Asian Pacific Journal of Tropical Biomedicine (2011)S249-S252 S249

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Document heading Nutritional value of gastropod spirata (Linnaeus, 1758) from Thazhanguda, Southeast coast of India

Periyasamy N*, Srinivasan M, Devanathan K, Balakrishnan S

Centre of Advanced Study in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai, Tamil Nadu-608 502, India

ARTICLE INFO ABSTRACT

Article history: Objective: i.e. BabyloniaTo study t hspiratae bioc heB.m ispiratacal compMethods:osition, , protein, carboB.hy dspiratarate, lipid, and amino Received 2 August 2011 acids from ( ). The gastropod ( ) were collected Received in revised form 2 September 2011 from the Thazhanguda, Southeast coast of India. The shell was broken and body muscles Accepted 25 September 2011 (expected digestive gland) were dried at 60 in an oven and used for biochemical analysis, such Available online 15 October 2011 曟 Results: as protein, carbohydrate, lipid, amino acid and molecular size of muscle protein. The results of proximate composition showed that the percentage of protein in the tissue was (53.86%), Keywords: followed by the carbohydrate content(16.85%). The amoB.u nspiratat of lip id was 9.30%, and totally 10 essential and nonessential amino acids were analyzed in and the content was 9.911 mg/ Proximate composition g. The molecular weight of the gastropod sample was low as 2 kDa oConclusions:n SDS-PAGE. FT-IR analysis Amino acid revealed the presence of aB.m ispiratano acids signals at different ranges. The results of FTIR this research showed that meat is a valuable food with high quality protein and well- SDS-PAGE balanced amino acids.

1. Introduction are suitable for human consumption, our knowledge on its nutritive value is fragmentary. The proximate composition varies widely and depends on several factors, like , Seafood is an excellent source of protein of high biological size, sex, maturity, season and feeding regimes. Information value. Finfish, shellfish and other aquatic organisms on daily dietary intake of nutrients, especially cholesterol, suitable for food and feed are of worldwide importance. is quite important for especially those with cardiovascular They are excellent sources of high quality proteins, which problems[3]. The malnutrition problems in our country are superior to those in meat and poultry. Man lives on land, can be overcomed by effective utilization of nutrient rich which occupies a quarter of the surface of the planet, and molluscan seafood. Malnutrition is considered as a serious takes most of his food from the land. Approximately 14% of problem which is faced by developing countries. In India the animal protein consumed by human beings comes from 20%-30% of the population does not get adequate nutrition. marine fisheries[1]. Biochemical assays and nutrients play a Proper exploitation of aquatic organisms through captures vital role in physical growth, development, maintenance of and culture fisheries will supply the balanced nutritious normal body function of physical activity and health. The food and malnutrition can be controlled. A balanced diet knowledge of the biochemical composition of any edible should provide around 60%-70% of total calories from organisms is extremely important since the nutritive value carbohydrates, preferable starch, about 10%-12% from is reflected in its biochemical contents. proteins and 20%-25% from fat. Generally, fish and shellfish meat is considered to be Cuddalore district in Tamil Nadu ranks fourth in fish highly i.e.nutritious, owing to its content of Omega-3 fatty landings, with 100 tons per day. It is interesting to note acids, , essential amino acids and proteins. In addition that after tsunami thereBabylonia is an incr espirataasing treB.nd spirataof gastropod to their dietary importance, proteins affect food texture, as population especially ( )[4]. It also do small peptides and amino acids contribute to food is usually found in the depth of 32 feet. Fisherman“ used to flavor[2]. Even though large amount of marine gastropods catch” this animal by using special net known as katcha valai . Its flesh and operculum has medicinal values and

also rich in nutrition. *Corresponding author: Periyasamy N. Centre of Advanced Study in Marine Biology, Infrared and Raman spectroscopy can be used to study Faculty of Marine Sciences, Annamalai University, Parangipettai -608 502, Tamil the vibration and rotational energies of molecules. In the Nadu - India. E- mail: [email protected] study of molecular vibration, infrared spectroscopy has contributed more to this field than Raman spectroscopy Periyasamy N et al./Asian Pacific Journal of Tropical Biomedicine (2011)S249-S252 S250 due to the rapid developments in infrared instrumentation. Infrared spectral analysis of biological materials was Molecular size of muscle protein was determineetd ualsing utilized to investigate their chemical constituents. These are SDS-PAGE gel following the method of Sambrro [11]. recognized even when the availability of material is less[5,6]. Glass plates were assembled and 20 mL of 15% resolved Electrophoresis of protein samples in the polyacrylamidegel gel was prepared and poured immediately to the notch is an indispensable analytical tool and in some cases, a plate. It was overlaid with butanol. After polymerization was preparative tool for the protein scientist. Electrophoresis completed, over lay was poured off and the top layer was can be used to separate and compare complex protein washed with deionized water. Then 8 mL of stack gel was mixtures, and to evaluate purity of a protein during physic overlaid. Approximate volume of 1*SDS gel loading buffer characteristics, such as sub-unit composition, isoelectric and sample was taken and was heated at 100 曟 for μ3 min. point, size and chaB.rg espirat. Then it was fixed in electrophoresis apparatus and 15 L of The gastropod ( ) meat is used for the preparation sample and marker (6.5 - 97.4 kDa) was loaded respectively of good recipes, such as stews, soups, salads, appetizers in the well. The gel was run and stained with Coomassie and hotpot mixed with other seafood. Because of health brilliant blue. consciousness, nowadays people are interested in taking more seafood in view of its relatively higher nutrition. The 3. Results present study was carried out to analyze the proximaB.te spiratacomposition and amino acid profiles from gastropod in 3.1. Estimation of protein, carbohydrate, lipid and amino . acids

2. Materials and methods TB.he spirata protein, carbohydrate and lipid contents were found 2.1. Preparation of experimental materials in and the proportions the them are 53.86%, 16.85% and 9.3%, respectively. The percentage composition B. spirata of essential and non-essential amino acids is shown in The gastropods °( ’ ) wer°e co’llected from Table 1. The ratio o f amB.i nspiratao acids (essential and non- Thazhanguda (Lat. 11 45 N; Long. 79 45 E) southeast essential) is 9.91 mg/g in . Among the amino acids coast of India. They were brought to the laboratory, the isoleucine ranked the first in the list by contribution of 3.081 animal shell was broken and body muscles (expected mg/g on dry matter basis and other amino acids were found digestive gland) were dried at 60 曟 in an oven and used for fluctuating between 0.012 and 3.081 mg/g (Figure 1, 2). biochemical analysis. 2.2. Estimation of total protein Table 1 B. spirata. Amino acid composition of the muscle of [7] The Folin - Ciocalteu phenol method was adopted for S. No. Amino acids Ratio (mg/g) the estimation of total protein in the tissue. 1 Isoleucine 3.081 2.3. Estimation of total carbohydrate 2 Valine 2.066 3 Lysine 1.012 4 Phenylalanine 1.011 The total carbohydreta tale content was estimated by using the procedure of Dubois [8] with phenol - sulphuric acid. 5 Leucine 0.889 2.4. Estimation of total lipid 6 Methionine 0.766 7 Proline 0.665 et 8 Tryptophan 0.221 The chloroform-methanol extraction procedure of Folch 9 0 112 al[9] Glutamic acid . was used for extracting lipid from various body parts. 10 Alanine 0.088 2.5. Estimation of amino acid Total 9.911 B. spirata The experimental gastropods ( ) were dried at 60 24 曟 for hours in an oven and the dried samples were finely 100 ground for estimating the amino acids in high performance liquid chromatography (HPLC) (Merc’k hitachedL-7400) 80 [10] following the method of Baker and Han s . 60

2.6. FTIR spectrophotometry ( fourier transform- 40 n t e s i y ( m V ) infrared spectrum analysis) I 20 B. spirata The lyophilized samples of (10 mg) were 0 100 0 5 10 15 20 25 30 35 40 45 mixed with mg of dried potassium bromide (kBr) and Retention time (min) compressed to prepare a salt disc. The disc was then read Figure 1. spectrophotometrically (Bio- Rad FTIR-40- model, USA). Standard graph amino acids. The frequencies of different components present in each sample were analyzed. 2.7. Determination of molecular size of muscle protein Periyasamy N et al./Asian Pacific Journal of Tropical Biomedicine (2011)S249-S252 S251

100

80 Biochemical studies are very important from the nutritional point of view. The biochemical constituents in 60 are known to vary with season, size of the animetcal, stage of maturity, temperature and availability of food . 40 n t e s i y ( m V )

I Protein is essential for the sustenance of life and accordingly 20 exists in the largest quantity of all nutrients to form the components of the human body[12]. The acceptability and 0 easy digestibility of fish proteins make it very valuable in 0 5 10 15 20 25 30 35 40 45 combating protein malnutrition, especially in children. The Retention time (min) Figure 2. B. spirata. protein of fish has a high biological value with its capacity of Estimation of amino acids in promoting grow. Fish occupy an important part in the world 3.2. FTIR spectral analysis protein supply, accounting for about 10% of the total protein supply. About 60% of the population in the developing countries depends on 40% or more of their total animal FTIR spectrum of the lyophilized sample showed that protein supplies from fish. The average protein content of 8 major peaks were shown at 2 860.27, 1 484.9-31 , 1 342.47, fish approximately ranges from 8 g to 23 g per 100 g of wet 1 218.14, 1 152.18, 933.63, 899.B.25 ,spirata 860.39 cm , whereas edible protein. the spectrum of the sample of showed that all The phylum is the biggest and has more remaining peaks were very close and at 3388.75, 2958.90, than 85 000 representatives. The molluscs are reported 2925 32 1643 07 1539 73 1402 05 673 97 615 76 536 99 40 . , -1 . , . , . , . , . , . and to contain a high amount of proCellonatein ran rotaging from % to 465.75 cm (Figure 3). 78%[13-15]. The protein ranges of fluctuate from [16] 42.33%-82 Cerithium.45% . K rrubusishna kumary reported that the protein 90 values in varied from 38.34%-65.93% in 6 3 2 5 . 80 . 39 59 57 65 [18] 3 9 males and . %- . % in females . The recent study . 9 3 8 9

70 4 7 . 8 6 0 reported by Devanathan and Srinivasan showed that the B. spirata [4] 60 9 3 48 68 .

1 3 4 2 nutritive values of protein level is %- % . 7 5 . 1 4 8 50 9 The protein value of the present investigation revealed 1 4 a n s m i t c e . .

4 6 5 B. spirata 7 3 9 7 7 6 8 . . . . T r 1 8

5 3 6 that the maximum level of protein content in is

40 . 2 7

. Onchidium verraculatum 1 2 8 % 6 7 3 6 1 5 0 7 2 6 1 5 3 9

. . 53 86 7 5 1 0 4 . %. The protein content in the .

30 1 5 2 0 5 2 8 6 0 . 9 0

. 14 21 11 34 1 6 4 3 1 0 8 3 2 is in mantle ( . %) and in foot ( . %), which is lower . 20 3 8 Pythia plicate 1 4 0 2

2 9 5 8 than in another pulmonate snail . Thivakaran

2 9 5 Littorina 10 observed the protein value (35.94%) of the 4000 3000 2000 1000 quadricentus[18] -1 . Tagore reported that the protein value in Wavenumbers (cm ) Thais biserials [19] et Figure 3. B. spirata. al ranges from 18.5%-43.61% . Ansellm The FTIR spectrum of lyophilized sample of [20] reported that in molluscs generally the carbohydrates 3.3. SDS-PAGE reserve may be utilized under unfavorable conditions and the great variation found in the tissue indicates that the level B. spirata of mobilized carbohydrate reserves may fluctuate widely and Crude protein sample of yielded 6 bands ranging rapidly in response to fluctuation in conditions of the protein from 2.0 kDa to 110.0 kDa with well defined. Gastropod of the animBabyloniaal and the zeylanicacarbohyd raB.te szeylanica (2.54%-2.94%) obtained sample was compared with standard protein molecular from male ( ). Carbohydrates weight marker 6.5 - 97.4 kDa (Figure 4). constitute only a minor percentage of total biochemical Lane 1. Standard molecular Lane 2. Crude protein profiles in composition. Carbohydrates in fishery products contain B. spirata weight marker no dietary fiber but only glucides, the majority of which consist of glycogen. They also contain traces of glucose, fructose, sucrose and other mono and disaccharides[12]. In Lane. 1 Lane. 2 16 65 110 kDa the present study the percentage of carbohydrates( . %) is in body tissue. When compared with the other gastPlicateropods 3 04 2 98 97.4 kDa 97.4 kDa plicaspecies, it was higher than that ( . % to . %) of . Nirmal found that the level of lipid was 10.38% B. zeylanica Pleuroploca trapezium[21] in and 1.97% in B. spirata. 30 kDa In the present study lipid content is 9.3% in . 45.5 kDa Thirumavalavan has reported that the range of lipid is 3% to B. spirata[22] 10% in . RRapanaajakum rapiformisar reported that the range is 0.85%-2.12% in male and 0.95%- 2.96% 20 kDa Chicoreus ramosus in females[23]. In the lipid values were 6.5 kDa assessed to be 2% in the foot muscle[24]. 4.5 kDa 2 kDa Biological value of protein is obviously reflected upon its essential amino acids concentration. In general, the shellfish have a balanced distribution of all essential amino acids Figure 4. B. spirata. The SDS-PAGE analysis of required by an adult per day. There are 20 amino acids found in fish pi.eroteins. Some of these are listed as essential amino acids, ., arginine, histidine, isoleucine, leucine, 4. Discussion lysine, methionine, phenylalanine, threonine, tryptophan and valine. The lowest level of any of essential amino acids Periyasamy N et al./Asian Pacific Journal of Tropical Biomedicine (2011)S249-S252 S252 Molecular cloning in a protein source, “which limits the utilizatio”n of that T, editor. . New York: Cold Spring Harbour [25] Laboratory Press; 2006. protein, makes it the first limB.i tispiratang amino acid . The Nutritional and functional properties present study showed that, in , isoleucine (3.081 [12] ofOkuzumi squid and M, cuttleFujii T.fish: 35thAnniversary of commemorative mg/g) is maximum, followed by valine (2.066 mg/g) and publication lysine (1.012 mg/g). Both essential and nonessential amino . Tokyo, Japan: National Association of Squid Processors; 2000, p. 223. acids are 9.Perna911 mg viridis/g. The total compoCrassostreasition of am madrasensisino acids in Meretrix meretrix [13] Jayabal R. Studies on (Linneaus) mollusca, molluscan Meretrix casta is 95.76%, et al 98.4% and 65.17%[26]. Babu [27] reported Bivaliva, Vener idea from Porto Novo water [Dissertation]. Tamil Nadu, India: Annamalai University; 1984. that the total essential an dBursa none sspinosasential amino acids are Hemifusus pugilinus [14] Anandakumar S. Stuids on (Born) mollusca 50.01 mg/g and 46.79%et i nal (mesogastropod) respectively. Sudhakar [28] found that shellfish, the hard gastropod (Velemidae) from portonova waters [Dissertation]. and soft shell crabs totally contributed to maximum amount Tamil Nadu, India: AnnamalaiScylla University; serrata 1986. of essential amino acid (51.09%), and the minimum was [15] Raja S. The edible crab and its fishery in Indo Pacific- region. A Review [Dissertation]. Tamil Nadu, India: in soft shell crabs (43.627Nertia%). Pa crepidularialpandi reported the amino acids (totally 6) content of is 68.506% and Annamalai University; 1981. 31.012% in four essential and two non essential amino acids [16] Patil DS,Cellana Mane rotaVH. Levels of proteinIndian fat J and Mar glycogen Sci in 11the respectively[29]. Electrophoresis on SDS-PAGE revealed a six limpet (Chemnitz). 1982; : 182-183. bands protein corresponding to the molecular weB.igh spiratat. The presence of six bands indicates that the protein in [17] Krishnakumary L. Ecological and biochemical studies with special reference to pollution on selected species of molluscs has simB.il aspiratar molecular weight. Whereas the FTIR spectrum from Bombay [Dissertation]. Bombay, India: University of of the sample a-l1so recorded the n-u1mber of peaks Bombay; 1985. lying between 465.75 cm andB. 3 3spirata88.75 cm . The present Littrnids Litrolna fuadricentua research showed that gastropod meat is a valuable [18] Thivakaran GH.Nodilittorina Studies on thePyramidalis food with high quality protein and well-balanced essential (Philippi) and away and Granimard amino acids. (1983) Pros Branlria; Cittornidae from the Tranqubar rocky shore South coast of India [Dissertation]. Tamil Nadu, Acknowledgements India: Annamalai University; 1988. Thais bierialis [19] Tagore J. StudiesThais bufoon the thaisides. 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