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The Effect of Starvation on the Biochemical Composition of the Digestive Gland, the Gonads and the Adductor Muscle of the Scallop Flexopecten Glaber

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The Effect of Starvation on the Biochemical Composition of the Digestive Gland, the Gonads and the Adductor Muscle of the Scallop Flexopecten Glaber Food and Nutrition Sciences, 2013, 4, 405-413 http://dx.doi.org/10.4236/fns.2013.44052 Published Online April 2013 (http://www.scirp.org/journal/fns) The Effect of Starvation on the Biochemical Composition of the Digestive Gland, the Gonads and the Adductor Muscle of the Scallop Flexopecten glaber Khaoula Telahigue1*, Tarek Hajji1,2, Imen Rabeh1, M’hamed El Cafsi1 1Research Unit of Physiology and Aquatic Environment, Faculty of Sciences of Tunis, University of Tunis El Manar, Tunis, Tunisia; 2High Institute of Biotechnology of Sidi Thabet, University of Manouba, Ariana, Tunisia. Email: *[email protected] Received February 13th, 2013; revised March 13th, 2013; accepted March 20th, 2013 Copyright © 2013 Khaoula Telahigue et al. This is an open access article distributed under the Creative Commons Attribution Li- cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT The effects of the starvation trial on the biochemical composition and the fatty acid dynamics in the triacylglycerol frac- tion of the digestive gland, gonads and adductor muscle of the scallop Flexopecten glaber were assessed. Results show that three weeks of food deprivation induce depletion of carbohydrates and a significant decrease in proteins and lipids. The noteworthy patterns recorded for the various classes of lipids were the increase of the amount of phosphatidyletha- nolamine against a strong decline of mono-diacylglycerols, triacylglycerol and phosphatidylserine classes in gonads. These results reflect the ability of Flexopecten glaber to remodel endogenous lipid classes in order to avoid the gonads deterioration. In the starved specimens, severe declines of the n-3 polyunsaturated fatty acid group were recorded in the triacylglycerol fraction of digestive gland and adductor muscle against the increase of this group in gonads. These re- sults confirm the role of triacylglycerol as a polyunsaturated fatty acids reservoir and pointed out to their mobilization from storage organs to the developing gonads during the food shortage trial. Examination of fatty acid data revealed that food deprivation lead Flexopecten glaber to invest in saving and accumulation of highly unsaturated fatty acids in go- nads. This applies mainly to the arachidonic acid (20:4n-6) and the docosahexaenoic acid (22:6n-3). Keywords: Starvation; Scallop; Chemical Composition; Organs; Fatty Acid 1. Introduction lagoon of Bizerte. The relatively low density of this spe- cie in its natural habitat and its interesting nutritional Bivalve mollusks experience nutritive stress, particularly values allowed us to consider F. glaber as a potential under critical environment conditions and during the pu- species for aquaculture [12]. In the case of bivalves, dif- rification process for the consumed species [1,2]. Under ferences in the response to food availability may be of a stressful conditions, bivalves’ survival depends on their great interest when considering the possibility of devel- metabolic reserves [3,4]. The relative contribution made oping their culture [13]. by lipids, carbohydrates and proteins towards the utiliza- The goals of this study is to gain information regarding tion of energy reserves during starvation depends on the the effects of nutritive stress on the biochemical compo- species and their stage of development [5,6]. According sition and the fatty acid dynamics in lipid classes of the F. to many authors, the digestive gland and the adductor glaber digestive gland, gonads and muscle. It has also muscle represent the principal energy storage organs in allowed to assess the relative importance of different pectinids [7-10]. Under a period of high energy demands, metabolites and the strategy adopted by F. glaber to a transfer of metabolites from the digestive gland and the modulate its endogenous metabolic reserves to overcome adductor muscle to the gonad was described in different the starvation trial. scallop species [4,9]. The scallop Flexopecten glaber is endemic Mediterra- 2. Material and Methods nean species [11]. In Tunisia, this bivalve occurs in the 2.1. Experimental Design and Animal Sampling *Corresponding author. A total of 96 wild adult Flexopecten glaber individuals Copyright © 2013 SciRes. FNS 406 The Effect of Starvation on the Biochemical Composition of the Digestive Gland, the Gonads and the Adductor Muscle of the Scallop Flexopecten glaber of mean shell height 43 ± 3 mm were collected by scuba 2.2.3. Analysis of Fatty Acids diving from the Bizerte lagoon in the North east of Tuni- After evaporation to dryness, lipid extracts and fractions sia (longitude 9˚48'E - 9˚56'E and latitude 37˚08'N - were trans-esterified according to the method of Cecchi 37˚14'N) in the spring season. Thirty six specimens were et al., [18]. Methyl nonadecanoate 19:0 (Sigma), which sacrificed upon arrival to the laboratory and considered didn’t exist in our samples, was added as internal stan- as initial condition or “T0” (time zero) group. The other dard. Separation of FAMEs was carried out on a HP specimens were divided into equal groups and placed in 6890 gas chromatograph with a split/splitless injector three tanks (20 individuals per tank). Each tank contained equipped with a flame ionization detector at 275˚C, and a 70 L of filtered and UV-treated seawater maintained at 30 m HP Innowax capillary column with an internal di- constant temperature (15˚C) and salinity (35 psu). A ameter of 250 µm and a film thickness of 0.25 µm. In- photoperiod of 12-h light/12-h dark was sustained over jector temperature was held at 250˚C. The oven was pro- the duration of the experiment. grammed to rise from 50˚C to 180˚C at a rate of 4˚C/min, During 3 weeks, samples were maintained unfed. A from 180˚C to 220˚C at 1.33˚C/min and to stabilize at gentile aeration was applied in tanks to ensure an ade- 220˚C for 7 minutes. Nitrogen was the carrier gas. Iden- quate supply of oxygen. Water was continuously filtrated tification of FAMEs was based on the comparison of and changed twice per week. their retention times with those of a mixture of methyl esters (SUPELCO PUFA-3). Fatty acid peaks were inte- 2.2. Chemical Analysis grated and analyzed using the HP chemstation software. 2.2.1. Gross Chemical Composition 2.3. Statistical Analysis Total lipids were extracted according to the method of Folch et al., [14] with the solvent mixture chloroform- To assess significant differences between means, data methanol (2:1, v/v) containing 0.01% buthylatedhydroxy was analyzed using the software R Version 2.1.2.1. ac- toluene (BHT) as an antioxidant. Carbohydrate and pro- cording to the One Way Analysis of Variance method tein contents were colorimetrically quantified as de- (ANOVA) method. In this respect, Duncan test was ap- scribed by Dubois et al., [15] and Lowry et al., [16] re- plied and differences were considered significant when p spectively. < 0.05. 2.2.2. Lipid Classes’ Separation 3. Results Lipid classes were separated using thin-layer chroma- 3.1. Gross Chemical Composition tography (TLC) with one dimensional double develop- ment as described in Olsen and Henderson [17]. Doses of Data related to the effects of food deprivation on the 500 µl of lipid extracts were separated on plates (20 × 20 biochemical composition of the digestive gland, the go- cm, silica gel 60, Merck, Germany) using hexane: diethyl nads and the adductor muscle of F. glaber are presented ether: glacial acetic acid (80:20:2, v/v) as developing in Table 1. We noticed that in control individuals (ini- solvent for neutral lipids and methyl acetate: isopropanol: tial), lipids were the major biochemical components in chloroform: methanol: 0.25% KCl (25:25:25:10:9, v/v) the digestive gland and gonads. Glycogen is mainly as developing solvent for polar fraction. Lipid classes found in adductor muscle reaching 29.5 mgg−1ww. were visualized under UV light after spraying with 0.1% During the fasting trial, up to 60% of proteins were mo- 2’-7’dichloro-fluorescein in absolute methanol. bilized in the studied organs. The highest loss of protein Table 1. The effect of the duration of food deprivation on the biochemical composition, proportions of neutral and polar lip- ids (percentage of total lipid), in the digestive gland, the gonads and the adductor muscle of Flexopecten glaber. Digestive gland Gonads Muscle Initial Unfed Initial Unfed Initial Unfed Proteins (mgg−1ww) 13.7 ± 0.5 3.8 ± 0.6 9.5 ± 1.2 3.6 ± 0.2 10.7 ± 0.2 4.2 ± 0.2 Glycogen (mgg−1ww) 3.6 ± 0.1 0.4 ± 0.1 4.0 ± 0.7 0.1 ± 0.0 29.5 ± 0.8 0.1 ± 0.0 Lipids (mgg−1ww) 59.5 ± 1.1 19.1 ± 0.8 48.0 ± 0.5 22.9 ± 0.6 28.2 ± 0.9 3.9 ± 0.9 Neutral lipids (%) 91.5 2.7 88.7 0.8 71.9 ± 2.6 67.7 ± 1.4 16.1 ± 0.2 35.5 ± 1.5 Polar lipids (%) 8.5 0.5 11.3 1.0 28.1 ± 0.2 32.3 ± 1.2 83.9 ± 0.9 64.5 ± 0.7 Results are mean ± standard deviation of triplicate analysis (n = 12) except for neutral and polar lipids (n = 6). Copyright © 2013 SciRes. FNS The Effect of Starvation on the Biochemical Composition of the Digestive Gland, the Gonads and the 407 Adductor Muscle of the Scallop Flexopecten glaber was recorded in the digestive gland with 72% of the ini- neutral lipids against an increase of the polar ones in the tial amount. Concerning glycogen, we noted that this digestive gland and the gonads. compound dropped to a complete depletion after 21 days in different examined tissues.
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