Comparative Studies On Muscle Lipid Fatty Acid Profiles Of Some Cultured Indian Carp Species

Comparative Studies On Muscle Lipid Fatty Acid Profiles Of Some Cultured Indian Carp Species

Kaur M, Sehgal G.K. and Sehgal H. S

M.sc, Department of Zoology, Punjab Agricultural University Ludhiana, Punjab.

Abstract

The Polyunsaturated fatty acids (PUFAs), mainly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), help in preventing coronary heart disease. These fatty acids play an important role in reducing the risk of colon, breast and prostate cancer. Although, carps form the major chunk of harvested fish worldwide, the information about their fatty acid profiles is meager.The present study was conducted to know the seasonal variations on muscle total lipid content (MTLC) and fatty acid profiles of three Indian carp species, Labeo. rohita(Ham.), Catla. catla (Ham.) and Cirrhinusmrigala (Ham.). The MTLC varied within the species of fish (1.06±0.02% in C. mrigala to 3.00±0.06% in C. catla) and season of the year. The minima were recorded during spring except C .mrigala (summer) and maxima during winter in all species. The total PUFAs highest in C. catla (23.21±3.67%) and minimum in C. mrigala (18.50±4.95%).The mean ratio of n3/n6 was highest (1.49) in C. catla and lowest (0.89) in C. mrigala.

1. Introduction

   Fishes are considered as high quality human food and a rich source of proteins (15-25%), minerals (Ca, P, Fe) and vitamins (A,D,E,K). Recently, interest has grown in fish and fish products as sources of polyunsaturated fatty acids (PUFAs), mainly of the n-3 family (also known as omega-3 fatty acids). Special attention is being paid to long chain omega-3 fatty acids (LC-PUFA) or highly unsaturated fatty acids (n-3 HUFA) i.e. eicosapentaenoic acid (EPA, C20:5n-3) and docosahexaenoic acid (DHA, C22:6n-3). This interest stems largely from those studies which have suggested that n-3 PUFAs may have an important

   role in prevention and management of cardiovascular diseases (Glomset,1985; Goodnight et al 1982; Kromkoutet al 1985; Norum and Drevon, 1986) and may even decrease the risk of cancer development at certain sites (Caroll and Bradon, 1984; Herold and Kinsella, 1986). It is important to maintain an appropriate balance of omega-3 and omega-6 fatty acid in the diet as these works together to promote healthbut the typical diet tends to contain 11 to 30 times more omega-6 fatty acids than omega-3. This imbalance is a significant factor in increasing the range of human diseases which respond to n-3 HUFAs (www. fsomega.com). Fishes vary in their ability to produce EPA and DHA endogenously.Variation in lipid and fatty acid composition between and within species, depending on factors, such as season, food availability, location, sex and age have been well documented by numerous authors (Exleret al 1975, Gorgun and

   Akpinar 2007; Rueda et al. 1997; Shearer 1994). Thus, carps being the most commonly cultured fish species in India, including Punjab, need to be investigated for their fatty acid profiles. The present study was, therefore, conducted with a concept to generate information on fatty acid profiles of carp species being cultured in Punjab. Carp(s) with highest proportion of LC-PUFA and the highest omega-3 to omega-6 fatty acids also needed to be identified for human consumption. Therefore, the aim of this paper is to work out details of the fatty acid profiles of muscle lipids of some cultured carp species and to study the seasonal differences in the fatty acid profiles of these carp species.

2. Material and Methods

   1. Sample collection

      Comparative studies on muscle total lipid content and fatty acid profiles of three carp fish species cultured in Punjab belonging to order Cypriniformes, family Cyprinidae were made. The fish species were three Indian Major Carps, rohu,Labeo. rohita(Hamilton); catla, Catla. catla (Homilton); mrigal, Cirrhinus. mrigala (Hamilton).Table-sized (> 500 g) fresh specimens of each of the three fish species were obtained from the local fish market during each season of the year.Each fish was scaled, finned, headed and gutted. The fish sample was then cleaned with tap water and 3 x 2.5 cm pieces of flesh were taken at random from threedifferent parts of fish body viz. above lateral line from both the sides. Bones were removed and the flesh was thoroughly mixed together to form a composite or representative sample of edible portion of the fish. Composite

      sample was packed in clean labeled ziplock polythene bags and stored at -250C until analyses.

   2. Lipid extraction and fatty acid analysis

      Total lipid (TL) content was determined by solvent extraction method as described by AOAC (2000).

      Fatty acid composition was determined by Gas Chromatography (GC) on M/s Nucon Engineers AIMIL Gas Chromatograph (solid state) model Nucon series 5700/5765 equipped with flame ionization detector fitted with SS column 1/8 outer diameter x 2M length, packed with 15% D.E.G.S on CHROMOSORB W.H.P, 80-100 mesh size. The

      conditions for the separation were oven temperature 2000C, injector temperature 2300C, detector temperature 2400C, hydrogen flow 30ml/min, air flow 300ml/min and nitrogen flow 40ml/min. Identification of peaks was done by comparison of their retention time with those of standard fatty acyl esters (M P Biomedicals Inc. USA). Relative concentration of fatty acid was calculated by use of an automatic integrator-Windows based AIMIL Ltd., DASTA 710 Gas Chromatograph Datastation software, version WinAcds 7.1.

   3. Statistical analysis

      One way and Multifactor ANOVA was used to determine the inter-specific and inter-seasonal differences in the total lipids and fatty acid profiles of the three species of fish. The analyses were performed using Microsoft EXCEL and STATGRAPHICS statistical packages.

3. Results and discussion

   In our study, Considerable variations in the muscle total lipid content (MTLC) during different seasons of the year.The MTLC varied from1.06±0.02 % in C. mrigaladuring summer and 3.00±0.06% in C. catladuring winter( Table 1). These values are close to those observed by Aggelousis and Lazos (1991) in the freshwater fishes of Greece (0.6-3.5%),Varjlinet al (2003) in D. vulgaris and C .conger (1.3 to 3.7%).Memonet al (2010) reported higher lipid content ranging from 0.85-18.32% in Indus river fish species. Total lipid content of carps varied with seasons. In all carp species, higher lipid content was observed in winter and lower in all carp species were observed in spring except for C. mrigala(which was observed in Summer). The study therefore, supports the observations of Rasoarahonaet al (2004) that lipid content was higher in colder than in warmer months. Guleret al (2011) also observed higher lipid content in winter.

   A comparison of fatty acid profiles of the presently studied fish species revealed that amongst the n-3 fatty acids, the maximum mean value of Len (1.99±0.85 %) was found in C. mrigalaand minimum (1.23±0.61%) in C. catla. The mean values of EPA were, however, comparable amongst C. catla (3.98±2.60%) and L. rohita (3.80±1.84%). In L. rohita,the EPA was minimum (2.20±2.19%)in spring andmaximum (6.46±5.74%) in winter. DHA was maximum (3.55±3.16%) in autumn and minimum (0.16±0.08%) in summer. EPA+DHA was maximum (9.96%) in winter and minimum (2.36%) in summer.InC. catla, EPA was maximum (10.61±10.29%) in autumn and minimum (1.73±0.91%) in spring. The DHA was minimum (0.18±0.18%) in summer and maximum (4.83±4.83%) in spring and EPA+DHA was maximum (12.79%) in autumn and minimum (2.01%) in summer. In C. mrigala,EPA was highest (9.74±5.21%) in summerand EPA+DHA was minimum (0.18%)in autumn and maximum (17.24%) in summer. Total n-3 PUFAs was maximum in L. rohita (12.98±9.79%) during autumnand minimum (8.42±6.76%) during spring. The mean total n-3 PUFA values were recorded in the order of C. catla(12.36±3.51%)>L. rohita (10.89±3.41%)>C. mrigala(8.51±2.12%)( Table 2,3,4).

   The fishes differed in respect of their n-6 PUFAcomposition also. L. rohitahad highest mean Lin content (10.20±2.54%) followed by C. catla (8.27±2.90%) andC. mrigala(6.58±3.45%). Lin was lowest in spring in all carp species and highest in autumn in C. catla (24.29±10.43%) and C. mrigala (18.99±13.69%).The AA was highest in C. mrigala

   (3.63±0.94%) followed by C. catla (2.15±0.90%), L. rohita(1.60±0.63%). The mean values of EDA also differed amongst different fish species viz.L. rohita (0.34±0.18%),C.mrigala (0.18±0.08%), and C. catla (0.05±0.04%). The mean total n-6 values were comparable, although highest values were observed in L. rohita (12.14±2.55%), followed by C. catla (11.31±3.12%) and C. catla(10.06±3.44%). The n-

   3/n-6 ratio differed in different fishes. The highestmean of n-3/n-6 (1.49±0.63) was found in C. catlaand the lowest (0.89±0.20) in C. mrigala. The ratio of n-3:n-6 PUFA contents ranged from 2.67 to

   12.61 in selected 12 marine fish species living in Turkish waters (Bayiret al 2006). Compared with freshwater fish, marine fish contains higher levels of n-3 PUFAs (Vlieg and Body,1988). Therefore, in our study, this ratio did not fall in these ranges in carp species.

   The most abundant monounsaturated fatty acid (MUFA) was oleic acid in all the carp species. The high level of oleic acid and Total MUFAs (33.31±4.95%) was found during autumn except L. rohitawhich was found in summer (1.80±1.80%).

   In the present study, major saturated fatty acid (SFA) was palmitic acid and other predominant SFAs were stearic acid in all carp species. Celiket al (2005) and Ozogulet al (2007) also reported similar results. In our study stearic acid was lower in spring (0 to 1.38%) than in winter (18.05% to 32.03%). Total SFAs was minimum (13.89% to 18.54%) in spring and maximum (47.01% to 63.69%) in winter in all the carp species.

   Table 1 Seasonal variations in muscle total lipid content (%) of different fish species.

   Season Fish species

   (Month/Year)

   Spring

   Labeorohita

   (rohu)

   Catlacatla

   (catla)

   Cirrhinusmrigala

   (mrigal)

   (April,2009) 2.01±0.03a 1.50±0.06a 1.15±0.23a

   Summer

   (July,2009) 2.05±0.07ab 2.29±0.04b 1.06±0.02a

   Autumn

   (October,2009) 2.22±0.04b 2.24±0.05b 1.82±0.05b

   Winter

   (January,2010) 2.44±0.02c 3.00±0.06c 2.17± 0.02b

   Mean 2.18±0.02 2.26±0.03 1.55±0.06

   Values with same alphabetic superscript in a column do not differ significantly (p>0.05).

   Table 2Seasonal differences in fatty acids of Labeo. rohita(Hamilton).

   Fatty Acid			Season			Mean
   	Spring	Summer		Autumn	Winter	(of the four seasons)
   	(April, 2009)	(July, 2009)		(October, 2009)	(January, 2010)
   (C18:3 n-3)		4.88±2.45a		1.02±0.79a	0.43±0.27a	1.58±0.65
   (C20:5 n-3)	3.49±2.79a	2.20±2.19 a		3.05±2.98 a	6.46±5.74 a	3.80±1.84
   (C22:5 n-3)	3.64±2.95 a	2.82±2.55 a		5.35±4.12 a	1.71±1.56 a	3.38±1.47
   (C22:6 n-3)	1.29±1.04 a	0.16±0.08 a		3.55±3.16 a	3.50±1.44 a	2.13±0.91
   n-3	8.42±6.76 a	10.06±2.38 a		12.98±9.79 a	12.11±6.21 a	10.89±3.41
   (C18:2 n-6)	0.76±0.48a	18.99±8.75a		11.62±3.44a	9.42±3.86 a	10.20±2.54
   (C20:4 n-6)	2.84±2.19 a	0.47±0.25 a		2.31±1.16 a	0.76±0.37 a	1.60±0.63
   (C20:2 n-6)	0.47±0.47 a	0.32±0.32 a		0.43±0.43 a	0.16±0.16 a	0.34±0.18
   n-6	4.06±2.37 a	19.78±8.87a		14.36±2.05a	10.35±3.98a	12.14±2.55
   n-3:n-6	2.08±0.85 a	0.73±0.28 a		1.03±0.80 a	1.21±0.56 a	1.26±0.33
   PUFAs	14.87±8.86 a	30.50±8.39 a		27.81±8.90 a	19.46±6.73 a	23.16±4.13
   (C16:1 n-7)	1.80±1.80 a	0.16±0.09 a		0.07±0.03 a	1.05±1.00 a	0.77±0.52
   (C18:1 n-9)		28.89±3.11b		9.73±7.21ab	7.71±3.88a	11.58±2.19
   MUFAs	1.80±1.80 a	29.06±3.17b		22.17±9.23ab	8.73±4.12ab	15.44±2.69
   (C10:0) (C11:0)	0.37±0.37 a	0.35±0.35 a 0.04±0.04 a		2.70±1.81 a	0.02±0.02 a	0.18±0.13 0.69±0.45
   (C12:0)	5.53±3.59a	0.07±0.07a	0.05±0.05a		0.21±0.07a	1.46±0.90
   (C14:0)	5.03±3.46a	0.24±0.16a	1.11±0.55a		2.16±1.07a	2.13±0.92
   (C15:0)	0.68±0.68a	0.11±0.11a	0.85±0.79a		0.38±0.19a	0.51±0.27
   (C16:0)	4.07±3.35a	24.42±4.43ab	32.63±2.30 b		22.33±10.54ab	20.86±3.03
   (C17:0)	2.57±1.97 a	1.49±1.12 a	1.54±1.39 a		3.34±3.34 a	2.24±1.07
   (C18:0)		4.34±0.99 a	4.48±2.29 a		18.05±11.61a	6.72±2.97
   (C20:0) SFAs	0.30±0.30 a 18.54±9.13a	0.12±0.12a 31.21±5.57a		43.80±2.36a	0.42±0.27 a 47.01±19.35 a	0.21±0.10 35.14±5.56

   Values are mean±S.E. and are presented as %age of total lipids; Values with same superscripts in a row do not differ significantly (p>0.05).

   Table 3Seasonal differences in fatty acids of Catla. catla(Hamilton).

   Fatty Acid	Season Spring Summer Autumn Winter	Mean (of the four
   	(April, 2009) (July, 2009) (October, 2009) (January, 2010)	seasons)
   (C18:3 n-3)	1.47±1.33a 2.81±2.02a 0.63±0.31a	1.23±0.61
   (C20:5 n-3)	1.73±0.91 a 1.83±0.91 a 10.61±10.29 a 1.75±0.76 a	3.98±2.60
   (C22:5 n-3)	7.51±3.69 a 3.52±2.39 a 2.51±2.30 a 2.07±1.23 a	3.90±1.28
   (C22:6 n-3)	4.83±4.83 a 0.18±0.18 a 2.19±2.0 a 1.93±1.28 a	2.28±1.35
   n-3	17.91±2.04 a 7.01±1.56 a 18.12±13.50 a 6.38±2.87 a	12.36±3.51
   (C18:2 n-6)	5.16±5.05a 24.29±10.43a 3.64±0.37 a	8.27±2.90
   (C20:4 n-6)	2.18±2.18 a 3.69±2.86 a 1.02±0.39 a 1.73±0.18 a	2.15±0.91
   (C20:2 n-6)	0.01±0.01 a 0.16±0.15 a 0.02±0.02 a	0.05±0.04
   n-6	2.18±2.18 a 12.19±6.71 a 25.46±10.29a 5.40±0.35 a	11.31±3.12
   n-3:n-6	1.12±1.12 a 2.22±1.75 a 1.42±1.29 a 1.21±0.53 a	1.49±0.63
   PUFAs	19.43±4.61 a 17.62±7.98 a 43.64±11.09 a 12.17±2.69 a	23.21±3.67
   (C16:1 n-7)	2.20±2.20 a 2.58±2.42 a 0.09±0.06 a 0.08±0.05 a	1.24±0.82
   (C18:1 n-9)	14.17±7.58 a 19.98±9.74a 12.57±3.35a	11.68±3.20

   MUFAs 2.20±2.20 a 16.75±7.89a 20.07±9.77a 12.65±3.39a 12.92±3.30

   (C10:0) 0.48±0.48 a 0.20±0.20 a 0.06±0.06 a 0.19±0.13

   (C11:0)	4.33±2.29 a		0.30±0.30 a		1.16±0.58
   (C12:0)	0.76±0.76 a	0.27±0.19 a	0.09±0.05 a	1.04±0.73 a	0.54±0.27
   (C14:0)		4.33±2.92 a	0.88±0.47 a	1.05±0.21 a	1.56±0.74
   (C15:0)		0.96±0.56 a	0.05±0.05 a	0.87±0.43 a	0.47±0.18
   (C16:0)	4.17±4.17 a	18.75±10.67a	16.52±7.54 a	25.11±3.90 a	16.14±3.56
   (C17:0)		0.10±0.10 a	7.38±7.38 a	0.07±0.04 a	1.89±1.84
   (C18:0)		7.48±5.08 a	4.38±2.28 a	32.03±3.68b	10.97±1.67
   (C20:0)	10.02±10.02a	0.27±0.23 a	0.04±0.04 a	0.21±0.12 a	2.64±2.50
   SFAs	19.88±14.90a	37.36±9.98 a	23.80±1.21 a	60.72±8.05 a	35.44±4.92

   Values are mean±S.E. and are presented as %age of total lipids; Values with same superscripts in a row do not differ significantly (p>0.05).

   Table 4 Seasonal differences in fatty acids of Cirrhinusmrigala(Hamilton).

   Fatty Acid	Season Spring Summer Autumn Winter				Mean (of the four
   	(April, 2009) (July, 2009) (October, 2009) (January, 2010)				seasons)
   (C18:3 n-3)	2.83±2.76a 2.22±1.18a 1.87±1.48a 1.04±0.48a				1.99±0.85
   (C20:5 n-3)	9.74±5.21 a 0.14±0.14 a 0.81±0.42 a				2.67±1.31
   (C22:5 n-3)	0.35±0.35 a 3.86±1.96 a 0.60±0.44 a 0.84±0.60 a				1.41±0.53
   (C22:6 n-3)	0.52±0.37 a 7.50±1.50b 0.04±0.02 a 1.79±0.77 a				2.46±0.43
   n-3	3.69±3.47 a 23.32±7.45b 2.64±1.87 a 4.38±1.01ab				8.51±2.12
   (C18:2 n-6)	3.20±1.55 a 18.99±13.69a 4.11±0.58 a				6.58±3.45
   (C20:4 n-6)	2.28±2.28 a 5.42±2.57 a 0.89±0.46 a 5.91±1.48a				3.63±0.94
   (C20:2 n-6) n-6	2.30±2.28 a	9.08±0.91 a	20.14±13.43a	8.72±1.82 a	0.18±0.08 10.06±3.44
   n-3:n-6	0.50±0.50 a	2.50±0.63b	0.12±0.02 a	0.44±0.10 a	0.89±0.20
   PUFAs	6.02±5.73 a	32.40±8.19 a	20.96±16.97 a	14.60±1.90 a	18.50±4.95
   (C16:1 n-7)	2.29±2.29 a	3.21±3.21 a	0.03±0.02 a	0.43±0.24 a	1.49±0.99
   (C18:1 n-9)	3.45±3.45 a	16.11±4.00a	33.27±4.94b	4.05±0.90 a	14.22±1.82
   MUFAs	5.74±5.74 a	19.33±1.70ab	33.31±4.95b	4.47±0.94 a	15.71±1.96
   (C10:0)	0.97±0.97a	1.53±0.77a	0.03±0.03a	2.00±0.71a	1.13±0.36
   (C11:0)	3.68±3.68a	0.88±0.88a	0.06±0.06a	0.21±0.13a	1.21±0.95
   (C12:0)	2.33±2.33 a	1.77±1.25 a	0.04±0.04 a	0.29±0.29 a	1.11±0.67
   (C14:0)		1.87±0.94ab	0.99±0.51ab	3.40±0.99b	1.57±0.36
   (C15:0)	1.64±1.61ab	0.19±0.19a	0.30±0.20ab	4.46±0.85b	1.65±0.46
   (C16:0)	3.87±3.46 a	34.17±3.18c	18.67±2.22b	35.26±2.56c	22.99±1.45
   (C17:0)		0.17±0.17a	0.07±0.04a	0.34±0.27a	0.14±0.08
   (C18:0)	1.38±1.38 a	3.71±0.41 a	20.73±8.10 a	19.48±2.11a	11.33±2.12
   (C20:0)	0.02±0.02a	0.79±0.79a	0.36±0.28a	0.30±0.29a	0.36±0.22
   SFAs	13.89±13.42a	44.49±4.51 ab	41.28±10.47ab	63.69±2.23 b	40.84±4.44

   Fatty Acid	Season Spring Summer Autumn Winter				Mean (of the four
   	(April, 2009) (July, 2009) (October, 2009) (January, 2010)				seasons)
   (C18:3 n-3)	2.83±2.76a 2.22±1.18a 1.87±1.48a 1.04±0.48a				1.99±0.85
   (C20:5 n-3)	9.74±5.21 a 0.14±0.14 a 0.81±0.42 a				2.67±1.31
   (C22:5 n-3)	0.35±0.35 a 3.86±1.96 a 0.60±0.44 a 0.84±0.60 a				1.41±0.53
   (C22:6 n-3)	0.52±0.37 a 7.50±1.50b 0.04±0.02 a 1.79±0.77 a				2.46±0.43
   n-3	3.69±3.47 a 23.32±7.45b 2.64±1.87 a 4.38±1.01ab				8.51±2.12
   (C18:2 n-6)	3.20±1.55 a 18.99±13.69a 4.11±0.58 a				6.58±3.45
   (C20:4 n-6)	2.28±2.28 a 5.42±2.57 a 0.89±0.46 a 5.91±1.48a				3.63±0.94
   (C20:2 n-6) n-6	2.30±2.28 a	9.08±0.91 a	20.14±13.43a	8.72±1.82 a	0.18±0.08
   n-3:n-6	0.50±0.50 a	2.50±0.63b	0.12±0.02 a	0.44±0.10 a	0.89±0.20
   PUFAs	6.02±5.73 a	32.40±8.19 a	20.96±16.97 a	14.60±1.90 a	18.50±4.95
   (C16:1 n-7)	2.29±2.29 a	3.21±3.21 a	0.03±0.02 a	0.43±0.24 a	1.49±0.99
   (C18:1 n-9)	3.45±3.45 a	16.11±4.00a	33.27±4.94b	4.05±0.90 a	14.22±1.82
   MUFAs	5.74±5.74 a	19.33±1.70ab	33.31±4.95b	4.47±0.94 a	15.71±1.96
   (C10:0)	0.97±0.97a	1.53±0.77a	0.03±0.03a	2.00±0.71a	1.13±0.36
   (C11:0)	3.68±3.68a	0.88±0.88a	0.06±0.06a	0.21±0.13a	1.21±0.95
   (C12:0)	2.33±2.33 a	1.77±1.25 a	0.04±0.04 a	0.29±0.29 a	1.11±0.67
   (C14:0)		1.87±0.94ab	0.99±0.51ab	3.40±0.99b	1.57±0.36
   (C15:0)	1.64±1.61ab	0.19±0.19a	0.30±0.20ab	4.46±0.85b	1.65±0.46
   (C16:0)	3.87±3.46 a	34.17±3.18c	18.67±2.22b	35.26±2.56c	22.99±1.45
   (C17:0)		0.17±0.17a	0.07±0.04a	0.34±0.27a	0.14±0.08
   (C18:0)	1.38±1.38 a	3.71±0.41 a	20.73±8.10 a	19.48±2.11a	11.33±2.12
   (C20:0)	0.02±0.02a	0.79±0.79a	0.36±0.28a	0.30±0.29a	0.36±0.22
   SFAs	13.89±13.42a	44.49±4.51 ab	41.28±10.47ab	63.69±2.23 b	40.84±4.44

   0.01±0.01 a 0.46±0.27 a 0.26±0.18 a

   Values are mean±S.E. and are presented as %age of total lipids; Values with same superscripts in a row do not differ significantly (p>0.05).

4. Conclusion

   Seasonal variations affected fatty acid composition of carp species. The MTLC was found maximum in C. catla during winter and minimum in

   C. mrigala during summer. Total PUFAs was obtained maximumin C. catla(43.64±11.09%) during autumn season and minimum in C. mrigala (6.02±5.73%)during spring season. The n-3:n-6 ratio may not have high nutritional value.

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