Blue Economy: A Road Map for the Future Planet New Millennium Graphics publication Copyright © Techno India University, West Bengal Copyright of individual chapters remains with the authors Blue Economy: A Road Map for the Future Planet ISBN: 978-81-951712-5-5 First Edition: March, 2021 Latest Edition Published by: New Millennium Graphics Kolaghat, Purba Medinipur, Pin – 721 134, W.B Article ** Saltmarsh Grass Enriched Diet for Tiger Shrimp Penaeus Monodon Pavel Biswas1, Sufia Zaman1, Prosenjit Pramanick1 and Abhijit Mitra2 Department of Oceanography, Techno India University West Bengal, Salt Lake Campus, Kolkata 700091, India 2Department of Marine Science, University of Calcutta, 35 B.C Road, Kolkata 700019, India Abstract The salt-marsh grass Porteresia coarctata was used in formulated diet as a major source of protein and astaxanthin for black tiger shrimp Penaeus monodon Significant differences in water quality (nitrate, dissolved oxygen and organic carbon) of the experimental pond and growth performance of the shrimps fed with P coarctata diet in terms of survival rate, condition index, weight and feed conversion ratio were observed (confirmed through ANOVA) The protein and astaxanthin level increased considerably in the cultured shrimps which may have a potential for aquaculture industry in future Introduction In recent times, aquaculture nutritionists are showing interests in developing technologies to induce shrimp pigmentation by feeding supplementary diets enriched with carotenoids (specially astaxanthin) either synthetically or naturally (Palafox et al.,, 2006, Boonyaratpalin et al., 2001, Cruz-Suarez et al., 2008b, Gocer et al., 2006, Latscha, 1989, Pan et al., 2001, Bjerkeng, 2008) Astaxanthin is naturally sourced from Phaffia rhodozyma, Dunaliella salina, Haematococcus pluvialis, Spirulina, Chnoospora minima, Isochrysis galbana, alalfa, Tagetes erecta, Capsicum annuum (Palafox et al., 2006, Vernon-Carter et al.,, 1996, Meyers and Latscha, 1997) Mangroves and coastal vegetations have also been documented as a rich source of protein (Mitra and Banerjee, 2006) and astaxanthin (Banerjee et al., 2009) Porteresia coarctata, a salt marsh grass is widely distributed in the mudflats of lower Gangetic delta region We tried an experiment to prepare shrimp 172 feed using this species with an aim to upgrade the ecological health of the culture pond as well as to observe the biological performance of the species Materials & Methods Selection of station Two culture ponds (control and experimental) were selected in Jharkhali Island of Indian Sundarbans owing to its vicinity to Matla river to initiate shrimp culture during January to April, 2016 Stocking and culture Seeds of Penaeus monodon purchased from hatchery were stocked in both the culture ponds on the same day and time The traditional feed was provided to the control pond and the specially formulated feed prepared from Porteresia was provided to the experimental pond in order to monitor the efficacy of two different feed types Analysis of shrimp biochemical parameters Astaxanthin in the shrimp tissue was estimated as per the standard spectrophotometric method (Schuep and Schierle 1995) and body pigmentation of the cultured shrimp was assessed (for each treatment) after boiling the shrimp for in water and comparing the orangered coloration with Roche SalmoFanTM color score The protein content of shrimp was estimated by Lowry’s method (Lowry et al., 1951) Analysis of shrimp biological parameters Sampling of shrimps was done at fortnightly interval during the 90 days culture period and the relevant biological variables were determined for each control and experimental ponds Condition Index was analyzed as per the expression: C.I = W/L3 x 100, where W = weight of the cultured species (in gm) and L = length of the cultured species (in cm) Percentage weight gain was documented by calculating the difference in weight from the average final weight with respect to the initial weight; weight gain = [(average individual final weight – average individual initial weight)/average individual initial weight] x 100 The survival rate was measured as percentage of the difference of stocking number and production volume (No.) at the end of the culture period Feed Conversion Ratio (FCR) is the weight of feed consumed per unit of body weight gain and was analyzed after the harvesting of 173 shrimps as per the expression: FCR = ∆f/∆b, where, ∆f = Change in feed biomass and ∆b = Change in body biomass of the cultured species Analysis of water quality parameters The surface water salinity was recorded by means of an optical refractometer (Atago, Japan).The dissolved oxygen (DO) was measured by means of a portable D.O meter The pH of surface water was recorded through a portable pH meter (Hanna, USA) The surface water temperature was recorded by using a digital thermometer A Secchi disc was used to measure the transparency of the water column Surface waters were analyzed for nutrient concentrations (nitrate, phosphate and silicate) following the standard spectrophotometric method (Strickland and Parsons, 1972) The phytopigment (chlorophyll a) concentration was analyzed by the method of Jeffrey and Humphrey (1975) Organic carbon content of pond bottom soil was estimated by the standard titration method (Walkey and Black, 1934) Statistical analysis ANOVA – Single factor was performed in order to find out the significant differences between the culture ponds in terms of all the water quality parameters as well as the biological performance and biochemical content of the culture species Results and Discussion The results of the experiment were highlighted in Figs 7.5.1(a) and (b) for biochemical parameters Astaxanthin concentration is reflected through the development of muscle color of the cultured species It has been reported that dietary carotenoids were responsible for less than 20% of flesh pigmentation in aquatic organisms (Torrisen et al., 1989, Storebakken and No, 1992) Astaxanthin content depends upon the dietary source of pigment, dosage level, duration of feeding, dietary composition and degree of carotenoid esterification (Meyers and Latscha, 1997, Bjerkeng, 2000, Buttle et al., 2001, Gomes et al., 2002, White et al., 2002) The present study showed a significant difference in astaxanthin content between the culture ponds (p < 0.01) as shown in Fig 7.5.1(a) Our results are in line with those observed in farmed P monodon (Latscha, 1989) 174 Astaxanthin (ppm) 18 16 14 12 10 C E Jan Feb M ar Fig.7.5.1(a): Monthly variations in astaxanthin content of cultured shrimps from control (C) and experimental (E) ponds 30 Protein (%) 25 20 C 15 E 10 Jan Feb Mar Fig 7.5.1(b): Monthly variations in protein content of cultured shrimps from control (C) and experimental (E) ponds Dietary protein and energy levels are known to influence the growth and body composition of fish (Lovell, 1989) Improper protein and energy levels in feed increases fish production cost and deteriorates water quality Insufficient energy in diets causes protein waste due to 175 the increase proportion of dietary protein used for energy and the produced ammonia can reduce the water quality (Phillips, 1972; Prather and Lovell, 1973; Shyong et al., 1998) Protein is the major growth promoting factor in feed The protein requirement of fish are influenced by various factors such as fish size, water temperature, feeding rate, availability and quality of natural foods and overall digestible energy content of diet (Satoh, 2000; Wilson, 2000) In the present study, protein level in shrimps was found to be higher in experimental pond in comparison to control pond fed with traditional feed (p < 0.01) Condition Index is a unique indicator of shrimp health The values were higher in experimental group than the control group as shown in Fig 7.5.2 (a) ANOVA results show a significant difference between the culture ponds (p < 0.01) This may be attributed to the growth in biomass of the species throughout the culture period Penaflorida and Golez (1996) also observed better condition in P monodon fed diet including 50 g/kg Kappaphycus alvarezii meal But in comparison to adult, P monodon post-larvae exhibited better growth in biomass by feeding diet supplemented with astaxanthin (Thongrod et al., 1995) However in another experiment no significant difference in condition index was observed by supplementing a diet containing natural astaxanthin (Boonyaratpalin et al., 2001, Gocer et al., 2006) 4.5 Condition Index 3.5 2.5 C E 1.5 0.5 Jan Feb Mar Fig 7.5.2 (a): Monthly variations in Condition Index of cultured shrimps from control (C) and experimental (E) ponds 176 Average final weight was higher in shrimps of experimental pond than the control group as shown in Figure 7.5.2 (b) and this difference was significant (p < 0.01) This may be due to better palatability and acceptability of Porteresia based feed by the culture species Similar results were obtained in P.monodon fed seaweed based feed (Penaflorida and Golez, 1996) 30 Weight (gms) 25 20 C 15 E 10 Jan Feb Mar Fig 7.5.2 (b): Monthly variations in weight of cultured shrimps from control (C) and experimental (E) ponds Colouration of shrimps depends on the dietary astaxanthin content In the present study colour score was better in experimental group than the control group (Fig 7.5.2 c) A significant difference in colour score was obtained between the culture ponds (p < 0.01) Similar results were obtained by feeding alfalfa meal to P monodon (Brun and Vidal, 2006) The effects of astaxanthin on survival rate of aquatic organisms have been controversial However, dietary phytobiotic component which may serve as source of pigment plays a vital role in the physiology of crustaceans (Harpaz and Schmalbach, 1986) In the present study survival rate was higher in experimental group as compared to the control group (Fig, 7.5.2 d) A significant difference was observed between the culture ponds (p < 0.01) This may be attributed to the physiological role of astaxanthin in combating the physical and 177 environmental stress conditions Another reason may be the effect of stocking density having a significant relation with the survival rate Our results tally with the works done by other researchers where dietary astaxanthin resulted in higher survival rate (Gocer et al., 2006, Chien and Jeng, 1992) 35 Color Score 30 25 20 C 15 E 10 Jan Feb Mar Fig 7.5.2 (c): Monthly variations in Roche Salmo Fancolor score of cultured shrimps from control (C) and experimental (E) ponds 58.40% C E 69.20% Fig 7.5.2 (d): Survival rate (%) of cultured shrimps from control (C) and experimental (E) ponds 178 FCR Feed Conversion Ratio (FCR) is the ratio between total feed biomass to that of the total species biomass In the present study, FCR was lower in experimental group as compared to the control group (Fig 7.5.2 e) indicating a better condition of the cultured species in the experimental pond The possible reason may be attributed to the minimum wastage of Porteresia based feed by the culture species It has been reported that seaweed based diet are also efficient in lowering of FCR (Penaflorida and Golez, 1996) In another experiment, a significant lowering of FCR was observed by adding Ulva in the diet of P monodon (Cruz-Suarez et al., 2008b) Da Silva and Barbosa (2008) also reported a better feed conversion with the inclusion of Hypnea cervicornis and Cryptonemia crenulata meals Significant statistical differences exist between the culture ponds (p < 0.01) 1.8 1.6 1.4 1.2 0.8 0.6 0.4 0.2 C E Jan Feb Mar Fig 7.5.2 (e): Monthly variations in FCR of cultured shrimps from control (C) and experimental (E) ponds In context to the water quality parameters, no significant differences were observed between the ponds except dissolved oxygen, nitrate and organic carbon load (Figs 7.5.3 a-j) This reveals the acceptance of Porteresia based feed by the culture species The significant variation in DO may be attributed to feed quality Traditional feed contains fish meal and trash fish dust which lowers the DO due to its utilization for oxidizing the residual matter Porteresia based feed on 179 Surface water temparature (C) the other hand generates very limited residue due to which DO remains almost unaltered (p < 0.01) The variation in nitrate may be due absence of leaching of feed ingredients (particularly the animal component of traditional feed) in experimental pond water and also the faecal matter that generates ammonia (Mitra and Choudhury, 1995) Similarly the excreta and feed residue are the major contributors to organic load in the pond bottom which caused such variation o 30.4 30.2 30 29.8 29.6 29.4 29.2 29 28.8 28.6 C E Jan Feb Mar Surface water salinity (psu) Fig 7.5.3 (a): Monthly variations in surface water temperature of control (C) and experimental (E) ponds at Jharkhali Island 30 25 20 C 15 E 10 Jan Feb Mar Fig 7.5.3 (b): Monthly variations in surface water salinity of control (C) and experimental (E) ponds at Jharkhali Island 180 8.4 8.2 pH 7.8 C 7.6 E 7.4 7.2 Jan Feb Mar Fig 7.5.3 (c): Monthly variations in surface water pH of control (C) and experimental (E) ponds at Jharkhali Island Transparency (cm) 30 25 20 C 15 E 10 Jan Feb Mar Fig 7.5.3 (d): Monthly variations in surface water transparency of control (C) and experimental (E) ponds at Jharkhali Island 181 -1 D.O (mg L ) C E Jan Feb Mar Fig 7.5.3 (e): Monthly variations in dissolved oxygen of control (C) and experimental (E) ponds at Jharkhali Island 25 -1 Nitrate (ug-at L ) 30 20 C 15 E 10 Jan Feb Mar Fig 7.5.3 (f): Monthly variations in nitrate concentration of control (C) and experimental (E) ponds at Jharkhali Island 182 -1 Phosphate (ug-at L ) 3.5 2.5 C 1.5 E 0.5 Jan Feb Mar -1 silicate (ug-at L ) Fig 7.5.3 (g): Monthly variations in phosphate concentration of control (C) and experimental (E) ponds at Jharkhali Island 90 80 70 60 50 40 30 20 10 C E Jan Feb Mar Fig 7.5.3 (h): Monthly variations in silicate concentration of control (C) and experimental (E) ponds at Jharkhali Island 183 -3 Chl a (mg m ) 2.5 C 1.5 E 0.5 Jan Feb Mar Fig 7.5.3 (i): Monthly variations in Chl a concentration of control (C) and experimental (E) ponds at Jharkhali Island Organic carbon (%) 1.5 C E 0.5 Jan Feb Mar Fig 7.5.3 (j): Monthly variations in organic carbon (%) of control (C) and experimental (E) ponds at Jharkhali Island 184 Conclusion In aquaculture industry, Penaeus monodon is regarded as a species of high consumer appeal The present study is a pathfinder for the unexploited salt-marsh grass vegetation of Indian Sundarbans as a unique reservoir of natural astaxanthin and protein The effect of adding Porteresia to the diets not only improved the growth performance of shrimps but also upgraded the ecological health of the pond This development is of considerable importance for commercial aquaculture practices as good results can be achieved by adding cheap source of protein and astaxanthin rather than expensive ingredients which usually enhances the feed cost However to keep the present venture sustainable an additional back-up nursery is suggested as part of conservation of this marshy vegetation 185 Suggested References Banerjee, K., Ray, D., Basu, S., Chakraborty, B and Mitra A 2009 A comparative study of astaxanthin level in mangrove species Proceedings of the National Academy of Sciences, Sect B, Vol 79 Pt II, 135-142 Bjerkeng, B 2008 Carotenoids in aquaculture: Fish and Crustaceans In Birkhauser Verlag Basel Ed Carotenoids 4, Natural Functions 237-254 Bjerkeng, B 2000 Carotenoid pigmentation in salmonid fishes – recent progress In: Avances en Nutricion Acuicola V – Memorias del Quinto Simposium Internacional de Nutricion Acuícola, Merida, Mexico, 19-22 Noviembre 2000 (Eds: Cruz-Suarez, L.E., RicqueMarie, D., Tapia-Salazar, M., Olvera-Novoa, M.A and CerecedoOlvera, R.), 5: 71-89 Universidad Autonoma de Nuevo Leon, 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Abstract The salt-marsh grass Porteresia coarctata was used in formulated diet as a major source of protein and astaxanthin for black tiger shrimp Penaeus monodon Significant differences in water quality... the shrimp for in water and comparing the orangered coloration with Roche SalmoFanTM color score The protein content of shrimp was estimated by Lowry’s method (Lowry et al., 1951) Analysis of shrimp. .. dissolved oxygen and organic carbon) of the experimental pond and growth performance of the shrimps fed with P coarctata diet in terms of survival rate, condition index, weight and feed conversion