ReseaRch PaPeR Journal of Agricultural and Marine Sciences Vol. 20 (2015): 62-65. Reveived 5 Aug. 2014 Accepted 19 Feb 2014 Improving the quality of tiger shrimp Penaeus monodon through dietary incorporation of algae as a source of natural pigment Kunal Mondal*, Subhra Bikash Bhattacharyya and Abhijit Mitra * Department of Marine Science, University of Calcutta, 35 B.C. Road, Kolkata - 700019, India . Kunal Mondal ( ) email: bottu.marine@ gmail.com. Introduction Inclusion of algae as dietary supplement in animals have been investigated previously as a source of pigment (Strand et al., 1998). The effects of dietary inclusion of algae have resulted in improved perfor- mance including better animal product quality (Moss, 1994; Penaflorida and Golez, 1996; Cruz-Suarez et al., 2000). To be more specific the commercial production of shrimps and prawns as an edible food product rep- resents one of the fastest growing areas of aquaculture (Rosenberry, 2005) with high consumer appeal and at- tractive market for shrimps due to their body coloura- tion or pigmentation which is a direct measure of its astaxanthin content. Carotenoid utilization by aquacul- ture species is well documented as it plays a regulatory role in providing antioxidant and pro-vitamin A activity, enhancing immune response, improving reproductive performance, growth, maturation and photo-protection (Howell and Matthews, 1991). They also help the spe- cies to resist environmental stress (Meyers, 1994). Algae and higher groups of plants are the major producers of carotenoids which comprises a family of over 600 nat- ural fat soluble pigments (Britton et al., 1981). Studies reveal that alternative utilization of plant pigments in formulated diets have improved the body pigmentation حتسني نوعية روبيان النمر )Penaeus monodon( إبستخدام الطحلب كمصدر طبيعي للصبغة يف الغذاء كونال موندال* وسوفرا بيكاش باتاشاريا وأهبيجيت ميرتا Abstract. Tiger shrimp is one of the major candidate species for export oriented aquaculture which dominates the seafood market in regions of European Union, Japan and USA. Carotenoid content in seafood has now become one of the important criteria in determining the quality of edible product. Recent trends in supplementing fish diets with natural pigment source are an alternative to the utilization of expensive synthetic pigments. In this context, green algae Enteromorpha intestinalis was selected as a source of natural pigment for inclusion in the diet of tiger shrimp Penaeus monodon. Astaxanthin being an important category of carotenoid pigment was monitored in shrimp muscle tissue during the feeding trial. Significant variation (p<0.05) was observed between the experimental groups as confirmed through ANOVA thus exhibiting higher astaxanthin content of shrimps (18.70 ± 4.48 ppm) fed with E. intestinalis incorporated diet as compared to control (15.80 ± 2.33 ppm). The present programme therefore emphasizes on the quality improvement of aquaculture product by dietary inclusion of algae as a natural pigment source. Keywords: Shrimp; aquaculture, feed, algae, pigment. املســتخلص: يعتــر روبيــان النمــر أحــد أهــم األنــواع املســتزرعة للتصديــر الــي هتيمــن علــى ســوق املأكــوالت البحريــة يف مناطــق االحتــاد األورويب واليابــان والواليــات املتحــدة األمريكيــة، وحاليــاً يعتــر حمتــوى الكاروتــن )Carotenoid( املوجــود يف املأكــوالت البحريــة أحــد أهــم املعايــر املســتخدمة يف حتديــد جــودة املنتجــات الصاحلــة لالســتهالك. ويتــم التوجــه حاليــاً الســتخدام الصبغــات الطبيعيــة كبديــل للصبغــات االصطناعيــة عاليــة التكلفــة املســتخدمة يف الغــذاء الســمكي، ويف هــذا الســياق مت اختيــار الطحلــب األخضــر )Enteromorpha intestinalis( كمصــدر طبيعــي للصبغــة إلدراجهــا يف الغــذاء املســتخدم لروبيــان النمــر )Penaeus monodon(. متــت متابعــة نســبة األستاكســانثن )Astaxanthin( والــذي يعتــر أحــد اهــم فئــات الكاريتينويــد )Carotenoid( يف األنســجة العضليــة للروبيــان خــالل جتــارب التغذيــة، وقــد لوحــظ وجــود اختــالف هــام )p >0.05( بــن اجملموعــات التجريبيــة كمــا أثبتــه اختبــار حتليــل التبايــن )ANOVA( والــذي يظهــر وجــود حمتويــات أعلــى لـــ Astaxanthin )18.70 ± 4.48 جــزء يف املليــون( يف الروبيــان الــذي تغــذى علــى وجبــات حتتــوي علــى E. intestinalis مقارنــة مــع الروبيــان املتحكــم يف غذائــه )15.80 ± 2.33 جــزء يف املليــون(، وبالتــايل فــإن الرنامــج الــذي مت اســتخدامه يثبــت حتســن جــودة منتجــات الكائنــات البحريــة املســتزرعة مــن خــالل إدراج الطحالــب كمصــدر طبيعــي للصبغــة يف غذائهــا. الكلمات املفتاحية: الربيان، االستزراع السمكي، أعالف األحياء املائية، الطحالب، الصبغة 63Research Article Mondal, Bhattacharyya, Mitra of farmed crustaceans, particularly penaeids in order to achieve better market price (Lorenz, 1998; Liao and Chien, 1994). Therefore the present work is an attempt to utilize Enteromorpha intestinalis as a natural dietary pigment source for farmed tiger shrimp (Penaeus monodon) in relation to its quality improvement. Materials and methods Collection of algae and preparation of experi- mental diets Live and healthy algae Enteromorpha intestinalis was collected from Bali Island (22º 04´ 35.17˝ N latitude and 88º 44´ 55.70˝ E longitude) of Indian Sundarbans during low tide. The collected material was rinsed in ambient water and then with distilled water, oven-dried at 50ºC and finally processed to make powder. Experimental diet was formulated through incorporation of algae (DietENT) at a level of 5%. Simultaneously a control diet (DietC) was also formulated to study the comparative performance (Table 1). Feeding trial A feeding trial was run at Bali Island (22º04´ 35.17˝N latitude and 88º 44´ 55.70˝ E longitude) of Sundarbans in grow-out ponds for 90 days of experimental duration (Fig. 1). Shrimp juveniles were procured from hatchery and stocked in experimental ponds at a density of 2 indi- viduals/m2. Experimental diets were randomly assigned, the culture species were fed twice daily and the uneaten feed was checked at regular intervals. Astaxanthin analysis The astaxanthin content in shrimp muscle tissue was analyzed according to the spectrophotometric method outlined by Schuep and Schierle (1995). Its value in % was converted to ppm level for easy interpretation of data. The body colouration of shrimps after boiling was compared by Roche SalmoFanTM colour score. Statistical analysis The collected data were finally subjected to one-way analysis of variance (ANOVA). All statistical calcula- tions were performed with SPSS 9.0 for Windows. Results and discussion The average astaxanthin content in muscle tissue was higher in shrimps fed with DietENT as compared to DietC (Table 2). A darker orange-red colouration was observed in shrimps fed with DietENT after boil- ing them in water for 5 minutes when compared with Roche SalmoFanTM colour score. The colour score was 30 for DietENT fed shrimps whereas a score of 27 was recorded from shrimps fed with DietC. ANOVA results showed significant variation (p<0.05) in average astax- anthin content which may be attributed to the capabil- ity of P. monodon to easily convert the fraction of algal astaxanthin into tissue astaxanthin. The Enteromorpha Figure 1. Map showing the P. monodon farming site at Bali Island of Indian Sundarbans. Table 1. Formulation of experimental diets. Ingredients Diet Control Diet Enteromorpha Fish meal 35 30 Soybean oil cake 11 11 Mustard oil cake 11 11 Rice polish 23 23 Wheat flour 16 16 oyster shell dust 2 2 Shark oil 2 2 Enteromorpha intestinalis (source of astaxanthin) 0 5 Table 2. Variations in astaxanthin content of P. monodon (ppm muscle tissue) fed with experimental diets. Days of culture (DOC) Diet Control Diet Enteromorpha 0 12.83 ± 0.22 13.02 ± 0.27 30 15.21 ± 0.22 17.62 ± 0.55 60 16.96 ± 0.53 20.71 ± 0.76 90 18.22 ± 1.23 23.47 ± 1.31 Average 15.80 ± 2.33b 18.70 ± 4.48a *Means with different letters(a,b) in a row differ significantly (p<0.05); values are means of three replicates 64 SQU Journal of Agricultural and Marine Sciences, 2015, Volume 19, Issue 1 Improving shrimp quality through dietary incorporation of natural algal astaxanthin sample selected for the investigation is found to contain 120.78 ppm astaxanthin as reported from the present study region (Mitra et al., 2013; Banerjee et al., 2009; Chakraborty and Santra, 2008). Carotenoid, particularly astaxanthin content of feed is one of the major factors influencing the colour development in animals (Moretti et al., 2006) but at the same time scientific knowledge about several factors like dietary pigment source, their dosage level, feeding duration, dietary composition and magnitude of carotenoid esterification is also required to identify these interaction processes (Meyers and Lats- cha, 1997; Bjerkeng, 2000; Buttle et al., 2001; Gomes et al., 2002; White et al., 2002). The present study showed significantly different as- taxanthin content of the farmed shrimp which are in agreement to the observations that crustaceans exhib- it strong tendency towards selection of specific carot- enoids at a specific rate for their metabolic absorption (Meyers and Latscha, 1997). Similar work conducted from the present study region reveal that P. monodon when fed with diet containing red algae Catenella repens at a level of 5% improved the body astaxanthin content (Banerjee et al., 2010). The search for natural astaxan- thin was not only limited to the algal resources, rather salt-marsh grass Porteresia coarctata was also tested as a natural dietary astaxanthin source in P. monodon feed with better results from the present geographical locale (Mitra et al., 2011, 2013). In continuation such natural carotenoid supply to the diet of shrimps has been stud- ied for P. japonicus and Litopenaeus vannamei too from different parts of the globe. The ingredients of natural origin that have been used in the diet are red yeast (Phaf- fia rhodozyma) and microalgae Dunaliella salina (Chien and Jeng, 1992); Chnoospora minima (Menasveta et al., 1993); Spirulina sp. (Liao et al., 1993; Chien and Shiau, 1998); Haematococcus pluvialis (Chien and Shiau, 1998) and Isochrysis galbana (Pan et al., 2001). An usual trend of marked increase in the body carotenoid content has been observed when organisms were fed with plant pig- ment source diets. For example, feed supplemented with 50 ppm algal material (Dunaliella salina) improved the body colouration of P. monodon (Boonyaratpalin et al., 2001). Three types of diet when provided to P. semisulca- tus containing natural carotenoid sources like red pep- per and marigold flower resulted in higher carotenoid accumulation in body tissues (Gocer et al., 2006). How- ever research findings from Mexico also reported that feed incorporated with cultivated green alga Ulva clath- rata significantly improved the body pigmentation of farmed shrimp L. vannamei (Cruz-Suarez et al., 2009). 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