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International Journal of Agriculture S c i e n c e s ISSN: 0975-3710 & E-ISSN: 0975-9107, Volume 7, Issue 9, 2015, pp.-671-677. Available online at http://www.bioinfopublication.org/jouarchive.php?opt=&jouid=BPJ0000217

TILAPIA AS FOOD FISH: ENHANCEMENT OF Ω-3 POLYUNSATURATED FATTY ACIDS IN TILAPIA (Oreochromis spp.)

CHAVAN B.R.1*, YAKUPITIYAGE A.2, ATAGUBA G.A., KAMBLE M.T.2 AND MEDHE S.V.2 1Marine Biological Research Station (Dr. BSKKV), Ratnagiri, (MS), 2Aquaculture and Aquatic Resource Management(AARM), Food, Agriculture and Bio-system Cluster (FABS), School of Environment, Resources and Development (SERD), Asian Institute of Technology, P.O. Box 4, Klong Luang, Pathumthani-12120, Thailand. *Corresponding author e-mail: [email protected]

Received: September 21, 2015; Revised: October 20, 2015; Accepted: October 21, 2015

Abstract- A plethora of literature exist regarding the health benefits of polyunsaturated fatty acids (PUFA) to man. Human evolution pa ssed through a stage where ω-3: ω-6 PUFA ratio was 1 but with modernization this ratio has decreased in favour of ω-6 PUFA with attendant increases in inflammatory diseases. This review therefore focused on the importance of tilapia as a food fish in human diets and a cursory look at the metabolism of PUFA as well as existing protocols for incorporation in the flesh of tilapia and other food animals with a view to understanding protocols for increased incorporation in tilapia flesh as well as expected benefits. Dietary supplementation ω-3 PUFA: docosahexaenoic acid and eicosa pentaenoic acid (DHA and EPA), as found in fish , have been shown to decrease plasma triacylglycerols in humans, which helps to prevent atherosclerosis and coronary heart disease. lipid sources in the diets of fish hold promise for sparing , which is currently a subject of debate between aqua culturists, scientists and conservationists. shows greater promise as a lipid feed constituent with ability to provide required ω-3 PUFA in muscle and plasma of fish with the possibility of transfer to man in the diets of fish.

Keywords- PUFA, ω-3, DHA, EPA, Tilapia.

Citation: Chavan B. R., et al. (2015) Tilapia as Food Fish: Enhancement of Ω-3 Polyunsaturated Fatty Acids in tilapia (Oreochromis spp.), International Journal of Agriculture Sciences, ISSN: 0975-3710 & E-ISSN: 0975-9107, Volume 7, Issue 9, pp.-671-677.

Copyright:Keywords Copyright©2015- Pearl millet, ChavanHalf-diallel, B.R., etGene al. This effects, is an open Pennisetumglaucum-access article distributed. under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.

Introduction is essential for many metabolic functions in vertebrates including man [7]. The Fish and fish products are recognized as an important part of the human diet and Food and Agriculture Organization [8] promotes a hike in the proportion of PUFA demand is expected to increase [1]. With declining wild fish production [2], fish to saturated fatty acids in diets as a means to avoid atherosclerosis and coronary farming now has a significant role in ensuring fish supplies. The Nile Tilapia, which heart disease. The quantity of saturated fatty acids in plasma can be reduced is also called the aquatic chicken, Oreochromis niloticus, is a common freshwater with the intake of fish oil which has a high concentration of highly unsaturated ω-3 found in most tropical and subtropical waters, with extremely fast growth [3]. fatty acids [9, 10] and prevent cardiovascular disease [10]. The risk of ventricular Tilapia farming makes a large and growing contribution to world fish supplies. fibrillation is considerably reduced in animals with dietary intake of long-chain n-3 However, it is important to maintain high lipid quality in the muscle of cultured tilapia PUFAs, as against saturated and monounsaturated [11]. The possibility of to provide a large amount of the health promoting n-3 PUFA for the consumer. the same effect in humans has been subject of controversy until recently [11], The positioning of two or more double bonds at the cis end of the molecule is with the proof of reduced ischemic stroke being associated with a greater than used as the criteria for defining polyunsaturated fatty acids (PUFA). The Omega once consumption of fish per month [12]. Generally, a meal with one fatty fish per (ω) symbol denotes the particular location of the initial double bond when week [13] or between one fish to greater than 5 fish per week [12] is ideal for enumerating the double bonds from the methyl end of the molecule of the fatty reducing risks associated with cardiovascular disease while higher intake will not acid. For instance: , in the n-6 family, is designated as C18:2n-6 to lead to further reduction in risk [13] indicate that it has 18 carbons and 2 double bonds, with the first double bond at the sixth carbon [4]. PUFA are classified mainly as either ω-3 PUFA or ω-6 Overview of Tilapia PUFA. Two fatty acids that are classified as dietary essential fatty acids include Tilapia is one of the most important species in 21st century aquaculture with linoleic acid (ω-6) (LA) and alfa- (ω-3) (LNA). They are classified as production spanning more than 100 countries as of the year 2000 [14] and essential because of the inability of man to synthesize them [4]. Arachidonic acid greater than 135 countries in 2014 [15]. Tilapia has become one of the most and indeed all PUFA, of the ω-6 series are considered as essential in the diets of abundantly produced fish in aquaculture [16]. Tilapia farming is now in a dynamic man and have been credited with inflammatory mediation and regulation [5]. AA state with worldwide expansion to satisfy the demand from both domestic and can either be consumed directly in the diet or formed within the body from linoleic international markets and to provide an affordable source of animal protein. acid (LA, 18: 2n-6), itself consumed in the diet (18: 2n-6 to 20: 4n-6). LA is Although several tilapia species are cultured worldwide, the most popular is the abundant in nearly all commonly available vegetable such as , Nile tilapia (Oreochromis niloticus). Nile Tilapia is extremely fast growing reaching sunflower , seed and oils [6]. harvest maturity of 1-2 kg in 8 to 10 months [17]. Tilapia is the second largest The long-chain n-3 polyunsaturated fatty acids, EPA, 20: 5n-3 and DHA, 22: 6n-3 farmed fish in the world (next to Carp) with production level of 4.5 million metric tons in 2014 and a projected production of about 4.72 million mt in 2015 [18].

International Journal of Agriculture Sciences ISSN: 0975-3710 & E-ISSN: 0975-9107, Volume 7, Issue 9, 2015 || Bioinfo Publications || 671 Tilapia as Food Fish: Enhancement of Ω-3 Polyunsaturated Fatty Acids in tilapia (Oreochromis spp.)

Tilapia is now a top priority in aquaculture because of its ability to efficiently use typically insoluble in water, have an even number of carbon atoms in a long-chain natural foods. Being herbivorous in nature, it thrives primarily on vegetation and to form carboxylic acids having carbon atoms ranging from 10 to 24 and either algae and is often stocked in canals and artificial lakes for algae and vegetation saturated or unsaturated [62]. The long chain can either be straight or branched control. Tilapia is resistant to handling and diseases if stress is not induced, and nomenclature (n or ω) is based on the number of carbon atoms from the reproduces easily in captivity, tolerant to wide range of environmental conditions, methyl end on which the double bond is first located [63]. Dietary lipids for fish grows relatively fast and can easily are bred [19]. Tilapia, also known as Aquatic should contain ω-3 and ω-6 fatty acids for enhanced growth [64, 65]. Lipids are chicken, St. Peter's fish, Nile perch and Hawaiian sunfish, is a member of the useful in fish diets because they serve as a protein sparing energy source, Cichlid family, with origins from Africa. Tilapia is very hardy and can thrive in for lipid soluble vitamins to aid their absorption, cellular membrane marine, brackish or fresh water. Tilapia ranges in skin color from brilliant golden constituents and precursors of hormones [61]. The addition of lipids in fish diets red, pale red, white, gray, gray-blue, dark blue to black [3]. also improves palatability, texture and [61, 66]. Nutritional Requirement of Tilapia Lipid inclusion in diets for hybrid tilapia has a minimum and maximum level of 5% Tilapia has specific requirements for nutrients such as amino acids from protein, and 12% , respectively [67] and can be increased to 18% with increased protein fats, minerals and vitamins. Appropriate mixtures of protein, carbohydrates, sparing effect, but must not reach the 24% level [26,68] due to reduced protein vitamins, minerals, fibre and lipids in Tilapia diet will elicit good growth sparing effect [68]. Although the protein efficiency ratio is significantly influenced performance [20] considering the protein to energy ratio which lies between 89 by the protein percentage in the diet, it was not influenced by P/E ratio or an and 108 [21,22]. Nutritional requirements differ according to species and life interaction of both factors in diets of hybrid tilapia [26]. The optimal P/E ratio in stage with minerals, vitamins, protein and lipid being highly demanded by fry and tilapia diets with lipid as source of energy range from 15 to 27 mgkJ g-1 fingerlings due to growth and organ development, while juvenile and sub-adult depending on the size of fish [69, 70, 71]. Lipid inclusion can be reduced from as fish require energy from and carbohydrates for maintenance and a little protein much as 8-12% for small sized tilapia to as low as 6 – 8% as the fish get larger to cater for growth even though it is at a reduced rate [23]. [72] provided carbohydrates are included to provide energy and spare protein Protein [73]. Tilapia has a dietary requirement for linoleic acid (LA, 18:2 n-6) and α- linolenic acid (α LNA, 18:3 n-3) due to inability to synthesize these fatty acids Considering the size and age of tilapia, optimum dietary protein levels vary from within the body hence they are designated essential dietary fatty acids [61]. 28% to 50% [24,25,20,26]. Essential amino acids should make up 60% or more Contradiction trails the reports of quantifiable essential requirements of of the protein [27], since tilapia require all 10 EAA’s as other fish [28] with non- tilapia [29, 61]. essential amino acids making up the percentage so as to spare energy for their synthesis [29]. Energy The energy content of feeds can be quantified at three different levels: the gross, Carbohydrate digestion and metabolism levels with the latter offering a greater advantage in Although there is no dietary requirement of carbohydrates determined for fish determining the energy content of feed since it gives the precise value of energy species [29, 30], its inclusion in the diet will spare the catabolism of proteins and that can be available for growth taking into account energy spent on metabolism lipids for energy and other intermediates [31]. It has been determined that the of proteins. Energy should be given priority in tilapia feed formulation with utilization of carbohydrate by tilapia is related to fibre content [32,33] and size inclusion of energy providing feedstuff such that energy is not a limiting factor that [34]. Inclusion of carbohydrates in the diet of tilapia at rates between 30 to 70% of will trigger the catabolism of protein to provide energy. Metabolizable energy the diet does not affect growth negatively and utilization is in the range of 35 to (ME) of feedstuff is a good index for selecting ingredients for feed formulation. 40% of digestible carbohydrate with larger tilapia utilizing better than small sized However, there is great difficulty in its determination of fish and other aquatic tilapia [34]. species; hence it is better to use digestible energy (DE) values or gross energy Vitamin and Mineral (GE) values if both ME and DE are unavailable. It has been reported that protein content and P/E ratio do not interact to affect PER of tilapia [26] but the optimal Vitamins and minerals are critical to proper nutrition in tilapia reared under dietary P/E ratio is related to the size of the fish [69, 70, 71] with the P/E ratio intensive condition. Water soluble vitamin requirements of tilapias that have been having an inverse relationship with fish size [74]. At high levels of dietary protein, studied are: Thiamine: 2.5 - 4.0 mg/kg [35, 36, 37], pantothenic acid: 6 – 10 a P/E ratio of 20.5 mgkJ g-1 is recommended [75] but a range of 22 – 24 mg kJ g-1 mg/kg [38, 39, 40, 41], pyridoxine: at 28% crude Protein (CP) diet level, 1.7 - 9.5 is adequate in such circumstance [76]. mg/kg, and at 32% CP level, 15 - 16.5 mg/kg [42], niacin: 26 - 121mg/kg [43], biotin: 0.06 mg/kg [44], choline: 1000 – 3000 mg/kg [45, 46], Inositol: 100 – 400 n-3polyunsaturated fatty acids mg/kg [47, 48], riboflavin: 5 – 6 mg/kg [49,50], folic acid: 0.5 - 1.0 mg/kg [51, 52], ω-3 polyunsaturated acids with the first double bond on the third carbon atom and ascorbic acid: 1.25 – 80 mg/kg [53, 54]. Fat-soluble vitamins for tilapia from the methyl end are termed omega-3 (ω-3) fatty acids and they are abundant concern only vitamin D and E [24]. Five minerals are required for tilapia namely; in fish and other marine animals [77,78,79]. Three ω-3 polyunsaturated fattyacids Calcium, phosphorous, magnesium, zinc and potassium have been quantified for includeα-linolenic acid (LNA, C18:3ω-3), eicosa pentaenoic acid (EPA, C20:5ω- their requirements in tilapia, by [25]. Fat-soluble vitamins whose requirements 3) and docosahexaenoic acid (DHA, C22:6ω-3). LNA can be obtained from have been determined for tilapia include Vitamin A: 5850 - 6970 IU/kg as retinyl vegetable sources while EPA and DHA are obtained from marine sources. The acetate [55] or 28.6 44 mg/kg as b-carotene in the presence of 84 IU/kg of dietary efficiency of mammals at converting LA and ALA to AA, DHA, and EPA is low vitamin A [55], vitamin D: 375 mg/kg as cholecalciferol [56] and : 10 – given the fact that synthesis of LA from is not possible therefore; 206 mg/kg [57, 58, 59]. Recommended levels of eight minerals: calcium, dietary intake of AA, DHA and EPA is important [77]. Lipids are a source of chromium, potassium, magnesium, iron, phosphorous, sodium and zinc are energy in human and animal diets and also supply a full complement of fatty presented by [60]. acids that have significant roles to play in physiological processes ranging from production of metabolic precursors to production of hormones and associated Lipid effect on neurological, reproductive and cardiovascular processes. Lipid requirement of fish is related to the weight (size) of the fish with 10% Polyunsaturated ω-3 fatty acids have been reported to mitigate cardiovascular inclusion in diets for fish less than 2 g and a final inclusion level of between 6 – disease risk through effects on factors such as a drop in concentration of 8% as rearing continues until harvest [61]. Lipids are biological compounds that plasma triacylglycerol, platelet clustering, and blood pressure [80], with vital comprise fatty acids or steroid group’s hence five classes that include roles in fetal and central nervous system development particularly DHA [81] phospholipids, triacylglycerides, sphingolipids, waxes and sterols. Fatty acids are

International Journal of Agriculture Sciences ISSN: 0975-3710 & E-ISSN: 0975-9107, Volume 7, Issue 9, 2015 || Bioinfo Publications || 672 Chavan B.R., Yakupitiyage A., Ataguba G.A., Kamble M.T. and Medhe S.V.

However, there is a need for balance in ratio between ω-6 and ω-3 fatty acid Table-1 PUFA (ω-3 and ω-6) composition of selected vegetable oils concentration in plasma otherwise deleterious effects can occur [82] including prothrombotic and proaggregatory effects, increased blood , vasospasm, ω-3 (α-linoleic) ω-6 (Linolenic) and vasoconstriction and reduced bleeding time [83]. To ensure this balance, the 6.6 21.6 ratio of intake of ω-6/ω-3 should be less than or equal to 10 for infants and a ratio Safflower oil 10 70-85 of 1/5 – 10 is ideal for adult nutrition [77]. Flaxseed oil 48-60 <20 Sources of PUFA Humans lack fatty acid desaturases to create double bonds on fatty acids further - 5-7 than the ninth carbon atom hence linoleic acid (18:2 ω-6 and linolenic acid (18:3 0.5 41-45 ω-3) are essential dietary components [84]. Vegetable oils and fish oil are widely <0.8 60-72 used as dietary sources of ω-3 fatty acids but microalgae and other marine sources are beginning to gain attention [85,86,87]. Both DHA and EPA (ω-3PUFA oil 4-10 48-58 [Fig-1], can either be obtained directly from the diet or synthesized within the Rice oil 1.1 39.1 body from precursors [6]. Linoleic acid (18:2n-6) and α-linolenic acid (18:3 ω-3) Wheat sprout oil 8 57 are precursors of longer chain PUFA that are synthesized by desaturation and Source: [89] elongation. Linoleic acid undergoes desaturation, elongation with the addition of

two carbon atoms and further desaturation to give rise to Arachidonic acid (20:4 Production of long chain PUFA by fish is made possible by the presence of a ω-6) while a-linolenic acid undergoes the same process steps to yield complete set of genes alongside expression of fatty acid desaturase and Eicosapentaenoic acid (EPA, 20:5 ω-3) with further elongation to yield elongase (fad and elovl) for the production of EPA and DHA from α-linolenic acid Docosahexaenoic acid (DHA, 22:6 ω-3). Desaturation is enhanced by the [95,96]. Marine fish on the other hand do not have a robust ability to convert enzymes Δ6 and Δ 5 desaturases [88]. linolenic acid into EPA or DHA [97] hence adequate dietary provision of linolenic H C COOH acid, DHA, EPA and arachidonic acid (AA) must be maintained if they are held in 3 captivity [98]. However, it has been demonstrated that herbivorous marine fish of the family Siganidaecan elongate PUFA. Siganus canaliculatus produces two Linolenic acid (C18:3ω-3) desaturases specific for Δ4 and Δ6/Δ5 positions as well as two elongases: Elovl4 and Elovl5 [99,100] while S. argusexpressedΔ-6 fatty acid desaturase in the liver H3C COOH and brain with the degree of expression being dependent on dietary intake of

lipids as well as fatty acid composition [97]. Eicosapentanoic acid (C20:5ω-3) Linoleic acid is abundant in sunflower oil, sesame oil and while linolenic acid is abundant in flaxseed oil. These vegetable oils can be used as H C energy sources in the diets of fish as demonstrated for the African catfish 3 COOH (Heterobranchus longifilis) [101] and common carp (Cyprinuscarpio) [102]. A more detailed description of PUFA content in food is reported in [Table-1]. Docosahexanoic acid (C22:6ω-3) Dietary PUFA’s and Man through the course of evolution H C The capacity of the human digestive system to utilize PUFA has undergone 3 COOH evolutionary changes over the period since human existence. Over the past 1000 years, the percentage of energy derived from ω-6 PUFA has increased while Linoleic acid (C18:2ω-6) contribution from ω-3 PUFA has decreased [103]. This has led to a shift in the ratio of ω-6/ω-3 in the human diet from less than 1 predicted for the Paleolithic H3C COOH population [104,105] to about 0.5 [103]. Over the past millennium, development of food engineering and agriculture have

led to changes in food intake and associated nutrient uptake. Total fat intake has Arachidonic acid (C20:4ω-6) increased over the past 1,500 years with associated increase in saturated fatty Fig-1. ω-3 Poply Unsaturated Fatty Acids acid and trans fatty acid intake [103]. Sources of fats in this regard include grain- oils are the major sources of ω-3 and ω-6 fatty acids. [Table-1] shows fed cattle, hydrogenated vegetable oils and oils from vegetable sources [104]. a list of vegetable oils and the percentage of ω-3 and ω-6 fatty acids they contain There is a strong effect of this dietary change on human health with evidence in in the form of α-linoleic acid and linolenic acids respectively. Sources of ω-3 fatty the United States. People of African descent are disproportionately affected by acids include nuts, , , vegetables, meat, , game, fish and inflammatory diseases [103] due to the change in diet with consumption of seafood [90,91]. Specific contribution of ω-3 to the diet comes in small quantities purified foods of origin having excess saturated and trans fatty acids with from some fruits, egg yolk, poultry, and meat, while fish is the main source of more ω-6 PUFA compared to ω-3 PUFA. EPA and DHA [92]. Vegetables provide a rich source of ω-6 fatty acids. Linoleic There is a correlation between feed consumed by animals and the output of acidcan be made available in diets with , safflower oil, sunflower oil, and PUFA in red meat with the trend being in favour of meat rich in ω-3 as against ω- soybean oil [93]. 6 PUFA [106] Consumption of red meat from grass fed animals has been found Metabolism of PUFA in Fish to increase dietary ω-3 PUFA intake which reflects in the concentration of long Freshwater fish can convert 18:2ω-6 and 18:3ω-3 to C20 and C22 homologues to chain PUFA in the plasma and platelets compared to consumers of meat from meet their needs [1]. Fish cultured using extensive culture system as well as wild animals fed concentrate feed [107]. The increase in consumption of red meat has fish have a fatty acid profile that suits human requirements with lower 18:2ω-6 led to a decrease in the ω-6/ω-3 PUFA ratio as well as reports of implication in content and high 18:3n-3, 20:5n-3 and 22:6n-3 hence a high ω-3/ω-6 PUFA ratio incidence of cardiovascular disease and colon . On the other hand, [94]. However, some reports show that wild fish contain more total PUFA than moderate consumption of lean red meat is ideal for providing dietary fatty acids farmed fish [78]. [108].

International Journal of Agriculture Sciences ISSN: 0975-3710 & E-ISSN: 0975-9107, Volume 7, Issue 9, 2015 || Bioinfo Publications || 673 Tilapia as Food Fish: Enhancementof Ω-3 Polyunsaturated Fatty Acids in tilapia (Oreochromis spp.)

Differences in PUFA content of fish flesh Marine fish flesh contains more ω-3 PUFA than freshwater fish lending credence Table-2 Quantity of n-3 PUFA in Fishes to the variation in ω-3 PUFA content as observed for various fish species in Sr. Fin fishes n-3 Reference [Table-2].The amount of n-3 PUFA in tilapia is 0.7g/100g of edible fish tissue. The no FAs consumption of phytoplankton by marine fish confers on them a greater tissue 1 Channel Catfish 0.3 [129] concentration of eicosa pentaenoic acid (EPA) and docosahexaenoic acids (DHA) [109]. The fatty acid content of food fish is affected by several factors 2 Carp 0.6 [129] including: season, feeding, species, sex, sexual maturity, size, place of capture 3 Brown Bulhead 0.5 [129] (for wild fish) and water temperature [110]. Seasonal food availability as well as Catfish food choices are important drivers of total lipid content of fish flesh while dietary 4 Sucker 0.6 [130] niche and variability of component dietary sources are secondary factors in this 5 Red Tilapia 0.12 [131] regard[111,112,113]. 6 Tilapia 0.7 [131] Protocols for increase of n-3 PUFA in human diet Both ω-3 (α-Linolenic acid – ALA) and ω-6 (Linoleic acid – LA) PUFA are 7 Whitefish 1.8 [130] essential dietary components for man since their synthesis is not possible within 8 Striped Bass 0.8 [132] the body. Grains like rapeseed, flaxseed and walnuts as well as leaves of wild 9 Freshwater Eel 0.62 [131] are rich in ALA while most other grains are rich in LA. The metabolic pathways of these fatty acids rely on the same enzymes: elongases and 10 Big Head Carp 0.3 [131] desaturases hence there is competition between the fatty acids leading to either 11 African Catfish 0.2 [131] more of arachidonic acid (AA) if ω-6 PUFA prevails or eicosa pentaenoic acid 12 Rohu 1.2 (EPA) if ω-3 PUFA prevail. Increased production of EPA as a result of a low ω- 13 Catla 2.2 6:ω-3 ratio increases the production of anti-inflammatory agents [114]. The

identification of ω-3 PUFA food sources that are acceptable to the public and 14 Mrigal 1.1 taken in the right quantities is the pivot to meeting ω-3 intake requirements [115]. 15 Silver Carp 1.1 [132]

Fish as a food source is rich in ω-3 PUFA hence several efforts to increase the 16 Grass carp 1.7 content of ω-3 PUFA in fish as food have been carried out. Wild fish tend to have 17 Calbasu 1.5 greater n-3 PUFAs and DHA/EPA ratios than cultured fish [78] however a feeding 18 Rainbow Trout 0.6 [132] strategy that involves the use of palm oil during culture and a reversion to fish oil (Steelhead) based finishing diet have been reported to successfully increase and 19 Burbot 0.2 [130] tocotrienols in tilapia [116], ω-3 PUFA levels in farmed gilthead sea bream and European sea bass [117, 118] and can be used as a quick method of restoring ω- 20 Snakehead 0.8 [131] 3 levels in tilapia just before harvest. Furthermore, supplementation of human diets with fish oil has been reported to produce a 35% decrease in plasma and reduced numbers of small dense lipoprotein lipase (LPL) with Fish fed corn oil and soybean oil based diets produced a high amount of ω-6 fatty evidence of genetic link in the production of LPL through increased levels of LPL acid in their carcass as well as polar lipid fraction of the liver while fish fed linseed mRNA [119]. oil produced more of ω-3 PUFA in the carcass as well as liver [122]. In addition, Alternatives to direct consumption of vegetable oil food sources lie in the feeding juvenile Pike Perch fed linseed oil based diets produced more ω-3 PUFA in of animals with these sources and subsequent intake of animal protein by man. phospholipids and compared to fish fed fish oil and soybean oil Leghorn pullets fed with diets containing flaxseed showed a marked increase in based diets with triglyceride ω-3:ω-6 ratio of 1.5 for fish fed linseed oil based diet ω-3 PUFA content of their eggs without alteration to cholesterol content of egg compared to 1.0 and 0.3 for fish oil and soybean oil respectively [123]. yolk and when humans consumed these eggs, there was a 33% increase in ω-3 Fortification of Tilapia with ω-3 PUFA PUFA with a increased ratio of ω-3: ω-6 PUFA in phospholipids obtained from Fish have the ability to synthesize long chain PUFA from plant oils without platelets of volunteers [120]. Similarly, addition of canola oil, and adversely lowering the levels of DHA and EPA in the flesh hence a significant linseed oil in diets for layers at 70 g/kg increased the ω-3 PUFA concentration advantage for man both in nutrition and economics of aquaculture. Use of plant from 1.2% of fatty acids in the egg yolk to 4.6, 6.3 and 7.8% respectively while oil sources in fish feeds will spare long chain ω-3 fatty acids in fish oil for only consumption by humans produced no significant increase in cholesterol and low vital physiological needs [124]. It has been reported that ω-3 PUFA, the density lipoprotein cholesterols (LDL-C) but resulted in a decrease of plasma DHA/EPA ratio in fats from wild fish are higher than the farmed fishes [78]. High triglycerides, production of less atherogenic LDL and decreased platelet levels of n-6 PUFA in farmed fish will mean a high ω-6:ω-3 ratio hence creating aggregation [115]. the possibility of inflammatory diseases for fish consumers. Test diet for littermate pairs of pigs containing 10 g/kg LA and 4 g/kg ALA with Supplementation of tilapia diet with linseed oil at rates of 5% and 7% has been addition of linseed led to a 56% increase in the ω-3 PUFA content of muscle, found to reduce ω-6:ω-3 ratio from 4.6:1 to 2.3:1 while ω-3 PUFA increased by 100% increase in EPA content and 35% increase in DHA content with 46% and 58% respectively [125]. Similarly, Feeding vegetable oil complemented concomitant increases in the fatty tissues, followed by a significant reduction in diets to tilapia O. niloticus has been reported to produce an increase in ω-6 the ω-6: ω-3 ratio from the average of 7 to 5 with high levels of ω-3 PUFA being PUFA content of muscle and liver over fish fed with fish oil. However, ω-3 PUFA observed in sausages after 6 months of frozen storage [121]. These results content of muscle and liver was not significantly different from soybean oil, show that pork has the potential to provide dietary ω-3 PUFA for humans in the linseed oil and fish oil diets with ω-6: ω-3 ratios of 1.31, 0.96 and 0.75 long run. Linseed oil is may be used as a substitute for fish oil in fish diets respectively in the muscle and 2.34, 1.24 and 0.72 respectively in the liver [126]. because it is rich in ALA, a substrate for synthesis of (n-3) PUFA with significant This further goes to show that linseed oil supplementation provides benefits close amount of LA hence an ω-3:ω-6 ratio of 3-4:1. A test of four various dietary lipid to fish oil and consumption of tilapia fed linseed as feed ingredients offers the sources in diets of Surubim (Pseudoplatystoma coruscans) revealed that the fatty advantage of ω-3 uptake by consumers with attendant benefits. acid composition of carcass lipids are affected by the lipid sources which included . , corn oil, soybean oil and linseed oil.

International Journal of Agriculture Sciences ISSN: 0975-3710 & E-ISSN: 0975-9107, Volume 7, Issue 9, 2015 || Bioinfo Publications || 674 Chavan B.R., Yakupitiyage A., Ataguba G.A., Kamble M.T. and Medhe S.V.

Alternative lipid sources for tilapia with the aim of incorporating PUFA in the [12] Siscovick D. S., Raghunathan T. E., King I., Weinmann S., Bovbjerg V.E., muscle have been reported by scientists. The use of (obtained from the Kushi L., Cobb L.A., Copass M.K., Psaty B.M., Lemaitre R., Retzlaff B. and tung Vernicia fordii in ) in diets of GIFT tilapia has resulted in Knopp R. H. (2000)American Journal of Clinical Nutrition, 71(1), 208-212. accumulation of the conjugated form of linolenic acid in processed fillets by up to [13] Lemaitre R.N., King I.B., Mozaffarian D., Kuller L.H., Tracy R.P. and 1.02% of total fatty acids with associated biosynthesis of conjugated linolenic acid Siscovick D.S. (2003) American Journal of Clinical Nutrition, 77,319 – 325. (CLA) isomers amounting to about 1.08% of fatty acid composition of fillet while EPA and DHA were 2.85 and 3.08% respectively [127]. Nile tilapia fed with [14] Fitzsimmons K. (2000) In: Fitzsimmons K. and Filho J. C. (Eds.) Proceeding groundnut leaf meal as against arahar leaf meal (Cajanascajan) and commercial from the Fifth International Symposium on Tilapia Aquaculture, 3-8. feed produced the highest PUFA content in muscle with the same trend also [15] FAO (2014) Food and Agriculture Organization of the United Nations, observed for EPA and DHA. The level of ω-3 PUFA in the muscle of fish fed Rome, p25. groundnut leaf meal was reported to be higher (14.15%) compared to 11.63% [16] Lovshin L. L. (1997) In: Anais do 1 Workshop International de Aquaculture, and 10.54% for an arahar leaf meal and commercial diets respectively while ω-6: October, 15-17, Sao Paulo, , 961-66. ω-3 ratio were 0.83, 0.71 and 0.68 for commercial feed, arahar leaf meal and groundnut leaf meal respectively [128]. [17] King M. (2000) The Institute of Food and Agricultural Sciences, University of Florida, 1-3.

[18] Tveterås R. (2014) Global aquaculture advocate November/December Conclusion 2014, 8 - 9, 11. Modern agriculture, with its dependence on grain diets, has led to an increase in total saturated fatty acids and ω-6 polyunsaturated fatty acids, linoleic and [19] Guerrero R. D. III. (1982) In: Pullin R.S.V. and Lowe-McConnell R.H. arachidonic acids in human diets. In the past century as a result of the industrial (eds.). ICLARM Conference Proceedings 7. International Center for Living revolution, with the emergence of agribusiness, processed foods, grain fattened Aquatic Resources Management, Manila, , 432. livestock, and hydrogenated vegetable fats, has further reduced the intake of ω-3 [20] Jauncey K. and Ross B. (1982) University of Sterling, Scotland, 1-95. fatty acids and increased ω-6 fatty acid intake as well as the ratio of ω-6 to ω-3 [21] Winfree R.A. and Stickney R.R. (1981) Journal of Nutrition, 111, 1001-1012. essential fatty acids in diets today. [22] Li Y., Bordinhon A. M., Davis D. A., Zhang W. and Zhu X. (2013) Recent studies suggest that excessive amounts of n-6 PUFA and a very high ω- Aquaculture International, 21, 1109 – 1119. 6/ω-3 ratio promotes inflammatory diseases while balancing or reducing the ratio of ω-6/ω- 3 fatty acids may decrease the risk of these diseases Thus, for good [23] Stickney R. R. (1996) World Aquaculture, 27(1), 45-50. health it is necessary to have a balance of ω-6/ω-3 fatty acids in the diet and in [24] El-Sayed A.F.M. and Temisha S. I. (1992) Aquaculture, 103, 55-63. our bodies. To overcome this modern nutritional problem, there is a need to [25] Davis A.T. and Stickney R. R. (1978) Transaction of American Fisheries enhance the content of ω-3PUFA in foods especially fish. Society, 107, 479-83. New finding may provide a new strategy for production of ω-3 PUFA-enriched [26] Santiago C. B. and Laron M. A. (1991) In: De Silva S.S. (ed) Special food stuff by developing new a economical protocol for enhancedω-3 PUFA Publication No.5, Asian Fisheries Society, Manila, the Philippines, 55-62. incorporation in human diet by producing ω-3 PUFA enriched Tilapia. This is because tilapia is affordable and can be produced easily in several environments. [27] Jauncey K. (2000) In: Tilapias: Biology and Exploitation, M.C.M. Beveridge and B.J. McAndrew (eds), Kluwer Academic Publisheires, 327-375. Finally, discovery of alternative lipid sources with attributes of economical feed [28] Santiago C. B. and Lovell R. T. (1988) The Journal of nutrition, 118(1212), formulation as well as capability to replace fish oil with enhancement of ω-3 1540-1546. PUFA in fish diets will provide the aquaculture industry a greater array of choices [29] Shiau S.Y. (2002) In: Webster C.D. and Lim C. (eds). Nutrient for feed ingredients for use in formulating feeds. Requirements and Feeding of Finfish for Aquaculture. CAB International, References 418pp. [1] Sargent J. R. and Tacon A. G. J. (1999) Proceedings of the Nutrition [30] NRC (1993) National Academies Press, Washington, D.C., 115pp. Society, 58, 377-383. [31] Steffens W. (1989) Chichester, West Sussex, England. [32] Anderson J., Jackson A.J., Matty A.J. and Capper B.S. (1984) Aquaculture, [2] Naylor R. L., Goldburg R. J., Primavera J., Kautsky N., Beveridge M. C. M., 37, 303 – 314. Clay J., Folke C., Lubchenco J., Mooney H. and Troell M. (2001) Nature, [33] Qiang J., Yang H., He J., Wang H., Zhu Z.X. and Xu P. (2014) Turkish 405(29), 1017-1022. Journal of Fisheries and Aquatic Sciences, 14, 515 - 525. [3] Popma T. and Masser M. (1999) SRAC Publication, No. 283. [34] Shiau, S.Y. (1997) Aquaculture, 151: 79-96. [4] Sigma Tau S. P. A. (2003) Scientific Department, 1-56. [35] Lim C. and Leamaster B. (1991) Journal of World Aquaculture Society, 22, 36A. [5] Calder P.C. (2006) Scandinavian Journal of Food and Nutrition, 50(S2), 54 [36] Lim C., Barros, M.M., Klesius P.H. and Shoemaker C.A. (2000). In : Book of –61. Abstracts. Aquaculture America 2000, New Orleans, Louisiana. Baton [6] Sargent J. R. (1997) British Journal of Nutrition, 78 (1), 5-I3. Rouge, Louisiana: World Aquaculture Society. [7] Simopoulos A. P. and Cleland L. G. (eds) (2003) World Review of Nutrition [37] Lim C., Yildrim-Aksoy M., Barros M.M. and Klesius P. (2011) Journal of the and Dietetics. (Basel, Karger), 92, 37-56. World Aquaculture Society, 42, 824–833. [38] Kasper C.S., White M.R. and Brown P.B. (2000) Journal of Nutrition, 130, [8] FAO (1977) FAO agriculture Series, 8, 1-60. 238–242. [9] Matson F.H. and Grundy S.M. (1985) Journal of Lipid Research, 26,194- [39] Roem A.J., Stickney, R.R. and Kohler C. C. (1991) Progressive Fish 202. Culturis,t 53, 216–219. [10] Fernandez M.L. and McNamara D.J. (1989) Metabolism, 381094-1102. [40] Soliman A. K. and Wilson R. P. (1992a) Aquaculture, 104,121 –126. [11] Gebauer S.K., Psota T.L., Harris W.S. and Kris-Etherton P.M. (2006) American Journal of Clinical Nutrition, 83(6), 1526 - 1535.

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