Utilization of mulberry leaves treated with seed powder cowpea, Vigna unguiculata (L) for feeding the fifth instar larvae of silkworm, Bombyx mori (L) (Race: PM x CSR2)

ABSTRACT
The present attempt was to screen the changes in the cocoon parameters; silk filament
parameters and activities of biochemical reactions catalyzed by the midgut enzymes fifth instsr larvae
of silkworm fed with mulberry leaves treated with aqueous solution of seed powder of Cowpeas
(Vigna unguiculata). The cowpea seed powder was dissolved in distilled water and diluted to 2.5%,
5%, 7.5%, and 10% concentrations. Fresh mulberry leaves were dipped in each concentration of
aqueous solution of cowpea seed powder for half an hour. 1000 ml solution was used for 100 grams of
mulberry leaves. Treated mulberry leaves were drained off completely and then used for feeding. The
mulberry leaves were fed five times per day at the rate of 100 grams per 100 larvae for each time.
Untreated group of larvae were feed with untreated mulberry leaves. Water treated group of larvae
were feed with water treated mulberry leaves. The experimental groups of larvae were feed with feed
separately with 2.5 percent cowpea treated; 5.00 percent cowpea treated; 7.5 percent cowpea treated
and 10.00 percent cowpea treated mulberry leaves. Treating the mulberry leaves with various
concentrations of aqueous solutions of cowpea seed powder and feeding to the fifth instar larvae of
multivoltine cross breed race of silkworm, Bombyx mori (L) was found reflected into significant
improvement in the weight of cocoon (31.862 Percentages); cocoon shell weight (52.336 percentages);
pupal weight (26.336 percentages) and shell ratio. There was significant improvement in the silk
filament length ( 00.323 percentages); silk filament weight ( 21.317 percentages) and denier scale of
silk filament. Midgut enzymes (Protease, Amylase, Trehalase, Sucrase and Urease) were found
influenced through treating mulberry leaves with cowpea seed powder and feeding to the fifth instar
larvae of multivoltine cross breed race of silkworm, Bombyx mori (L) (Race: PM x CSR2). The
contents of cowpea seeds may be associated with improvement in the growth and development
through increased pattern of protein turn over and overall metabolism.

References
[1] Kellner O, Kakizaki S, Matsuoka M, Yoshu T. XXIV. On the physiology of the silk
worm. By Alexander pringle jameson and william ringrose gelston atkins. Landw
Versuchs-stationen 1887; 33: 381.
[2] Mahmood R, Jan MT, Khan MI. Effect of nitrogen (farmyard manure + urea) treated
mulberry trees on the larval development and cocoon weight of silkworm, Bombyx
mori L. Asian J Plant Sci 2002; 2: 93-4.
[3] Ravikumar C. Western ghat as a bivoltine region prospects, challenges and strategies
for its development. Indian Silk 1988; 26:3 9-54.
[4] Sengupta K, Singh BD, Mustafij C. Nutrition of silkworm. Bombyx mori L.I. Studies on
the enrichment of mulberry leaf with various sugars, proteins, amino acids and vitamins
for vigorous growth of the worm and increased cocoon crop production. Indian J Sci1972; 11: 11-27.
[5] Javed H, Gondal MH. Effect of food supplementation by N and Ascorbic Acid on larval
mortality of silkworm (Bombyx moriL.). Asian J Plant Sci 2002;1:556-7.
[6] Kanekatsu R. Amylase in the digestive juice of silkworm larvae, Bombyx mori. J Seric
Sci 1972; 41: 445-51.
[7] Kanekatsu R. Studies on further properties for an alkaline amylase in the digestive juice
of silkworm, Bombyx mori. J Fac Text Sci Technol 1978; 76: 1-21.
[8] Eguchi M, Iwamoto A. Alkaline protease in the midgut tissue and digestive fluid of
silkworm, Bombyx mori. Insect Biochem 1976; 6: 491-6.
[9] Sumida M, Yuan XL, Matsubara F. Sucrase activity and its kinetic properties in
pertrophic membrans, and in membrane-bound and soluble fractions of midgut in
silkworm, Bombyx mori. Comp Biochem Physiol A 1994; 108: 255-64.
[10] Abraham EG, Nagaraju J, Datta RK. Chemical studies of amylases in the
silkworm, Bombyx mori L.: Comparative analysis in diapause and nondiapause strains.
Insect Biochem Mol Biol 1992; 22: 867-73.
[11] Blade, S. F., Shetty, S. V. R., Terao, T. & Singh, B. B. (1997) Recent developments in
cowpea cropping systems research. In Singh, B.B., Mohan Raj, D. R., Dashiell, K. E. &
Jackai, L. E. N. (Eds.) Advances in Cowpea Research. International Institute of Tropical
Agriculture and Japan International Research Center for Agricultural Sciences.
[12] Kanekatsu R, Ichimura H, Hori M. Distribution and developmental changes in midgut
sucrase activity of the silkworm,Bombyx mori. J Seric Sci Japan 1989; 58: 517-23.
[13] Kanekatsu R, Satoh M, Kodaira R, Miyashita T. Midgut sucrase-1 (suc-1) of the
silkworm, Bombyx mori: Geneties and changes in the activities during the pupal}adult
development. J Seric Sci Japan 1993; 62: 13-9.
[14] LANGYINTUO, A. S., LOWENBERG-DEBOER, J., FAYE, M., LAMBERT, D.,
IBRO, G., MOUSSA, B., KERGNA, A., KUSHWAHA, S., MUSA, S. & NTOUKAM,
G. (2003) Cowpea supply and demand in West and Central Africa. Field Crops
Research, 82, 215-231.
[15] Sumida M, Yuan X L, Mari YI. Mori H, Matsubara F. Changes in kinetic parameters
and total activity of midgut sucrase in the silkworm, Bombyx mori during larval
pupal}adult development. Comp Biochem Physiol B 1990; 96: 605-611.
[16] Asakawa H, Hamano K. Enzymatic properties of digestive amylase isozymes in
silkworms, Bombyx mori L. J Seric Sci Japan 1994; 63: 13-20.
[17] Singh B. “Improving the production and utilization of cowpea as food and fodder”.
Field Crops Research 2003; 84: 169-50.
[18] Scott C. “Sub-Saharan Africa news in brief: 25 March-9 April”. Science and
Development Network. Available from:http://www.scidev.net/en/sub-suharanafrica/news/sub-saharan-africa-news-in-brief-25-march-9-april.html. [Last accessed on 2008 Apr 10].
[19] Shaw M. “100 Most Protein Rich Vegetarian Foods”. SmarterFitter Blog. Available
from:http://smarterfitter.com/blog/2007/10/28/100-most-protein-rich-vegetarian-foods/.
[Last accessed on 2007 Oct 28].
[20] Singh, B.; Ajeigbe, H. A.; Tarawali, S. A.; Fernandez-Rivera, S.; Abubakar, M. (2003).
“Improving the production and utilization of cowpea as food and fodder”. Field Crops
Research 84: 169-150. doi:10.1016/S0378-4290(03)00148-5.
[21] Bernfeld P. Enzymes of carbohydrate metabolism: Amylases, á and â. Methods in
Enzymology In: Colowick SP, Kaplan NO, editors. New York: Academic Press; 1955;1: 149-58.
[22] Ishaaya, Swirski E. Invertase and amylase activity in the armoured scales
Chrysomphalus aordun and Aonidiella auranti. J Insect Physiol 1970; 16: 1599-606.
[23] Dahlman DL. Purification and properties of trehalase from tobacco hornworm larvae. J
Insect Physiol 1971; 17: 1677-87.
[24] Eguchi M, Iwamoto A. Comparison of three alkaline proteases from digestive fluid of
the silkworm, Bombyx mori. L. Comp Biochem Physiol B 1982; 71: 663-8.
[25] Sarangi SK. Alkaline protease in the midgut of the silkworm, Bombyx mori L: Changes
during metamorphosis. Proc Indian Acad Sci (Anim. Sci.) 1985;9 4: 567-72.
[26] Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin
phenol reagent. J Biol Chem 1951; 193: 265- 75.
[27] Hiware CJ. Effect of fortification of Mulberry leaves with homeopathic drug Nux
Vomica on Bombyx Mori. L. Homeopathy 2006; 95: 148-50.
[28] Hirayama C, Konno K, Shinbo H. The pathway of ammonia assimilation in the
silkworm Bombyx mori. J Insect Physiol 1997; 43: 959- 64.
[29] Rosenthal G. A. , Hughes C, Janzen DH. l-Canavanine, a dietary source for the seed
predator Caryedes brasiliensis (Bruchidae). Science 1982; 217: 353-5.
[30] Rosenthal GA, The interrelationship of canavanine and ureas in seeds of the Lotidae. J
Exp Bot 1974; 25: 609-13.
[31] Liu CH, Hays VW, Svec HJ, Catron DV, Ashton GC, Speer VC. The fate of urea in
growing pigs. J Nutr 1955;78:57-72.
[32] Rose WC, Dekker EE. Urea as a source of nitrogen for the biosynthesis of amino acids.
J Biol Chem 1956; 203: 107-21.
[33] SHARMA, H. C. (1998) Bionomics, host plant resistance, and management of the
legume pod borer, Maruca vitrata. Crop Protection, 17, 373-386.
[34] Snyderman SE, Holt LE, Dancis J, Roitman E, Boyer A, Balis ME. “Uuessential”
nitrogen: A limiting factor in human growth. J Nutr 1962; 78: 57-72.
[35] Grimson RE, Bowland JP, Milligan LP. Use of nitrogen- 15 labelled urea to study urea
utilization by pigs. Can J Anim Sci 1971; 51: 103-10.
[36] Richards P. Nutritional potential of nitrogen recycling in man. Am J Clin Nutr 1972; 25:615-25.
[37] Okumura J, Tanaka H, Muramatsu T. Incorporation of 15Nurea in chicks. Jpn J Poult
Sci 1979; 16: 45-8.
[38] Levenson SM, Crowly LV, Moriwitz RE, Malm OJ. The metabolism of carbon-labeled
urea in the germfree rats. J Biol Chem 1959; 234: 2061-2.
[39] PERRINO, P., LAGHETTI, G., SPAGNOLETTI ZEULI, P. L. & MONTI, L.M. (1993)
Diversification of cowpea in the Mediterranean and other centres of cultivation. Genetic
resources and crop evolution, 40, 121-132
[40] PHILLIPS, R. D., MCWATTERS, K. H., CHINNAN, M. S., HUNG, Y. C.,
BEUCHAT, L. R., SEFA-DEDEH, S., SAKYI-DAWSON, E., NGODDY, P.,
NNANYELUGO, D. & ENWERE, J. (2003) Utilization of cowpeas for human
food. Field Crops Res., 82, 193-213.
[41] Deguchi E, Niiyama M, Kagota K, Namioka S. Role of intestinal flora on incorporation
of 15N from dietary, 15N-urea, 15Ndiammonium citrate into tissue proteins in pigs. J
Nutr 1978; 108: 1572-9.
[42] Hirayama C. Effect of mulberry leaf powder addition in artificial diet on the excretion
of nitrogenous products and utilization of nitrogen in the silkworm, Bombyx mori (in
Japanese with English summary). J Sericult Sci Japan 1994; 63: 206-13.
[43] Krajewska B, van Eldik R, Brindell M (13 August 2012). “Temperature- and pressuredependent stopped-flow kinetic studies of jack bean urease. Implications for the
catalytic mechanism”. JBIC Journal of Biological Inorganic Chemistry 17(7): 1123-
1134.
[44] Lindroth RL, Barman MA, Weisbra AV. Nutritient deficiencies and the gypsy moth, L.
dispar: Effects on larval performance and detoxification enzyme activities. J Insect
Physiol 1991; 37: 45-52.
[45] Vyjayanthi N, Subramanyam MV. E!ect of fenvalerate- 20EC on sericigenous insects. I.
Food utilization in the late-age larva of the silkworm, Bombyx mori L. Ecotoxicol Ecol2002; 53: 206-11.
[46] Christopher MS, Mathavan S. Regulation of digestive enzyme activity in the larva of
Catopsilia crocale (Lepidoptera). J Insect Physiol 1985; 31: 217-21.
[47] Ito T, Arai N. Amino acid requirements in Bombyx mori. J. Insect Physiol 1966; 23:861-9.
[48] Dadd RH. Proteolytic activity of the midgut in relation to feeding in the beetles,
Tenebrio molitor and Ditiscus marinalis L. J Exp Biol 1956; 33: 311-24.
[49] Hamano K, Mukaiyama F. Some properties of digestive fluid proteases in the
silkworm, Bombyx mori, with reference to the relation between dissociation degree and
nutritive value of some proteins. J Sericult Sci Japan 1970; 39: 371-6.
[50] Jadhav G, Kallapur VL. Influence of age, sex and feeding on the protease activity of
certain tissues of fifth instar silkworm, Bombyx mori. Entomon 1988; 13: 289-93.
[51] Azuma M, Yamashita O. Cellular localization and proposed function of midgut
trehalase in silkworm larva, Bombyx mori. Tissue Cell 1985; 17: 539-51.
[52] Hasegawa K, Yamashita O. Mode d′action de I′hormone de diapause dans le
metabolisme glucidique de ver aand soie, Bombyx mori L. Ann Endocrinol 1970; 31:631-6.
[53] Nath BS. Changes in carbohydrate metabolism in haemolymph and fatbody of the
silkworm, Bombyx mori L., exposed to organophosphorus insecticides. Pestic Biochem
Physiol 2000; 68: 127-37.
[54] RANGEL, A., DOMONT, G. B., PEDROSA, C. & FERREIRA, S. T. (2003)
Functional properties of purified vicilins from cowpea (Vigna unguiculata) and pea
(Pisum sativum) and cowpea protein isolate. Journal of agricultural and food
chemistry, 51, 5792-5797.
[55] Singaravelu G, Sumathi S, Prabu P, Jagapriya L. Biological activity of azadirachtin on
certain reproductive aspects of female moth of Bombyx mori L. Toxicol Int 2004; 11:27-31.
[56] Zimmer M (Apr 2000). “Molecular mechanics evaluation of the proposed mechanisms
for the degradation of urea by urease”. J Biomol Struct Dyn. 17(5): 787-97.
doi:10.1080/07391102.2000.10506568. PMID 10798524.

Download all article in PDF

WSN 40 (2016) 147-162