water spinach planted for 42 d in T2–T4. The changes in dissolved oxygen (DO) levels were similar among
treatments gradually decreased during the experiment, ranging from
Performances T2 T3 T4
9.34 to 6.61 mg L−1. Water temperature in all tanks decreased
Stocking
throughout the experiment and ranged from 27.5 to 22.7 °C. pH tended
Biomass (g tank−1) 2300.0 ± 0.0 2300.0 ± 0.0 2300.0 ± 0.0
Density (g m−2) 153.3 ± 0.0 153.3 ± 0.0 153.3 ± 0.0 to be relatively stable and ranged between 7.78 and 8.48. The changes of
pH appeared to be more stable in T3 and T4 though the values increased
Harvesting in all treatments at the beginning of the experiment, peaking at week 3,
Biomass (g tank−1) 5161.3 ± 99.8Aa 4963.7 ± 42.4Aa 4229.0 ± 68.3Bb then decreased. Water transparency in each tank decreased over
Note: T2, T3 and T4 represent different treatment tanks as shown in Table 1. time (range: 32.9 to 10.9 cm). T1 and T2 had significantly higher
82 L. Liu et al. / Aquaculture 418–419 (2014) 79–86

a 12.00
b 4.00

3.50
10.00 T1 T1
3.00 T2
T2
8.00

TP (mg L-1)
TN (mg L-1)

T3 2.50 T3
T4 T4
6.00 2.00

1.50
4.00
1.00
2.00
0.50

0.00 0.00
1 2 3 4 5 6 1 2 3 4 5 6
Week Week

c d 0.60
1.50
T1
1.40 0.50
T1 T2
1.30

Nitrate-N (mg L-1)


T2 T3
0.40
1.20
TAN (mg L-1)

T3 T4
1.10 T4 0.30
1.00
0.90 0.20

0.80
0.10
0.70
0.60 0.00
1 2 3 4 5 6 1 2 3 4 5 6
Week Week
e f 250
400

350
T1 200 T1
Chlorophyll a (ug L-1)

300 T2 T2
TSS (mg L-1)

250 T3 150 T3
T4 T4
200
100
150

100
50
50

0 0
1 2 3 4 5 6 1 2 3 4 5 6
Week Week

g 0.6

T1
0.5 T2
T3
0.4
T4
N
W
0.02

0.01

0
1 2 3 4 5 6
L. Liu et al. / Aquaculture 418–419 (2014) 79–86 83

transparency at the end of the experiment as compared to T3 and T4 a


14.00
(P b 0.05).
The concentration of total nitrogen (TN) and total phosphorus (TP) 12.00
increased consistently in all treatments during the culture period, T1
10.00
except for slight decline from week 5 (Fig. 1a and b). Treatment T1 T2

FV-15 (mL)
had the highest TN (P b 0.05) and TP levels at the end of experiment, 8.00 T3
while TP and TN levels in late stage were consistently lower in T4, the
T4
one obtaining 80% feed as compared to the other treatments 6.00
(P b 0.05). The concentration of TAN tended to increase in each tank
4.00
during the initial stage of the experiment, and then decreased
(Fig. 1c). TAN levels in T1 and T2 at harvest were significantly higher 2.00
than treatments where starch was added in T3 and T4 (P b 0.05). The
NO2-N concentration increased sharply in T1 and T2, but not in T3 and 0.00
T4 (Fig. 1g). The final NO2-N values for T3 and T4 were lower than the 1 2 3 4 5 6
Week
initial values, and were also extremely significantly lower than those
for T1 and T2 (P b 0.01). The levels of NO3-N were relatively constant
throughout the experiment but were higher in T3 and T4 than in b
80.00
other treatments (Fig. 1d). In comparison with other treatments, Chl a
level in T1, the treatment without fish, increased rapidly in the first 70.00
T1
5 weeks, then decreased (Fig. 1e), but remained significantly higher 60.00
T2
than in other treatments at the end of the experiment (P b 0.05). TSS

SVI (mL g-1)


50.00 T3
concentration in each tank increased during the experiment (Fig. 1f),
and was significantly higher in T3 than in T1 and T2 at the end of the 40.00 T4
experiment (P b 0.05).
30.00

3.3. Biofloc parameters 20.00

10.00
The changes in the volume of bioflocs and SVI in each treatment
throughout the experiment are shown in Fig. 2. The volume of bioflocs 0.00
increased in all treatments throughout the experiment (Fig. 2a). In 1 2 3 4 5 6
Week
comparison with the treatments that were not supplemented with
starch, the volume of bioflocs was consistently higher in T3 (P b 0.01) Fig. 2. a. Variations of biofloc volume after 15 min sedimentation (FV-15) in each
and T4 (P b 0.05). The SVI levels increased in all treatments throughout treatment during the experiment. Error bars indicate S.E. Note: T1, T2, T3 and T4 represent
the experiment, with the exception of a slight decrease in the later different treatment tanks as shown in Fig. 1. b. Variations of sludge volume index (SVI) in
each treatment during the experiment. Error bars indicate S.E. Note: T1, T2, T3 and T4
stages. SVI (70.67 mL g−1) was significantly higher in T3 at the end
represent different treatment tanks as shown in Fig. 1.
of experimental period (Fig. 2b) compared with other treatments
(P b 0.01). In contrast, The SVI value was lowest in T1 and was
significantly lower than in T2 (P b 0.05) but not in T4. The nutrient had a beneficial effect on shrimp survival (Gao et al., 2012). Water
composition of the bioflocs collected from all tanks at the end of the quality, particularly NO2-N and TAN levels, are generally thought to be
experiment is given in Table 4. The mean values for crude protein the primary limiting factor of shrimp survival (Barbieri, 2010;
content in T3 and T4 were 24.0% and 18.1% respectively, which were Santacruz-Reyes and Chien, 2012; Tseng and Chen, 2004; Xian et al.,
significantly higher than those in other treatments (P b 0.01). 2011). In the present study, high shrimp mortality in T1 and T2 might
have resulted from significantly higher NO2-N and TAN levels in water
3.4. Economic return column of tanks. The survival of shrimp was similar between T1 and
T2, indicating that the IMTA model used in this study did not have a
The total revenue and total net return were significantly higher in T3 significant effect on NO2-N and TAN levels resulting in shrimp mortality,
than in T2 and T4 (Pb 0.05), and highly significantly different from those an observation consistent with Yuan et al. (2010).
in T1 (P b 0.01) (Table 3), which was similar to BCR. In comparison with
T1, BCR and total net return were significantly higher in T2, T3 and T4 4.2. Production performance
(P b 0.01).
Shrimp tended to grow more slowly (SGR) in the IMTA treatments
4. Discussion than in monoculture. Since feed input to all treatments, except T4, was
limited to an equal amount and shrimp survival was higher in T2 and
4.1. Shrimp survival T3 than in monoculture, food competition might have occurred in T2
and T3 both among shrimp and between shrimp and spotted scat,
Shrimp survival was higher in treatment groups with starch which might be a major reason for retarded growth of shrimp in T2
addition, suggesting that the biofloc process resulting from the addition and T3. Consistent with the present observations, Yuan et al. (2010)

Fig. 1. a. Variations of total nitrogen (TN) in each treatment during the experiment (T1: Single shrimp tank system with 100% of the feeding amount; T2: Shrimp-spotted scat-water
spinach tank system with 100% of the feeding amount; T3: Shrimp-spotted scat-water spinach tank system with 100% of the feeding amount and starch supplementation; T4: Shrimp-
spotted scat-water spinach tank system with 80% of the feeding amount and starch supplementation). Error bars indicate S.E. b. Variations of total phosphorus (TP) in each treatment
during the experiment. Error bars indicate S.E. Note: T1, T2, T3 and T4 represent different treatment tanks as shown in Fig. 1. c. Variations of total ammonia nitrogen (TAN) in each
treatment during the experiment. Error bars indicate S.E. Note: T1, T2, T3 and T4 represent different treatment tanks as shown in Fig. 1. d. Variations of nitrate-N in each treatment during
the experiment. Error bars indicate S.E. Note: T1, T2, T3 and T4 represent different treatment tanks as shown in Fig. 1. e. Variations of chlorophyll a (Chl a) in each treatment during the
experiment. Error bars indicate S.E. Note: T1, T2, T3 and T4 represent different treatment tanks as shown in Fig. 1. f. Variations of total suspended solids (TSS) in each treatment during the
experiment. Error bars indicate S.E. Note: T1, T2, T3 and T4 represent different treatment tanks as shown in Fig. 1. g. Variations of nitrite-N in each treatment during the experiment. Error
bars indicate S.E. Note: T1, T2, T3 and T4 represent different treatment tanks as shown in Fig. 1.
84 L. Liu et al. / Aquaculture 418–419 (2014) 79–86

Table 3
Economic performance of T1–T4 in the experiment (Unit: US$).

Parameters Treatment

T1 T2 T3 T4

Fixed cost
Electricity 4.84 ± 0.00 4.84 ± 0.00 4.84 ± 0.00 4.84 ± 0.00
Feed for culturing temporarily 2.50 ± 0.00 2.50 ± 0.00 2.50 ± 0.00 2.50 ± 0.00
Seed 4.26 ± 0.00 4.26 ± 0.00 4.26 ± 0.00 4.26 ± 0.00
Subtotal 11.60 ± 0.00 11.60 ± 0.00 11.60 ± 0.00 11.60 ± 0.00

Variable cost
Scat juveniles 14.63 ± 0.00 14.63 ± 0.00 14.63 ± 0.00
Feed 4.94 ± 0.00 4.94 ± 0.00 4.94 ± 0.00 3.95 ± 0.00
Maize starch 1.25 ± 0.00 1.25 ± 0.00
Polystyrene foam board 1.51 ± 0.00 1.51 ± 0.00 1.51 ± 0.00
Water spinach 1.11 ± 0.00 1.11 ± 0.00 1.11 ± 0.00
Subtotal 4.94 ± 0.00 22.19 ± 0.00 23.44 ± 0.00 22.45 ± 0.00
Total 16.54 ± 0.00 33.79 ± 0.00 35.04 ± 0.00 34.05 ± 0.00
Cost of working capital 0.33 ± 0.00 0.68 ± 0.00 0.70 ± 0.00 0.68 ± 0.00
Total inputs 16.87 ± 0.00 34.47 ± 0.00 35.74 ± 0.00 34.73 ± 0.00

Financial return
Shrimp sale 16.97 ± 0.40Bb 17.68 ± 0.12Bb 28.72 ± 2.24Aa 22.49 ± 1.47ABb
Scat sale 25.75 ± 2.08a 23.18 ± 0.63a 21.56 ± 2.12a
Water spinach sale 2.50 ± 0.05Aa 2.40 ± 0.02Aa 2.05 ± 0.02Bb
Total revenue 16.97 ± 0.40Bc 45.93 ± 2.19Ab 54.30 ± 2.06Aa 46.10 ± 1.78Ab
Total net return 0.10 ± 0.40Bc 11.46 ± 2.19Ab 18.56 ± 2.06Aa 11.37 ± 1.78Ab
BCR 0.01 ± 0.01Bc 0.33 ± 0.06Ab 0.52 ± 0.06Aa 0.33 ± 0.05Ab

Note: T1, T2, T3 and T4 represent different treatment tanks as shown in Table 1.

reported that shrimp grew significantly faster in monoculture than sufficiently high for the survival and growth of L. vannamei (Aquacop
in shrimp–tilapia polyculture tanks given equal amounts of feed. et al., 1988; McGraw et al., 2001). The gradual decrease in DO levels
Interestingly, shrimp may also consume naturally produced food was probably due to increasing biological demand associated with
organisms such as phytoplankton (Muangkeow et al., 2007). The an increase in total biomass and the accumulation of organic matter.
significantly higher level of Chl a in monoculture at harvest suggested pH did not vary significantly among all treatments and were well within
that the biomass of phytoplankton was significantly higher than in tolerant range of white shrimp. The decrease of pH in the later stage of
IMTA treatments, thereby offering an alternative food source for shrimp. the experiment might have resulted from the accumulation of organic
In addition, shrimp in monoculture grew faster than in T4, probably due matter or due to nitrification that took place along the growing season.
to the increased feed availability. This also explains why shrimp growth The consistent decrease of transparency during the experimental period
was lowest in T4 among the IMTA treatments. Bioflocs produced by was likely a result of the increase in accumulation of suspended solids.
the application of BFT is a proven food source for cultured species, The increase in biofloc production in T3 and T4 because of starch
and results in a decreased requirement for supplemental feeding addition probably exacerbated the decrease in transparency in these
(Avnimelech, 2007; Burford et al., 2004; Kuhn et al., 2009; Li et al., tanks.
2013). Further research is needed to evaluate the role that biofloc The higher TP and TN levels in T1 at the end of experiment suggested
plays in replacing feed as a food source for shrimp and spotted scat. that IMTA model used in the experiment was effective at reducing
Shrimp yields and FCR were better in treatments that received starch the nutrients and hence can help alleviate environmental pollution
supplementation, and the results were similar to those between concerns. The lower TP and TN levels associated with the presence of
IMTA treatments and monoculture. Despite lower SGR in IMTA tanks spotted scat probably resulted from progressive grazing of spotted
compared with monoculture, the presence of spotted scat and higher scat on uneaten feed and organic matter. Likewise, water spinach
shrimp survival resulted in significantly higher total production and utilized nutrients in the water column for their growth, thereby
lower combined overall FCR in IMTA treatments. reducing the concentrations in the rearing system. Li et al. (2011)
reported that planting water spinach (I. aquatic) on floating beds in
4.3. Water quality aquaculture wastewater significantly reduced TP and TN levels. Further
research is needed to investigate the actual effects of water spinach on
Mean water temperature at dawn was 24.6 ± 0.7 °C during the nutrient removal, and to evaluate the optimal biomass for nutrient
experimental period. Such a low water temperature might have had removal and accumulation of N and P in water spinach.
some negative effects on shrimp growth and survival (Wyban et al., Avnimelech (1999) reported that the addition of carbonaceous
1995). Conversely, DO levels observed at dawn were above 6 mg L−1 substrates led to a significant reduction in the accumulation of
and though levels did decrease slightly over time, they remained ammonium in tanks, which may explain the difference of TAN between
treatments with and without starch addition in our study. Nitrite
reduction could be due to nitrification, as corroborated by the rise of
nitrate concentration, as well as by denitrification. Aerobic denitrifiers
Table 4
significantly reduce nitrite-N by denitrification under aerobic and C/N
Mean proximate chemical composition of the biofloc in each treatment.
conditions (Kim et al., 2008; Liao et al., 2006). The aerobic denitrifying
Composition (%) T1 T2 T3 T4 bacteria can utilize organic carbon sources for their growth under
Crude protein 8.7 ± 1.5 Cc 9.3 ± 1.8Cc 24.0 ± 1.2Aa 18.1 ± 0.9Bb aerobic conditions, and produce N2 from nitrate and nitrite in culture
Crude lipid 0.8 ± 0.2Bb 0.6 ± 0.1Bb 2.5 ± 0.4Aa 1.9 ± 0.3ABa water column by denitrification, which might have been the reason
Crude fiber 1.3 ± 0.2Bb 1.1 ± 0.1Bb 4.5 ± 0.8Aa 3.9 ± 0.5 Aa for the extremely low nitrite-N levels in the treatments with starch
Note: T1, T2, T3 and T4 represent different treatment tanks as shown in Table 1. addition throughout the experiment.
L. Liu et al. / Aquaculture 418–419 (2014) 79–86 85

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