Growth performance of spiny lobster Panulirus ornatus in land-based Integrated Multi-Trophic Aquaculture (IMTA) system

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Growth performance of spiny lobster Panulirus ornatus in land-based Integrated Multi-Trophic Aquaculture (IMTA) system Milennie Vialantine Sumbing*, Sujjat Al-Azad, Abentin Estim & Saleem Mustafa Borneo Marine Research Institute, Universiti Malaysia Sabah, Jalan UMS, 88400, Kota Kinabalu, Sabah, MALAYSIA. *Corresponding author. E-Mail: milennie.bibie@yahoo.com.my; Tel: +60-88-32000; Fax: +60-88-320261 Received: 21 March 2016 Revised: 8 April 2016 Accepted: 13 May 2016 In press: 20 May 2016 Online: 30 June 2016 Keywords: Integrated Multi-Trophic Aquaculture; Recirculating system; Flow-through system; Spiny lobster A b s t r a c t Aquaculture waste in the form of particulate organic matter and inorganic nutrients produces negative impacts when released to the environment. Integrated Multi-Trophic Aquaculture (IMTA) system is gaining importance for reasons of its environmental compatibility and quality of yield. The purpose of this study is to compare growth performance of a popular specis of spiny lobster in two different rearing systems. These were modeled based on recirculating system (RS) and flow-through system (FTS). Spiny lobster (Panulirus ornatus), sea cucumber (Holothuria scabra) and seaweed (Kappaphycus alvarezii) were used in the trials in both these systems. Water flow rate was maintained at 0.08 ± 0.1 L/sec. The stocking rate was 5 specimens / tank for spiny lobster (mean weight of 151.44 ± 7.14 g) and sea cucumber (mean weight of 32.16 ± 1.40 g), while mean initial biomass for seaweed was 500.65 ± 1.76 g/tank. Trials were conducted for 10 weeks. The results indicated that the SGR of lobster was not significantly different (p>0.05) in FTS (0.125 % day -1 ) and in RS (0.096 % day -1 ). There was no significant difference (p>0.05) in the survival of spiny lobster in FTS and RS. The survival rate of spiny lobster was 93.3 % and 80.0 % in FTS and RS, respectively. The inorganic nutrients, namely ammonia (NH 3 ), nitrite (NO 2 ), nitrate (NO 3 ) and phosphate (PO 4 ) were significantly higher (p<0.05) in RS than in FTS. Evidently, the FTS is the better option for culture in terms of efficiency of water quality remediation and growth of the stocked species. Transactions on Science and Technology 2016 Introduction Integrated Multi-Trophic Aquaculture (IMTA) refers to farming of different aquaculture species together in a way that the system comprises fed aquaculture, organic extractive and inorganic extractive species in the same production module to reduce organic and inorganic wastes (Neori et al., 2004). In an IMTA system, the organic matter comprising uneaten feed and faeces from fed aquaculture (for example, fish or shrimp) is consumed by the deposit feeders (for example, sea cucumber). In addition, the inorganic nutrients, mainly nitrogen and phosphorus are absorbed by seaweed. Incorporation of species from different trophic levels helped in utilizing the waste from one species as a feed or fertilizer for other species, and this balance system ensures better water quality within the environment. The aim of IMTA system is to approach long-term environmental sustainability and profitability. There is a growing number of publications suggesting the feasibility of Sumbing et al., 2016. Transactions on Science and Technology. 3(1-2), 143-149

Sumbing et al., 2016. Transactions on Science and Technology. 3(1-2), 143-149 144 IMTA modules in different regions. Interesting findings on Gilt-head bream (Sparus aurata), abalone (Haliotis discus hannai) and seaweed (Ulva lactuca) published by Neori et al. (2000) emphasized the sustainability element in the IMTA. Spiny lobster was selected because it is one of the most valuable seafood and marketable species in Sabah. This species is marketed locally and is also exported to other countries such as Hong Kong and Taiwan. The price of this species averages RM70/kg in the wholesale local market and is as high as RM115/kg in the export trade to Hong Kong (Biunsing and Lin, 2004). Among the various spiny lobster species in the Indo-west Pacific region, Panulirus ornatus and Panulirus homarus are the most preferred ones for aquaculture because of high demand, market value and availability of wild seeds (Clive, 2010). Spiny lobster fed trash fish grows fast. Lobster waste comprising uneaten feed and excrement, can adversely affect water quality, especially when ammonia concentration is high (Vinh and Huong, 2009). To address this problem, spiny lobster was chosen as the first trophic level in IMTA system. The study was designed to compare the growth and survival of spiny lobster, P. ornatus, in recirculating and flow-through systems, as well as to observe the dynamics of water quality parameters in the two systems. Methodology Three aquaculture species (spiny lobster, sea cucumber and seaweed) formed the basis of trial modeled on IMTA concept. One experimental module was designed as flow-through system (FTS) and the other as recirculating water (RS) system that consisted of (1 m 1 m 1 m) 1000L water capacity tank for spiny lobster and two (1.3 m 0.8 m 0.4 m) 500L tank for both sea cucumber and seaweed. In the flow-through system (Figure 1), seawater was directly supplied to spiny lobster tank. Effluent from spiny lobster tank was drained into the sea cucumber tank through 25 mm outlet pipe. Water was pumped to seaweed tank from where it drains out. Sequence of water flow was much the same in the RS (Figure 2) except that the water from the seaweed tank was made to flow back to the spiny lobster tank. The tanks were aerated continuously to ensure adequate dissolved oxygen (DO). Figure 1. Recirculating IMTA system Figure 2. Flow-through IMTA system

Sumbing et al., 2016. Transactions on Science and Technology. 3(1-2), 143-149 145 Spiny lobster unit Spiny lobster specimens were obtained from Pulau Banggi Kudat, Sabah. The stocking density was 5 specimens/tank (average body weight of 151.44 ± 7.14 g). PVC pipe of 10 cm diameter was provided to serve as a as shelter. Lobsters were fed trash fish with 3% of the body weight twice daily at 8:00 h and 16:00 h. Feed intake was recorded. Tanks were siphoned every morning to clean the uneaten feed. Analysis of water sample In-situ water quality parameters such as dissolved oxygen (mg/l), ph, temperature ( C) and salinity (ppt) recorded daily by using YSI Multi-parameter Pro Plus. Ex-situ water quality parameters such as total ammonium nitrogen (mg/l), nitrate (mg/l) and phosphate (mg/l) and total suspended solid (mg/l), were analysed every 15 days interval using standard analysis technique (Parson et al., 1984). Collection of data Measurement of each spiny lobster started from initial day of stocking and at 15-day interval. Body weight (g) and carapace length (mm) of spiny lobster was measured. Specific growth rate (SGR) and survival were calculated according to the following formula (Tuan, 2012): SGR (% day -1 ) = (ln W e ln W s ) n 100 W e = Final weight of targeted species W s = Initial weight of targeted species n = Period of culture (days) Survival (%) = x 100 Statistical analysis Data pertaining to weight gain, specific growth rate (SGR), survival and all the water quality parameters were analyzed using T-test in SPSS software version 21.0 in accordance with the design of the experiment. Results Growth of spiny lobster was observed uniform in RS and FTS during first two weeks. Growth gradually increased in FTS (Figure 3). Finally, at the end of the experiment the average weight of 164.731 g was observed in FTS, which was not significantly different (P>0.05) from the average weight of 161.04 g obtained in RS.

Sumbing et al., 2016. Transactions on Science and Technology. 3(1-2), 143-149 146 Survival (%) Figure 3. Mean body weight of Panulirus ornatus lobster in recirculating system and flow-through system Consequently, the specific growth rate of 0.125 % day -1 of spiny lobster in FTS was not significantly different (P>0.05) than the specific growth rate of 0.096 % day -1 in RS (Figure 4) Specific growth rate (% day - 1 ) 0.25 0.2 0.15 0.1 0.05 0 2 4 6 8 10 Cultivation period (weeks) RS FTS Figure 4. Specific growth rate of spiny lobster in recirculating system and flow-through system within 10 weeks cultivation. The percentage survival of 93.3 % and 80.0 % was observed in FTS and RS respectively (Figure 5), which was not significantly different (P>0.05). 100 90 RS 80 70 0 5 10 Cultivation period (weeks) Figure 5. Survival rate of spiny lobster in recirculating system and flow-through system

Sumbing et al., 2016. Transactions on Science and Technology. 3(1-2), 143-149 147 Range of water quality parameters in the spiny lobster tank from both FTS and RS were shown in Table 1 and 2. Little variations were observed in In-situ water quality parameters (Table 1). Table 1. Range of In-situ water quality parameters recorded in spiny lobster tank during the experiment period Systems DO (mg/l) ph Temperature ( C) Salinity (psu) Recirculating 5.25 7.54 8.05 8.27 28.00 30.70 31.12 32.92 Flow-through 5.12 7.32 7.92 8.15 30.00 31.20 30.60 31.42 The T- test analysis showed that the concentrations of NH 3 + NH 4, NO 2 -, NO 3 - and PO 4 - in both the culture systems were significantly different (P<0.05). RS recorded the highest concentration of NH 3 + NH 4, NO 2 -, NO 3 - and PO 4 - during the experimental period. Table 2: Mean (± SD) concentration of NH 3 + NH 4, NO 2 -, NO 3 - and PO 4 - in the two culture systems during 10 weeks experiment Systems Ammonia (mg/l) Nitrite (mg/l) Nitrate (mg/l) Phosphate (mg/l) Recirculating 0.401 ± 0.209 3.639 ± 1.499 2.836 ± 1.220 0.089 ± 0.331 Flow-through 0.120 ± 0.050 0.201 ± 0.037 0.347 ± 0.112 0.024 ± 0.005 Discussion Growth performance and specific growth rate of spiny lobster and survival rate were better in FTS compared to RS. However, there were no significant differences in both growth and survival between the two systems. Although FTS appeared to provide better growth performance compared to RS, P. ornatus appears to have advantages in aquaculture with regard to its tolerance to environmental fluctuations (Dennis et al., 1997). Appropriate husbandry, type of rearing system and feeding are the required parameters in the culture of spiny lobster (Tuan, 2012). Mortality was relatively high in RS and this could be due to cannibalism that occurs just after the lobster molting. The requirement for type of shelter required by juvenile lobsters may be different and change with age (Kittaka and Booth, 1994). The provision of shelter can minimize the interaction during molting. Observations made on the effect of stocking density and shelter in rearing system of spiny lobster were earlier carried out by Tuan (2012). The off-bottom shelter (hanging plastic table) setting resulted in significant differences in survival that was 86.2 % compared to on-bottom shelter (plastic pipes) in which survival of 76.7%, during 60 days of the experiment. The type of shelter does not affect the specific growth rate of spiny lobster which was 0.48 % day -1 and 0.47 % day -1 using shelter of hanging plastic table and plastic pipes respectively. This was substantially higher than that observed in the present study. In-situ water quality problems were not observed during this experiment. Lobsters are poikilotherm (cold blooded) animals and their growth rate is directly correlated with the temperature of the environment (Hartnoll, 2001). Jones and Shanks (2009) conducted an experiment on juvenile P.

Sumbing et al., 2016. Transactions on Science and Technology. 3(1-2), 143-149 148 ornatus where specimens were grown in tanks at five different temperatures (19, 22, 25 and 28 and 31 C). Growth was found to be significantly affected by temperature (P<0.01), and maximal growth occurred at 25 31 C (Jones and Shanks, 2009). In the present study, temperatures recorded in RS (28.00 30.70 C) and FTS (30.00 31.20 C) were in range of 25 31 C. The concentration of NH 3 + NH - 4, NO - - - 2, NO 3 and PO 4 were lower in flow-through system compared to recirculating system. Flow-through system received continuous seawater supply through the tank and effluents drain out to the environment after it is filtered by seaweed. A study has been conducted for spiny lobster grown in shrimp pond water in northern Australia and the data were compared with the tank system with filtered sea water. The spiny lobster was noticed to be tolerant of turbid water with elevated levels of suspended solids and nutrient loading including high productivity of both phytoplankton and zooplankton species (Jones and Shanks, 2008). In a recirculation system, high level of ammonia may be acceptable to the spiny lobster (0.401 ± 0.209 mg/l). Ammonia levels more than 10 mg/l have been shown to inhibit growth in crustaceans (Chen and Lin, 1992). In this system, the water is recirculating, thus allowing the operation of seaweed as a biofilter for removal of dissolved inorganic nutrients such as ammonia that are harmful to the cultured species. Seaweed can function as a biological filter by virtue of its ability to absorb and utilize the nitrogenous during photosynthesis which can minimize the water consumption and pollution impact on the environment (Chopin et al., 2001). Recirculating IMTA system allows limitation of water consumption in landbased aquaculture. Conclusion Growth performance of P.ornatus was not significantly different in flow-through system and recirculating system, but survival was higher in FTS, suggesting the preference of the lobster for flowing water or favorable water quality parameters. The concentrations of total ammonium nitrogen, nitrite, nitrate and phosphate in FTS were significantly lower than the water quality parameters observed in RS. With further investigations it will be possible to design better systems, including the types of species and their biomass, for still better yield. Acknowledgements We are grateful for the financial support provided by Ministry of Education Malaysia through Niche Research Grant Scheme (NRGS 0001). References [1] Chen, J. C & Lin, C. Y. (1992). Effect of ammonia on growth of Penaeus penicillatus juveniles. Comparative Biochemistry and Physiology Part C: Comparative Pharmacology, 101(3), 443-447. [2] Chopin, T., Buschmann, A. H., Halling, C. Troell, M., Kautsky, N., Neori, A., Kraemer, G. P., Zertuche- Gonzalez, J. A., Yarish, C. & Neefus, C. (2001). Integrating seaweeds into marine aquaculture systems: a key towards sustainability. Journal of Phycology, 37, 975-986 [3] Clive, M. J. (2010). Tropical spiny lobster aquaculture development in Vietnam, Indonesia and Australia. Journal of the Marine Biological Association of India, 52(2), 304 315.

Sumbing et al., 2016. Transactions on Science and Technology. 3(1-2), 143-149 149 [4] Dennis, D. M., Skewes, T. & Pitcher, C. R. (1997). Habitat use and growth of juvenile ornaterock lobsters, Panulirus ornatus (Fabricus, 1798), in Torres Strait, Australia. Marine and Freshwater Research, 48(8), 663-670. [5] Hartnoll, R. G. (2001). Growth in Crustacea - twenty years on. Hydrobiologia, 449, 111 122. [6] Jones, C. & Shanks, S. (2008). Grow-out of tropical rock lobster in shrimp pond conditions in Australia. The lobster Newsletter, 21(2), 18-21. [7] Jones, C. & Shanks, S. (2009). Requirement for the aquaculture for Panulirus ornatus in Australia. Proceedings of an International Symposium on Spiny Lobster Aquaculture in the Asia-Pacific Region. Nha Trang, Vietnam. 9-10 December 2008. [8] Kittaka, J. & Booth, J. D. (1994). Prospectus for aquaculture. In: Phillips, B. F., Cobb, J. S. and Kittaka, J. (Eds). Spiny lobster management. Oxford: Blackwell Scientific Publication. [9] Neori, A., Shpigel, M. & Ben-Ezra, D. (2000). A sustainable integrated system for culture of fish, seaweed and abalone. Aquaculture, 186(3-4), 279-291. [10] Neori, A., Chopin, T., Troell, M., Buschmann, A., Kraemer, G., Halling. C., Shipigel. M & Yarish, C. (2004). Integrated aquaculture: rationale, evolution and state of the art emphasizing seaweed biofiltration in modern mariculture. Aquaculture, 231(1-4), 361 391. [11] Parsons, T. R., Maita, Y. & Lalli, C. M. (1984). A Manual of Chemical and Biological Methods for Seawater Analysis. Oxford: Pergamon Press. [12] Biusing, R. E. & Lin, C. F. (2004). Status of Spiny lobster resources in Sabah, Malaysia. Seminar Paper, Department of Fisheries Sabah (http://www.fishdept.sabah.gov.my/?q=en/download/74). Accessed on April 29, 2016. [13] Tuan, L. A. (2012). Spiny lobster (Panulirus ornatus) reared in concrete tanks using lab made diet: Effect of stocking density and shelter settings. Proceedings of the International Fisheries Symposium (IFS 2012). 6-8 December, 2012. Can Tho City, Viet Nam. pp 3-9. [14] Vinh, L. T. & Huong, L. L. (2008). Improving environmental quality for Panulirus ornatus lobster aquaculture in Van Phong Bay, Vietnam, by combined culture with Perna viridis mussels. International Symposium on Spiny Lobster Aquaculture in the Asia-Pacific Region. 9-10 December 2008, Nha Trang, Vietnam.