International Journal of Pure and Applied Sciences and Technology

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1 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), pp International Journal of Pure and Applied Sciences and Technology ISSN Available online at Research Paper Role of Food Additives on Young Age Silkworm (Bombyxmori.L) Rearing Pallavi 1,*, M. Muthuswami 2, R.N. Bhaskar 3 and V. Naveen 4 1,3,4 College of Sericulture, Chintamani University of Agricultural Sciences, Bangalore, Karnataka, India 2 Department of Seed Science and Technology, Tamil Nadu Agricultural University, Coimbatore (T.N), India * Corresponding author, (pallavireddy214@gmail.com) (Received: ; Accepted: ) Abstract: The overall development of silkworm larvae depends on getting the required food which can be easily digested and assimilated in the body tissues properly for the silk synthesis. Keeping this in view, an attempt has been made to increase the growth of young silkworm Bombyx mori. This is ensured through supplementation of food additives, intern biochemical constituents of worm. The results of the experiment revealed that, food additives were found effective in increasing the protein, carbohydrate and lipid content in Chawki worms. Among silkworm hybrids viz, (PM x CSR2, CSR2 xcsr4, APM1 x APS8, APM2 x APS12 and (CSR6 x CSR26) x (CSR2 x CSR27)), the highest protein content of 19.91mg, 21.44mg, 19.83mg, 19.71mg and 24.03mg was recorded on soya flour dusted 10g/kg of respectively, compare to control (17.63mg, 18.02mg,17.58mg, 17.51mg and 22.72mg). The same trend was noticed in lipid content of all the hybrids. Further it is very much reflected that, even the highest carbohydrate content in the body of young silkworm was recorded on supplementation of corn 10g / kg of followed by corn flour 5g / kg of in all the silkworm breeds. Keywords: Young age silkworm, Food additives, Silk synthesis, Protein, Carbohydrate, Lipid 1. Introduction: Sericulture activities are agro-based and the industrial sector. The agro-based part involves mulberry cultivation and silkworm rearing. In Silkworm rearing, Chawki is considered as one of the important stage from which a successful cocoon crop is ensured. The first three instars are highly sensitive and require at most care for their growth and development. Chawki rearing needs optimum environmental conditions which can sustain

2 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), the outbreak of silkworm disease avoiding crop failures during late age rearing (Bedford, 1944; Jolly, 1986). Various factors have shown to affect the success of silkworm cocoon production including environment (37%), mulberry leaf quality (38.2%), rearing technique (9.3%), silkworm race (4.2%), silkworm egg quality (3.1%) and others (8.2%) (Mastsumara, 1975; MukulDeka, 2011) Owing to the high cost of artificial food, introduction of these techniques in India is very limited. Hence an economically cheaper technique which could still improve cocoon character and production is needed (JOCV, 1980). Fortification of mulberry with soybean products fulfilled the above need. Quantitative and qualitative supplementation of protein plays a critical role in influencing the growth and development of the insect. Elevation of dietary protein to an optional level (Horieet al., 1971) and supplementation of low nutritive proteins with their limiting amino acids (Ito and Arai, 1965) have found to accelerate the growth of the silkworm. Ito (1980) stated that rich sources of dietary proteins like soyaprotein are known to promote growth and to improve the economic characters of the silkworm. Horie and Watanabe (1983) showed that the supplementation of soybean protein increased the protein and amino acid content in the larval haemolymph of the silkworm B.mori. These are also known as fortification agents or supplementary nutrients when added to the normal diet. They enriched the nutritional value of the diet, making it more useful from the nutritional point of view (Bajpeyiet al., 1991). Krishnan et al. (1995) showed that the hydrolyzed soyaprotein (P-soyatase) supplementation decreased the larval duration, increased the accumulation of haemolymph protein (SP-1: female specific protein and SP-2; an arlyphorin), larval weight and cocoon characters. Keeping in view the above facts, an attempt was made to increase the growth of young age silkworm and cocoon yield using suitable mulberry variety and food additives. Hence, to fulfil the above needs, an experiment was designed to find out suitable food supplement/ additives for young age silkworm and estimation of biochemical constituents of chawki worms. 2. Materials and Methods: 2.1 Silkworm rearing Preparation for rearing Before commencement of rearing, the rearing room and appliances were thoroughly cleaned and the floor was washed with five per cent bleaching powder solution. The whole room was disinfected by spraying with 2.5 per cent Sanitech in 0.5 per cent slacked lime solution (Dandinet al., 2003) Breeds The silkworm breeds and hybrids/cross breeds used for various experiments are, PM x CSR 2, APM 1 x APS 8, APM 2 x APS 12, CSR2 x CSR4 and (CSR6 x CSR26) x (CSR2 x CSR27)

3 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), Incubation of eggs The layings procured and incubated by adopting standard incubation and dark treatment practices (Krishnaswami, 1979) Brushing and rearing The Newly hatched larvae were placed in the disinfected rearing trays. The tender mulberry from V1 variety cut into small squares of 0.25 cm 2 and sprinkled over the egg cards. The neonate larvae crawled over the tender and started feeding. Later, cards were tapped on to the rearing bed to brush the remaining worms.bed cleaning was done once during first instar and twice during second instar. (Jolly, 1986) Experiment on food supplements / additives for young age silkworm Studies were carried out on enriching the mulberry with soya flour, corn flour, horse gram flour to improve the silkworm nutrition. Experiments were conducted with five silkworm hybrids viz., PM x CSR2, CSR2x CSR4, APM1 x APS8, APM2 x APS12, and(csr6 x CSR26) x (CSR2 x CSR27). In each treatment 100 larvae of second instar were used and dusted with soya flour, corn flour, horse gram flour in two 10g / kg of and 5g / kg of as first feed in addition to artificial diet (Serinutrid) and normal control. This experiment was conducted in Completely Randomised Design with three replications. Factorial Completely Randomised Design (FCRD) as described by Panse and Sukhatme (1957). Treatment details T 1 Soya flour 10 g / kg of T 2 Soya flour 5 g / kg of T 3 Corn flour 10 g / kg of T 4 Corn flour 5 g / kg of T 5 Horse gram flour 10 g / kg of T 6 Horse gram flour 5 g / kg of T 7 Artificialdiet (Serinutrid) T 8 Control The silkworms were reared as per the improved rearing techniques (Dandin et al., 2003). The newly hatched worms were brushed on semi-synthetic diet in three replications under controlled temperature (28 0 C) and humidity (75-85%) till second moult. The semi-synthetic diet was made into elongated ribbon shaped strips of 2cm width and 2mm thickness. The diet feeding was done once in each instar, suitable leaf controls were also reared. The biochemical constituents viz., protein, carbohydrate and lipid content was estimated as per standard procedure

4 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), Estimation of biochemical constituents of second instar reared under feed additives Estimation of total proteins of second instar silkworm One gram of second instar larvae was taken for total protein estimation. The total protein was precipitated by homogenising 1g of silkworm in 0.3N perchloric acid (HClO 4 ). The precipitation was centrifuged at 2000 rpm for five minutes. The supernatant was dissolved in 0.5 N NaOH and boiled for 10 minutes (Rajashekaragouda, 1991). Total protein was estimated as suggested by Lowry et al. (1951) Estimation of total carbohydrate in second instar silkworm Full grown second instar larvae (50 numbers) were sampled and ground with pestle and mortar using 0.3 N perchloric acid solution. One gram from the homogenate was transferred to centrifuge tube and centrifuged at 2000 rpm for 2 minutes, allowed to settle for 10 minutes in the bath and again centrifuged for 10 minutes at 5000 rpm. Supernatant was collected and made to volume of 25 ml using distilled water. One ml of this solution was taken in test tube and 5 ml of anthrone reagent was added and boiled for 30 minutes and allowed to cool. Absorbance was observed at 640nm Estimation of total lipid content in second instar larvae About a gram of crushed larva was homogenised in ethanol, chloroform (1:2) mixture. The homogenised extract was heated in a water bath at 60 0 C for 5 minutes. It was subsequently centrifuged at 5000 rpm for 5 minutes. Supernatant was collected and transferred to separating funnel with 3% sodium chloride. Lower portion of the liquid in the separating funnel was collected and subjected to evaporation. The resultant residue was dissolved in diethyl ether and subjected to evaporation. The final residue expresses the total lipid content in milligram / gram of body weight. 3. Results and Discussion: Investigations were carried out on the influence efficiency of certain feed additives and its effect on biochemical constituents revealed significant results. Supplementation of feed additives Biochemical analysis Table 1. Protein content (mg) in young age silkworm B. mori reared on feed additives Treatment Protein content (mg) in silkworm PM x CSR2 CSR2 x CSR4 APM1 x APS8 APM2 x APS12 (CSR6 x CSR26) x (CSR2 x CSR27) Soya flour10g / kg of 19.91a 21.44a 19.83a 19.71a 24.03a Soya flour 5g /kg of 19.61c 21.19c 19.53b 19.43b 23.75c

5 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), Corn flour10g / kg of 19.19d 19.18d 19.08c 19.00c 23.45d Corn flour 5g /kg of 19.01e 19.03e 18.96d 18.87d 23.09e Horse gram flour10g / kg of Horse gram flour 5g /kg of 18.07f 18.57f 18.05e 17.92e 22.86f 17.92g 18.25g 17.89f 17.81f 22.76g Artificial diet 19.86b 21.36b 19.83a 19.72a 23.86b Control 17.63h 18.02h 17.58g 17.51g 22.72h SEd CD (0.05) Figures followed by similar letters are not different statistically at 5% level Table 2. Carbohydrate content (mg) in young age silkworm B.mori reared on feed additives Treatment Carbohydrate content (mg) in silkworm hybrids PM x CSR2 CSR2 x CSR4 APM1 x APS8 APM2 x APS12 (CSR6 x CSR26) x (CSR2 x CSR27) Soya flour10g / kg of 3.51c 4.21c 3.48c 3.43c 4.65c Soya flour 5g / kg of 3.38e 4.04e 3.36d 3.30d 4.54d Corn flour10g / kg of 3.93a 4.47a 3.89a 3.80a 4.96a Corn flour 5g / kg of 3.61b 4.31b 3.57b 3.50b 4.75b Horse gram flour10g / kg of Horse gram flour 5g / kg of 3.39e 3.58f 3.36d 3.29d 4.14e 3.06f 3.42g 3.00e 2.91e 3.84f Artificial diet 3.44d 4.14d 3.38d 3.45c 4.60c Control 2.97g 3.18h 2.91f 2.81f 3.66g SEd CD (0.05) Figures followed by similar letters are not different statistically at 5% level

6 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), Table 3.Lipid content (mg) in young age silkworm B.mori reared on feed additives Treatment Lipid content (mg) in silkworm hybrids PM x CSR2 CSR2 x CSR4 APM1 x APS8 APM2 x APS12 (CSR6 x CSR26) x (CSR2 x CSR27) Soya flour10g / kg of 13.97a 14.35a 13.84a 13.75a 16.81a Soya flour 5g / kg of 13.59c 14.05b 13.56c 13.44c 16.37b Corn flour10g / kg of 13.14d 13.89c 13.10d 13.01d 16.10c Corn flour 5g /kg of 12.73e 13.70d 12.69e 12.62e 15.87e Horse gram flour10g / kg of Horse gram flour 5g /kg of 11.64f 12.04e 11.52f 11.43f 12.45f 11.46g 11.87f 11.41g 11.33g 10.05g Artificial diet 13.88b 14.17ab 13.80b 13.68b 15.95d Control 11.13h 11.65g 11.02h 11.04h 9.83h SEd CD (0.05) Figures followed by similar letters are not different statistically at 5% level Biochemical analysis was done to estimate the biochemical constituents of young age silkworm, in order to find out the effect of different feed additives on protein, carbohydrate and lipid content of young age silkworm. The results showed that all the treatments were effective in increasing the protein content, among all the treatments, soya 10g / kg of recorded the highest protein content of mg, mg, mg, mg and mg in PM x CSR2, CSR2 x CSR4, APM1 x APS8, APM2 x APS12 and (CSR6 x CSR26) x (CSR2 x CSR27) respectively (Table-1), followed by artificial diet (Fig 1). Earlier, Subburathinamet al. (1992) reported that soya flour supplementation of 2g / dfl increased the protein profile. Sekar (1995) reported that the total protein in posterior silk gland and haemolymph showed maximum value, when the soya flour concentration was used at 12.5g / kg of mulberry, and opined that a concentration of 10g / kg was ideal for cross breeds while the same feeding at 12.5g / kg alone was optimum for L x NB18. Horie and Watanabe (1983) showed that the supplementation of soya bean protein increased the protein and amino acid content in the larval haemolymph of the silkworm. The data showed that the highest carbohydrate content was recorded on supplementation of corn flour 10g / kg of followed by corn flour 5g / kg of in respect of all the breeds (Table 2 Fig.2). This is may be due to the fact that corn flour contains more carbohydrate content and also infant silkworm needs more carbohydrate for their growth and development. Further the results on lipid content were also exhibited significant results. However highest lipid content was recorded on soya flour

7 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), g / kg of followed by artificial diet with respect of all the breeds (Table 3 &Fig 3).

8 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), Conclusions: From this study it can be inferred that, all the food additives were found effective in increasing the protein and lipid content in second instar larvae. Among treatments, the highest protein content of 19.91mg, 18.02mg, 19.83mg, 19.71mg and 24.03mg was recorded on soya 10g / kg of in PM x CSR2, CSR2 xcsr4, APM1 x APS8, APM2 x APS12 and (CSR6 x CSR26) x (CSR2 x CSR27) respectively, followed by artificial diet.19.86, 21.36, 19.83, and mg/ g larval weight. The highest lipid content was recorded on soya flour 10g / kg of followed by artificial diet with respect of the breeds. The highest carbohydrate content was recorded on supplementation of corn flour 10g / kg of followed by corn flour 5g / kg of in respect of all the breeds. References [1] E.C.G. Bedford, Continuous rearing of silkworm, J. Ent. Soc. S. Afr., 7 (1944) [2] C.M. Bajapeyi, R.N. Singh and K. Thangavelu, Supplementary nutrients to increase silk production., Indian Silk, 30 (1991) [3] S. B. Dandin, Jayant Jayaswal and K. Giridhar, Handbook of sericulture technologies, Central Silk Board, Bangalore, [4] Y. Horie and K. Watanabe, Design of the composition of the artificial diet for the silkworm bombyxmori by linear programming method: application of ingredients of feed for domesticated animals and fowls, Bull. Seric. Expt. Stn., 26 (1983) [5] M. S. Jolly, Pebrine and its control, Bulletin No. 5., CSR&TI, Mysore, [6] Japan Overseas Co-operation Volunteers, Text Book of Tropical Sericulture, Tokyo, (1980), 387. [7] Y. Horie, K. Watanabe and Shinohara, Effect of dietary composition on growth, silk glands and components in haemolymph of the silkworm, Acta. Sericologia, Japan, 78(1971)

9 Int. J. Pure Appl. Sci. Technol., 7(2) (2011), [8] Y. Horie and K. Watanabe, Effect of various levels of dietary protein and supplementation with limiting amino acids on growth, haemolymph components and uric acid excretion in silkworm, bombyxmori, J. Insect Physiol., 29 (1983) [9] T. Ito and N. Arai, Nutrition of the silkworm, b.mori., VIII, amino acid requirements and nutritive effects of various proteins, Bulletin of Sericulture Experimental station, Japan, 19 (1965) [10] T. Ito, Dietary requirements of the silkworm, bombyxmori L. and its artificial diets., XVI Int. Cong. Entomol. Abr., Tokyo, Japan, (1980) 448. [11] S. Krishnaswami, M. N. Narasimhanna, S. K. Suryanarayana and S. Kumararaj, Manual on sericulture-2, Silkworm Rearing, FAO, Rome, (1973) [12] M. Krishnan, K.M. Subburathinam and S. Janarthan, Effect of hydrolyzed protein (P- Soyatose) on haemolymph protein profile, larval and pupal characters of the silkworm, bombyxmori L. (lepidoptera:bombycidae), Sericologia, 35 (1995) [13] O.H. Lowry, N.J. Rosebrough, A. L. Farr and R. J. Randall, Protein measurement with the folin phenol reagent., J. Biol. Chem., 193(1951), [14] Mukul Deka, Saranga Dutta and Dipali Devi, Impact of feeding of samiacynthiaricini boisduval (red variety) (lepidopera: saturnidae) in respect of larval growth and spinning, Int. J. Pure Appl. Sci. Technol., 5(2) (2011), [15] V. G. Panse and P. V. Sukhatme, Statistical Methods for Agricultural Workers, ICAR Publication, New Delhi, [16] R. Rajashekaragouda, Studies on methods to increase silk yield of bombyxmori L., Ph.D. Thesis, TNAU, Coimbatore, [17] K. Sekar, Foliar supplementation of soya flour for improving the silk yield of mulberry silkworm, bombyxmori L. M.Sc. Thesis, Tamil Nadu Agricultural University, Coimbatore, [18] K. M. Subburathinam, S. Janarathanan and M. Krishnan, Nutritional response of mulberry silkworm to different levels of soya bean protein, Abstract of National Conference of Mulberry Sericulture Research held on Dec , 1992, CSR&TI, Mysore.

NUTRITIVE CONTENTS OF DIFFERENT VARIETIES OF MULBERRYLEAVES

NUTRITIVE CONTENTS OF DIFFERENT VARIETIES OF MULBERRYLEAVES Anshul Srivastava and Vadamalai Elangovan Department of Applied Animal Sciences, Babasaheb Bhimrao Ambedkar University (A Central University)