National Academy Science Letters ISSN 0250-541X Natl. Acad. Sci. Lett. DOI 10.1007/s40009-013-0137-z
Your article is protected by copyright and all rights are held exclusively by The National Academy of Sciences, India. This e-offprint is for personal use only and shall not be selfarchived in electronic repositories. If you wish
to self-archive your article, please use the accepted manuscript version for posting on your own website. You may further deposit the accepted manuscript version in any repository, provided it is only made publicly available 12 months after official publication or later and provided acknowledgement is given to the original source of publication and a link is inserted to the published article on Springer's website. The link must be accompanied by the following text: "The final publication is available at link.springer.com”.
Author's personal copy Natl. Acad. Sci. Lett. DOI 10.1007/s40009-013-0137-z
Received: 17 September 2012 / Revised: 14 February 2013 / Accepted: 1 April 2013 Ó The National Academy of Sciences, India 2013
Abstract Molluscs are delicious and protein rich food among the sea foods. The gastropods in the coastline could form an important source of food, raw material for village industries, indigenous medicine, etc., and it is widely used as a cheaper food source for coastal area people. Due to the importance and abundance of the commercial gastropod species, Babylonia zeylanica that commonly occur in intertidal areas of Indian coasts; this study investigates the nutritive value of B. zeylanica. Identification and quantification of B. zeylanica were done by using standard methodology. The results of the present study provides not only the information about the composition but also recommended the consumption of this whelk B. zeylanica since they are rich in minerals, vitamins,. It could also be added that the consumption of marine gastropod is a nutritional assurance to millions of malnourished hungry people. The malnutrition problem in our country can be overcome by effective utilization of nutrient rich molluscan seafood. Overall, B. zeylanica was similar or superior to other marine shellfish and hence B. zeylanica could be utilized as an alternative meat source in human diet.
Introduction Humans and shellfish have been inextricably linked for millennia, not only as an important animal protein source, providing many millions of livelihood means and food security at large, but also from an evolutionary viewpoint. Fatty acids are the fundamental structural components of practically all forms of lipids. From the point of view of human health, it is thus important to increase the consumption of fish and other marine organisms which are rich in polyunsaturated fatty acids of the n-3 family and poor in polyunsaturated fatty acids of n-6 family . Vitamins are organic compounds representing a minor fraction in the day today life. Based on solubility, vitamins are differentiated as fat soluble A, D, E and K and water soluble B and C. Mollusc is a good dietary source of vitamin B complex. Minerals are required for the normal life processes of all animals including Molluscs. The edible and commercially important species B. zeylanica is abundantly distributed in the southeast coast of India [2, 3]. The common species of Babylon snail (Babylonia areolata) are marketed alive in the fish markets of Philippines and Thailand .
Keywords Fatty acids Á Amino acids Á Vitamins Á Minerals Á B. zeylanica Materials and Methods
K. Kesavan (&) Á A. Murugan Centre of Advanced Study in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai 608 502, Tamil Nadu, India e-mail: firstname.lastname@example.org V. Venkatesan Central Marine Fisheries Research Institute, Cochin 682018, Kerala, India
Random samples of 35–50 specimens of Babylon snail B. zeylanica were collected from Cuddalore fishing harbor which is one of the major fish landing centre in Tamil Nadu, Southeast coast of India situated at (latitude 11°440 N; longitude 79°460 E) were brought to the laboratory; the animals with size group ranging from 50 to 65 mm in length were selected for biochemical studies. The biochemical analysis was carried out in the entire body of the snails. The body parts were dried at constant
Author's personal copy K. Kesavan et al.
temperature of 60 °C for 24 h in a hot air oven. Then the dried meat was powdered and the required quantity was taken for the biochemical analysis. The estimation of amino acids, fatty acids, vitamins and minerals was done by following the standard procedures. Estimation of Amino Acid The collected gastropod samples were dried at 60 °C for 24 h in an oven and they were packed in airtight polyethylene covers and kept in desiccators. The oven-dried samples were finely grounded before estimating amino acid profile. Amino acids were estimated in HPLC—Lachrom e merck in SPD—10A VP detector. Fatty Acid Analysis For fatty acid analysis, the samples (foot and mantle) were homogenized with chloroform:methanol (2:1 v/v) mixture and the samples were extracted using the method . After the fat was extracted, it was esterified with 1 % H2SO4 and fatty acid methyl esters were prepared by following the procedure . Identification and quantification of fatty acids were done using gas chromatography (Hewlett Packard 5890 model). Estimation of Vitamins The fat soluble vitamins A, D, E and K and the water soluble vitamins B1, B2, B6, B12 and C were analysed in the HPLC (Merck Hitachi L-74000) following the method . The folic acid was estimated by following the calorimetric procedure . The pyridoxine, pantothenic acid and vitamin B12 were estimated by following methods suggested in USP NF 2000 Asian edition. Estimation of Minerals The samples were oven dried at 60 °C for 24 h and used for the estimation of mineral content. Sample Preparation and Derivatization 500 mg of dried sample was digested by microwave sample preparation system (Anton Paar Multiwave 3000) using an acid mixture containing nitric acid and perchloric acid (3:1 V/V). The residues were dissolved in 2 N hydrochloric acid and filtered through Whatman No. 1 filter paper and the volume was made up to 25 ml with deionized water in a standard flask. The clear solution was used to measure the concentration of different minerals. Minerals such as sodium, potassium and calcium were analyzed using digital flame photo meter (Model CL 22 D)
pre-calibrated with respective standards. Magnesium, phosphorus, iron, zinc, copper, manganese, nickel and cobalt determinations were performed by optical emissionspectrophotometer (Perkin Elmer Model Optima 2100 DV). The trace minerals were quantified on the basis of peak areas and comparison with a calibration curve obtained with the corresponding standards.
Results and Discussion Amino Acids Results of the amino acid estimation are presented in Table 1 Total amino acid content was 95.82 % and the percentage of unidentified amino acids was 4.18 %. The result of the present study showed that the percentage of essential amino acids (EAA) was more (77.85 %) than that of non-essential amino acids (NEAA, 18.04 %). EAA determined were arginine, histidine, isoleucine, leucine, methionine, phenylalanine, threonine and of which methionine constituted the maximum percentage (19.54 %) and Isoleucine the minimum concentration (3.45 %). The contribution of arginine, histidine, leucine, phenylalanine, threonine were also in considerable amount. The non-essential amino acid detected was alanine, asparagine, glutamic acid, glutamine, glycine and cystine. In this study glutamine 5.54 % which is found maximum and glutamic acid 0.46 % were minimum contributors in the NEAA. Fatty Acids In the present study, 24 individual fatty acids were identified. Among them, saturated fatty acids (SFA) were more (63.51 %) and most of which were 16:0 (22.32 %) and 18:0 (15.86 %) (Table 2).The monounsaturated fatty acids (MUFA) were the next common fatty acids (27.68 %) with the higher levels of C20:1n-7 (6.93 %) and 16:1x7c (5.16 %). The Poly unsaturated fatty acids (PUFA), which occupied the third position, contributed 8.78 % of total fatty acids, represented by C20:4n-12 (4 x-6) (4.14 %) and 20:2 (5.05 %) which together accounted for PUFA. Omega-6 and omega-3 fatty acids accounted for 6.8 and 1.98 % of the total PUFA. Vitamins Among the vitamins detected fat soluble vitamins such as A (retinol 134.78 IU), D (calciferol in traces), E (tocopherols 14.55 mg), K (phylloquinone 1.131 mg) and water soluble vitamins C (ascorbic acid 15.33 mg), B6 (pyroxidine 0.113 mg) and B12 (cobalamins 0.003 mg) were observed (Table 3).
Author's personal copy Nutritive Profile of B.zeylanica Table 1 List of amino acids present in B. zeylanica
Table 2 Fatty acid profile of B. zeylanica
Saturated fatty acids (SFA)
Glutamic acid Glutamine
C18:0 ISO C18:1 ISO H
Mono unsaturated fatty acids (MUFA)
Minerals The quantity of minerals present in muscle tissue varied significantly. Totally five macro minerals and two micro minerals were detected. Among the macro minerals, calcium (124.5 mg) and magnesium (34.5 mg) were present at higher and lower levels, while other macro-minerals sodium (0.1441 mg) potassium (0.3043 mg) and iron (10.1 mg) were found in lower levels. The micro minerals such as and zinc (12.3 mg) and copper (0.1121 mg) were also detected in this species (Table 4). This study is to evaluate the biochemical constituents such as amino acids, fatty acids, vitamins and minerals in the spiral Babylon B. zeylanica. Pronounced seasonal changes in the biochemical constituents have already been reported in various gastropods [9–12]. Increases in biochemical constituents in B. zeylanica were noted from December to February and maximum during March and April due to maturation of gonads when gonads were seen fully matured. Biochemical constituents decreased slowly from July to September. Estimation of amino acids revealed that the total amino acid content was 95.82 % and the unidentified amino acids 4.18 %. The result of the present study also showed that the percentage of EAA was more (77.85 %) than that of NEAA (18.04 %). Among the EAA, methionine contributed the highest percentage (19.54 %) and isoleucine the lowest concentration (3.45 %). The non-essential amino acid detected was alanine, asparagine, glutamic acid, glutamine, glycine, cystine. In this study glutamine (5.54 %) is found to be maximum and glutamic acid (0.46 %) found to be minimum contributors in the NEAA. Most of the
Poly unsaturated fatty acids (PUFA) 21.
NEAA were denatured by acid hydrolysis of proteins. In previous study from the Portonovo waters three species of bivalve Molluscs Amusium pleuronectes (Linnaeus) Meretrix meretrix (Linnaeus) and Katelysia opima (Gmelin) were analysed for free amino acid and protein amino acid. Among the free amino acids, alanine, glutamic acid, glycine, proline, aspartic acid, phenylalanine and threonine showed high concentrations. In this study, the same results of phenylalanine, methionine, alanine and glutamic acid were revealed . The amino acid content in the bivalve Molluscs Perna viridis, Crassostrea madrasensis and Meretrix casta. Totally 18 amino acid were recorded. The total amino acid in the P. viridis was 95.76 % among them essential amino acid 47.28 %, in C. madrasensis total amino acid 98.4 % among them essential amino acid 54.52 % and in the M. casta least amount was found to be 65.17 % among them essential amino acid 38.17 % .
Author's personal copy K. Kesavan et al. Table 3 Vitamins profile of B. zeylanica
Composition (mg/100 g)
Recommended dietary intake for male (age 19–70)a
C (ascorbic acid)
Table 4 Minerals profile of B. zeylanica S. No.
Composition (mg/100 g)
Micro minerals 6.
The total amino acid content of G. tumidum was reported and from that the essential amino acids contents was (EAA) were 20.77 % and NEAA were 22.20 %. Among them, phenylalanine was 1.10, 1.19, 1.00 % and methionine was 1.02, 1.04 and 1.00 % in mantle, viscera and foot, respectively which forms the major EAA. Alanine 1.09, 1.10, 1.02 % and glutamic acid 0.99, 1.08, 0.86 % contributed as major NEAA. Lysine in EAA and serine in NEAA showed the lowest concentration . In the present study the total amino acid content was 95.82 % and the unidentified amino acids 4.18 %. The result of the present study also showed that the percentage of EAA was more (77.85 %) than that of NEAA (18.04 %). In neuroscience, glutamate is an important neurotransmitter which plays a key role in long-term potentiation and is important for learning and memory and in the present study it is recorded as 5.54 %. So intake of this animal (B. zeylanica) may help neurological transmission as reported . In the present study, 24 individual fatty acids were identified. Among them, SFA were more (63.51 %) and most of which were 16:0 (22.32 %) and 18:0 (10.36 %). The MUFA were the next common fatty acids (27.68 %) with the higher levels of C20:1n-7 (6.93 %) and 16:1x7c (5.16 %). The PUFA, which occupied the third position, contributed 8.78 % of total fatty acids, represented by C20:4n-12 (4.14 % %) and 20:2 (5.05 %) which together
accounted for PUFA. Omega-6 and omega-3 fatty acids accounted for 6.8 and 1.98 % of the total PUFA. In the freeze dried and frozen samples of green lipped mussel, Perna canaliculus, reported that, 30 individual fatty acids and polyunsaturated fatty acids (40–41 % of total fatty acids) were found to be dominant . Likewise in abalones (both wild and cultured), the saturated, MUFA and PUFA contributed 31–32, 19–22 and 47–49 % respectively . In the present study also, more or less the same MUFA content was found. But there is a drastic difference between the levels of SFA and PUFA. PUFA tends to reduce the blood cholesterol levels and is considered a ‘‘good’’ fat. In the present study arachidonic acid (C 20:4) contributed 4.14 % of the total PUFA content. Since, B. zeylanica is reported to be having 4.14 % of arachidonic acid; it could be considered as a good source for this fatty acid. In the present study the levels of fatty acids such as C22:3x3 and 20:4x6 fatty acids were found to be 1.98 and 4.14 % respectively and hence B. zeylanica could be considered as a good source of these polyunsaturated fatty acids and could be used as good seafood and also in aquaculture for formulating the feed. x3FA deficiency was reported in patients with neuropathy in an interesting review article ; low DHA levels in patients suffering from schizophrenia, depression, dementia, parkinsonism and other behavioral disorders . They described that in some of the cases x3FA supplementation had positive effects on the neurological symptoms. Hence dietary intake of omega-3 fatty acids is helpful in pronouncing less inflammatory responses towards bronchial asthma, lupus erythematosus multiple sclerosis, psoriasis and kidney diseases and also inhibit the development of cancer cells. In this study the level of x3FA has been estimated to be 1.98 %. So, intake of this animal B. zeylanica may be of help in curing neurological symptoms. In the present study, the level of linoleic acid was recorded as 1.64 %. In future the consumption of B. zeylanica may fulfill the daily requirements of linoleic acid, thereby preventing skin changes. The minimal alpha
Author's personal copy Nutritive Profile of B.zeylanica
linoleic acid requirement for a 7-year-old girl is 0.54 % . Therefore, this species could be better to utilize for edible purpose. The second type of fat is MUFA. These fats are often referred to as ‘‘good’’ fats because studies have shown that they help in reducing blood cholesterol levels and protect against heart disease. The suggested fats are all monounsaturated or polyunsaturated fats because of their benefits to health. In the present study MUFA constituted 27.68 % of the total fatty acids. Among them C20:1n-7 (6.93 %) and 16:1x7c (5.16 %) were dominant. Gastropods were found to contain 18:1 major fatty acid [22, 23]. Oleic acid (18:1) contributed more than 10 % in Chlamys tehuecha . In the present investigation B. zeylanica had 2.28 % of 18:1x9c acid. But earlier studies suggested that MUFA content was observed 23 % in the freeze dried and frozen green lipped mussel P. canaliculus . In the present study, SFA such as lauric acid, tridecanoic, myristic, pentadecanoic, palmitic, heptadecanoic, stearic, nonadecanoic, arachidic, heneicosanoic, behenic, tritricosanoin and lingoceroyl acid were found in B. zeylanica and collectively the SFA were present in much greater quantity than unsaturated ones. Among the SFA, stearic acid occurred in the largest quantity. The water soluble esters of lauric acid and of palmitic acid, Tween 20 (polyoxyethylene sorbitan monolaurate), G2144 and Tween 40 (polyoxyethylene sorbitan monopalmitate) exhibited appreciable bacteriostatic and bactericidal activity against tubercle bacilli in concentrations of 0.01–0.001 %, but esters of stearic and oleic acid (Tween 60 polyoxyethylene sorbitan monostearate) and Tween 80 (PSM, polyoxyethylene sorbitan monooleate) were found inhibitory only at higher concentrations . In the present study the percentages of lauric and palmitic acid were found to 1.64 and 1.02 %. That way B. zeylanica could be a better alternative source (against tubercle bacilli and bactericidal effect), since it contains both palmitic and lauric acids. In previous studies 36 individual fatty acids that were identified in the wedge clam Donax cuneatus. Among them, the SFA were the dominant fatty acids (35.28 %) and most of which were 18:0 (15.68 %) and 14:0 (7.15 %). The MUFA were the next most common fatty acids (26.57 %) with the higher levels of 16:1x7c (12.71 %) and 18:1x9c (11.18 %). The PUFA occupying the third position contributed 11.72 % of total fatty acids, represented by 20:4 (6.75 %) and 18:2 (2.41 %) together accounted for about 90 % of the PUFA. At the same time, the omega-6 and omega-3 fatty acids accounted for 10.74 and 0.48 % of the total PUFA . In this study, only seven fatty acids were recorded. Among them, three were saturated fatty acid, one was unsaturated fatty acid and three were polyunsaturated fatty acids. In the present study, 24 individual fatty acids
were identified and most of the important acids were also recorded as of the previous study. The results of the present study provide not only information about the fatty acid composition, but also suggest the consumption of this Spiral Babylon B. zeylanica since it is rich in stearic, linoleic, omega-3 and arachidonic acids. It could also be added that the consumption of marine Molluscs is a nutritional insurance to millions of malnourished hungry people. The malnutrition problem in our country can be overcome by effective utilization of nutrient rich molluscan seafood. Further, the presence of 22:3x3 and 18:2x6 fatty acids in this Indian Babylon adds more value through the possibility of utilizing this whelk in the aquaculture feed industry also, as a feed ingredient. Vitamins are essential for the normal growth and development of a multicellular organism. These nutrients facilitate the chemical reactions that produce among other things, skin, bone, and muscle. Even minor deficiencies may cause permanent damage. For the most part, vitamins are obtained with food, but a few are obtained by other means. Microorganisms in the intestine commonly known as ‘‘gut flora’’ produce vitamin K and biotin, while one form of vitamin D is synthesized in the skin with the help of the natural ultraviolet wavelength of sunlight. Vitamins A, D, and B12 are stored in significant amounts in the human body, mainly in the liver. Well-known human vitamin deficiencies involve vitamin C (scurvy), vitamin K (Bleeding diathesis), vitamin A (night-blindness). In B. zeylanica among the vitamins detected fat soluble vitamins such as A, E and K were found in high levels than the prescribed levels  for human dietary intake whereas in water soluble vitamins B12 was found higher than that of given level. The other vitamins were observed in optimal level and in nutrition point it is good to utilize this species as a supplement in the daily intake for vitamin malnutrition and to prevent these diseases. Minerals also constitute important components of hormones, enzymes and enzyme activators in human nutrition . Mineral deficiencies can cause biochemical, structural and functional pathologies which depend on several factors, including the duration and degree of mineral deprivation. The present study ascertained the levels of nutritionally significant minerals and their variations in B. zeylanica. The quantity of minerals present in muscle tissue varied significantly. Totally five macro minerals and two micro minerals were detected. The Molluscs are a good source of calcium. Some types of seafood are excellent sources of dietary calcium with values varying from 5 to 200 mg per 100 g. Calcium is the key constituent of bones and teeth; essential for vital metabolic processes such as nerve function, muscle contraction, and blood clotting.
Author's personal copy K. Kesavan et al.
Deficiency may lead to osteomalacia, osteoporosis, rickets and tetany. In the present study, the calcium was more in B. zeylanica and it constituted (124.5 mg per 100 g dry weight) of total minerals analyzed. Magnesium is needed for functioning of approximately 90 enzymes. Deficiency can occur gradually, leading to: anxiety; fatigue; insomnia; muscular problems; nausea; premenstrual problems . Magnesium was observed (34.5 mg) as the second highest of all the minerals in the present study. Among the micro minerals zinc (12.3 mg) was recorded as maximum. Zinc is needed for functioning of many over 200 enzymes for a strong immune system. Deficiency may lead to lesions on the skin, oesophagus and cornea, retarded growth of children.
Conclusions It is proved yet again that the shellfish, particularly Molluscs are excellent source of amino acids, fatty acids, vitamins and minerals. To meet the requirements of the ever increasing population of India and to commensurate the catch with the demand, non-conventional sources like ivory shell B. zeylanica can very well be used. Acknowledgments Authors are thankful to the Dean and Director, CAS in Marine Biology, Faculty of Marine Sciences and authorities of Annamalai for providing necessary facilities.
References 1. Burr ML (1989) Fish and the cardiovascular system. Prog Food Nutr Sci 13:291–316 2. Satyamurthi ST (1952) The mollusca of Krusadai Island (on the Gulf of Mannar) I Amphineura and Gastropoda. Bull Madras Gov Mus 1(2):1–6 3. Rao NVS (2003) Indian sea shells 1 Polyplacophora and Gastropods. Zoological Survey of India, Kolkata, pp 20–337 4. Srimukda B, Chaweepack S, Nupin V (2005) Commercial production of spotted Babylon (Babylonia areolata Link, 1807) seeds. Technical paper no 24/2005. Chanthaburi Coastal Fisheries Research and Development Center, Coastal Fisheries Research and Development Bureau,Department of Fisheries Ministry of Agriculture and Cooperatives. Bangkok, p 34 5. Bligh EG, Dyer WJ (1959) A rapid method for total lipid extraction and purification. Can J Biochem Physiol 37(8):911–917 6. AOAC (1995) Official methods of analysis, 16th edn. Association of Official Analytical Chemists, Washington DC 7. Sadasivam S, Manickam A (1996) Biochemical methods, 2nd edn. New Age International (P) Ltd Publisher, New Delhi, pp 179–186 8. Sethi PD (1997) Quantitative analysis of drugs in pharmaceutical formulation, 3rd edn. CBS. Publishers and distributors, New Delhi, pp 589–590 9. Stella C (1995) Studies on the taxonomy and eco-biology of Chicoreus species (Class Gastropoda Family Muricidae) from
123 View publication stats
Parangipettai waters southeast coast of India. PhD thesis, Annamalai University, Chidambaram, p 195 Sanjeevi SB (2001) Studies on ecobiology of the spider conch Lambis lambis (Gastropoda Prosobranchia Strombidae) from Mandapam waters south east cost of India. PhD thesis, Annamalai University, Chidambaram, p 160 Arularasan S (2010) Studies on eco-biology of the dog conch Strombus canarium (Linnaeus 1758) (Gastropoda Prosobranchia Strombidae) from the Gulf of Mannar marine biosphere reserve south east coast of India. PhD thesis, Annamalai University, Chidambaram, p 180 Palpandi C (2010) Studies on mollusca (biology biochemistry and heavy metal accumulation) in Nertia (Dostia) crepidularia Lamarck1822 from mangroves of Vellar estuary south east coast of India. PhD thesis, Annamalai University, Chidambaram Kalyani M (1991) Amino acid composition of Amussium pleronectes (Linnaeus) Katelysia opima (Gmelin) and Meretrix meretrix (Linnaeus) (Mollusca: Bivalvia) from Porto Novo waters India. In: Proceedings of Tenth International Malacological Congress (Tubingen 1989), pp 163–166 Baskar DA (2002) Nutritional evaluation of molluscan seafood. PhD thesis, Annamalai University, Chidambaram, p 129 Babu A, Venkatesan V, Rajagopal S (2012) Biochemical composition of different body parts of Gafrarium tumidum (Roding, 1798) from Mandapam South East Coast of India. Afr J Biotechnol 11(7):1700–1704 Sapolsky R (2005) Biology and human behavior: the neurological origins of individuality, 2nd edn. The Teaching Company Guide Book, Chantilly, pp 19–20 Murphy KJ, Mann NJ, Sinclair AJ (2003) Fatty acid and sterol composition of frozen and freeze-dried New Zealand green lipped mussel (Perna canaliculus) from three sites in New Zealand. Asia Pac J Clin Nutr 12(1):50–60 Dunstan GA, Baillie HA, Barrett SM, Volkman JK (1996) Effect of diet on the lipid composition of wild and cultured abalone. Aquaculture 140:115–127 Holman RT (1998) The slow discovery of the importance of omega 3 essential fatty acids in human health. J Nutr 128:427–433 Yoshida S, Sato A, Okuyama H (1998) Pathophysiological effect of dietary essential fatty acid balance on neural systems. Jpn J Pharmcol 77:11–22 Holman RT, Johnson SB, Hatch TF (1982) A case of human linolenic acid deficiency involving neurological abnormalities. Am J Clin Nutr 35:617–623 Ackman RG, Hooper SN, Ke PJ (1971) The distribution of saturated and isoprenoid fatty acids in the lipids of three species of Molluscs, Littorina littorea, Crassostrea virginica and Venus mercenaria. Comp Biochem Physiol 39B:579–587 Johns RB, Nichols PD, Perry GJ (1980) Fatty acid components of nine species of molluscs of the littoral zone from Australian waters. Comp Biochem Physiol 62B:207–214 Pollero RJ, Maria R, Brenner R (1979) Seasonal changes of the lipids of the molluscs Chlamys tehuelcha Catedra de Bioquimica Instituto de Fisiologia Facultad de ciencias medicas Universidad National de La plata Argentina and centro National Patagonica Madryn Argentina. Comp Biochem Physiol 64A:257–263 Dubois RJ (1947) The effect of lipids and serum albumin on bacterial growth. J Exp Med 85:9–22 Shanmugam A, Palpandi C, Sambasivam S (2007) Some valuable fatty acids exposed from wedge clam Donax cuneatus (Linnaeus). Afr J Biochem Res 1(2):14–18 DRI (2001) Dietary reference intake: vitamins. The National Academics, Washington, DC