JOURNAL OF THE WORLD AQUACULTURE SOCIETY

Vol. 39, No. 4 August, 2008

Improvement of Water Quality by the Macroalga, Gracilaria lemaneiformis (Rhodophyta), near Aquaculture Efuent Outlets
YONGJIAN XU1
Faculty of Life Science and Biotechnology, Ningbo University, Ningbo 315211 China

JIANGUANG FANG

AND

QISHENG TANG

Yellow Sea Fisheries Research Institute, CAFS, Qingdao 266071 China

JUNDA LIN
Department of Biology Sciences, Florida Institute of Technology, Melbourne, Florida 32901 USA

GUANZONG LE

AND

LV LIAO

Faculty of Life Science and Biotechnology, Ningbo University, Ningbo 315211 China

Abstract
Raft culture of the seaweed, Gracilaria lemaneiformis (Bory) Dawson, was used as biolter for purifying efuent and removing nutrients from the nearby animal aquaculture. The results showed that the cultivated seaweed grew well outside efuent discharge outlets for growout ponds and a hatchery plant. The biomass (fresh weight) of Gracilaria from the 0.5-ha area outside the growout ponds increased about 60 times, from 80 to 4750 g/m during the 65 d of experiment (with the daily special growth rate [SGR] of 3.87%/d); the yield was 30.4 tons and 93.97 kg N and 12.81 kg P were removed; the seaweed biomass from the 0.2-ha area outside the hatchery outlet increased 37.5 times, from ,100 to 3600 g/m (with SGR of 3.4%/d); the yield was 10.4 tons and 31.7 kg N and 4.33 kg P were removed. Several water quality parameters such as dissolved oxygen, secchi disk depth, and pH value in the algal cultivation area were substantially higher than those in the noncultivation zone, whereas chemical oxygen demand, dissolved inorganic nitrogen, and dissolved inorganic phosphorous at both areas were similar.

Rapid development of intensive aquaculture in coastal areas throughout the world has raised increasing concerns on its environmental impacts (e.g., Wu 1995; Mazzola et al. 1999). Organic and inorganic inputs of sh culture feed have lead to a substantial increase of organic matter and nutrient loading in coastal waters. In general, 5295% of the nitrogen and about 85% of the phosphorus input into a marine sh culture system as feed may be lost into the environment through feed wastage, sh excretion, and feces production, which may lead to eutrophication, harmful algal blooms (HAB), and anoxia (Wu 1995). In China, efuent from shrimp culture ponds often exceeds the purify-

Corresponding author.

ing capacity of local coastal sea or bay and accelerates the eutrophication and results in HAB (Sun et al. 1990; Sheng and Shi 1996). Annually, about 4.4 billion tons of efuent, including 17,000 tons of organic matter, from 8000-ha shrimp culture ponds in Yantai, Shandong Province, China, were discharged (Zhai 1996). The negative economic and environmental impacts of monospecic (especially of high trophic level) aquaculture operations are being recognized and their sustainability questioned (Naylor et al. 2000; Chopin et al. 2001; Neori et al. 2004). In recent years, there has been increasing emphasis on developing sustainable approaches to coastal aquaculture (Folke and Kautsky 1989; Wurts 2000; Frankic and Hershner 2003; Neori et al. 2004). Polyculture of different trophic levels is the basis of environmentally friendly

Copyright by the World Aquaculture Society 2008

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aquaculture (Chopin et al. 2001; Neori et al. 2004). By integrating fed mariculture (e.g., sh and shrimp) with inorganic and organic extractive mariculture (seaweed and lter-feeding bivalve), the waste of one resource consumer becomes a resource (fertilizer or food) for the other in the system. Such a balanced ecosystem approach provides mutual benets to cocultured organisms, economic diversication by producing value-added products, nutrient bioremediation capacity, and environmental sustainability (Chopin et al. 2001; Schuenhoff et al. 2003; Neori et al. 2004). In the past several decades, large-scale suspended seaweed farming has been commercialized (Tseng and Fei 1987; Petrell et al. 1993; Troell et al. 1997; Chopin et al. 1999; Neori et al. 2004). However, there are only few species of macroalgae that are suitably cocultured with animals, especially in ponds. The genus Gracilaria (Rhodophyta) has been demonstrated to be an attractive candidate for intensive culture because of its value as human food and its ability to achieve high yields and to produce commercially valuable products (Chopin et al. 2001; Fei 2004; Neori et al. 2004). The edible red alga, Gracilaria lemaneiformis, occurs naturally in coastal areas of Shandong Peninsula, north China. This species has been successfully introduced to subtropical coastal waters in south China where it is also cultivated widely (Fei et al. 2000). In this study, we investigated the algal growth and bioremediation potential near efuent outlets of animal culture ponds and a hatchery plant. Materials and Methods Study Site The study was carried out in Xiangshan Bay (29299320N, 121339590E), Zhejiang Province, China. Raft culture of the red alga, G. lemaneiformis, was established at two sites. The rst one was a 0.5-ha (50 3 100 m) area in the bay outside an outlet (PM) that discharged the efuent (directly to the bay without treatment) from about 15-ha aquaculture growout ponds where several fed animals (Pacic white shrimp, Litopenaeus vannamei; mud crab, Scylla serrata; swimming crab, Portunus trituberculatus; and

crab, Nibea miichthioides) and ltered animals (cockle, Tegillarca granosa Linnaeus; clam, Cyclina sinensis; and razor clam, Sinonovacula constricta) were cultured. Another 0.2-ha (40 3 50 m) algae cultivation area was similarly established in the bay outside another outlet (HM) that discharged efuent from a hatchery of the shrimp, L. vannamei, and the crab, P. trituberculatus. The untreated aquaculture efuent was discharged directly to the bay during ebb tide of spring tide and water depth in the area during low tide is about 2 m. The bay area has a regular semidiurnal tide of about 1.2- to 1.5-m amplitude with the tidal ow of about 0.30.5 cm/sec. Cultivation of Gracilaria The alga, G. lemaneiformis, provided by the Hutoudu Aquaculture Company in Xiangshan Bay was inserted into polyethylene ropes that were supported by the rafts. A suspension culture unit was composed of four rafts, each in turn was composed of eight ropes. Each rope (about 10 m long and 6 cm diameter) was inserted with a piece of alga (about 15 g at the PM site and 20 g at the HM site) every 15 20 cm. Distance between adjacent ropes was 4050 cm and between adjacent units was 2 3 m. Twenty units (with 640 ropes and 500 kg algal seedling) and 8 units (with 256 ropes and 250 kg algal seedling) were established at the PM and HM sites, respectively. Ten (at the PM site) and ve (at the HM site) ropes with Gracilaria were randomly selected and weighted every 15 d between March 2 and May 7, 2006. The weights of the polyethylene ropes without the algae were the same before and after the study. Average daily specic growth rates (SGRs) were calculated as SGR 5 (100 ln (Wt/W0))/t, where W0 is initial wet weight and Wt is the wet weight at day t (Lobban and Harrison 1994; Yang et al. 2006). Water Quality Sampling For both growout pond (PM) and hatchery (HM) sites, two transects were similarly established (Fig. 1). Transects A and B radiated from the efuent outlet, with Transect A passed through the algal cultivation area, whereas Transect B did not. Sampling stations were set

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FIGURE 1. Map of the study area. For both growout pond (PM) and hatchery (HM) sites, four transects were similarly established. Transects A and B radiated from the efuent outlet; Transect A passed through the algal cultivation area, whereas Transect B did not. Sampling stations were set up every 1525 m along each transect, with a total of seven stations along Transects A (with A11A17 and A21A27 for PM and HM sites, respectively) and B (with B11B17 and B21B27 for PM and HM sites, respectively).

up every 1525 m along each transect, with a total of seven stations along Transects A (with A11A17 and A21A27 for PM and HM sites, respectively) and B (with B11B17 and B21 B27 for PM and HM sites, respectively). Between March 2 and May 7, 2006, water samples were taken every 78 d at Transects A and B (total 10 times) during efuent discharge when tide ow ebbed. During each sampling, several water quality parameters such as water temperature (WT), salinity (S), secchi disk depth (SD), pH, dissolved oxygen (DO), chemical oxygen demand (COD), dissolved inorganic nitrogen (DIN; NH3N + NO2N + NO3N), and dissolved inorganic phosphorous (DIP) were measured. WT, S, SD, pH, and DO of surface water were measured using YSI-85 (YSI, Marion, MA, USA) in the eld; COD was estimated following the JGOFS Protocol No. 19 (JGOFS International Project Ofce 1994). NO3N, NO2N, NH4N, and PO4P were determined with a Skalar San and Plus Flow Analyzer (Skalar, Delft, Holland). Samples were collected and placed into a foam box with ice and then transported to the laboratory for immediate analysis.

10 SGR-PM SGR-HM 8 Density-PM Density-HM

SGR (%/d)

6 2 4 1 2 0

0 0 15 30 45 65

Time (Day)
FIGURE 2. Changes of the daily specic growth rate (SGR) and unit density of the red alga, Gracilaria lemaneaformis, at the growout pond (PM) and hatchery (HM) sites during the 65-d study.

Density (Kg/m)

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Statistical Analysis Results were analyzed with the software Sigmaplot 8.0 and SPSS 12.0. Mean and standard deviation of each treatment (n 5 3) were calculated. The corresponding stations of Transects A and B within each site (PM or HM) were compared using Students t test. Results Growth of Gracilaria Changes in SGR and density of the seaweed, G. lemaneiformis, over the study period are shown in Figure 2. The patterns were similar
Site of pond outlet (PM) 1.2 1

between the two sites (PM and HM). The average SGRs during the 65-d study period were 3.87%/d and 3.40%/d at the PM and HM sites, respectively. The densities increased from 80 to 4750 g/m and from ,100 to 3600 g/m at the PM and HM sites, respectively. The biomass (fresh weight) increased 59.4 and 37.5 times (Fig. 2), and the yield was 30.4 and 10.4 tons at the PM and HM sites, respectively. Environmental Parameters During the study period, WT varied from 13.2 to 20.7 C with an average of 18.8 C. WT at the
Site of hatchery outlet (HM) 1.2 1

SD (m)

SD (m)
7

0.8 0.6 0.4 0.2 0

11 13 17 B1 3 A 15 B1 1 B1 5 A A A B1

0.8 0.6 0.4 0.2 0

A 25 27 A 23 B2 5 B2 1 A A B2 3 B2 7 5 B2 B2 7 B2 7 21

14 12

12 10

DO (mg/L)

DO (mg/L)

10 8 6 4 2 0
B1 1 B1 5 A 11 A 13 A 15 A 17 B1 3 B1 7

8 6 4 2 0
21 23 A 27 A 25 B2 3 B2 3 B2 1 A A B2 5

8.5 8.4

8.5 8.4

pH

8.3 8.2 8.1

1 A 11 A 15 A 17 B1 3 5 B1 7 A 13 B1 B1

pH

8.3 8.2 8.1 8

25 27 A

Sampling Sites

Sampling Sites

FIGURE 3. Average (6SD) of secchi disk depth (SD), pH, and dissolved oxygen (DO) in the surface water at the growout pond (PM) and hatchery (HM) sites during the study period. d represents AV1 (from March 2 to April 15, 2006); s represents AV2 (from April 16 to May 7, 2006).

B2 1

21

23

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HM site was slightly higher than that at the PM site because of the input of heated wastewater from the hatchery plant. Salinity was stable, ranged from 24.7 to 26.6&. SD, DO, and pH values from March 2 to April 15 (AV1) were similar between A and B transects at both PM and HM sites (P . 0.05) (Fig. 3). However, between April 15, when the seaweed density reached about 2000 g/m, and May 7 (AV2), each parameter was signicantly higher at Transect A (within the algal culture area) than at Transect B (no algal culture) for both PM and

HM sites (P , 0.05) but similar between the corresponding stations within both the PM and the HM sites (P . 0.05) (Fig. 3). COD, DIN, and DIP at both Transects A and B were similar between AV1 and AV2 (P . 0.05), as well as between the corresponding stations within both PM and HM sites (P . 0.05) (Fig. 4). Discussion Species of the genus Gracilaria are commonly cultured for food, commercial products, as well as bioremediation (Troell et al. 1997,

Site of Pond outlet (PM) 3 3

Site of Pond outlet (HM)

COD (mg/L)

COD (mg/L)

0
11 13 17 B1 1 B1 3 B1 5 B1 7 15 A A A A

0
21 23 A 27 B2 1 25 3 B2 5 B2 5 B2 5 B2 A B2 B2 7 B2 7 A A 7

Dissolved inorganic nitrogen (mol/L)

50 40 30 20
3 11 A 13 A 15 17 B1 1 B1 5 B1 7 B1 A A

Dissolved inorganic nitrogen (mol/L)

60

50

40

30

20
21 23 27 A 25 B2 1 A A A B2 B2 3 3

Dissolved inorganic phosphate (mol/L)

1.6 1.2 0.8 0.4 0

15 17 3 11 A 13 B1 1 B1 7 B1 5 B1 A A A

Dissolved inorganic phosphate (mol/L)

2 1.6 1.2 0.8 0.4 0

21 23 25 27 A A A A

Sampling Sites

Sampling Sites

FIGURE 4. Average (6SD) of dissolved inorganic nitrogen, dissolved inorganic phosphorous, and chemical oxygen demand (COD) in the surface water at the growout pond and hatchery sites during the study period. d represents AV1 (from March 2 to April 15, 2006); s represents AV2 (from April 16 to May 7, 2006).

B2 1

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1999; Chopin et al. 2001; Fei 2004; Neori et al. 2004). Gracilaria grows fast and can uptake large quantities of nutrient from the water. Troell et al. (1997, 1999) estimated that 1 ha of another cultured red alga, Gracilaria chilensis, had the potential to remove at least 5% of DIN and 27% of DIP released from a sh farm. In the two studies conducted in Jiaozhou Bay, north China, 1 ha of the alga, G. lemaneiformis, cultivated at the sh and bivalve culture area yielded an estimated 70 tons (0.22 tons N and 0.03 tons P removal) (Zhou et al. 2006) and 258 tons (1.02 tons N removal) of fresh weight annually (Yang et al. 2006). In the present study, 0.5 (PM) and 0.2 ha (HM) of the culture area yield an estimated 30.4 and 10.4 tons of fresh weight, respectively, resulting in a production of about 5060 tons/ha in the 65-d study period. The alga, G. lemaneiformis, can grow all year round in the Xiangshan Bay area (it does not grow well in the winter and summer in north and south China, respectively). Seaweed culture outside the aquaculture discharge outlets may be an effective way of treating the efuent as well as enhancing seaweed growth. The seaweed cultivation can also improve other aspects of water quality, such as increasing DO and SD (Fig. 2). Even though the efciency of organic uptake by the seaweed is low (Xu et al. 2004), its photosynthesis produces DO that accelerates decomposition of organics. On the other hand, high-density raft culture of Gracilaria impedes the water circulation and may decrease COD in the water column. Several species of Gracilaria can produce oxygen under low light conditions, such as in rainy days, and remediate anoxia (Xu et al. 2004). Methods for treating efuents from mariculture systems with macroalgae were initiated in the mid-1970s and developed in recent years (Neori et al. 1996, 2000; Neori and Shpigel 1999; Troell et al. 1999; Chow et al. 2001; Nelson et al. 2001; Neori et al. 2004; Zhou et al. 2006). However, how to achieve an appropriate balance of animal/algae ratio (biomass or density) is still a challenge. Troell et al. (1997, 1999) suggested a biomass ratio of animal to seaweed to be 1:4. Yang et al. (2006) speculated that the sh/seaweed biomass ratio at approxi-

mately 1:1 was optimal for efcient nutrient uptake and for maintaining low nutrient levels. Many factors affect the optimal ratio. For example, changes in temperature affect growth and excretion rates of both animals and seaweed. Trophic levels of animals (e.g., lter-feeding species vs. carnivorous species) would have a profound impact on the waste production. In addition, both the economic factors and the capacity of nutrient bioremediation by seaweed should be considered (Chopin et al. 2001; Neori et al. 2004). Acknowledgments This research was nanced by the National Basic Research Program of China (no. 2006CB 400608), Ningbo Science and Technology Bureau (2005A610025; 2006A610087), and Young College Faculty Foundation of Zhejiang Province (2005757). This research was also sponsored by K.C.Wong Magna Fund in Ningbo University. The manuscript was greatly improved by the constructive comments of several reviewers. Literature Cited
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