Silkworm Probiotics

Mulberry (Morus species) leaf is the solo food and source of nutrition for the silkworm,
Bombyx mori L. due to the presence of morin (Tribhuwan, et al., 1989). Nutritional study on
silkworm is an essential prerequisite for its proper commercial exploitation. Nutrition of
silkworm is sole factor which almost individually augment quality and quantity of silk (Laskar
and Datta, 2000). Feeding of nutritionally enriched leaves provided better growth and
development of silkworms as well as gain in economic characters of cocoons (Krishnaswami et
al., 1971). Effects of diets with different nutrients such as proteins, amino acids, carbohydrates,
hormones, vitamins and minerals on the better production of cocoon crops have been reported
earlier by Narasimhamurthy and Govindappa (1988), Bose and Majumdar (1989), Islam and
Khan (1993), Reddy et al. (1994), Khan and Saha (1997a,b), Faruki (1998), Nirwani et al. (1998)
and Nakamura (2000). The dietary nutritional management influences directly the quality and
quantity of silk production (Shyamaia and Bhat, 1968; Hugar et al., 1977; Benchamin and Jolly,
1986; Tayade et al., 1988 and Murgan et al., 1998). Studies on nutritional ecology of an insect
are very important for its commercial exploitation (Scriber and Slasky, 1981). Eestimation of
rate of ingestion, digestibility and conversion efficiency of food, so also growth rate of the
animal etc, were carried out by Englemann, 1966. The principles of the major insect nutritional
requirements for growth and reproduction have been established since 1940s through the studies
on reprentative of the major insect group (Dadd, 1973; Davis, 1968; House, 1965; House, 1974;
Nation, 2001; Simpson and Raubenheimer, 1995; Slansky and Scrieber, 1985, Genc H. Phaon.,
2002).
The nutrition, particularly as it relates to the physiology of digestion, is the most
fundamental and important challenges in the sericulture.. According to Kellner, 1887 the
silkworm digests albumin, fat and carbohydrate except cellulose. Mulberry leaves of high
yielding varieties cultivated under poor soil nutrient management with low nutritive value, when
fed to larvae of highly productive silkworm breeds, it will lead to inferior quality cocoon
production.
The silkworm, Bombyx mori L. is a holometabolus insect that passes through four
distinct stages in its life cycle. The silkworm feeds only in the larval stage, which extends for a

period of about 25-26 days. The larval stage in important for the fact that the silk produced at the
end of this stage is dependent on the quality and quantity of silk, mulberry leaves play a very
important role in the larval life of silkworm, Bombyx mori L. Benchamin and Jolly (1986) are
also of the same opinion that the quality of mulberry leaves influences the growth of the
silkworms that help in the production of good cocoon crops. The leaves with low nutritive value,
when fed to the silkworm will lead to the production of inferior quality cocoons. The imported
physiological factor in silkworm growth, silk productivity is nutrition.
Importance of the research on effect of different fortification agents in silkworm nutrition
can be judged from the principle of co-operating supplements (House, 1996), which states that
the substitution sources of nutrients cooperating with the commonly recognized food stuff of the
species, essential to fulfill the nutritional requirements in many insects. Supplementary nutrients
are chemicals which, when added to normal food makes it more useful (Bajpeyi et al., 1991). In
recent years, several attempts have been made to fortify the leaves with different beneficial
nutrients and combination of nutrients (Rajegowda, 2002) to enhance the quality of cocoon crop.
the silkworm, . Mulberry leaf supplemented with spirulina as a feed to Bombyx mori L.
(Lepidoptera: Bombycidae) orally found to be effective in enhancing the larval and cocoon
characters (Venkataramana, 2003). Sometimes, nonessential amino acids motivate growth,
because of the optimization of the nutrient balance and the good organization of the biochemical
pathways concerned in the synthesis of the nonessential amino acids. For example, alanine and
glycine or serine is necessary for optimal growth in Bombyx mori (Nation, 2001). Some amino
acids are important in morphogenesis. It has been shown that tyrosine is crucial for cuticular
sclerotization and tryptophan for the synthesis of visual screening pigments. Some other amino
acids are known to be neurotransmitters such as -aminobutyric acid and glutamate. Proline is
essential for development and as energy sources for Culex spp. and several Diptera species.
Aspartic acid and glutamic acid are important for Phormia sp. and Bombyx mori L. (Chapman,
1998).
Proteins and especially free amino acids play important roles in insect development.
Presence of high levels of amino acids in insects indicates that besides their role in protein
synthesis, they have additional functions related to neural transmission, detoxification, synthesis
of phospholipids, energy production and morphogenetic processes (Chen, 1985). Rodwell (1993)

reported that amino acids and their derivatives participate in intracellular functions as diverse as
nerve transmission, regulation of cell growth and the biosynthesis of various compounds in
silkworm.
The requirements of amino acids in silkworm were confirmed by a number of
experiments (Inokuchi and Ito, 1972; Bose and Majumdar, 1989; Quader et al., 1994; Khan and
Saha, 1995). It has been determined that 10 amino acids viz., arginine, histidine, isoleusine,
leusine, glysine, methionine, phenylalanine, threonine, tryptophan and valine are essential for
silkworm nutrition (Arai and Ito, 1964; Inokuchi et al., 1967; Ito, 1967; Akter and Arghar, 1972).
Bose et al. (1989) stated that amino acids are one of the most important constituents in
silkworms that require 18 amino acids for their adequate nutrition. It is thus obvious that the
study of larval growth is an important aspect of research for the insects that are economically
important. Successful cocoon crops in sericulture depend mostly on a healthy larval growth.
Bombyx mori L. encounters various constraints like attack from pathogen in the
environment and accumulation of toxic phytochemicals from feed itself. Supplementation of
some important nutrient/ factors is one valuable strategy to increase the disease tolerance for
better survival of the silkworm (Chandrakala et al., 2007). The nutrition background of the larvae
influences greatly on the resulting pupa, adults and silk production. The leaves of superior
quality enhance the chances of good cocoon crop. The dietary nutritional management influences
directly on quality and quantity of silk production in Bombyx mori L. (Muragan, et al., 1998).
These supplements when added to the mulberry enrich nutrition value of diet making it more
useful from the nutritional point of view (Bhargava et al., 2008).
The first attempt was made to use homeopathic drugs by Hiware (2006) and Hiware
(2005a, b) also shows that silkworm Bombyx mori L. larvae were fed on mulberry leaves treated
with the homeopathic drug, Nux vomica mother tincture, Chelidonium mother tincture and
Phytolacca berry mother tincture shows better results on biological characters of silkworm
Bombyx mori L.
Considering the great bio-diversity of Indian flora, phytochemical research is done to
provide potent radio modifiers and anticancer agents as well as formulations for eco-friendly
agriculture and sericulture. To this end, a new polysaccharide obtained from the Indian medicinal

plant, Tinospora cordifolia has been found to possess impressive immunomodulatory (Indian
Patent 183805) and radio protective properties. The tissue culture technique has been evoked for
its optimized generation. A cheap moulting phytohormone (MH) preparation that provides better
quality of silk in a short time has been developed from an indigenous natural source. The MH
formulation was prepared by a continuous extraction technique using an indigenous plant that is
widely growing in the costal region of India. Application of the formulation at a very low
concentration (20 ppm) assists in faster and uniform spinning of the silkworms leading to better
silk productivity with minimum loss of silk. The MH technology has been developed up to a
scale of 5 kg of the plant. The product is given to Central Sericulture Research and Training
Institute, Mysore for free distribution to silk farmers (Chattopadhyay, 2010).
The substances which are regulating the growth and yield in plants are called plant
growth regulators. It has been suggested that the plant growth regulators such as gibberellins,
auxins, cytokinines, abscissic acid and ethylene participate decisively in complex bioprocesses.
Administration of plant growth regulators are reported to influence upon the developmental
process of insects (Carlisle et al., 1963). Panitz, (1967) has found a specific effect of gibberlic
acid GA3 upon the activity of the genome exposed by puffs in the polytene chromosomes of
larvae of midge, Acricotopus indicus. Supp-lementation of GA3 increased larval duration, larval
weight, decreased fecundity and hatching in the bollworm, Heliothis virescence (Guerra, 1970),
increased larval and pupal periods, emergence percentage and decreased pupal weight longevity,
fecundity and hatching in Spodopter littoralis (Salama and Sharaby, 1972); absorption of plant
materials in phytophygous insects influences development and physiology increased fecundity
and viable eggs in the grass hopper, Aulocara eliotti (Neumam, 1982). It has been reported that
the treatment with chloremphenicol increases the larval weight and shell ratio in the silkworm, B.
mori (Krishnaswami, 1978). Feeding of mulberry leaves sprayed with GA3 to IV instar larvae
was reported to increase the larval weight and cocoon weight in the silkworm, Bombyx mori L.
(Kamada and Ito 1984), Pai et al., (1986) have reported that topical application with
Paraminobenzoic acid causes a significant reduction in hatchability of eggs of NB18 race of the
silkworm, Bombyx mori L.
Magadum and Hooli (1989, 1991 a, b) have reported that topical application with IAA,
IBA, GA3 and IPA in different larval stadium of pure Mysore multivoltine breed of the

silkworm, B. mori, resulted in a significant increase in the larval weight, silk gland weight,
cocoon weight and its shell weight in bivoltine silkworm, Bombyx mori L. (Hugar and Kaliwal,
1997). Goudar and Kaliwal (2001a,b) have reported that plant growth regulators 2, 4-dichlorophenoxy acetic acid (2, 4-D) and naphthoxyacetic acid (NOA) enhance the economic traits like
larval weight, silk gland weight, cocoon weight, filament length and denier in the silkworm,
Bombyx mori L. presences of different types of bacteria in the gut have been reported (Roy et al,
2000 and Kodama, 2001). Most of the species belonging to Genus Streptococcus are found to be
pathogenic to Bombyx mori L. larvae while bacteria from Genus Pediococccus, Leuconostoc and
Lactobacillus didnt produce any infected silkworm. The precise mechanism of beneficial effect
on host or interaction among the different bacterial strains present as micro-flora is not known
(Steinhaus, 1949 and Kodama, 2001). Different species of lactic acid bacteria have been
extensively studied (Bruno et al., 1993; Bernet Camard et al., 1997; Gibson et al 1998; Kodama,
2001) and found to be beneficial as probiotics (Fuller, 1991; Bernet Camard, 1997; Sakamoto et
al; 2001). Therefore it was planned to study the effect of a lactic acid bacteria namely
Lactobacillus plantarum a known probiotics on economic parameters of Bombyx mori L.
Recently plant extracts particularly weeds are being used extensively in sericulture to
increase the silk cocoon and egg yield of Bombyx mori L. Rajashekhargouda (1991). As the
weeds are easily available and help to bring down the cost of production. It is seen that, there are
very scanty attempts made by using the plants product fortification for improving the status of
sericulture industry. An attempt made by Hiware and Bhalerao (2008) find out the effect of
extract prepared from four medicinal plants viz. Phyllanthus niruri, Tephrosia purpurea,
Phyllanthus emblica, and Phyllanthus amarus on the biological parameters of silkworm Bombyx
mori L. In recent years many attempts have been made to fortify mulberry leaf with nutrients of
botanical extracts so as to improve the leaf quality and feeding efficiency of silkworm which in
turn will help to increase the cocoon production and silk quality (Bajpeyi, 1991; Das et al.,1993;
Shivakumar et al., 1996; Kim et al., 1997; Patil et al., 1997a,b; Krishnaprasad et al., 2001;
Hipparagi et al; 2003; Jeyapaul et al., 2003; Sabitha et al., 2003; Sujatha and Purushothamrao,
2003; Sujatha et al., 2003a,b; Venkataramana et al., 2003). Treatment of mulberry leaf with
botanicals can increase the silkworm productivity leading to economic gains. The dietary
nutritional management influences directly the quality and quantity of silk production in Bombyx
mori L. (Benchamin and Jolly, 1986; Murugan et al., 1998).

Rajesh (2011) done a current experiment on mulberry leaves were supplemented with goose
berry extract to study its effect on growth and silk production of silkworm. He shows that the
mature larval weight did not differ significantly among the treatments. Gain in larval weight was
significantly higher in control (without water), other treatments on par with each other. Pupal
weight was significantly higher in the 20% solution treatment, other treatments being on par with
each other. Goose berry (Phyllanthus emblica L.) is one of the richest sources of ascorbic acid.
Barthakur and Arnold (1991) estimated that the berry contain 5889 ppm ascorbic acid. Apart
from this the berry contain sugars, various minerals, amino acids and vitamins etc all of which
play important role in the nutrition of silkworm and silk production (Shyamaia and Bhat, 1965;
Subburathinam and Sulochana Chetty, 1990; Ito, 1960 and Ito, 1961).
Among various technologies to improve the cocoon productivity, application of juvenile
hormone (JH) on silkworm is practiced in several silk producing countries. They tend to keep the
Fifth instar silkworm young for a specified period by extending the larval period and increase the
silk secretion. It postpones the spinning stage by few hours. Altosid, R394, Manta and ZR512 are
some of the commercial synthetic JH analogues used in countries like Japan and China.
Synthetic JH analogues are hitherto not available for use by sericulturists in India. Moreover, the
import of the chemical is cost prohibitive. The crystalline material of the moulting hormone
secreted by the prothoracic gland was first isolated from Bombyx mori L. pupae and was named
ecdysone (Butenandt et al., 1954). Many plants contain a variety of chemicals with moulting
hormone activity in insects. The ready isolation from plants in contrast to the extremely poor
yield from insects and other sources makes it possible to produce large amounts of active
substances for biological testing. Over 100 ecdysteroids have been structurally identified from a
wide range of plant species (Lafont and Horn., 1989).
Chandramohan (2011) also reported in Department of Sericulture, Tamil Nadu
Agricultural University, Coimbatore has screened several plant products and short listed fewer
plants possessing JH activity, (Phytojuvenoid) against silkworms. Extraction procedures were
standardized and a novel botanical formulation was developed. Comparing the growth of
silkworm to different phases of moon, the new formulation has been named ILAMATHI
(young moon) as it keeps the larvae young. The botanical formulation was found to increase the
larval duration, cocoon weight and there by yield of 25.16 percent and also an increase in cocoon

and over control.

D. lablab is a leguminous plant, found in India, is a seasonal dicotyledonous legume. It is
commonly called as Indian bean. For the fulfillment of need of dietary proteins, the population of
the subtropics, being predominantly vegetarian, looks to legumes like D. lablab as it is having
more protein content. It is also called as poor man bean as it is cheap when compared with other
beans. Extracts of D. lablab seeds were found to be mitogenic properties (Aurich et al., 1971). Its
effect on silkworm is studied by Saravanan Manjula et al., (2010). Kim et al., (1997) reported the
effects of methanol extract of Achyranthes japonica on the development stages of silkworm,
Bombyx mori L. Administration of methanolic extract of this plant at 6 hours before the
appearance of early mature larvae enhanced remarkably the pupation ratio and cocoon quality
besides shortening mounting duration. Murugan et al., (1999) shows aqueous of botanicals such
as Tridax procumbens, Lantana camera, Clerodendron sp. and Croton sparsiflours stimulated the
growth and development of mulberry silkworm, Bombyx mori L. (PM NB4D2). Krishnaprasad
et al., (2001) reported that mulberry leaf supplemented with potato leaf extract once during 3and
4instar and twice during 5instar recorded superior larval weight, lower larval mortality and larval
duration higher cocoon weight, shell weight and shell ratio.
Hipparagi et al., (2003) showed the effects of filed spray of Lantana camara, Tridex procumbens
and Parthenium hysterophorus extract at 30 and 40 % concentration during rainy, winter and
summer seasons on silkworm growth and development. The results indicated that Parthenium (30
% aqueous) produced more fecundity while Lantana (30 %) and Tridex (40 %) results more
cocoon yield. The effects of botanical extracts were found to be superior during rainy season
than winter and summer seasons on silkworm growth and development. The supplementation of
the Spirulina (100 ppm) with mulberry feed to silkworm larvae (NB4D2 race) during 5instar
significantly enhanced the pupation rate, cocoon weight, shell weight and shell as compared to
control.
Sujatha and Purushothamrao (2003) studied the effect of Eucalyptus globules leaf extract
(0.1, 1.0, 2.0 and 5.0%) when fed to silkworm along with mulberry leaf. The total rearing period
was reduced in all the concentration of E. globules. The economical character such as cocoon
weight, shell weight, shell ratio and filament length showed improvement in 15 concentration of

Eucalyptus leaf extract. Sujatha et al., (2003b) reported the effects of leaf of Azadirachta indica
and Vitex negundo when supplemented to mulberry feeding of silkworm. There was significant
improvement in shell ratio and filament length at 2.0% concentration of Vitex leaf extracts. But
cocoon weight decreased in comparison to control.
Sabitha et al., (2003) reported that phytoecdysteroid treated silkworm during the 5instar
completed spinning in 14 to15 hours as compared to control (32 to 38 hours). There was no
adverse effect of phytoecdysteroid on cocoon characters. Jeypaul et al., (2003) reported that food
assimilation rate, conversion efficiencies were significantly higher in silkworm fed on mulberry
leaf supplemented with Coffea Arabica leaf extract at 1:25 concentration. The treatment recorded
significantly higher cocoon weight, pupal weight and shell weight.
Padmalatha et al., (2005) also study the effect of four ferns such as Nephrolepis
auriculata, Christrella parasitica, Decranopteris linearis and Pityurogramma calomelanes on the
quantitative traits and energetic as of B. mori L. The impact of ferns to increase in the larval
weight was noticed among fern extracts treated batches. The mean of the observations of three
concentrations of each fern showed improvement in larval for N. auriculata, C. parasitica, D.
linearis and P. calomelanes, respectively over control (0.493 g / animal). Significant differences
in economic parameters of silkworm existed in relation to plant extracts. N. auriculata extracts at
0.1 0.2 and 0.4 per cent concentrations recorded higher cocoon weight, shell weight, pupal
weight, and shell percentage. Leucas aspera plant is a medically important and easily available
plant. In view of these facts, the growth promoting effect of L. aspera on feeding, growth and
bio-chemical changes of Bombyx mori L. were done (Senthil Nathan and Sangeetha, 2002).
It is reported that humoral immune system has efficient self-defense mechanism against
infection through induction of synbiotics (combination of prebiotic and prebiotic) one of the
promising alternatives for passive immunization and simultaneously growth and development of
the silkworm. Scientists of Silkworm Pathology has reported that feed supplement (Oral
immunization) of probiotics contain, Lactobacillus acidophillus controlled bacterial disease
88.62 % and other diseases - 55.25 % during unfavourable seasons (June - September) in
multivoltine breeds, Nistari(M), M12(w) and M6DP(c). The combination of probiotics and
prebiotics in a synbiotic preparation, has not been studied in silkworm earlier that might improve

the survival of the beneficial bacteria enhancing their effects might be additive or even
synergistic. Therefore, effort has been taken for making a synbiotic preparation, combination of
probiotic and prebiotic to study the synergistic effect of the live microorganisms to control
silkworm diseases, thereby increases the cocoon productivity.
The digestive system is home to more than 500 different types of bacteria. They help
keep the intestines healthy and assist in digesting food. They are also believed to help the
immune system (Martin, 2001). These friendly organisms also help fight bacteria that cause
diarrhea.
Silkworms are attacked frequently by pathogens due to failure of cellular defense system.
Humoral immune system has efficient self-defense mechanism against infection through
induction of defense molecules. These defence molecules include phenoloxidases, clotting
factors, complement factors, lectins, protease inhibitors, antimicrobial proteins, Toll receptors,
and other humoral factors found mainly in hemolymph plasma and hemocytes. These
components, which together compose the innate immune system, defend invertebrate from
invading bacterial, fungal and viral pathogens. According to Charles (2004) lower animals do not
have well developed humoral immunity and under such circumstances vaccine development may
not be of much use and in these lower animals immuno-stimulation could be achieved easily
through Probiotics.
In recent years attempts have been made in sericulture with feed supplement of nutrients, such
as proteins, carbohydrates, amino acids, vitamins, hormones, and antibiotics etc. for better
performance of good quality of cocoons (Sannappa, 2002).and up gradation of genes mediating
innate immunity. Prebiotic promote the bifidobacterial growth. As innate immunity is an
important defense system in B.mori , the non-pathogenic bacteria enhanced the immunity factors
and reduced the susceptibility to bacterial pathogenic infections in B.mori.
There is suggestive evidence that several probiotic strains with the prebiotic, oligofructose are
useful in boosting the immune response. Indirect evidence has been obtained in studies aimed at
preventing acute infectious disease (nosocomial diarrhoea in children, influenza episodes in
winter) and studies that tested antibody responses to vaccines. Nowadays, the microbes
Lactobacilli and Bifido bacteria are widely used in probiotic therapy. These bacteria are

producing lactic acid that constitutes a major part of the normal intestinal microflora in all
animals. Hence, the present attempt is a very good approach on B.mori to strengthen the host
immunity to resist the microbial pathogenic attack and to promote good cocoon yield.
As innate immunity is an important defense system in B.mori (Ponnuvel and Yamakawa,
2002) the non-pathogenic bacteria enhanced the immunity factors and reduced the susceptibility
to bacterial pathogenic infections in B.mori. Scientists of Silkworm Pathology Section has
already studied control of silkworm diseases through induction of different immunogens and it
has been found that individual induction of Proline, Nicotinic acid and Ascorbic acid are very
much effective for immunization to control the silkworm diseases to the tune of 67.33 - 84.22%
in general and bacterial disease, 88.80 - 90.25 %, thereby increases the cocoon productivity. Oral
immunization with Lactobacillus acidophillus has effected satisfactory result when mortality%
was recorded significantly (P< 0.1) lower (27.0 %) compare to micronutrient supplement. The
combination of probiotics and prebiotics in a synbiotic has not been studied. This combination
might improve the survival of the bacteria crossing the upper part of the gastrointestinal tract,
thereby enhancing their effects in the large bowel. In addition, their effects might be additive or
even synergistic (Am J Clin Nutr., 2000). Therefore, effort has been taken for making a synbiotic
preparation, combination of probiotic and prebiotic to study the synergistic effect of the live
microorganisms to control silkworm diseases, thereby increases the cocoon productivity.
Bifidobacterium bifidum, known as B. bifidum, is a strain of bacteria commonly used as a
probiotic. Probiotics are living microorganisms that simulate the beneficial bacteria found in the
gastrointestinal tract of humans. B. bifidum is one of many bacterial strains that occur naturally
in the gut flora, living in the colon. The inclusion of B. bifidum in a dietary regimen provides the
host with a variety of following benefits (Erica Wickham, 2003).
The lactobacillus plantarum is a probiotic which improves the cocoon production of mulberry
silkworm Bombyx mori (Singh et al., 2005). Certain probiotic bacteria inhibit the growth of
microbes. Streptomyces noursei are probiotic microbes which prove the antibacterial activity and
good eco-friendly management of silkworm diseases. Streptomyces noursei isolated from
silkworm breeds revealed their antibiotic potential against a range of Gram positive and Gram
negative bacteria and it was found to inhibit the germination of conidia of entomopathogens B.
bassiana and M. anisopliae in vitro (Mohanraj, 2007). Probiotic applications of S. noursei have

10

resulted in increase of endogenous actinomycetes population by 123.08 and 141.86 %,

respectively in PM and CSR2. Application of probiotics has paved way for eco-friendly
silkworm disease management (Subramanian et al., 2009).Recently efficacy of yeast as a nutrient
supplement had been tried in sericulture. Probiotic supplementations improved commercial
characteristics, disease resistance and protein in the silkworm, Bombyx mori L. Feed
supplementation not only enhanced economic and nutritional parameters but also prevent
bacterial infection in B.mori (Amala Rani et. al.,2011)..
Mode of action: Prebiotics affect intestinal bacteria by increasing the numbers of beneficial
anaerobic bacteria and decreasing the population of potentially pathogenic microorganisms.
Probiotics affect the intestinal ecosystem by stimulating mucosal immune mechanisms and by
stimulating non-immune mechanisms through antagonism / competition with potential
pathogens. These phenomena are thought to mediate most beneficial effects, including reduction
of the incidence and severity of diarrhea, which is one of the most widely recognized uses for
probiotics.
in mulberry silkworm (Bombyx mori ) presence of different types of bacteria in the gut have
been reported (Roy et al, 2000 and Kodama,2001).Most of the species belonging to Genus
Streptococcus are found to be pathogenic to Bombyx mori larvae while bacteria from Genus
Pediococccus, Leuconostoc and Lactobacillus didnt produce any infected silkworm. The precise
mechanism of beneficial effect on host or interaction among the different bacterial strains present
as micro-flora is not known (Steinhaus, 1949 and Kodama, 2001).
Different species of lactic acid bacteria have been extensively studied (Bruno et
al .,1993; Bernet-Camard et al.,1997 ;Gibson et al 1998; Kodama,2001) and found to be
beneficial as probiotics (Fuller, 1991;Bernet Camard, 1997;Sakamoto et al ; 2001).Therefore it
was planned to study the effect of a lactic acid bacteria namely Lactobacillus plantarum a known
probiotics on economic charecters. Silkworms fed on supplemented leaves dipped in different
lower and higher concentrations of three secondary metabolites to all the larval instars four times
in a day. Lower concentrations of pectin, amino acid mixture and proline proved to be
considerably effective showing higher shell ratio, weight of shell and cocoon was also noticed to
be higher, reliability was significantly higher and lower renditta found to be superior in lower

11

concentrations of secondary metabolites than the higher concentrations. These findings reported
that economic traits was noticed and found to be superior in lower concentration of secondary
metabolites, which performed best results amongst the treatments and natural diet, which showed
that significantly, improved the silk production and they can recommend as leaf supplementation
for increasing farmers income as well.

Vitamins
Studies of Ito (1978) determined that generally vitamins present in the mulberry leaves satisfy
minimum needs of silkworm but the amount of vitamins present in mulberry leaves varies on the
basis of environmental conditions, usage of fertilizers in field and mulberry varieties and other
field practices. Sengupta et al. (1972) showed that B. mori requires specific essential sugars,
amino acids, proteins and vitamins for its normal growth, survival and also for the silkgland
growth. Akhtar and Asghar (1972) found that vitamins and mineral salts played an important role
in the nutrition of silkworm. The effect of vitamin supplementation on the growth of B. mori
have been investigated by many researchers (Majumdar and Medda, 1975; Bhattacharyya and
Medda, 1981; Das and Medda, 1988; Faruki, 1990, 1998, 2005; Babu et al., 1992; Faruki et al.,
1992; Khan and Saha, 1996; Nirwani and Kaliwal, 1996, 1998; Mosallanejad, 2002; Etebari et
al., 2004; Rajabi et al., 2006a,b). Keeping the importance of vitamins on silkworm nutrition in
mind, following review was accomplished in order to determinate enrichment efficacy of
mulberry leaves by vitamins. A list of vitamin quoted in international research papers are showed
in Table 1.
Etebari et al. (2004)

Location
Iran

Sarkar et al. (1995)

Bangladesh

El-karaksy and Idriss (1990)

Egypt

Chauhan and Singh (1992)

Babu et al. (1992)
Gomma et al. (1977)
Das and Medda (1998)
Majumdar and Medda (1975)
Bhattacharya and Medda (1981)
Rajabi et al. (2006b)

India

Faruki (2005)

Derivative

Vitamin
Vitamin C

India

Cyanocobalmin (B12)

Iran

Pyridoxine (B6)

Bangladesh

Rajabi et al. (2006a)

Iran

Riboflavin (B2)

Nirwani and Kaliwal (1998)

Khan and Saha (2003)

India

Thiamine (B1)

12

Faruki (1998)
Nirwani and Kaliwal (1996)
Faruki and Khan (1992)
Khan and Faruki (1990)
Khan and Saha (1996)
Etebari and Matindoost (2004)
Saha and Khan (1996)
Etebari and Matindoost (2005)

Bangladesh

Folic acid (B9)

Bangladesh

PABA

Bangladesh

Fe-Plus

Iran

Niacin (B3)

Iran

Multi-vitamin compounds

Muniandy et al. (1995)

India

Saha and Khan (1996)

Bangladesh

Evangelista et al. (1997)

Thianomin

India

Brazil

Sarkar et al. (1995)

Bangladesh

B-complex

Mosallanejad (2002)

Iran

Vitamin E

Recently, much research has been done on the diet supplementation of mulberry leaves fed to
silkworms. These supplementations include vitamins such as ascorbic acid, thiamin, niacin, folic
acid and multi-vitamins (Etebari 2002; Nirwani and Kaliwal 1996, 1998; Saha and Khan 1996;
Etebari et al., 2004). Although some of the compounds have shown significant results,
enrichment has not always caused the improvement of biological characteristics of the silkworm.
In different studies, increase of supplement compound or increase of the period of its use show
negative effects that could be due to hypervitaminosis. Etebari et al., (2004) reported the yield
decrease, when ascorbic acid concentration is enhanced in silkworm diet. Saha and Khan (1996)
reported the same effects from multi-vitamins. When silkworm larvae were treated with different
concentrations of nicotinamide, high mortality was observed, although this vitamin is essential
for Bombyx mori L. (Etebari 2002). Dosage of this vitamin is very determinative for normal
growth of Mediterranean fruit fly. Chang and Li (2004) reported that nutritional interactions exist
between vitamin B3 and other groups of vitamin B.
Ascorbic acid

ascorbic acid decreases reactive oxygen species and oxidative pressure, and, as a result, the
absorption of nutritious substances in the midgut would increase (Felton and Summers, 1993).
Ascorbic acid shows a particular behaviour as it is very susceptible to degradation, especially
when in solution, and/or exposed to light, oxygen, and free radicals. Ito (1961) recorded

13

relationship of ascorbic acid supplementation and growth of silkworm. The absence of ascorbic
acid in the diet of first and second instar larvae postponed growth and development of silkworm.
There is enough Vitamin C in mulberry leaves and ascorbic acid content of growing larvae is
dependent on amount of this vitamin in diet. Supplementation of mulberry leaves more than any
other vitamin ascorbic acid has been used (Etebari et al., 2004). Several research demonstrated
phagostimulatory effect of ascorbic acid for insects (Ito, 1978; Dobzhenok, 1974). In silkworm a
gustatory stimulating activity have been observed to some extent (Ito, 1961). Gomma et al.
(1977) observed that ascorbic acid significantly increased the weight of silkworm larvae. Babu et
al. (1992) observed that the first and second instar larvae reared on 1.5 % ascorbic acid enriched
mulberry leaves resulted in higher silk filament length, weight and denier values. Sengupta et al.
(1972) reported that silk production increased with 1% ascorbic acid in the diet of silkworm.
Etebari et al. (2004) demonstrated that feeding on mulberry leaves enriched with ascorbic acid at
3% concentration decreased larval weight due to hypervitaminosis. Chauhan and Singh (1992)
showed that 1% concentration of ascorbic acid could increase the number of eggs in the
silkworm. Although its lower concentration the leaves in the first and second generation also did
not have positive effects on the fecundity in the silkworm.

Vitamin B Complex

The vitamin B complex is traditionally made up of 10 members (listed below) that differ in their
biological actions, although many participate in energy production from carbohydrates and fats.
The optimal levels of essential vitamins such as biotin,
choline, pyridoxine, panthotinate, inositol, riboflavin, thiamine, nicotinic acid have been
determined by Horie and Ito (1963, 1965) (Table 2).
Riboflavin (B2)
Riboflavin is important in promoting the release of energy from carbohydrates, fats and proteins
i.e. in the metabolic pathway for ATP production. The enrichment of mulberry leaves with

14

riboflavin at 77 ppm enhanced certain economic characters of silkworm, and improved silk
production in north climatic conditions of Iran (Rajabi et al., 2006a). Male cocoon weight (1.195
g) was greater at 77 ppm while female cocoon weight (1.622 g) was greater at 127ppm.
Maximum male pupal weight was recorded at 37 ppm (0.895 g) compared to 127 ppm for the
female (1.169 g). Male and female shell weight (0.311 and 0.318 g) had significant increase at 77
ppm compared to control (0.276 and 0.277 g). Male and female cocoon shell percentage reached
their maximum at 77 ppm treatment, which were 26.06 % and 21.46 % respectively. Average of
50 egg weight, number of eggs for every female and hatchability percentage did not show
significant difference among the treatments and the control. Similar improvements were not
obtained in Natanz, in the center of Iran, place which has dry climatic conditions.
Folic acid (B9)
Folic acid plays a major role in cellular metabolism including the synthesis of some of the
components of DNA and pigment precursor (National Research Council (U.S), 1987, Chapman,
1998). Yosuhiro and Sholchi (1971) noted that the silkworm growth decreased when folic acid
was eliminated from artificial diet. Nirwani and Kaliwal (1996) determined that folic acid has
phagostimulatory effects with significant increase in female and male cocoon weight and shell
weight.
Para-amino benzoic acid (PABA) is a growth regulator and represents one of the forms of
folic acid. PABA is one of the substances belonging to the vitamin B-complex group and
supports vital function in insects and especially in silkworm (Pai et al., 1988). PABA
supplementation has no significant effects on adult weight whereas it caused deleterious effects
on their length (P<0.001) and wing-span (P<0.001) (Faruki and Khan, 1992). Fe-PLUS (ferrus
fumarate+folic acid) supplementation significantly increased larval, pupal and adult weight in
comparison with controls with lowest and highest growths obtained at the concentrations of 0.32
and 0.64 %, respectively (Khan and Saha, 1996). Larval and pupal periods decreased at lower
doses (0.08 and 0.16 %) while they increased after exposure to higher doses (0.32 and 0.64 %).
Fertility increased significantly in all treatments when compared to control except for 0.64 %
concentration (Khan and Saha, 1996).
Pyridoxine (B6)

15

Pyridoxine is necessary for the proper functioning of over 60 enzymes that participate in
amino acid metabolism. It is also involved in carbohydrate and fat metabolism. Without
pyridoxine or its derivatives no larva reached the third instar under aseptic condition. Pyridoxine
is important in protein metabolism and its deficiency in mammals results in decrease in
phosphorylases (National Research Council (U.S), 1987). Faruki (2005) reported that mulberry
leaf fortification by pyrol (Pyridoxine hydrochloride HCl.25) in various concentrations from the
third instar significantly reduced the fecundity. In this experiment it was found the lowest
number of eggs was produced at the lowest concentration (10 g /ml) and at higher
concentrations it remains lower than in the control. Banerjee and Khan (1992) observed that
vitamin B6 enhances the oviposition of the silkworm infected by Bacillus thuringiensis but the
rate was lower than the control. Faruki (2005) reported that higher concentration of vitamins
reduced the fecundity and fertility of silkworm. Further, the percent reproduction control (PRC)
in lower concentrations was more than higher concentrations (Faruki, 2005). The reproductive
success in lower concentrations was prominent with compared with the higher concentrations
(Faruki, 2005; Etebari and Matindoost, 2005). Rajabi et al (2006b) showed that in north climatic
conditions of Iran, larval weight reached a maximum 2.601 g at the end of 5th instar. Effective
rate of rearing (ERR) was higher (75.33 %) at 100 ppm concentration compared with other
concentrations (10, 500 and 1000 ppm). Larval and silk indices reached their maximum at 100
ppm concentration while pupal and adult indices reached their maximum at 1000 ppm
concentration in male and female. Larval duration was longer (622.5 h) in treatments against
control (604.5 h). Treatment differences were recorded also in respect of other economic
characteristics (Rajabi et al., 2006b).
Nicotinic acid (B3, Niacin)
Niacin is important for the release of energy from carbohydrates and fats, the metabolism of
proteins and production of several hormones (National Research Council (U.S), 1987). Horie and
Ito (1965) showed the required level of niacin for silkworm is highly regulated to the most
appropriate level of 33 g/l of dry weight and the increase of niacin reduced the larval weight.
Horie (1995) showed a reduction of requirement pattern with increasing larval weight. Niacin
caused significant deleterious effects on larval growth. Cocoon parameters such as cocoon
weight, pupal weight and cocoon shell weight also showed significant decrease in all treatments

16

(Etebari and Matindoost, 2004). Furthermore, mulberry leaf enrichment with nicotinamide (10,
20 and 30 g/l) from first instar caused intensive mortality in the larval growth and only 1.2%
larvae could reach the 5th instar. High doses (20 and 30 g/l) of nicotinamide killed all larvae
before entering the 5th instar (Etebari and Matindoost, 2004). Horie and Ito (1965) reported that
the analogues of niacin, 4-acetyl pyridine interrupts the larval growth when added to mulberry
leaves and acted as an antimetabolite. Niacin with 0.5 g/l acted as an antifeedant for silkworm
larvae and decreased their metabolism (Etebari and Matindoost, 2004).

Thiamine (B1)
Thiamine is important for energy metabolism (National Research Council (U.S), 1987). Nirwani
and Kaliwal (1998) reported that the weight of larvae and silk glands in all the thiamine fed
groups had not shown any significant changes. On the other hand, larval duration, cocoon
weight, shell weight and fecundity increased significantly. In opposition with what observed for
ascorbic acid and folic acid, it has been found that thiamine has no phagostimulatory effect on
silkworm (Horie and Ito, 1963; Nirwani and Kaliwal, 1996). Faruki (1998) reported that the
thiamine derivative thianomin enhanced the growth of silkworm larvae, pupae and adults at all
concentrations used (50, 100, 500 and 1000 ppm). Mulberry leaf enrichment with thianomin
increased the growth indices such as larval, pupal and adult weight. Silk index increased too in
all treatments. Thianomin increased cocoon characters such as cocoon weight, shell weight,
cocoon shell length and breadth significantly (P< 0.05).
Cyanocobalamin (B12, Cobalamin)
Cyanocobalamin plays important role in silkworm because it is a co-factor of propionate
metabolism which is important substrate for biosynthesis of juvenile hormone in insects
(Halarnkar and Blomquist, 1989). Vitamin B12 does not occur in mulberry leaves but
considerable amount of this vitamin was observed in larvae and pupae. It is believed that
actinomycetes in the gut lumen produce vitamin B12. Das and Medda (1998) reported that
supplementation of mulberry leaves with B12 vitamin could increase the synthesis of nucleic
acid and proteins in the silkglands of the silkworm.

17

Pantothenic acid (B5)

Pantothenic acid is the precursor of coenzyme A that is vital for the metabolism of
carbohydrates, the synthesis and degradation of fats, the synthesis of sterols and the resultant
steroid hormones, and the synthesis of many other important compounds (National Research
Council (U.S), 1987).
Biotin (B8, Vitamin H)
Biotin has an important role in carbohydrate and fat metabolism. It has been showed that biotin
is one of the essential vitamins for the silkworm B. mori (Horie et al., 1966). It has important
role in the synthesis of fatty acids in the silkworm and it is confirmed that minimal optimal level
of biotin for growth and survival of the silkworm was much lower than those of other vitamins
including nicotinic acid, pantothenic acid and pyridoxine (Horie and Ito, 1965). It is identical
with the minimal threshold for alteration of fatty acid composition (Horie and Nakasone, 1968).
However, to the best of our knowledge, research reports on enrichment mulberry leaves with
biotin are not available.
Choline and inositol
Choline is traditionally not a vitamin; however it was identified as part of vitamin B complex.
Inositol is an important part of signaling mechanism that transmits information from outside to
the inside of cells. It is generally considered dispensable in insect diets. Choline and inositol are
required by silkworm in higher level because they are lipogen substrate, also involved in the
production of cell membranes. However, research reports on enrichment mulberry leaves with
choline and inositol do not seem to be available.
Multi-vitamin compounds
Saha and Khan (1996) described the extensive effects of multi-vitamin compounds as diet
factors on growth interruption and the decrease of cocoon economical characteristics. It is
showed that multi-vitamin and mineral compounds could increase the food intake, growth and
conversion efficiency of silkworm (Muniandy et al ., 1995). Evangelista et al. (1997) reported
that the larval and cocoon weight increase under multi-vitamin compound treatment, but did not

18

have any positive effects on cocoon shell weight. Feeding with multi-vitamins in the larval stage
adversely affected the hatchability of eggs. Multi-vitamins even though could increase some
biological characteristics in silkworm, did not influence the economical and yield contributing
parameters. Etebari and Matindoost (2005) reported that feeding of silkworm on mulberry leaves
enriched by with multi-vitamins from 4th instar increased female cocoon shell weight in 2.5%
concentration, while female pupal weight increased in 1% concentration. Male and female shell
ratio did not increase compared to controls
Fat-soluble vitamins

Fat-soluble vitamins consist of the A, D, E and K vitamins. Among these, enrichment of

mulberry leaves with vitamin A, D or K for silkworm do not seem to have been studied.
Vitamin E
-tocopherol is slightly effective in increasing the number of eggs laid by moths and -carotene
has also some growth-promoting effect (Ito, 1978). Enrichment of mulberry leaves with E
vitamin did not have significant effect on food consumption in silkworm larvae (Mosallanejad,
2002).
Antibiotics
Supplementation of antibiotics and some botanicals to mulberry leaves were attempted by a few
workers to increase the survival in silkworms previous reports indicated that Choromycetin
treatment increased resistance to disease (Murthy and Sreenivasaya, 1953)
This review summarizes data showing that enrichment of mulberry leaves with various
vitamins have different effects on economic traits and biological parameters of the silkworm.
However, reported effects depend also on weather condition, larval stage treated, type of vitamin,
varieties of mulberry and silkworm race studied. This possibly indicates the need for elaborating
comprehensive studies on the subject. It is advisable that we keep in mind the negative effects of
enrichment beside its positive effects on economic traits and biological parameters. Moreover,

19

each intervention on the natural content of silkworm food should take into account also costs,
environmental safety and large scale feasibility.

References

Akhtar M, Asghar A. Nutritional requirement of silkworm Bombyx mori. Pakistan J. Zool. 4:

101-107, 1972.
Babu M, Swamy MT, Rao PK, Rao MS. Effect of ascorbic acid-enriched mulberry leaves on
rearing of Bombyx mori L. Indian J. Seric. 31: 11-114, 1992.
Banerjee SK, Khan AR. Effect of vitamin B6 on the fecundity of Bacillus thuringiensis var.
kurstaki treated Bombyx mori L. Bangladesh J. Zool. 20: 361-362, 1992.
Bhattacharyya A, Medda AK. Effect of cyanocobalmine and cobalt chloride on glycogen of
silkgland of Bombyx mori L.Nistari race. Sci. Cult. 77: 268-270, 1981.
Chapman, R. F. 1998. The Insect Structure and Function, Cambridge Univ. Press, Cambridge.
Chapman, R. F. The Insect Structure and Function, Cambridge University Press, Cambridge.
1998.
Chauhan T, Singh K. Studies on the effect of ascorbic acid on the fecundity in the mulberry
silkworm. Sericologia 32: 567-574, 1992.
Das S, Medda A. Effect of cyanocobalamine on protein and nucleic acid contents of ovary of
silkworm, Bombyx mori L., during larval, pupal and adult stages of development. Insect Sci.
Appl. 9: 641-646, 1988.
Dobzhenok NV. The effect of ascorbic acid on the physiological condition of the codling moth
and its resistance to fungus and bacterial infection. Zakhist Roslin. 19: 3-7, 1974.

20

El-karaksy, I. R. and M. Idriss. Ascorbic acid enhances the silk yield of the mulberry silkworm,
Bombyx mori L. J. Appl. Entomol. 109: 81-86, 1990.
Etebari K, Ebadi R, Matindoost L. Effect of feeding mulberrys enriched leaves with ascorbic
acid on some biological, biochemical and economical characteristics of silkworm Bombyx mori
L. Int. J. Indust. Entomol. 8: 8187, 2004.
Etebari K, Matindoost L. Application of multi-vitamins as nutrients on biological and
economical characteristics of silkworm Bombyx mori L. J. Asia-Pacefic Entomol. 8: 1-6, 2005.
Etebari K, Matindoost L. Effects of hypervitaminosis of vitamin B3 on silkworm biology. J.
Biosci. 29: 417422, 2004.
Etebari, K. and L. Matindoost. 2005. Application of multivitamins as supplementary nutrients on
bio economical characteristics of Bombyx mori. J. Asia- Pacific Ent., 8(1): 1- 6.
Evangelista A., Carvalho AD, Takahashi R, De Carvalho AD. Performance of silkworm
(Bombyx mori L.) fed with vitamin and mineral supplement. Rev. Agricul. Pirac. 72: 199-204,
1997.
Faruki SI, Khan AR, Mannan A. Fecundity and fertility of the silkworm, Bombyx mori L. fed on
mulberry leaves supplemented with para-amino benzoic acid. Bangladesh J. Zool. 20: 351-353,
1992.
Faruki SI. Effect of pyridoxine on the reproduction of the mulberry silkworm, Bombyx mori L.
(Lepidoptera: Bombycidae). Inv. Surv. J. 2: 28-31, 2005.
Faruki SI. Nutritive effects of thianomin enriched mulberry leaves on the silkworm, Bombyx
mori L. Univ. J. Zool. Rajshahi Univ. 17: 39-44, 1998.
Faruki SI. The effect of para-amino benzoic acid on Bombyx mori L. (Lepidpotera; Bombycidae)
M.Sc. Thesis, Rajshahi University, Bangladesh, 1990.
Felton GW, Summers CB. Potential role of ascorbic oxidase as a plant defense protein against
insect herbivory. J. Chem. Ecol. 19: 1553-1568, 1993.

21

Ganga, G. and Sulochana Chetty. 1991. Introduction to Sericulture. 2nd edn. Oxford and IBH
Pub. Comp. Pvt. Ltd. New Delhi. pp: 26-35.
Gomma AA, El-Shaaraway MF, Salam YS, Rizk MA. Effect of dietary constituents on the
biology of silkworm, Bombyx mori L. II. Vitamins. Z. Ang. Ent. 64: 231-240, 1977.
Halarnkar PP, Blomquist G. Comparative aspects of propionate metabolism. Comp. Biochem.
Physiol. 92B: 227-231, 1989.
Harizanis, C. P. 2004. Manual of Sericulture Silkworm Rearing and Mulberry Cultivation.
Athens, pp:22.
Horie Y, Ito T. Nutrition of the silkworm, Bombyx mori-X. Vitamin B requirments and the
effects of several analogues. J. Insect Physiol. 11: 15851593, 1965.
Horie Y, Ito T. Vitamin requirements of the silkworm. Nature 197: 98-99, 1963.
Horie Y, Nakasone S. Effect of dietary biotin on fatty acid composition of the silkworm, Bombyx
mori L. J. Insect. Physiol, 14: 1381-1387, 1968.
Horie Y, Watanabe K, Ito T. Nutrition of silkworm, Bombyx mori L. XIV. Further studies on the
requirements for B vitamins. Bull. Seric. Exp. Stn Japan, 20393-409, 1966.
Horie Y. Recent advances of nutritional physiology and artificial diet of the silkworm, in Japan.
Korean J. Seri. Sci. 37: 235243, 1995.
Ito T. Effect of dietary ascorbic acid on the silkworm, Bombyx mori L. Nature, 192: 951-952,
1961.
Ito T. Silkworm Nutrition; in the Silkworm an Important Labratoary Tool. Tazima, Y. (Ed), pp.
121-157, Kodansha Ltd, Tokyo, 1978.
Ito, T. 1978. Silkworm nutrition: Silkworm an important laboratory tool. Tazima, Y. edn
Kodansha Ltd., Tokyo. 121-157.
Ito, T. 1983. Nutrition of silkworm and the artificial diet (In Japanese). Jpn. Seric. J. Press,

22

Tokyo. Jpn., Pp: 292.

Joshi, K. L. 1985. Studies on growth index for Eri-silkworm, Philosamia ricini. Sericologia., 24:
21-27.
Kabila, V., K.M. Subburathinam and J.S. Chetty. 1994. Growth and economic characters of
Bombyx mori on feed enriched with neutralized aspartic acid. Ind. J. Seric., 33: 80-81.
Khan AR, Faruki SI. Growth and development of Bombyx mori L. on feed supplemented with
para-amino benzoic acid. Univ. J. Zool. Rajshahi Univ. 9: 47-53, 1990.
Khan AR, Saha BN. Nutritive effects of Fe-PLUS (ferrus fumarate + acid folic) on the
silkworm, Bombyx mori L. Bangladesh J. zool. 24: 199-203, 1996.
Khan AR, Saha BN. Nutritive effects of thiamine on the silkworm, Bombyx mori. Bangladesh. J.
Zool. 31: 169-176, 2003.
Kirimura, J. 1962. Studies on amino acid composition and chemical structure of silk protein by
microbiological determination. Bull. of Seric. Expt. Stn., 17: 447-522.
Legay JM. Recent advances in silkworm nutrition. Ann. Rev. Ent. 3: 75-86, 1958.
Legay, J. M. 1958. Recent advances in silkworm nutrition. Ann. Rev. Ent., 3: 75-86.
Lu, S. L and Z.D. Jiang. 1988. Absorption and utilization of amino acids in mulberry leaves by
Bombyx mori L. Acta Sericologia Sancta., 14: 198-204.
Majumdar AC, Medda AK. Studies on the thyroxine and vitamin B12 induced changes in the life
cycles of silkworms. Indian J. Physiol. Appl. Sci. 29: 1-13, 1975.
Mosallanejad H, Bagheri Zonus E, Nouzari J, Talebi, M. Effect of feeding the first to third instar
larvae of silkworm (Bombyx mori) with mulberry leaves enriched with vitamin E on some
reproductive characteristics. Proceeding of 15th Iranian Plant Protection Congress. September,
2002. Kermanshah. P. 167, 2002.
Muniandy S, Sheela M, Nirmala S. Effect of vitamins and minerals (Filibon) on food intake,

23

growth and conversion efficiency in Bombyx mori. Environ. Ecol. 13: 433-435, 1995.
National Research Council (U.S). Vitamin tolerance of animals. National Research Council
(U.S.). Subcommittee on Vitamin Tolerance.; NetLibrary, Inc.National Academy Press,
Washington DC, pp 96, 1987.
Nirwani RB, Kaliwal BB. Effect of Folic acid on economic traits and the change of some
metabolic substances of bivoltine silkworm, Bombyx mori L. Korean J. Seric. Sci. 38: 118-123,
1996.
Nirwani RB, Kaliwal BB. Effect of thiamine on commercial traits and biochemical contents of
the fat body and haemolymph in the silkworm Bombyx mori L. Sericologia 38: 639-646, 1998.
Pai IK, Hedge SN, Krishnamurthy NB. PABA induced genotoxic effects in silkworm, Bombyx
mori. Proc. Int. Sym. On nature of genetic variation in man (XII Ann. Conf. of EMSI),
Hydarabad, p. 54, 1988.
Radjabi, R., R. Ebad, S.Z. Mirhoseini and S. Nair. 2009. Effects of feeding alanine enriched
leaves on economic characters of Bombyx mori. Formosan Entomologist. 29:73-78.
Radjabi, R.; Ebadi, R.; Mirhoseini, S.Z. and S. Nair. 2010. Effect of leaves enrichment with
amino acid supplementary nutrients on Bombyx mori L. Acad. J. of Ent., 3(1): 45-51.
Rahmathulla, V. K.; Priyabrata, D.S.; Ramesh, M. and R.K. Rajan. 2007. Growth rate pattern and
economic traits of Bombyx mori under influence of folic acid administration. J. Appl. Sci. Envn.
Manage., 11(4); 81-84.
Rajabi R, Ebadi R, Fazilati M, Mirhoseini SZ. Nutritive effects of mulberry leaves enrichment
with riboflavin vitamin on bio-economic characters of silkworm, Bombyx mori L. 9th Arab
Congress of Plant Protection. 19-23 November, Damascus, Syria, 2006a.
Rajabi R, Ebadi R, Fazilati M, Mirhoseini SZ. The effects of mulberry leaves enrichment with
pyridoxine-HCl on economic traits and biological parameters of silkworm Bombyx mori L. 17th
Iranian Plant Protection Congress, December 2006, Teheran, P391, 2006b.

24

Saha BN, Khan AR. The growth and development of the silkworm, Bombyx mori L. on feed
supplemented with nicotinic acid. Bangladesh J. Life Sci. 1: 103-109, 1996.
Sarkar A, Haque M, Rab M, Absar N. Effect of feeding mulberry (Morus sp.) leaves
supplemented with different nutrients to silkworm, Bombyx mori L. Curr. Sci. 69: 185-188,
1995.
Seki K, Oshikane K. Res. Reports. Fac. Textile and Sericulture, Shinshu University, 1959.
Seki, K., and K. Oshikane. 1959. Res. Rept. Fac. Text. and Seric., Shinshu Univ., China.
Sengupta K, Singh BD, Mustafi JC. 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 protection. Indian J. Seric. 11: 11-27, 1972.
Sengupta, K., Singh, B.D. and J.C. Mustafi. 1972. Nutrition of Bombyx mori. Studies on
enrichment of leaf with sugars, proteins, amino acids and vitamins for vigorous growth of
silkworm and increased cocoon crop production. Ind. J. Seric., 11: 11-27.
Tazima, Y. 1978. Silkworm: An Important Laboratory Tools, Kondansha Ltd, Tokyo.
Trivedy, K., Nair, K.S.; Ramesh, M.; Gopal, N.; and S.N. Kumar. 2003. New Semi-synthetic diet
Nutrid A technology for rearing young instar silkworm in India. Ind. J. Seric., 42(2): 158161.
Yasuhiro H, Sholchi N. Effects of levels of biosynthesis of fatty acids and carbohydrates in a diet
on the biosynthesis of fatty acids in larvae of silkworm. J. insect Physiol. 19: 1441-1450, 1971.

25