Received: 16 November 2016 | Revised: 24 January 2017 | Accepted: 26 February 2017

DOI: 10.1111/are.13335

ORIGINAL ARTICLE

Growth parameters of whiteleg shrimp Litopenaeus vannamei

and red seaweed Gracilaria corticata in integrated culturing
method under zero water exchange system

Hojjatollah Fourooghifard1 | Abbas Matinfar2 | Mohammad Seddiq Mortazavi1 |

Kiuomars Roohani Ghadikolaee1 | Maryam Mirbakhsh3

1
Persian Gulf and Oman Sea Ecological
Research Center, Iranian Fisheries Sciences
Research Institute, Agricultural Research,
Education and Extension Organization
(AREEO), Bandar Abbas, Iran
2
Iranian Fisheries Sciences Research
Institute, Tehran, Iran
3
Iran Shrimp Research Center, Bushehr, Iran
Correspondence
Hojjatollah Fourooghifard, Persian Gulf and
Oman Sea Ecological Research Center,
Iranian Fisheries Research Institute,
Agricultural Research, Education and
Extension Organization (AREEO), Bandar
Abbas, Iran.
Email: fourooghifard@yahoo.com
Funding information
Iranian Fisheries Research Institute; Persian
Gulf and Oman Sea Ecological Research
Center, Iran.
Abstract
Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gra-
cilaria corticata were measured using integrated culturing method under zero-water
exchange system in a 45-day period. A 2 9 3 factorial design was used with two
levels of shrimp stocking densities and three levels of seaweed weight densities. G.
corticata was cultured on a net tied to a round polyethylene frame. Culture tanks
were filled with 750-L filtered seawater. A 40-W compact fluorescent lamp was
hung over each tank to provide adequate and sufficient light for seaweed growth.
Growth parameters of shrimp and seaweed such as specific growth rate (SGR),
weight gained (WG) and average daily growth (ADG) were computed based on the
initial and final weight of shrimp and seaweed. The maximum and minimum SGR of
L. vannamei (1.97 and 1.69%/day) were observed in treatment S1A3 (25 shrimp/m2
and 400 g seaweed/m2) and S2A1 (50 shrimp/m2 without seaweed) respectively.
The best survival rate (94.67
1.33%), WG (129.9
2.9%) and feed conversion
ratio (1.67
0.04) were also observed in treatment S1A3. The SGR of G.corticata in
the treatment S1A3 (1.97
0.00%/day) was significantly higher, compared to
others. Strong positive correlations were obtained between the density of G. corti-
cata and the growth parameters of L. vannamei. The red seaweed G. corticata could
boost the growth parameters, survival rate and total production of L. vannamei
under zero-water exchange system.

KEYWORDS
FCR, Gracilaria corticata, Litopenaeus vannamei, specific growth rate, stocking density, survival
rate

1 | INTRODUCTION
Farming of whiteleg shrimp Litopenaeus vannamei has recorded rapid
prevalence in almost every coastal province of Iran. The leading
shrimp farms in Iranian provinces are located in Sistan and Baluches-
tan, Bushehr, Hormozgan, Khuzestan and Golestan (Baghaei & Suda-
gar, 2013; Kalbassi, Abdollahzadeh & Salari-Joo, 2013). A greater
portion of the feed eaten by shrimps are excreted as metabolic
waste which adds inorganic nutrients and organic matter to the
shrimp ponds’ water and sediment (Attasat, Wanichpongpan & Ruen-
glertpanyakul, 2013). Uneaten feed released into the natural environ-
ment, precipitates eutrophication in the environment (Neori, Cohen
& Gordin, 1991) and acute toxicity in the body of aquatic animals
€nnb€ack, Halling, Kautsky
(Neori, Shpigel & Ben-Ezra, 2000; Troell, Ro
& Buschmann, 1999). In traditional method, water exchange is used
for maintaining good quality of pond water. Effluents from shrimp

Aquaculture Research. 2017;1–8. wileyonlinelibrary.com/journal/are © 2017 John Wiley & Sons Ltd | 1
2 |
ponds may result in high pollution of coastal water body. Estimation
of nutrients and sediment released into coastal waterways from
shrimp ponds indicated that most of these materials originate from
exogenous feed (Briggs & Funge-Smith, 1994; Hopkins, Sandifer &
Browdy, 1995). The environmental impacts of untreated effluent
have heightened concerns about the sustainability of shrimp farming.
The treatment of the wastewater must be considered as a part of
the whole culture system (Hopkins et al., 1995). The decrease in
exchange rate of water has been studied globally to limit the export
of waste and reduce impact on coastal environment (Hopkins et al.,
1995; Sandifer & Hopkins, 1996). One economical and feasible
method that has been studied in recent years is, the use of seaweed
to maintain excellent quality of pond water in integrated culturing of
shrimps and seaweeds. It is anticipated that nutrient should be
absorbed by seaweed (Buschmann, Troell, Kautsky & Kautsky, 1996;
Neori et al., 1991; Seema & Jayasankar, 2005). Some species of sea-
weeds such as Ulva, Porphyra and Gracilaria have the capacity to
effectively reduce the nutrient load in effluents and assist in main-
taining water quality at acceptable levels (Neori et al., 2004). Sea-
weeds transform the available nutrients such as nitrogen and
phosphorus into composites necessary for their development, and
improve water quality for cultivated animals as well as reduce nutri-
ent concentration in the environment (Troell et al., 2003). Culturing
of juvenile shrimp L. vannamei and green seaweed Ulva clathrata in
outdoor tanks (2000 L) for 45 days indicated that water turbidity in
co-cultured tanks was lower than that in control group but survival
rate was similar among the experimental groups (>80%), and the Ulva
intake by shrimp improved the artificial feed conversion ratio as well
as the growth rate (Cruz-Suarez et al., 2010). Experiments conducted
on the growth rate of Pacific white shrimp L. vannamei in a biofloc
system co-cultured with green seaweed Ulva lactuca disclosed that
the mean final weight of the shrimp was higher in the biofloc sea-
weed (Brito et al., 2014). Integrated culturing of L. vannamei and the
red seaweed Kappaphycus alvarezii in floating cages revealed that
there was no negative interferences between co-cultured shrimp and
seaweed, and there was also no significant difference between
monoculture and polyculture for shrimp weight gain, survival rate
and feed conversion (Lombardi, de Almeida Marques, Pereira, Bar-
reto & de Paula, 2006). Successful integrated culturing of seaweed
and shrimp is primarily related to the concentration of nutrients,
type of cultivation and water flow (Buschmann, Troell & Kautsky,
2001). Growth rates of Gracilaria, high concentrations of nitrogen
tissue and phosphorus and good accumulated production of agar
obtained from the seaweed cultivated close to the fish farm indi-
cated that both economic and environmental advantages can be
achieved by integrating the cultivation of Gracilaria with fish farming
(Troell et al., 1997). The use of seaweeds as biofilters in intensive
mariculture systems showed that Gracilaria co-cultivated with salmon
accelerated production rates (Troell et al., 1999). The productivity of
G. chilensis close to salmon farms, as well as its nitrogen removal and
photosynthetic performance, indicated that Gracilaria growth perfor-
mance was always higher on the suspended cultures close to the sal-
mon cages (Abreu et al., 2009). Integrated culture of tiger shrimp
FOUROOGHIFARD ET AL.
(Penaeus monodon) with G. verrucosa resulted in the highest survival
rate (88.66
9.46), which was 24.3% higher than the controls.
Therefore, the presence of seaweeds promotes more survival rate of
tiger shrimp (Izzati, 2011). The objective of this study was to evalu-
ate the effects of integrated culture of G. corticata on shrimp perfor-
mance, such as survival rate, growth parameters and final production
of L. vannamei with different shrimp and seaweed densities under
zero-water exchange system.
2 | MATERIALS AND METHODS
2.1 | Design of experiment
This study was conducted for a period of 45 days between August
and October 2014 at Persian Gulf and Oman Sea Ecological
Research Center (PGOSERI). A 2 9 3 factorial design was utilized
with two levels of shrimp stocking densities (S1 and S2 with 25 and
50 shrimp/m2) as first factor and three levels of seaweed weight
densities (A1, A2 and A3 with 0, 200 and 400 g/m2) as second factor.
Two control treatments (C1 and C2) of a monoculture of seaweed
with a density of 200 g/m2 (with and without adding shrimp feed)
were also considered. All treatments had three replications. Experi-
ments were carried out in 24 round polyethylene tanks (r = 57 cm,
h = 100 cm) in covered saloon. Tanks were filled with 750 L of fil-
tered seawater. A 40-W compact fluorescent lamp was hung over
each tank to provide adequate and sufficient light for seaweed
growth. A neutral photoperiod was used which was 12 hours light
followed by 12 hr of darkness (12:12, L:D) (Yarish, Redmond & Kim,
2012). Each tank was aerated by two pieces of cylinder air stone
with an aeration power of 5 L/min.
2.2 | Culturing of shrimp and seaweed
The biomass of wild red seaweed G. corticata was collected from
coastal areas of Bandar-e Lengeh (26°330 29″N 54°520 50″E), Iran.
Juvenile L. vannamei (5.82
0.11 g) was obtained from a shrimp
farm located in Tiab shrimp farming site. Shrimps were stocked in a
5-ton fibre glass tanks for a one-week acclimation period, and the
seaweed was washed in the laboratory with seawater filtered by
rapid sand filter. Each clump of seaweed was rinse and carefully
inspected to completely eradicate encrusting organisms and trans-
ported to another tank filled with filtered seawater. G. corticata was
cultured on a net tied to a round polyethylene frame. Each net unit
had about 50 mesh intersections. Gracilaria seedlings were tied to
these places using soft plastic tie (Juanich, 1988). All frames had
three pods to support and hold nets 20 cm above the bottom. Prior
to fastening of the seaweed to the frames, initial weight of all
frames was determined and recorded on special sheet belonging to
each treatment. Twenty seedlings of 10 g each were fastened to
each net in treatments with 200 g seaweed (C1, C2, S1A2 and S2A2)
and forty seedlings of 10 g each were for treatments with 400 g
seaweed (S1A3 and S2A3). Initial weight of shrimps and their biomass
were determined by an A&D digital scale with a precision of 0.01 g,
FOUROOGHIFARD ET AL. | 3

prior to stocking in the culturing tanks. Shrimps were stocked at a 3 | RESULTS

density of 25 and 50 shrimp/m2, respectively, and were fed with
local commercial shrimp feed (Hormoz dam Shrimp grower Feed No. 3.1 | Water quality parameters
2, contained 35% protein), four times daily (06:00, 12:00, 18:00 and
Water temperature ranged from 30.4 to 35.8°C in the morning and
22:00 hours), which dispersed directly to each tank based on the
30.4 to 35.8°C in the afternoon, during the experimental period.
shrimp feeding table (Clifford, 1992). There was no exchange of
There was no significant difference between water temperature in
water and no water added during the culturing period.
the morning and in the afternoon. DO ranged from 5.1 to 6.36 mg/L
in the morning and from 5.63 to 6.56 mg/L in the afternoon. The
2.3 | Shrimp and seaweed biometry maximum and minimum fluctuations of pH (7.9–8.3 and 7.3–8.7)
were found in treatments S2A3 and S1A3 respectively. No signifi-
Shrimp and seaweed growth were estimated fortnightly; 15 shrimps
cant difference was observed between water temperature and pH in
were captured from each tank and their weight was determined.
the morning and in the afternoon. A significant difference was
Growth of seaweed was calculated by subtracting the total weight of
observed between pH and DO in various treatments. Shrimp density
seaweed and frame from the initial frame weight and at the end of the
had a significant effect on pH and DO in the morning and in the
experiments, all shrimps and seaweeds were harvested and weighed
afternoon, but the seaweed density did not have any significant
separately to obtain the total production for each treatment.
effect on these parameters; there was no significant interaction
effect between shrimp and seaweed densities on water quality
2.4 | Measurements parameters (Table 1). The minimum and maximum concentrations of
total ammonium (27.7
3 and 388.3
19 lg/L), nitrite
Water temperature, dissolved oxygen concentration (DO), pH (two
(4230
137 and 11822
305 lg/L), nitrate (44236
749 and
times per day, 6:00 and 14.00) and salinity (at 20 cm below the water
92437
2895 lg/L) and phosphate (2437
24.2 and
surface) were measured every 3 days during the experimental period.
6106
156.8 lg/L) were observed in treatments S1A3 and S2A1
Water temperature and dissolved oxygen were measured using a porta-
respectively. Shrimp and seaweed densities significantly affect the
ble oxygen meter (WTW, OXI 330i) with precisions of 0.1°C and
concentration of total ammonia, nitrite, nitrate and phosphate in
0.1 mg/L respectively. Water pH was measured with a portable pH
water (p < .05) (Table 1).
meter (WTW, pH 330i) with a precision of 0.01 pH unit. Salinity was
measured by an Atago Refractometer with a precision of 0.5 PPT. Total
ammonia, nitrite, nitrate and phosphate were measured by spectropho- 3.2 | Growth parameters of L. vannamei
tometric method (Cecil 3041 Spectrophotometer) with a precision of
Behaviour observation of L. vannamei during the experimental per-
1 lg/L (Strickland & Parsons, 1972). Sediments were collected from
iod showed that seedlings of G.corticata could act as a good shel-
each tank by siphoning, weighted and dried in an oven (at 70°C for
ter for L. vannamei. An equal growth in L. vannamei was observed
48 hr) and weighted by digital scale with a precision of 0.01 g.
in all treatments during the experimental period (Figure 1). The
maximum and minimum SGR of L. vannamei (1.97 and 1.69%/day)
2.5 | Growth parameters of shrimp and seaweed, was observed in treatment S1A3 (25 shrimp/m2 and 400 g sea-
survival rate and FCR weed/m2) and S2A1 (50 shrimp/m2 without seaweed) respectively
(Table 1). Results showed a density-dependent effect of shrimp
The following formulas were used to calculate the growth para-
and seaweed on ADG, SGR, FCR and survival rate of L. vannamei.

meters of shrimp and seaweed (Arnason, €rnsson, Steinarsson &
Bjo
The interaction effect of Shrimp and seaweed densities on all
Oddgeirsson, 2009; Ricker, 1975). Specific growth rate (SGR) (% per
growth parameters of L. vannamei, in exception of survival rate,
day) = 100(lnW2  lnW1)/(t2  t1), where W1 and W2 are the
was significant (Table 1) (p < .05). Strong positive and negative
weights of the shrimp and seaweed measured at times t1 and t2.
correlations were found between seaweed and shrimp densities
Weight gained (WG) (%) = 100(W2 – W1)/W1, where W1 and W2 are
and the growth parameters of L. vannamei respectively (p < .01)
the initial weight and final weight respectively. Average daily growth
(Table 3).
(ADG) = (W2  W1)/days, where W1 and W2 are the initial weight
The maximum survival rate (94.67
1.3%), maximum WG
and final weight respectively. Feed conversion ratio (FCR) = Total
(129.9
2.9%), maximum final mean weight (14.13
0.8 g), the
feed intake/total weight gain of the shrimp during the growth per-
minimum survival rate (51.3
1.3), minimum WG (10.10
3.1%)
iod. At the end of the experimental period, data were analysed using
and minimum final mean weight (12.51
0.82) were observed in
SPSS 22.0 software through the application of parametric tests. Data
treatments S1A3 (25 shrimp/m2) and treatment S2A1 (50 shrimp/m2)
were treated statistically by two-way analysis of variance (ANOVA)
respectively. The minimum and maximum FCR (1.67
0.04 and
with densities of shrimp and seaweed as fixed factors and shrimp
25.02
5.94) were obtained in treatments S2A1 and S1A3 respec-
production and growth parameters, as dependent variables. Signifi-
tively (Table 2). There was significant difference between FCR and
cant differences between the treatment means were compared by
survival rate in treatment S1A3 and other treatments (p < .05).
Duncan test. Differences were considered significant at p < .05.
 4
|

T A B L E 1 Effects of shrimp and seaweed densities on water quality parameters, growth parameters of Litopenaeus vannamei, and Gracilaria corticata and FCR in zero-water exchange system
during a 45-day culturing period (mean
SE)
S1 (25 shrimp per m2) S2 (50 shrimp per m2) Significancea
Shrimp density
Seaweed density (0 g) A1 (200 g) A2 (400 g) A3 (0 g) A1 (200 g) A2 (400 g) A3 SD AD SD 3 AD
Water quality variables
Temperature (°C) at 6 a.m. 30.4–35.8 30.4–35.8 30.5–35.8 30.5–35.8 30.5–35.8 30.7–35.8 NS NS NS
Temperature (°C) at 4 p.m. 30.7–35.8 30.6–35.8 30.7–35.8 30.8–35.8 30.8–35.8 30.7–35.8 NS NS NS
pH at 6 a.m. (n = 45)b 7.84–8.32 7.93–8.32 7.93–8.30 7.23–8.08 7.40–8.09 7.36–8.10 ** NS NS
pH at 4 p.m. (n = 45)b 7.82–8.31 7.91–8.31 7.92–8.30 7.18–8.08 7.33–8.13 7.29–8.70 ** NS NS
DO (mg/L) at 6 a.m. 5.41–6.29 5.42–6.29 5.42–6.36 5.10–6.31 5.28–6.22 5.29–6.29 ** NS NS
DO (mg/L) at 4 p.m. 5.76–6.56 5.79–6.49 5.73–6.48 5.85–6.44 5.67–6.43 5.63–6.56 ** NS NS
b
Salinity (PPT) (n = 45) 37.00–43.50 37.0–43.5 37.00–43.50 37.00–43.50 37.00–43.50 37.00–43.50 NS NS NS
Total Ammonium (lg/L)c 201
7 39.5
3 27.7
3 388.3
19 231
9.2 190
4.9 ** ** NS
Nitrite (lg/L)a (n = 3)c 6545
155 5153
31.8 4230
137 11822
305 9057
49 7615
247 ** ** **
Nitrate (lg/L)a (n = 3)c 57321
2551 48980
565.0 44236
749 92437
2895 86029
1397 78951
2207 ** ** NS
a c
Phosphate (lg/L) (n = 3) 3381
158 2894
28.6 2437
24.2 6106
156.8 5204
57 4371
88 ** ** **
Sediment dry weight (g)c 86
1.5 71
1.8 57.7
0.6 192
3 179
2.6 162
1.9 ** ** **
Shrimp
Final weight (g per shrimp) (n = 60) 12.6
0.2 13.4
0.2 14.1
0.2 12.5
0.2 13.2
0.2 13.4
0.2 ** ** **
ADG (g/day) 0.15
0.0 0.17
0.0 0.18
0.0 0.15
0.0 0.16
0.0 0.17
0.0 ** ** **
WG (%) 76.52
8.2 102.1
3.4 129.9
2.9 10.10
3.1 51.6
5.7 64.3
5.5 ** ** **
SGR (%/day) (n = 3) 1.72
0.02 1.85
0.02 1.97
0.0 1.7
0.01 1.83
0.02 1.85
0.0 ** ** **
Survival rate (n = 3) 81.3
3.5 88.0
2.3 94.7
1.3 51.3
1.3 64.7
2.7 71.3
2.4 ** ** NS
FCR 2.89
0.3 2.12
0.07 1.67
0.04 25.02
5.9 4.31
.45 3.42
0.28 ** ** **
Seaweed
Weight gained (%) 73.67
5.5 64.83
2.1 23.50
3.01 14.92
1.9 ** ** **
SGR (%/day) (n = 3) – 1.23
0.07 1.11
0.03 – 0.47
0.05 0.31
0.04 ** NS NS
a
Results from two-way ANOVA; SD, shrimp density; AD, Seaweed density; SD 9 AD, Shrimp density 9 Seaweed density interaction.
b
During the culturing period.
c
End of experimental period
**Significant at the .01 level
FOUROOGHIFARD
ET AL.
FOUROOGHIFARD ET AL. | 5

16 S1 A1 700 C1

Mean body weight (g)

S1 A2 600
14 C2
S1 A3
500

Weight (g)
12 S2 A1 S1A2
400
S2 A2 S1A3
10
S2 A3 300
8 S2A2
200
100 S2A3
6
4 0
1 15 30 45 0 15 30 45
Rearing time (days) Culturing time (days)

F I G U R E 2 Growth of Gracilaria corticata integrated culture with

F I G U R E 1 Weight growth of Litopenaeus vannamei in different L. vannamei in different shrimp and seaweed density (mean
SE)
shrimp and seaweed density (mean
SE)

6106
156.8 lg/L) were obtained in treatments S1A3 (25 shrimp/m2

3.3 | Growth parameters of G. corticata and 400 g seaweed/m2) and S2A3 (50 shrimp/m2 without any sea-
weed), respectively, and it can be said that seaweed densities signifi-
Growth trend of G. corticata has been shown in Figure 2; the SGR
cantly affect the concentration of total ammonia, nitrite, nitrate and
of G. corticata in treatments S1A2 (1.22
0.07%/day) and S1A3
phosphate in water (p < .05) (Table 1). A study on the biofiltering
(1.11
0.03%/day) was significantly higher than the other treat-
ability of seaweed in decreasing inorganic nitrogen of shrimp-cultur-
ments (p < .01), but there was no significant difference between the
ing ponds revealed that G. manilaensis was able to reduce up to
two (p > .05). All the seaweeds in treatment C2 (200 g/m2 with the
83.65% of NH4+, 33.33% NO2 and 68.42% NO3 after 24 hr
addition of shrimp feed to the tanks) decomposed gradually because
(Shukri & Surif, 2011). According to the results of this study, it can
of sedimentation of the feed and growth of fungi on their thalli
be inferred that an increase in the density of G. corticata led to
(Table 4). A significant negative correlation was found between
enhance the water quality and decrease waste pollution released
shrimp density and growth parameters of G. corticata (Table 5).
from the shrimp and increase the growth parameters of L. vannamei
in integrated culture with seaweed. The minimum and maximum sed-
iments (57.7
0.6 and 192
3 g dry weight) were also obtained in
4 | DISCUSSION treatments S1A3 and S2A3, respectively, increasing in the density of
G. corticata led to improve survival rate, reduce the wastage of the
According to the results, the best performance of L. vannmei (maxi-
feed as sediments and reduce the FCR. In fact, most of the feed was
mum survival rate, SGR, WG and minimum FCR) was observed in
wasted and increased sediments in treatment S2A1, because of bad
treatment S1A3 (25 shrimp/m2 and 400 g seaweed/m2) (Tables 1
water quality and low survival rate of L. vannmei (Table 1). The
and 2). The maximum and minimum SGR (1.97
0.00 and
increase in the density of shrimp led to a decrease in the SGR of
1.69
0.01%/day) was observed in treatments S1A3 and S2A1 (50
L. vannmei. There was no study on integrated culture of the whiteleg
shrimp/m2 and without seaweed) respectively. Strong positive corre-
shrimp L. vannamei and G. corticata under zero-water exchange sys-
lation was found between seaweed densities and the growth param-
tem. Some studies have focused on other species and culturing or
eters of L. vannamei (Table 3). In this study, the minimum and
management system such as cage size, stocking density or feed
maximum of total ammonium (27.7
3 and 388.3
19 lg/L),
source. The effect of the seaweed Caulerpa sertularioides on the
nitrite (4230
137 and 11822
305 lg/L), nitrate (44236
749
growth and survival of captured yellowleg shrimp Penaeus californien-
and 92437
2895 lg/L) and phosphate (2437
24.2 and
sis indicated that shrimp growth and survival rate in the presence of

T A B L E 2 Experimental data of
Treatment Initial biomass (g) Final biomass (g) Survival rate (%) Feed intake (g) FCR
production and feed conversion
ratio of L. vannamei integrated S1 A1 145.5
0.29 256.9
12.35 a
81.3
3.5 a
314.8 2.89
0.29a
cultured with G. corticata under S1 A2 145.7
0.19 294.4
4.86 b
88.0
2.3 b
314.8 2.12
0.07a
zero-water exchange system S1 A3 145.5
0.07 334.4
4.32c 94.7
1.3b 314.8 1.67
0.04b
(mean
SE)
S2 A1 291.6
0.26 321.1
9.29 c
51. 3
1. 3 c
629.6 25.0
5.94c
S2 A2 290.2
0.12 439.9
16.86d 66.7
2.7d 629.6 4.31
0.45d
S2 A3 290.4
0.12 477.2
16.27 d
71.3
2.4 d
629.6 3.42
0.28d

Superscript words in same column indicate homogeneous subsets as determined by Duncan’s test
(a = .05).
6 | FOUROOGHIFARD ET AL.

T A B L E 3 Correlation between
Correlations
seaweed and shrimp densities and
ADG WG SGR Survival rate FCR growth parameters, survival rate
Shrimp density and food conversion ratio (FCR) of
L. vannamei co-cultured with G.
Pearson correlation .230 .790a .294 .844a .484b
corticata under zero-water
Sig. (twotailed) .359 .000 .236 .000 .042
exchange system (mean
SE)
N 18 18 18 18 18
Seaweed density
Pearson Correlation .901a .570b .887a .461 .519b
Sig. (two-tailed) .000 .014 .000 .054 .027
N 18 18 18 18 18
a
Correlation is significant at the .01 level (two-tailed).
b
Correlation is significant at the .05 level (two-tailed).

T A B L E 4 Growth parameters of
Treatment Initial weight Final weight ADG (g/day) Weight gain (%) SGR (%/day)
G. corticata co-cultured with L.
C1 200
0.0 57.3
4.34 3.17
0.1a 71.33
2.2 2.79
0.17a vannamei in different densities of
C2 200
0.0 0.0
0.0 4.44
0.0 b
100.00
0.0 0.00
0.00b shrimp and seaweed under zero-
S1A2 200
0.0 347.3
11.1 3.27
0.3c 73.67
5.5 1.22
0.07c water exchange system
(mean
SE)
S1A3 400
0.0 659.3
8.4 5.76
0.2 d
64.83
2.1 1.11
0.03c
S2A2 200
0.0 247.0
6.1 1.04
0.1e 23.50
3.01 0.47
0.05d
S2A3 400
0.0 459.7
7.5 1.33
0.2 e
14.92
1.9 0.31
0.04d

Superscript words in same column indicate homogeneous subsets as determined by Duncan’s test
(a = .05).

water; therefore, the co-culture of G. manilaensis with shrimp has

T A B L E 5 Correlation between seaweed and shrimp densities and been suggested to improve water quality and reduce waste pollution
growth parameters of G. corticata under zero-water exchange released from the shrimp (Shukri & Surif, 2011). Study on Shrimp
system
and Ulva co-culture indicated that the combination of co-cultured
Seaweed density Seaweed biomass SGR WG micro-algae and artificial feed can improve the growth rate of L. van-
Shrimp density namei significantly, and it has been suggested that Ulva sp. may act
Pearson correlation .488 .969a .967a as a nutritional supplement and/or improve the utilization of nutri-
Sig. (two-tailed) .107 .000 .000 ents from the artificial feed (Cruz-Suarez et al., 2010). In contrast,
N 12 12 12 the polyculture of the whiteleg shrimp L. vannamei and the Philip-
Seaweed density pines seaweed Kappaphycus alvarezii in floating cages indicated that
Pearson correlation .854 a
.170 .168 after 103 days of growth, there were no negative interferences

Sig. (two-tailed) .000 .597 .601

N 12 12
a
Correlation is significant at the .01 level (two-tailed).
seaweed were 3.98
0.4 g and 100%, respectively, after 10 weeks,
whereas in the absence of seaweed were 0.46
0.4 g and
68.7
12% respectively. Significant differences
between growth rate and survival rate of shrimp in the absence and
 rdova,
presence of seaweed (Porchas-Cornejo, Antonio Martınez Co
 n Barajas, Naranjo Paramo & Portillo Clark, 1999). A study
Magallo
on the biofiltering ability of G. manilaensisi in reducing inorganic
nitrogen waste of shrimp culture showed that G. manilaensis could
mitigate the inorganic nitrogen concentration in the aquaculture
between the co-cultured shrimp and seaweed; in addition, there was
no significant difference between monoculture and polyculture for
12
shrimp weight gain, survival rate and feed conversion ratio.(Lombardi
et al., 2006). It was observed in the course of this study that cultur-
ing of G. corticata on net in shrimp-culturing tanks might also act as
an artificial substrate that can disperse the shrimp from the bottom
of the tank onto the seaweed substrate and thus reduce the nega-
tive effect of high stocking density on shrimp growth in the tanks. A
were found study on the effects of artificial substrates on the growth, survival
and spatial distribution of L. vannamei in intensive culture condition
indicated that both the average weight and survival rate in the tanks
with artificial substrates were significantly higher than those in the
control tanks; moreover, the weight and survival rate of L. vannamei
increased when the number of artificial substrates was increased.
(Zhang, Lin, Huang, Wang & Xu, 2010)
FOUROOGHIFARD ET AL.
In this study, the best growth rate of G. corticata was obtained in
the treatments S1A2 and S1A3 (with 25 shrimp and 400 g seaweed/m2)
(Table 4). The productivity of species in a mixed culture system is
dependent on the growth performance of both species in the system;
thus, environmental conditions have to be optimal for both species to
generate the highest production (Qian, Wu, Wu & Xie, 1996). A signifi-
cant negative correlation was found between shrimp density and
growth parameters of G. corticata (Table 5); an increase in the density
of shrimp can lead to increase in turbidity and decrease in the amount
of light, which can limit the growth of algae. The seaweeds in treat-
ment C2 (200 g seaweed/m2 with addition of shrimp feed to the
tanks) decomposed gradually during the culturing time. The sedimen-
tation of shrimp feed led to rot of the thalli of G. corticata because
they were covered by mould. In other treatments, the feed that set-
tled on seaweed’s thalli was eaten by shrimps and were cleaned in the
process. The maximum SGR of G.corticata in treatments was observed
in treatments S1A2 (1.22
0.07%/day) and S1A3 (1.11
0.03%/day).
Similarly, the growth rate of 1.11%/day was obtained for red seaweed
Kappaphycus alvarezii in co-culture with the whiteleg shrimp L. van-
namei in floating cages (Lombardi et al., 2006). Different SGR has been
obtained for G.corticata cultured in fibre glass tanks in different sea-
sons; SGR of 1.46
0.11, 3.74
0.31, 1.5
0.28 and 3.11

0.04%/day were obtained for G.corticata, respectively, in winter,
spring, summer and autumn seasons (Akbari, Aftabsavar, Malakouti,
Tamadoni & Ejlali, 2004).
Negative SGR (2.79
0.17) in G. corticata cultured in treatment
C1 (seawater without any shrimp feed or fertilizer) indicated that there
was no adequate dissolved nutrient in natural seawater for the growth
of G. corticata. Study on the effects of urea and TMRL1 media on the
growth rate of G.corticata indicated the growth rates of 1.87
0.54,
0.26
0.26 and 1.04
0.37 for TMRL media, urea and seawater
without any fertilizer, respectively (Fourooghifard, Tazikeh & Akbari,
2005). In contrast, growth rates of 3.72 to 7.17%/day were obtained
for red seaweed Kappaphycus alvarezii cultured inside a 4 9 4 m float-
ing net cage of Lates calcarifer (Hurtado-Ponce, 1992).
5 | CONCLUSION
Based on the higher survival rate, higher WG, higher final mean
weight of L. vannamei, minimum FCR and also higher SGR of G. corti-
cata in treatments S1A3 (treatments with 25 shrimp and 400 g sea-
weed/m2), we conclude that G. corticata with a density of
400 g seaweed/m2 and 25shrimp/m2 could be suitable for inte-
grated aquaculture of L. vannamei and G. corticata.
Increasing in the density of G. corticata led to improve survival
rate, reduce the wastage of the feed as sediments and reduce the
FCR. The red seaweed G. corticata can act as an artificial substrate
that reduce the negative effect of high stocking density on shrimp
growth in the tanks and boost growth rate parameters, survival rate
and total production of L. vannamei under zero-water exchange
1
Tung Kong Marine Research Laboratory.
| 7
system. Negative SGR in G. corticata cultured in treatment C1 (sea-
water without any shrimp feed or fertilizer) indicated that shrimp-
culturing activity provide adequate dissolved nutrient for the growth
of G. corticata in culturing tanks.
ACKNOWLEDGMENTS
The authors wish to thank Mr. Rabiei and Mrs. Moezzi for laboratory
assistance in the course of this research work. This study was sup-
ported by the Iranian Fisheries Research Institute, Persian Gulf and
Oman Sea Ecological Research Center, Iran.
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How to cite this article: Fourooghifard H, Matinfar A,
Mortazavi MS, Roohani Ghadikolaee K, Mirbakhsh M. Growth
parameters of whiteleg shrimp Litopenaeus vannamei and red
seaweed Gracilaria corticata in integrated culturing method
under zero water exchange system. Aquac Res. 2017;00:
1–8. https://doi.org/10.1111/are.13335