Squids and their by-products are widely used as pre-spawning feeds in marine fish hatcheries. Therefore, we studied the fatty acid composition of mantle tissue, arms, fins, and gonads of the marine cephalopod, Loligo vulgaris, from a broodstock nutrition point of view. Docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids represented nearly half (>45%) of the total fatty acids, with DHA the most abundant fatty acid in all investigated tissues. The fatty acid profile of the gonads differed significantly from those of the mantle, arms, and fins; EPA was present in a sig- nificantly higher proportion (20%), perhaps indicating its important role in reproduction. Results indicate that Loligo vulgaris and its by-products can be an important nutritional component of broodstock feeds, serving as a major source of DHA and EPA

Ozkizilcik, Sureyya

Salman, Alp

Salman, Yesim

Abstract Squids and their by-products are widely used as pre-spawning feeds in marine fish hatcheries. Therefore, we studied the fatty acid composition of mantle tissue, arms, fins, and gonads of the marine cephalopod, Loligo vulgaris, from a broodstock nutrition point of view. Docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids represented nearly half (&gt;45%) of the total fatty acids, with DHA the most abundant fatty acid in all investigated tissues. The fatty acid profile of the gonads differed significantly from those of the mantle, arms, and fins; EPA was present in a significantly higher proportion (20%), perhaps indicating its important role in reproduction. Results indicate that Loligo vulgaris and its by-products can be an important nutritional component of broodstock feeds, serving as a major source of DHA and EPA

Yesim Salman

Alp Salman

Sureyya Ozkizilcik

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The Fatty Acid Profile of the Marine Cephalopod Loligo vulgaris

eVols at University of Hawaii at Manoa

The Open Access Israeli Journal of Aquaculture – BamidgehAs from January 2010 The Israeli Journal of Aquaculture - Bamidgeh (IJA) will be published  exclusively  as  an  on-line  Open  Access  (OA) quarterly  accessible  by  all AquacultureHub  (http://www.aquaculturehub.org)  members  and  registered  individuals and institutions.  Please visit our website (http://siamb.org.il) for free registration form, further information and instructions.  This transformation from a subscription printed version to an on-line OA journal, aims at supporting the concept that scientific peer-reviewed publications should be made available to all, including those with limited resources. The OA IJA does not enforce author or subscription fees and will endeavor to obtain alternative sources of income to support this policy for as long as possible.Editor-in-ChiefDan Mires Editorial BoardSheenan Harpaz Agricultural Research OrganizationBeit Dagan, IsraelZvi Yaron Dept. of ZoologyTel Aviv UniversityTel Aviv, IsraelAngelo Colorni National Center for Mariculture, IOLREilat, IsraelRina Chakrabarti Aqua Research LabDept. of ZoologyUniversity of DelhiIngrid Lupatsch Swansea UniversitySingleton Park, Swansea, UKJaap van Rijn The Hebrew University Faculty of AgricultureIsraelSpencer Malecha Dept. of Human Nutrition, Food and Animal SciencesUniversity of HawaiiDaniel Golani The Hebrew University of JerusalemJerusalem, IsraelEmilio Tibaldi Udine UniversityUdine, ItalyCopy Editor Ellen Rosenberg Published under auspices ofThe Society of Israeli Aquaculture and Marine Biotechnology (SIAMB), University of Hawaii at Manoa Library andUniversity of Hawaii Aquaculture Program in association withAquacultureHub http://www.aquaculturehub.org              ISSN 0792 - 156X Israeli Journal of Aquaculture - BAMIGDEH.PUBLISHER: Israeli Journal of Aquaculture - BAMIGDEH -Kibbutz Ein Hamifratz, Mobile Post 25210, ISRAELPhone: + 972 52 3965809http://siamb.org.il The Israeli Journal of Aquaculture – Bamidgeh  59(3), 2007, 133-136. 133The Fatty Acid Profile of the Marine Cephalopod Loligo vulgarisYesim Salman1, Alp Salman2* and Sureyya Ozkizilcik31 Department of Chemistry, Faculty of Science, Ege University, 35100 Bornova Izmir, Turkey2 Department of Hydrobiology, Faculty of Fisheries, Ege University, 35100 Bornova Izmir,Turkey3 Ust Mandalya Sitesi No. 32A, Bodrum Mugla, Turkey(Received 25.11.06, Accepted 28.1.07)Key words: Loligo vulgaris, squid fatty acid profileAbstractSquids and their by-products are widely used as pre-spawning feeds in marine fish hatcheries.Therefore, we studied the fatty acid composition of mantle tissue, arms, fins, and gonads of themarine cephalopod, Loligo vulgaris, from a broodstock nutrition point of view. Docosahexaenoic(DHA) and eicosapentaenoic (EPA) acids represented nearly half (>45%) of the total fatty acids,with DHA the most abundant fatty acid in all investigated tissues. The fatty acid profile of thegonads differed significantly from those of the mantle, arms, and fins; EPA was present in a sig-nificantly higher proportion (20%), perhaps indicating its important role in reproduction. Resultsindicate that Loligo vulgaris and its by-products can be an important nutritional component ofbroodstock feeds, serving as a major source of DHA and EPA.IntroductionMost cephalopod species are important ele-ments in the marine food chain (Caddy,1983). The marine cephalopod, Loligo vul-garis, is one of the most sought after seafoods in the Mediterranean Sea. In marine fishaquaculture, it is common to use cepa-holopods as broodstock feeds (Zohar et al.,1995). Most marine fish hatcheries feed squidto bass and bream broodstock during vitello-genesis and spawning. Although total bodylipids represent only a small percentage oftheir dry weight (Navarro and Villanueva,2000; Passi et al., 2002), saponifiable lipidscontain significant n-3 fatty acids (Ferrara etal., 2001). Docosahexaenoic acid (DHA) andeicosapentaenoic acid (EPA) are crucial nutri-ents for the survival and growth of eggs andlarvae of most marine fish (Sargent, 1995).Zohar et al. (1995) found that including squidoil in feeds for bream (Sparus aurata) brood-stock increased egg and larvae survival.During processing of squid for human con-sumption, the tentacles and mantles are usu-ally saved for packaging while the gonads and*  Corresponding author. E-mail: alp.salman@ege.edu.tr134internal organs are discarded. Most studiesconcerning the fatty acid profiles of squidinvolve mantle tissues (Sinanoglou andMeimaroglou, 1998; Okuzumi and Fujii, 2000;Ferrara et al., 2001; Phillips et al., 2002).There have been few studies of the fatty acidcompositions of cephalopods (Navarro andVillanueva, 2000, 2003) and seasonalchanges in their fatty acid profiles (Ozyurt etal, 2006). The purpose of this study was todetermine differences in fatty acids in severaltissues of L. vulgaris as an indication of thepotential use of squid by-products in marinefish aquaculture. Materials and MethodsCollection of samples. Four specimens of L.vulgaris were obtained from trawlers at the localfisheries dock in Izmir in May 2005 and trans-ferred to the laboratory on ice. Upon arrival, thespecimens were weighed and the mantle length(ML) was measured. Sex and gonadal develop-ment were determined according to Lipinski(1979). Three specimens were stage 4 males(15.5 and 17.4 mm ML, 96.8 and 101.5 g) andone was a stage 4 female with a developedgonad (18.0 mm ML, 186.7 g).Analytical methods. Mantle tissue, arms,fins, and gonads were preserved indichloromethane, then 1-2 g of each werehomogenized with a glass pestle. Similaramounts were placed on aluminum foil anddried at 65°C for approximately 48 h to deter-mine the moisture content. Lipid and fatty acid analyses. Total lipidswere extracted according to Bligh and Dyer(1959) using dichloromethane:methanol:water (2:2:1 v/v/v; DCM). Approximately 1-2 gof tissue was homogenized with a glass pes-tle in a DCM mixture in three replicates. DCMwas used since it is less toxic to humans thanchloroform in laboratory conditions. Extractedlipids were dried under a stream of nitrogen ina glass vial. Two ml of hexane containing 2%HCl was added and the vial was sealed with aflame. Trans-sterification was carried out inboiling water for 1 h. Fatty acid methyl esters(FAME) were extracted with hexane-waterpartitioning and analyzed using a FisonsInstruments gas chromatography equippedwith a PSS (Split-Splitless) injector and aflame ionization detector with a J&W DB-23(code 122-2332) column (30 m length, 0.25mm I.D.), coated with 0.25 µm film. The injec-tion split was 50 ml/min and helium was usedas the carrier gas (70 kPa). The flame ioniza-tion detection (FID) was set at 100 kPa air and50 kPa hydrogen pressures. The injector anddetector temperatures were set at 290°C and300°C, respectively. The temperature was setto start at 155°C (1 min) and increase at2°C/min until reaching the final 240°C.ResultsThe moisture contents of mantle tissue, arms,and fins were 76.1%, 76.2%, and 79.1%,respectively. The total fatty acids of the lipidsin all three tissues were dominated by PUFA.DHA and EPA constituted approximately 45%of the FAME in all tissues (Table 1). Saturatedfatty acids were mostly represented by 16:0that formed almost one third of all FAME.Stearic and myristic acids were present inmuch lesser percentages. Mantle tissue,arms, and fins were almost identical in termsof total fatty acids. Gonads, on the other hand,contained significantly higher levels of EPAand arachidonic acid (p≤0.05) and lower per-centages of palmitic and docosahexaenoicacid (p≤0.05).DiscussionThe total lipid content of many cephalopodshas been documented in detail (Okuzumi andFujii, 2000; Phillips et al., 2002; Passi et al.,2002). Total lipids in squids and cuttlefishvaries 1-2 % on a wet weight basis (Okuzumiand Fujii, 2000). Phospholipids are the maincomponents of lipid moieties of saponifiablelipids in several squid species (Passi et al.,2002), representing 62-84% of the total lipids(Okuzumi and Fujii, 2000). We found that thetotal fatty acids of the studied tissues aredominated by DHA and EPA. Although noother study compares the fatty acid composi-tions of different tissues, similar values werereported by Navarro and Villanueva (2000)and Passi et al. (2002) for L. vulgaris.In Sepia officinalis, Sinanoglou andMeimaroglou (1998) found that the total DHASalman et al.content of neutral and polar lipids is 30.69% ofthe FAME while Ozyurt et al. (2006) found thatDHA was 23.9-29.5% of the total lipidsdepending on the season. In our study, sam-ples were collected in the spring when DHAseems to reach its maximum. Seasonal varia-tions may influence fatty acid compositions.The lowest DHA level was found in thegonads, mostly of males in stage 4. Navarroand Villanueva (2003) found that total lipidscontained 29.3% DHA in L. vulgaris hatch-lings. In our study, DHA represented 26.4% ofthe FAME. Presumably, more DHA accumu-lates during the late gonad development.The EPA level ranged 14.2-20.3%. Passiet al. (2002) reported that EPA represented14.1% of neutral lipids and 21.8% of polarlipids in L. vulgaris. Our values coincide withthose reported by Navarro and Villanueva(2000) for L. vulgaris hatchlings. Palmitic acid ranged 24.6-30.5%. Althoughless desired by nutritionists, palmitic acid isoften abundant in marine lipids at approxi-mately 20% of the total fatty acids of cephalo-pod tissues (Passi et al., 2002; Phillips et al.,2002; Ozyurt et al., 2006).135Fatty acid profile of the squid, Loligo vulgarisFatty acid Mantle Arm Fin GonadC 14:0 MA 4.4±0.5 2.5±0.06 2.4±0.18 1.7±0.18C 15:0 0.60±0.02 0.6±0.06 0.6±0.03 0.4±0.01C 16:0 PA 28.3±0.62 27.7±0.87 30.5±3.55 24.6±0.16C 16:1 n7 0.9±0.08 0.6±0.13 0.6±0.07 0.4±0.09C 18:0 SA 4.8±1.32 6.1±0.36 5.8±0.16 5.8±0.11C 18:1 n9 4.4±1.36 2.1±0.49 2.3±0.28 1.8±0.01C 18:1 n7 1.6±0.2 1.7±0.27 1.6±0.32 1.5±0.33C 18:2 n6 0.2±0.02 0.3±0.13 0.2±0.09 0.3±0.04C 20:1 n9 2.0±0.04 2.3±0.14 1.8±0.20 3.5±0.21C 20:4 n6 AA 0.9±0.11 1.2±0.18 1.3±0.13 1.6±0.13C 20:5 n3 EPA 14.6±0.14 15.0±0.28 14.2±0.31 20.3±0.81C 22:1 n9 0.8±0.26 0.5±0.06 0.5±0.03 0.5±0.04C 22:5 n6 0.3±0.03 0.4±0.05 0.5±0.04 0.4±0.03C 22:5 n3 0.5±0.08 0.8±0.22 0.6±0.22 0.5±0.03C 22:6 n3 DHA 32.5±2.63 33.4±0.75 30.9±1.84 26.4±2.57C 24:1 n9 0.3±0.03 0.2±0.04 0.2±0.07 0.2±0.04∑ Saturated FA (%) 37.2±0.54 36.8±0.53 39.5±3.60 32.6±0.45∑ Unsaturated (%) 59.0±0.70 58.5±0.72 54.7±2.26 57.4±2.19Unidentified (%) 3.8±1.25 3.6±0.86 5.6±2.05 9.7±1.76∑ n3 FA (%) 47.7±2.34 49.6±0.73 46.3±2.06 47.2±1.88∑ n6 FA (%) 1.1±0.15 1.9±0.34 1.5±0.10 1.7±0.40∑ monoenes (%) 10.2±1.62 7.4±0.64 6.8±0.64 7.9±0.13Table 1. Fatty acid composition (%) of mantle, arms, fins, and gonad (n = 3).MA = myristic acid; PA = palmitic acid; SA = stearic acid; AA = arachidonic acid; EPA = eicos-apentaenoic acid; DHA = docosahexaenoic acid136From the broodstock nutrition point ofview, L. vulgaris can be an important sourceof DHA and EPA that are critical for egg andlarvae development of marine fish. Hence, theby-products of squid processing has importantmerit in aquaculture nutrition. ReferencesBligh E.G. and W.J. Dyer, 1959. A rapidmethod of total lipid extraction and purifica-tion. Can. J. Biochem. Physiol., 37:911-931.Caddy J.F. (ed.), 1983. The cephalopods:factors relevant to their population dynamicsand to the assessment and management ofstock. pp. 416-452. In: Advances inAssessment of World CephalopodResources. FAO Fish. Tech. Rep., no. 231.Ferrara P., Fabietti F., Delise M., BoccaA.P. and E. Funari, 2001. Alkylphenolic com-pounds in edible mollusks of the Adriatic Sea(Italy). Environ. Sci. Technol., 35:3109-3112.Lipinski M.R., 1979. Universal maturity scalefor the commercially-important squids(Cephalopoda: Teuthoidea): the results ofmaturity classification of the Illex illecebrosus(Le Sueur, 1821) population for the years1973-1977. Int. Commission North AtlanticFish., no. 5364:1-40.Navarro J.C. and R. Villanueva, 2000. Lipidand fatty acid composition of early stages ofcephalopods: an approach to their lipidrequirements. Aquaculture, 183:161-177.Navarro J.C. and R. Villanueva, 2003. Thefatty acid composition of Octopus vulgarisparalarvae reared with live and inert food:deviation from their natural fatty acid profile.Aquaculture, 219:613-631.Okuzumi M. and T. Fujii, 2000. 35thAnniversary Commemorative PublicationNutritional and Functional Properties of Squidand Cuttlefish. Natl. Cooperative Assoc.Squid Processors, Japan.Ozyurt G., Duysak O., Akamca E. and C.Tureli, 2006. Seasonal changes of fatty acidsof cuttlefish Sepia officinalis L. (Mollusca:Cephalopoda) in the north easternMediterranean Sea. Food Chem., 95:382-385.Passi S., Cataudella S., Di Marco P., DeSimone F. and L. Rastrelli, 2002. Fatty acidcomposition and antioxidant levels in muscletissue of different Mediterranean marinespecies of fish and shellfish. J. Agric. FoodChem., 50:7314-7322.Phillips K.L., Nichols P.D. and G.D.Jackson, 2002. Lipid and fatty acid composi-tion of the mantle and digestive gland of fourSouthern Ocean squid species: implication forfood web studies. Antarctic Sci., 14:212-220.Sargent J.R., 1995. Origins and functions ofegg lipids: nutritional implications. pp. 353-372. In: N. Bromage, R. Roberts (eds.).Broodstock Management and Egg and LarvalQuality. Blackwell Sci.Sinanoglou V.J. and S.M. Meimaroglou,1998. Fatty acid of neutral and polar lipids of(edible) Mediterranean cephalopods. FoodRes. Int., 31:467-473.Zohar Y., Harel M., Hassin S. and A.Tandler, 1995. Gilt-head sea bream (Sparusaurata). pp. 94-117. In: N. Bromage, R.Roberts (eds.). Broodstock Management andEgg and Larval Quality. Blackwell Sci.Salman et al.

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The Fatty Acid Profile of the Marine Cephalopod Loligo vulgaris

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