UGSemsterSyllabus Zoology 6Sem8A619Zoology English SilkWarmbreeding PDF

BS619ZOO-DSE(B)-E
B.Sc.
THIRD YEAR SEMESTER-VI
ZOOLOGY
SILKWORM BREEDING AND GRAINAGE
“We may forgo material benefits of civilization, but we cannot forgo our right
and opportunity to reap the benefits of the highest education to the fullest extent
as the education is the greatest material benefit”
Dr. B. R. Ambedkar
Dr. B. R. AMBEDKAR OPEN UNIVERSITY

HYDERABAD
2020
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COURSE TEAM
Course Development Team Course Development Team (CBCS)

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Prof.A.Purushotham Rao Dr.Ponna Srinivas
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Dr. B. R. Ambedkar Open University, Hyderabad.

First Edition: 2020
Copyright © 2020, Dr. B. R. Ambedkar Open University, Hyderabad, Telangana.
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from the University.
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PREFACE
This course material is prepared as per the revised syllabus of Under Graduate Courses in Zoology taking
into account the recommendations of the Expert Committee. The chief purpose is to uplift and bring about
parity between this course material and the CBCS syllabus pattern of all the universities. Certain topics
with reference to competitive examinations are included in the syllabus. Recent findings on certain
aspects of structures and physiology are incorporated. To create interest in the learner and grasp the
subject the format adopted is self instructional in a simple and lucid language.
This book deals with the topics in Zoology included in the syllabus for the Third Year, of the B.Sc Course
offered by Dr. B. R. Ambedkar Open University. These topics cover the ―Elective‖ area of the subject to
be studied in the Third Year, sixth semester of the Three Year Degree Course in Science. The syllabus for
the sake of convenience is divided into 4 blocks, each of with 3 units. Each block is framed to cover a
specified area of the subject. The units are prepared by experts in accordance with a format so designed so
as to enable the student to read and understand the topics without much difficulty. Each unit begins with
contents followed by the objectives of the lesson, major topics covered and ends with model questions
intended to test the student‘s comprehension of subjects matter.
The objective of this book is to explain, the ―Silkworm Breeding and Grainage’’. Block I is
allocated to Silkworm Breeding to explain Introduction to Silkworm Breeding, Selection Methods,
Heterosis and Hybrid Vigour. Block II is for Grainage-I to detail Parental Races, Breeding stations,
Selection of seed cocoons. Block III is allocated to Grainage-II to describe Grainage, Silkworm Seed
Production, Egg Preservation and Hibernation; Block IV is allotted to Grainage-III to explain Acid
Treatment, By products and Economics. These are followed by Model question paper.
Every care is taken to follow the guidelines evolved by expert committee and at the same time to present
the subjects, topic and sub-topic wise for the easy reading and understanding of the student.
Improvement is a constant and continuous effort. We appreciate the constructive suggestions for further
improvements.
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CONTENTS
Block/Unit Title Page
BLOCK- I SILKWORM BREEDING

Unit –1 : Introduction to Silkworm Breeding 05-12
Unit –2 : Selection Methods 13-24
Unit- 3 : Heterosis and Hybrid Vigour 25-28
BLOCK -II GRAINAGE-I

Unit –4 : Parental Races 29-34
Unit –5 : Breeding Stations 35-42
Unit –6 : Selection of Seed Cocoons 43-47
BLOCK- III GRAINAGE-II
Unit –7 : Grainage 48-57

Unit –8 : Silkworm Seed Production 58-66
Unit –9 : Egg Preservation and Hibernation 67-71
BLOCK -IV GRAINAGE-III
Unit –10 : Acid Treatment 72-78

Unit –11 : By Products 79-83
Unit –12 : Economics 84-87
MODEL QUESTION PAPER 88
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UNIT-1 SILKWORM BREEDING
Contents
1.0 Objectives
1.1 Introduction
1.2 Silkworm Breeding
1.2.1 Inbreeding and Out breeding concepts
1.2.2 Objectives of silkworm breeding techniques
1.2.3 Different types of breeding methods
1.2.4 Line breeding, Cross breeding and Mutation breeding
1.3 Summary
1.4 Check your Progress Answers
1.5 Model Examination Questions
1.0 OBJECTIVES
After studying this unit, you will be able to:
 learns about silkworm breeding.
 understands differences between inbreeding and out breeding
 understands types of breeding methods.
1.1 INTRODUCTION
Silkworms were first domesticated in China over 5,000 years ago. Since then, the silk production capacity
of the species has increased nearly tenfold through artificial selection and an intensive breeding process.
The silkworm is an organism wherein the principles of genetics and breeding are applied in order to
harvest the maximum output. It is the second following maize in exploiting the principles of heterosis and
cross breeding. Traditional silkworm breeding has been focused on achieving higher production with
better quality while improving survival, reproduction and physiological characteristics.
1.2 SILKWORM BREEDING
Early history of Indian silkworm breeds

The goal of silkworm breeding is to attarnmaximum productivity in yield and quality. This goal is
achieved by bringing genetic improvement through combination of desired genes by crossing two
selected pure stocks of the silkworm followed by selection. Though, sericulture is introduced as
commercial venture way back in late 17"'sentury, silkworm breeding was initiated only in 1920s.
Basically polyvoltine culture prevailed up to the 50s and indigenous polyvoltine breeds like Pure Mysore
and C.Nichi in South India, Nistari in West Bengal, Sarupat and Moria in the North East were reared.
Though, these breeds were very well adapted and popular in respective regions, their productivity and
quality of silk was strikingly low with very high renditta.
Silkworm breeding is aimed at the overall improvement of silkworms from a commercial point of view.
The major objectives are improving fecundity (the egg-laying capacity of a breed), the health of larvae,
quantity of cocoon and silk production, and disease resistance. Healthy larvae lead to a healthy cocoon
crop. Health is dependent on factors such as better pupation rate, fewer dead larvae in the
mountage, shorter larval duration (this lessens the chance of infection) and bluish-tinged fifth-instar
larvae (which are healthier than the reddish-brown ones). Quantity of cocoon and silk produced are
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directly related to the pupation rate and larval weight. Healthier larvae have greater pupation rates and
cocoon weights. Quality of cocoon and silk depends on a number of factors, including genetics.
1.2.1 Inbreeding and Out breeding Concepts
Inbreeding is the mating of individuals or organisms that are closely related through common ancestry,
as opposed to out breeding, which is the mating of unrelated organisms. Inbreeding is useful in the
retention of desirable characteristics or the elimination of undesirable ones, but it often results in
decreased vigour, size, and fertility of the offspring because of the combined effect of harmful genes that
were recessive in both parents. It results in a decline in survival and reproduction (reproductive fitness),
known as inbreeding depression, in most species of plants and animals. Both inbreeding and hybridization
are forms of nonrandom mating or selective mating, but operate in opposite ways. Inbreeding is a kind of
genetic assortative mating as compared with phenotypic assortative mating of hybridization. While gene
frequencies do not change on the whole, genotypic frequencies do change toward the production of more
homozygotes and fewer heterozygotes. Thus, any change in the population mean as a result of inbreeding
must be related to a difference in genotypic value between homozygote and heterozygote.
If Mo and Mf indicate the original and subsequent population means, respectively, the result from
inbreeding is expressed as follows:
Mf = Mo – 2hpqF , where p and q represent gene frequencies of a population for a given character, F is
the inbreeding coefficient and h is the value of the heterozygote.
If the value of heterozygote, h, is zero, then there is no change in the population mean. That is, there is no
dominance and inbreeding shows no effect on the means. When ―h‖ does have a value, there is a change
in the population mean with inbreeding. This change is a consequence of dominance with the direction of
change toward the most recessive allele. In general, the characters that are important for fitness of the
individual will show a reduction due to inbreeding. Those characters unrelated to fitness will show little
or no inbreeding depression. When inbreeding is accompanied by directional selection, the resulting
families will be similar phenotypically, and total genetic variance among them will be decreased.
Since the self–fertilization in the silkworm is not possible the most intense form of inbreeding is full– sib
mating. Less intense forms of inbreeding can be obtained from cousin, second cousin and other types of
family mating. The relative effects can be seen in the inbreeding coefficient, F, in consecutive
generations.
In practical terms, the breeding expectation is that inbreeding leads to isolation of homozygous lines of
generally reduced vigor. The reduction in vigor is not in a direct relationship with generation, but
generally parallels the reduction in heterozygosity. Inbreeding can be expedited by selecting for the
ultimate inbred characters, if they are defined. This system of mating will exhaust its effect in a few
generations and has little impact thereafter. Many authors, on analyzing Bombyx mori L. breeding
methods, assume that the manifestation of inbred depression depends mainly on biological features,
geographic and genetic origin of the utilized initial population. For example, monovoltine populations
is less resistant to depression compared to bi- and polyvoltine ones due to their weaker adaptability to
unfavourable environmental conditions. At the same time inbreeding in combination with intensive
systematic selection led to creation of highly productive lines of Bombyx mori L.
Out breeding refers to matings between individuals from different populations, subspecies, or species.
Out breeding can result in a decline in reproductive fitness known as out breeding depression, but this is
less common than inbreeding depression. Inbreeding in small populations typically increases extinction
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risk, especially for species that do not normally inbreed. Out breeding between populations with
chromosomal incompatibilities or those that are adapted to different environmental conditions can also
increase extinction risk. Restoring gene flow between isolated populations can reverse inbreeding
depression.
It includes out-crossing, cross-breeding, and interspecific hybridization.
 Out-crossing: It is the mating between animals of the same breed, but not having common
ancestors for 4−5 generations. It is usually used for animals, which have below average
productivity and growth rate. It helps to overcome inbreeding depression
 Cross-breeding: It is the mating between a superior male of one breed with a superior female of
another breed. Superior qualities of both the breeds combine and this is known as hybrid vigour.
The progeny so formed is called a hybrid. A hybrid may be used as it is or may be further
subjected to inbreeding.
Example: Hisardale sheep is a hybrid of Bikaneri ewes and Marino rams.
 Interspecific Hybridization: Males and females of different, but related species are mated.
Progeny has desirable features of both the species.
Example: Mule is an interspecific hybrid of donkey and horse. It is swifter and stronger than
donkeys and more disease resistant than a horse.
Inbreeding Outbreeding
Mating of more closely related individuals. Mating of unrelated animals.
If mating is between animals of same breed then
Mating is between animals of the same breed
there should be no common ancestors for 4-6
for 4-6 generations.
generations.
Advantage is that it produces hybrids with
Advantage is that it increases homozygosity
desirable characters like better lactation period
and so is used for developing pure lines.
and high milk productions.
Disadvantage is that it causes inbreeding
It causes outbreeding depression because of
depression because of which there is decline in
which there is decline in reproductive fitness.
survival and reproduction.
Mating is between same species. Mating is between different species.
1.2.2 Objectives of silkworm breeding-techniques
The main objectives of silkworm breeding are

 Improving fecundity.
 Improving healthiness of larvae.
 Improving quantity of cocoon and silk production.
 Improving quality of cocoon and silk production, for specific purposes based on cocoon and silk
production
 For disease resistance etc.
i. Fecundity: It refers to the egg laying capacity of a breed. It is a very important factor since
commercial sericulture is strongly dependent on silkworm egg.
ii. Healthiness of larvae: Healthy larvae lead to healthy cocoon crop. Healthiness is dependent on
factors such as better pupation, rate, less number of dead larvae in the mountage, shorter larval
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duration (shorter the larval duration lesser the chances of infection) and bluish tinged fifth instar
larvae ( it is observed that bluish colored fifth instar larvae are healthier than the reddish brown
ones).
iii. Quantity of cocoon and silk: Quantity of cocoon produced is directly related to the pupation rate
and larval weight. Healthier the larvae more will be the pupation rate and cocoon weight.
iv. Quality of cocoon and silk: This depends upon a number of factors including genetic factors.
v. Specific purposes: Apart from commercial purpose advanced countries are giving attention to
specific breed development for specific purposes like sericin production, sex limited breeds,
thin/thick filament production etc.
vi. Disease resistance breeding: The major reason for crop losses is pathogen infection. Efforts are
vogue to evolve breeds which are tolerant or resistant to various pathogens.
Fig.1.1 Different Breeds
According to Lea(1993) there are at least five quantitative characters that are of interest to silkworm
breeders.
1. First is egg color , which is determined in a combination of shell color(maternal origin , genes
normally colourless to semi-transparent to opaque chorion and mutant yellow (ye) and green Gre,
etc.) and serosa color (embryonic origin, genes normally dark brown and mutants white we–
alleles, pink pe, red re, brown b–alleles etc.). Pigments appear in the serosa cells of the
hibernating egg on the second day after egg–laying, and the final colouring is reached in three or
four days. In the non-hibernating egg, the serosa remains colourless (the npnd gene).
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2. Second are larval markings which are normally p+ and p, S-, Ze – alleles with various markings
to plain p without any markings. The gene p is in multiple allelic series condition.
3. Third is blood colors are determined by two major independent genes, yellow Y and white + y,
and yellow inhibitors I/Is , plus red haemolymph rb.
4. Fourth is cocoon colors which are normally white. Green and yellow are also popular. A variety
of colored cocoons can be obtained by combining cocoon color genes: C-alleles - +c, golden
yellow C, light yellow Cd, inner layer yellow Ci. Dominant white I inhibits the action of Y, and
thus is colorless when YC; flesh colored F when combined as genotype YCF; pink Pk when
YCFPk and greens can be Ga, Gb, Gc.
5. Fifth is cocoon shape. This is heritable, but it is difficult to designate a gene to a certain shape due
to an almost continuous quantitative–like variation of the various topological factors involved.
The shape is commonly elliptic. More spherical cocoons are found in Chinese varieties, more
peanut–shaped in Japanese and more peaked cocoons in tropical varieties. The cocoon shape
inheritance is controlled by polygenic additive gene actions. There is also a wide range of
variation in size and surface texture, with double cocoon semi-dominant over single.
1.2.3 Different types of breeding methods
Methods of breeding sex limited for egg color, larval marking and cocoon color races:
It is well known that the main reason to use sex - limited silkworm lines is in order to make easier, and
more effective silkworm egg production, especially the sex discrimination. The first attempts to create
sex-limited silkworm breeds were made.
The first silkworm strain, having marked female larvae and plain male was created by Tazima (1944) by
treatment of newly laid silkworm eggs with X–rays and translocation of the P+ allele and its connection
with the sex W chromosome. Later, using the same method Hasimoto(1948) managed to create a new
sex-limited breed having zebra female and plain male larvae. By this method as initial material were used
sex limited races, crossed with other races, having plain larvae or yellow eggs. After that the hybrid
population was maintained by batch rearing for 4 generations and with inside batch mating (inbreeding).
On the 5th generation an inter batch crossing was made, followed again by inbreeding in the 6th and 7th
generations. At the 8th generation the two inbred lines were crossed and the new breed was created.
Using this method Petkov(1995) selected 2 sex limited for egg color lines (T15/4 and HT-215/38) and 5
sex limited for larval marking lines (B2/6,BTV-2/64,TBV-2/24,HB-2/22 and TV-3/2).
Sex-limited cocoon color:
Female cocoon is yellow and male white. It is easily for sex-discrimination and the male cocoon can
separately be reeled to produce high grade silk. According to Chen (2002), CY2c (Chinese race) and
JY628 are one of this type of commercial varieties. Their pure line performance are 1.543 g and 1.694g in
cocoon weight, 0.388 and 0.419g in shell weight, 25.15 and 24.71% in shell ratio, 83.09 and 93.05% in
survival rate of larva-pupa respectively. The performance of corresponding hybrid was close to the
control variety, and its male cocoon produced much better quality silk.
Method of breeding sex limited for larval marking races as analogues of pure lines of approved
commercial hybrids:
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In the recent time the efforts of silkworm breeding at Sericulture Experiment Station in Vratza, Bulgaria
were directed to create lines/hybrids having higher tolerance to adverse rearing conditions. The reasons
were the necessity to intensify the cocoon production in Bulgaria by adoption of summer and autumn
rearing as well as the possibilities to export silkworm eggs to some tropical countries.
As a result a new hardy silkworm four-way commercial hybrid (Hesa 1x KK) x (Gergana2 xVesletz2)
was selected, authorized by the government in 1999 and widely adopted in the field during the period
2000-2003.
Since the breeding methodology was one the same in all the 4 lines, and the following is an example only
with the breeding of the line Iva 1:
Step 1. Initial population: cross between females of Nig 1(sex limited for larval marking-female marked,
male plain, cocoons of Japanese type- elongated with constriction)and males of KK(having plain
larvae, cocoons of Japanese type elongated with high constriction).
Step 2. Backcrosses for 9 generations, using females of the above initial population, mated with selected
males of the line KK and mass rearing.
Step 3. Backcrosses for other 3 generations, but with batch rearing.
Step 4. New silkworm line: Iva 1 ,maintained every year by batch rearing and using selected female
individuals only, mated with selected males of the line KK.
1.2.4 Line breeding, cross breeding and mutation breeding
i. Line breeding: It is a form of inbreeding that involves selection of mates on the basis of their
relationships to a certain superior ancestor. The backcross (crossing a first-generation hybrid with
one of the parental types) is a common method of inbreeding. There is no clear distinction between
the two terms, but line breeding may encompass crosses between individuals and their descendants
or two cousins. This method can be used to increase a particular animal's contribution to the
population. While line breeding is less likely to cause problems in the first generation than does
inbreeding, over time, line breeding can reduce the genetic diversity of a population and cause
problems related to a too-small gene pool that may include an increased prevalence of genetic
disorders and inbreeding depression.
ii. Cross breeding: Out crossing is where two unrelated individuals are crossed to produce progeny. In
out crossing, unless there is verifiable genetic information, one may find that all individuals are
distantly related to an ancient progenitor. If the trait carries throughout a population, all individuals
can have this trait. This is called the founder effect.
iii. Mutation breeding:

The utilization of silkworm gene mutation in commercial production is widely noticed by silkworm
breeders. First of all, some of the mutations in economical characters can be directly utilized in
silkworm breeding. The polyphagous mutation had been utilized to breed silkworm varieties
adaptable to artificial diet; the trimolter mutation was utilized to breed trimoult silkworm varieties to
produce especially fine silk; the non-glutinous mutation was used to produce natural loose eggs.
Secondly, some of the morphological characters have multiple functions which also positively effect
on the economical characters, therefore, it is possible to be directly utilized in new variety breeding.
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Experiment had showed that K gene had positive effect on feed efficiency and heterosis; PS and
black pupa (bp) also showed the potential to increase feed efficiency; the introduction of non-scale
mutation on the wing (nlw) can greatly improve the silkworm seed production condition. Thirdly, sex
control can raise single sex larvae, such as to rear only male silkworm in commercial cocoon
production.
Increases in the efficiency of silk production (the basic product of the sericulture industry) requires the
employment of scientific methods of breeding, provision of standard conditions for adequate nutrition and
maintenance of environmental temperature, adequate humidity, light, sanitation, etc. It also involves the
use of genetics and breeding for accumulation of desirable genetic factors in commercial species of
silkworm. These days, conservation of genetic resources is one of the most important goals of breeding
science.
Check Your Progress

Note: a) Write your answer in the space given below.
b) Compare your answer given at the end of the unit.
1. Silkworms were first domesticated in _____________over 5,000 years ago.

2. __________________ refers to the egg laying capacity of a breed.
3. Mating of unrelated animals is_____________________________.
4. Mating of more closely related individuals is_________________________________
1.3 SUMMARY
Silkworm breeding is aimed at the overall improvement of silkworms from a commercial point of view.
Inbreeding is the mating of individuals or organisms that are closely related through common ancestry, as
opposed to out breeding, which is the mating of unrelated organisms.
Out breeding refers to matings between individuals from different populations, subspecies, or species. Out
breeding can result in a decline in reproductive fitness known as out breeding depression, but this is less
common than inbreeding depression.
Female cocoon is yellow and male white. It is easily for sex-discrimination and the male cocoon can
separately be reeled to produce high grade silk.
In the recent time the efforts of silkworm breeding at Sericulture Experiment Station in Vratza, Bulgaria
were directed to create lines/hybrids having higher tolerance to adverse rearing conditions.
Line breeding: It is a form of inbreeding that involves selection of mates on the basis of their relationships
to a certain superior ancestor.
Out crossing is where two unrelated individuals are crossed to produce progeny.
Mutation breeding utilization of silkworm gene mutation in commercial production is widely noticed by
silkworm breeders.
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1.4 CHECK YOUR PROGRESS ANSWERS
1. China.
2. Fecundity.
3. Out breeding.
4. Inbreeding.
1.5 MODEL EXAMINATION QUESTIONS
I. Answer the following in about 30 lines.
1. Detail about inbreeding and out breeding.

2. Write about concepts of breeding.
3. Write about different types of breeding methods.
II. Answer the following in 10 lines.
1. Write about inbreeding.

2. Write about out breeding.
3. Write about line breeding.
4. Write about cross breeding.
5. Write about mutation breeding.
6. Compare inbreeding and out breeding.
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UNIT-2 SELECTION METHODS
Contents
2.0 Objectives
2.1 Introduction
2.2 Selection Methods
2.2.1 Individual and Mass Selection
2.2.2 Fixation of characters
2.2.3 Formation of new breeds
2.2.4 Authorization of new breeds
2.3 Summary
2.4 Check your progress answers
2.0 OBJECTIVES
 learn the importance of selection methods.
 understands about individual and mass selection.
 know about methods of breeding.
 understands about formation of new breeds.
2.1 INTRODUCTION
Silkworm germplasm in different countries of the world, an approximate information indicate that there
are more than 4000 silkworm germplasm accessions available in different countries. A very recent
compilation of silkworm genetic stocks indicate that there are around 3000 genotypes of Bombyx mori at
the global level, which includes mutants, parthenoclones, polyploids and geographical races. In fact much
of the genetic diversity of Bombyx mori is derived from the inbred lines of land races and elite stocks
evolved by the silkworm breeders and also from hybridisation of different geographical races; mainly the
Japanese, Chinese, European and tropical races, which are distinct for several economic characters.
2.2 SELECTION METHODS
The bivoltine and univoltine races produce high quantity of good quality silk, whereas the multivoltine
races are hardy, tolerant to pathogen load and thereby resistant to diseases compared to the bivoltines but
produce low amount of poor quality silk. Thus, these geographical races are very valuable genetic stocks
for further improvement of silkworm races and evolution of superior breeds of B. mori. Apart from a rich
biodiversity of geographical races, there are also a large number of mutants. The silkworm genetic stocks
include more than 500 mutants for a variety of characters viz., serosal colours; larval and adult
integument colours; skin markings and body shapes; cocoon colours and shapes; physiological traits such
as diapause, number of larval moults and timing of larval maturity; food habits and biochemical features
such as digestive amylase, blood and egg esterases, larval integument esterase, alkaline and acid
phosphatases; haemolymph proteins; silk production and fibroin secretion; homeoproteins and body plan
determination etc. and the various mutants, gene locus and phenotype were documented. Thus, the
geographical races, mutants and the elite breeders stock constitute the major portion of the present day
silkworm germplasm at the global level apart from the parthenoclones, triploid, polyploids and wild
relatives of Bombyx and Bombycidae
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2.2.1 Individual and Mass Selection
Methodology for obtaining the data and calculation of the main breeding characters values:
Qualitative characters
 Egg serosa color: It is determined visually on all silkworm layings, produced from each
accession. The color is green, gray, brown, mixed.
 Egg chorion color: It is determined visually on 3 replicates having 200 eggs or 2 layings each.
The color is white, yellow and mixed.
 Larval markings: It is determined visually on the 5th-7th day of the 5th instar on the all larvae
reared per each accession. The larval markings are plain, normal marking and zebra.
 Cocoon shape: It is determined visually after harvesting and floss removal , on the whole amount
of good quality cocoons produced per each accession. Cocoon shape is oval, elongated oval, oval
with constriction, elongated, elongated with constriction, spindle.
 Cocoon color: It is determined visually after harvesting and floss removal on the whole amount
of good quality cocoons produced per each accession. Cocoon color is white and colored.
 Cocoon size: It is determined on random sample of 100 good quality cocoons .Cocoon size is big,
medium, small.
 Cocoon nature of grains: It is determined on random sample of 100 good quality cocoons. It is
fine, medium, coarse and flossy. As main qualitative characters for silkworm genetic resources at
SESVratza, Bulgaria characterization were accepted only egg serosa color and chorion color,
larval markings, cocoon shape and color.
Quantitative characters
 Hatchability in %: It is determined on 6 layings per accession and on each laying, selected for
incubation in P3 category pure lines. It is calculated on the 3rd day after hatching by the
following formula.
Number of normal eggs in the layings - Number of unhatched eggs in the layings
X 100
Number of normal eggs in the layings
 Larval duration in hr.: The beginning is day and hour of larval brushing, the end is day and
hour when the feeding is stopped and larvae mounted.
 5th instar duration in hr.: The beginning is day and hour of the first feeding after the 4th moult,
the end is day and hour when the feeding is stopped and larvae mounted.
 Pupation rate in %: It is calculated by the formula:

Number of cocoons with alive pupa/number of larvae counted x 100.
 Number of normal eggs in the laying: It is determined on random sample of 15 layings, after
20th October of the current year.
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 Weight of the normal eggs in the laying in gr: It is determined on random sample of 15 layings,
after 20th October of the current year.
 Abnormal eggs in the laying in %: It is calculated by the formula :number of abnormal eggs in
the laying/number of normal eggs in the laying x 100.
 Number of normal eggs per 1 gr: It is determined on 3 replicates, having 0.5 g loosing eggs
each.
 Fresh cocoon grades in %: It is determined immediately after cocoon harvesting and floss
removal. The cocoons are assorted in good quality(having alive pupae),double cocoons and
unreelable cocoons. After the assorting all the three categories are weighed.
 Fresh cocoon weight, and shell weight in gr: There are two methods:1.All good quality
cocoons/shells in the laying/replicate are weighed and after that divided by their number.2. A
random sample consisted of 30 female and 30 male good quality cocoons/shells is taken and after
weighting their weight is divided by the number. In the silkworm breeding the measuring is
individual for each cocoon/shell.
 Shell percentage: It is calculated by the formula:

Shell weight
X 100
Cocoon weight
 Fresh cocoon yield by one box of eggs (20000 eggs) in kg.: It is calculated by the following
formula: fresh cocoon yield by laying/replicate in kg x 20000 x (hatchability x 0.01)/ number of
larvae counted.
 Filament length in m.: It is determined on a random sample of 30 good quality cocoons after
single cocoon reeling test.
One revolution on epprouvette = 1.125mts. or 400 revolutions on epprouvette = 450mts
 Filament weight in gr: After the cocoon reeling the filament is dried to constant weight and
weighed.
 Filament size in denier: It is calculated by the formula:

Filament weight in gr.
X 9000
Filment length in mts
 Reelability in %: It is calculated by the formula:

Reeled cocoon number
 X 100
Number of ends feeding
 Raw silk percentage: The formula used is:

filament weight/dry cocoon weight x 100.
 Raw silk yield by one box of silkworm eggs: The following formula is used:
Fresh cocoon yield by 1 box of eggs x(good quality cocoons percentage x 0.01) x (percentage of dried cocoons
obtained from the fresh cocoons after drying to constant weight x 0.01) x (raw silk percentage x 0.01).
15
 Consumption index (CI) = dry weight of the mulberry leaf ingested in mg during the feeding
period/feeding period duration in days x average dry weight of one larva during the feeding
period in mg.
 Growth rate (GR) =body gain during the feeding period/ feeding period duration in days x
average dry weight of one larva during the feeding period in mg.
 Leaf ingestibility (AI) = dry weight of the mulberry leaf ingested in mg/mulberry leaf supplied
in mg dry weight x 100.
 Food digestibility (AD)= dry weight of the mulberry leaf ingested in mg – dry weight of the
excrements in mg/ dry weight of the mulberry leaf ingested in mg x 100.
 Efficiency for conversion of the food supplied in products (ECS) = dry weight of the products
obtained ,namely body gain during the feeding period or cocoon shell or pupa or eggs / mulberry
leaf supplied in mg dry weight x 100.
 Efficiency of conversion of the food ingested in products (ECI) = dry weight of the products
obtained ,namely body gain during the feeding period or cocoon shell or pupa or eggs / dry
weight of the mulberry leaf ingested in mg during the feeding period x 100.
 Efficiency of conversion of the food digested in products (ECD) = dry weight of the products
obtained ,namely body gain during the feeding period or cocoon shell or pupa or eggs / dry
weight of the mulberry leaf ingested in mg – dry weight of the excrements in mg x 100.
2.2.2 Fixation of characters
a. Characterization and evaluation of silkworm genetic resources:
Most of the silkworm accessions have gray or/and green egg serosa color, white and/or yellow egg
chorion color, marked or/and plain larvae, elongated with or without constriction or oval cocoon shape.
The breeds with white cocoons are prevailing. The data regarding the 5th instar duration in the pure lines
show that it vary from 180 h in the breed Gergana 2 to 218 h in the breed AS.
Most of the silkworm accessions manifest comparatively high hatchability. 80% of the silkworm
accessions have pupation rate higher than 85%, namely 26% of the strains have pupation rate from 85 to
90%, 39% have pupation rate from 90 to 95% , and 15% of the breeds manifest pupation rate higher than
95%.
Since the data for pupation rate were obtained by rearing of silkworms under optimal conditions they do
not give correct information about the survivability of the races under adverse environment.
Under the standard rearing regime most of the accessions manifested cocoon weight from 1.8 to 2g –43
%, and 35% –cocoon weight higher than 2g. 75% of the races have shell weight higher than 0.350g,
namely from 0.350 g to 0.400g – 35%,from 0.400g to 0.450g – 26%,from 0.450 to 0.500g – 8% and
higher than 0.500g – 6%. 70% of the accessions displayed shell ratio higher than 19% and 29%– shell
ratio higher than 21%.
93% of silkworm accessions have cocoon yield by one box of eggs higher than 25kg. Most of the races
(37%) manifested cocoon yield from 30 to 35kg, 29% -cocoon yield from 35 to 40kg and 5% -from 40 to
45kg.
16
57% of the accessions manifested cocoon filament length higher than 900 m , 17 % -from 900 to 1000 m,
21% - from 1000 to 1100 m,12% - from 1100 to 1200m, 6% - from 1200 to 1300m and 1% from 1300 to
1400m.
Vassileva and Tzenov(2001) tested 31 accessions from silkworm genetic resources for their ability for
parthenogenetic development. The results showed that the highest parthenogenetic eggs hatchability (over
20%) was detected in the races ShV, TV, Hesa 2,Siria 2, Mysore 1,70-42, MNB and China.
Tzenov(1996) studied the fecundity in 16 highly productive silkworm races and detected that in the
Japanese type races the moth emergence percentage was the highest in Kom 1(93.32%) and the lowest in
AS(61.88%). In the Chinese type races the moth emergence was the highest in ShV(88.04%), Hebar
2(83.18%) and TV(92.51%) and the lowest in Merefa 2(55.24%).In average the Chinese type races have
higher values of the traits number and weight of normal eggs in the laying than the Japanese type.
Other important traits are those characterizing the food ingestion, digestion and utilization. From his
study with the same 16 silkworm races, the tropical polyvoltine Bonde 517 and their hybrids
Tzenov(1996) detected that there were significant differences between the races regarding the amount of
dry mulberry leaves supplied during the whole fifth larval instar. The highest amount of leaf was supplied
to the race AS(5.227 g dry matter per 1 larva during the 5th instar) and the lowest amount–in multivoltine
Bonde 517–only 3.170g.
In the Japanese type races the amount of food ingested was the highest in Vratza 51(3.688 g) and the
lowest in Hebar 1(3.137g). In the Chinese type races the food ingested was the highest in Vratza 54
(3.784g) and the lowest in Hebar 2(2.858g).
The food ingested and digested was the lowest in polivoltine race- 0.943 and 0.347g respectively. The
data manifested that most of the hybrids studied expressed different in degree, positive heterosis for the
amount of food ingested and digested. The leaf ingestibility was the highest in the Chinese type race
Vratza 54 (73.53 %) and the lowest in the polivoltine race Bonde 517 – only 29.75 %. The highest food
digestibility was detected in the tropical race (36.80%), Super 1 (31.74 %), and Vratza 53(30.04%). The
food digestibility was the lowest in Hebar 2(25.72%), KS (26.07%) and Merefa 2 (26.65%).
Most of the hybrids studied showed different in degree positive heterosis for the leaf ingestibility, but the
heterosis manifested for the food digestibility was low or negative.
The consumption index was the highest in Vratza 51(0.829 mg),Vratza 53(0.825 mg),Vratza 52(0.827
mg),Vratza 54(0.809 mg), Bonde 517 (0.823 mg) and the lowest in Super 1(0.718 mg),Vratza 35(0.732
mg), AS (0.744 mg) and Merefa 2 (0.741 mg). Unlike the other races who have high leaf ingestion and
consumption index, the polivoltine race combined high consumption index with low food intake. As
regards the growth rate character values the inter-racial differences were negligible.
The polivoltine race manifested very high growth rate (0.267 mg) .The efficiency of conversion of the
food supplied(ECS) for body gain was the highest in Super1(17.83%), Merefa 2(17.37%), Vratza
35(17.16%) and the lowest in Hebar 2(13.53%) and Bonde 517 (9.65 %).
The utilization of the food supplied for silk shell was the highest in Super1 (7.65%),Vratza 54
(8.18%),Vratza 35(7.87%) and the lowest in Hebar 2(6.54%) and Bonde 517(2.08%). The utilization of
the food supplied for eggs was the highest in Super 1(2.62%), Hesa 2(2.66%) and the lowest in AS
(2.18%) and Hebar 2(1.84 %).
17
The data for efficiencies of utilization of the food ingested and digested, manifested that in most of the
cases the races having comparatively high leaf ingestion and digestion showed lower food utilization and
vice versa. The heterosis expression in F1 was low or negative.
b. Molecular characterisation of silkworm germplasm:
There is no comprehensive method available for genetic characterisation of the silkworm gene pool at the
molecular level. However, the well known molecular markers viz., Restriction Fragment Length
Polymorphic (RFLP) markers, Minisatellite DNA markers, Microsatellite DNA markers, Random
Amplification of Polymorphic DNA (RAPD) markers, Inter Simple Sequence Repeat (ISSR) markers and
Amplified Fragment Length Polymorphic (AFLP) markers provide valuable information to identify
duplicate accessions apart from trait based mapping and molecular tags. Molecular characterisation helps
to identify the genetic distinctness of a race / breed / stock and thereby helps to eliminate duplicates and
reduce the cost of germplasm maintenance and volume of work. Molecular markers that are linked to
metric and resistance traits and their subsequent mapping in silkworm is essential. Also these genetic
markers can be applied periodically to monitor changes in heterogeneity and heterozygosity in the
accessions, as they are routinely reproduced for maintenance.
2.2.3 Formation of new breeds
The first step in the silkworm breeding process is the primary evaluation of the initial breeding material
(breeds, lines, hybrids etc.). Usually the following evaluation is made:
 Phenotypic evaluation of the main qualitative and quantitative breeding characters.

 Phenotypic variation of the values of the quantitative characters.
 Heritability of the main quantitative characters.
 Correlation between the main quantitative breeding characters.
 Regressions between the main quantitative breeding characters.
For breeding new silkworm races the following different methods are used:
Segregation from foreign F1 hybrids:

By this method in Bulgaria were selected most of the older silkworm breeds , like Vratza 1,Vratza
2,Vratza 3,Vratza 4, Vratza 5, Vratza 6 etc.
Generally the method is as follows:

 Initial population: J-124 x C-122(Japanese F 1hybrid).
 Selection of larvae, having markings and elongate shaped cocoons in F2.
 Mating the selected moths.
 Batch rearing, selection for larval markings, cocoon shape, texture, main quantitative characters
for 11-16 generations.
 New breed: Vratza 1
Separation from foreign F1 hybrids:

By this method the pure line KK was selected in Bulgaria. It is common that in the F1 hybrids can be
found some individuals from the mother pure line due to some mistakes in the sex discrimination, leading
to mating within the pure line.
The method is the following:
18
 Initial population: South Korean hybrid.
 Selection of cocoons, having elongated with constriction shape and mating between them.
 Inbreeding for 4 generations, batch rearing, selection for cocoon shape and main quantitative
characters.
 Auto breeding for 9 generations, selection for cocoon shape and main quantitative characters.
 New breed: KK.
Using populations of Japanese races:

By this method in Bulgaria were selected many breeds like 157 K, Vratza 7,Super 1,Super 2, Hesa 1,
Hesa 2, Gergana 1, Gergana 2,Vesletz 1,Vesletz 2, Kom 1, Kom 2 etc. As an example we will give the
breeding of the line Super 1/11by Petkov (1976):
 Initial population: J 124.

 Selection of 6 layings as initial parents for future sub-lines.
 Inbreeding for 7-8 generations, batch rearing, selection for larval marking, cocoon shape, texture,
main quantitative characters.
 Checking the combining ability of each line by crossing with the race of Chinese type 157 K
(general combining ability) and rejection 2 of the lines due to poor combining ability.
 Inbreeding for another 7-8 generations, batch rearing, selection for larval marking, cocoon shape,
texture, main quantitative characters, with simultaneous checking the combining ability of each
line with 3 other promising races of Chinese type(specific combining ability).Some of the sub-
lines are rejected due to inbreeding depression.
 It was detected that the best specific combining ability had the line Super 1/11.
 New race: Super 1/11, ready for direct hybridization.
Other example is the breeding of pure lines Hebar 1/18 and Hebar 2/1(Petkov,1976) by the following
method:
 Initial population : Kinshu.
 Selection of 3 sub-lines.
 Inbreeding for 3 generations, batch rearing, selection for larval marking, cocoon shape, texture,
main quantitative characters.
 Rejection 2 of the sub-lines.
 Inbreeding in another 3 generations, batch rearing, selection for larval marking, cocoon shape,
texture, main quantitative characters.
 New breed : Hebar 1/18.
Since the breeding methodology was one the same in all the 4 lines, and the following is an example only
with the breeding of the line Iva 1.
Step 1. Initial population: cross between females of Nig 1(sex limited for larval marking-female marked,
male plain, cocoons of Japanese type- elongated with constriction)and males of KK (having plain
larvae, cocoons of Japanese type elongated with high constriction).
Step 2. Backcrosses for 9 generations, using females of the above initial population, mated with selected
males of the line KK and mass rearing.
Step 3. Backcrosses for other 3 generations, but with batch rearing.
Step 4. New silkworm line: Iva 1 ,maintained every year by batch rearing and using selected female
individuals only, mated with selected males of the line KK.
19
Practically the selection was made only with the female individuals of Iva 1, using selected males of KK.
In this case the basic pure line (KK) was maintained in big volume, allowing to make the necessary
selection and the sex limited analogue Iva 1was maintained in lower volume(8-10 batches during rearing).
By using 11 backcrosses, combined with proper selection methods were created 4 new silkworm sex-
limited for larval markings pure lines, having similar qualitative and quantitative characters values to
those in the original pure lines of the recognized by the government and widely adopted in the field
commercial hybrid (Hesa1xKK) x (Gergana2xVesletz2).
Breeding transgenic silkworm: In Japan Tamura (2000) succeeded in generation of the transgenic
silkworm in NIAS. Since exogenous protein gene could be integrated into genome by using of the
technique, it will result in an increase in number of the genetic stock of silkworm.
Fig.2.1 Process for generating transgenic or genome edited silkworms
2.2.4 Authorization of new breeds
Races developed by CSR&TI, Mysore

1. C.Nichi
2. Traditional polyvoltine hybrid - Pure Mysore x C.Nichi
3. Traditional polyvoltine x bivoltine hybrid - Pure Mysore x NB4D2
Polyvoltine breeds and hybrids of the past
20
Attempts made during the 60s to improve the indigenous polyvoltine breeds by introducing the exotic
bivoltine genes has resulted in the evolution of a number of improved polyvoltine breeds namely, Kolar
Gold, Kollegal Jawan, Mysore Princess, Hosa Mysore, Tamil Nadu White, A4E, MBDIY Dl4b etc.
4. Early polyvoltine pure races - Kolar Gold
5. Kollegal Jawan
6. Mysore Princess
7. Hosa Mysore
8. Tamil Nadu White
9. Hosa Mysorex NB18
Though, the exploitation of the cross breeds was realized as early as 1920, it is only during 70s,
development of- improved rearing technology paved the way for introduction of polyvoltine x bivoltine
hybrids using the bivoltine breeds evolved during 70s. This has resulted in the linear improvement of
cocoon yield and silk content, as a result renditta was brought down considerably. During 80s, new
breeding approaches were initiated to evolve polyvoltines with short larval duration, high silk content and
better quality silk. Breeds like, MY 1, RD 1, P2D I were developed. By crossing these races with
bivoltines namely, NB 1B and NB4D2 as male components, three hybrids were developed and authorized
in 1995.
Improved polyvoltine races

10. MY1 X NB1B
11. RD1 X NB18
12. P2D1X NBIB
Productive polyvoltine x bivoltine hybrids

During the 90s, new polyvoltine breeds like BL23 and BL2l were evolved using the indigenous and
exotic genetic resources through hybridization and selection. The hybrids BL23 x NB4D2 for rainfed
areas and BL24 x NB4D2 for irrigated areas were tested with the farmers on large scale. These hybrids
were authorized at the national level in 1991 . The new productive polyvoltine breed was crossed with
bivoltine and a new hybrid combination, BL43 x NB4D2 (Kapila) with better productivity and
quantitative traits than PM x NB4D2 was identified. The hybrid was authorized by Central Silk Board for
commercial exploitationin2002.
Productive Cross Breed (Polyvoltine hybrid) for irrigated zone

Recently, for the irrigated area, one more highly productive polyvoltine breed, BL67 with better
productivity and quality traits than Pure Mysore was developed.
The new breed was crossed with bivoltine and the new polyvoltine x bivoltine hybrid. Cauvery (BL67 x
CSR101) has been identified with better productivity traits, high silk recovery with 6.5-7.0 renditta and
"A" grade silk. The hybrid is being tested on large scale with the farmers of Karnataka, Tamil Nadu and
Andhra Pradesh. The hybrid recorded an average cocoon yield of 54 kg/100 dfls. The cocoons fetched
higher rate of Rs. l5-20 per kg with A grade silk when compared to PM x NB4D2.
Polyvoltine hybrid for rain-fed zone

To replace the existing PM x C.Nichi in rain-fed areas, a new polyvoltine x polyvolrine hybrid. Varuna
(8L24 x C.Nichi) with high survival and better productivity traits has been developed. The average
cocoon yield is 3lkg/l00dfls and renditta of 10-11 as compared to 23kg/100dfls and,12-13 renditta
realised for the popular hybrid, PM x C.Nichi. cocoons of the hybrid is fetching Rs.8- l0/- more per kg as
compared to the control hybrid, PM x C.Nichi,
21
Bivoltine breeds and hybrids of the past
During 70s, efforts were made to develop bivoltine breeds by extraction of lines from the single and
double hybrids of exotic origin. A number of productive bivoltine breeds like NB7, NB18, NB4D2 at
Mysore, KA, KB at Kalimpong, SH6, YS3 at Dehradun, J112, J122, C110, C108 at Kashmir (exotic
collection) were evolved.
Bivoltine hybrids
Bivoltine x bivoltine hybrids like KA x NN6D, KA x NB4D2, NB7 x NB 18, CC I x NB4D2, CA2 x
NB4D2 were found highly promising initially. However, this could not make a significant dent because
polyvoltine x bivoltine hybrids gained stupendous popularity among the farmers. Accordingly, the
bivoltines were used only as male components for cross breed preparation and PM x N84D2 became very
popular in South andNistarixN84D2 in West Bengal.
New productive bivoltine breeds and hybrids

The bivoltine breeds developed earlier could not be popularized as the shell ratio realized at the
commercial level was around 18-19% only which is slightly higher than cross breeds. Hence, efforts were
made under JICA, an Indo-Japanese collaborative project, to evolve hybrids with high survival and
cocoon shell ratio (above 23%) coupled with better reeling traits. As a result, five hybrids namely, CSR2
x CSR4, CSR2 x CSR5, CSR3 x CSR6, CSR12 x CSR6 and CSRI6 x CSR17 were identified as highly
productive hybrids and were authorized by CSB for commercial exploitation during the years 1991-1999.
The hybrids viz., CSR2 x CSR4 and CSR2 X CSR5 (shell ratio >23.0% and raw silk % 19-20) were
authorized (1997) and are being exploited commercially on a large scale at farmers level during
favourable months (Sep-Feb). These hybrids recorded an average cocoon yield of 65kg/100dfls. Both the
hybrids and their reciprocals recorded renditta on an average of 6.0 and produced quality silk of 2A to 4A
grade.
Three productive hybrids, CSR3 x CSR6, CSRl2 x CSR6 and CSR16 x CSR17 and their reciprocal
crosses (raw silk percentage :18-20 and, 2A to 3A grade silk) were authorized during 1999 for
commercial exploitation during favourable months. The hybrids, CSR3 x CSR6 and CSR16 x CSR 17
recorded an average cocoon yield of 65kg/100 dfls.
Robust bivoltine breeds and hybrid

The hot climatic conditions of tropics prevailing particularly in summer are not conducive to rear high
yielding bivoltine hybrids. Considering the importance of developing robust breeds for rearing especially
during unfavourable season of the year, CSR18 and CSR19 were evolved. By utilising these robust
breeds, the hybrid combination, CSR18 x CSR19 was developed which was authorized by CSB in 1998
for commercial exploitation throughout the year. The hybrid recorded an average yield of 50 kg per 100
dfls during summer months.
Productive bivoltine breed and hybrid for special denier

To cater to the need of thin denier for the production of finer fabric, directional breeding has been carried
out and a new productive bivoltine breed, CSR48 has been evolved with longer filament length (>1500
m) and thin denier (2d). By utilising the thin denier breed, CSR48 a productive hybrid , CSR48 x CSR4
has been identified for longer filament length of more than 1400 m with thin denier of less than2.4.
Thin denier hybrid, CSR48 x CSR4

Productive hybrid with longer filament, less denier and size deviation. No difference in yield/100 dfls
when compared with CSR2 x CSR4. Good raw material for the manufacture of Zari, Chiffon and
georgette fabric.
22
Sex-limited breeds-A boon for egg producers
To overcome these drawbacks, two new sex-limited breeds, CSR8(SL) and Nandi (CSR2-SL) have been
developed which produces yellow coloured female and white coloured male cocoons. This is a boon to
the grainages as it is not only helping in easy sex separation process but also saves considerable time,
labour and money.
Similarly, in place of normal CSR2 as male component with PM, the new breed "Nandi (CSR2-SL)" with
sex-limited for cocoon colour was developed to facilitate easy sex separation based on cocoon colour at
grainages (yellow cocoon : female and white cocoon : male) .
Double hybrid for high egg recovery and cocoon crop stability
Owing to the existence of negative correlation between high cocoon shell ratio and low pupation rate in
pure races, the handling of these pure races needs more care and attention by seed cocoon farmers.
Keeping this in mind, the double hybrid, (CSR6 x CSR26) x (CSR2 x CSR27) has been selected for
commercialization.
Bivoltine breed and hybrid lbr sub-optimal conditions

By introgression the amylase genes from the popular polyvoltine breeds, Pure Mysore and Nistari into
CSR2 and CSR5 two breeds namely, GEN3 and GEN2 were evolved. By utilising these breeds, the
hybrid, GEN3 x GEN2 with higher amylase activity has been developed.
Check Your Progress

1. In silkworm ____________ germplasm accessions are available.

2. One revolution on epprouvette is____________________
3. Expand RFLP ______________________________________________________________
2.7 SUMMARY
Silkworm germplasm in different countries of the world, an approximate information indicate that there
are more than 4000 silkworm germplasm accessions available in different countries.
The bivoltine and univoltine races produce high quantity of good quality silk, whereas the multivoltine
races are hardy, tolerant to pathogen load and thereby resistant to diseases compared to the bivoltines but
produce low amount of poor quality silk.
Egg chorion color is determined visually on 3 replicates having 200 eggs or 2 layings each. The color is
white, yellow and mixed.
Cocoon size is determined on random sample of 100 good quality cocoons .Cocoon size is big, medium,
small.
Pupation rate in % is calculated by the formula:

Number of cocoons with alive pupa/number of larvae counted x 100.
Fresh cocoon yield by one box of eggs (20000 eggs) in kg. is calculated by the following formula: fresh
cocoon yield by laying/replicate in kg x 20000 x (hatchability x 0.01)/ number of larvae counted.
23
Filament weight in gr is the one found after the cocoon reeling the filament is dried to constant weight
and weighed.
Most of the silkworm accessions manifest comparatively high hatchability. 80% of the silkworm
accessions have pupation rate higher than 85%, namely 26% of the strains have pupation rate from 85 to
90%, 39% have pupation rate from 90 to 95% , and 15% of the breeds manifest pupation rate higher than
95%.
There is no comprehensive method available for genetic characterisation of the silkworm gene pool at the
molecular level.
1. 4000.
2. 1.125 mts.
3. Restriction Fragment Length Polymorphic.
I. Answer the following in about 30 lines each.
1. Write about methodology for obtaining qualitative characters for calculation of breeding.
2. Write about methodology for obtaining quantitative characters for calculation of breeding.
3. Detail about formation of new breeds.
4. Write about characterization and evaluation of silkworm genetic resources.
5. Detail about new breeds developed by CSR&TI.
6. Write about authorization of breed.
II. Answer the following in about 10 lines each.
1. Transgenic silkworm.
2. Four lines of breeding methodology.
3. Molecular characterization of silkworm breeding.
4. Draw transgenic process of silkworm.
24
UNIT-3 HETEROSIS AND HYBRID VIGOUR
Contents
3.0 Objectives
3.1 Introduction
3.2 Heterosis
3.3 Hybrid Vigour
3.4 Summary
3.5 Check Your Progress Answers
3.0 OBJECTIVES

 learns the importance of heterosis.
 understands about hybrid vigour.
3.1 INTRODUCTION
Heterosis, hybrid vigor, or out breeding enhancement is the improved or increased function of any
biological quality in a hybrid offspring. An offspring is heterotic if its traits are enhanced as a result of
mixing the genetic contributions of its parents. These effects can be due to Mendelian or non-Mendelian
inheritance. Hybrid vigour (or hybrid vigor) is the improved activity and survival of hybrid offspring.
3.2 HETEROSIS
According to Lea (1993) the phenomena of heterosis is the reverse of inbreeding and is defined as a
restoration of vigor, often to a point exceeding the performance of the parents. The less related the strains
are, the more heterotic the reaction is. This can be demonstrated in crosses of inbreeds with one, or no
parents in common. The increase in heterosis corresponded to the reduction in common parentage.
Several explanations or theories have been proposed to explain heterosis. The first is the dominant
favorable gene hypothesis which begins with the idea that there are deleterious genes that build up in
random mating populations. These are not expressed, however due to the masking effect of the dominant
alleles. When inbreeding is imposed on this population, the deleterious genes are unmasked, and
weakened individual lines appear. However, when two inbred pure lines are crossed, the dominant
favorable alleles suppress the appearance of the effect of the recessives, and a superior line results. This
explanation does not exclude the possibility of complementary gene action.
Heterosis is defined as the ‗extra vigour‘ or superiority shown by the F1 progeny over either of its parents
of the mid parental value. It is given by the formula.
F1 - Mid parental value F1 - Better parental value

x 100 or x 100
Mid parental value Better parental value
The phenomenon of heterosis was first observed and explained by Shull in 1990 in silkworm. The
progenies obtained by crossing two inbred lines showed better performance and extra vigour. The factors
determining heterosis are: genetic relationship, compatibility and origin.
The theoretical explanation of the phenomenon of heterosis gives four theories as given below.
25
i. Genes control characters: Genes are the functional units of chromosomes. Certain characters are
determined by single genes while others are determined by more than one gene. The output of
single gene controlled characters is less. The resultant effect of multiple genes can be improved by
their optimum combination.
ii. Theory of dominance: It suggests that out of the various allele combination the homozygous
dominant is stronger. That is out of ‗AA‘ and ‗aA‘ and ‗aa‘ AA will have stronger effect.
iii. Theory of over dominance: It says that the heterozygous dominant allele will have a stronger
effect. That is to say the crossing of a homozygous recessive parent with a homozygous dominant
parent out of the various combinations the heterozygous dominant combination will be stronger.
iv. Epistasis: It is the interaction between genes. It takes place when the action one gene is modified y
one or several other genes, which are sometimes called modifier genes. The gene whose phenotype
is expressed is said to be epistatic, while the phenotype altered or suppressed is said to be
hypostatic.
Though the above theories are different they are not mutually exclusive.
The phenomenon of heterosis if fully exploited in silkworm and maize breeding. In the case all other
animal breeding, pure line selection method is made use of. Hence those breeding programs are deprived
of the beneficial effects of heterosis. It was Toyama (1909) who discovered heterosis in silkworm. Till
then pure line selection was the sole breeding method in silkworm. Since the finding of heterosis,
Japanese scientists conducted trials by crossing pure line in order to harvest the luxuriousness, robustness
and superiority in production. During the late 20th century Japanese cocoon and silk production increased
manifold by the terminal cross breeding strategy.
Heterosis in Antheraea assama Westwood in respect of oviposition is observed over mid parental value
only in respect of four crosses viz., N × W1, N × W2, Y × W1, and Y × W2 and there is no inbreeding
depression in oviposition rate until F3 generation. The cocoon weight and silk content in all the six crosses
exhibit significant heterosis over both mid parental and better parental values. In the F3 generation
significant inbreeding depression is noted for both the characters and the values are at par to the mid
parental value. Commercial exploitation of hybrid vigour therefore should be limited up to F2 generation.
3.3 HYBRID VIGOUR
Hybrid vigour refers to positive heterosis. It is the extra vigour or improvement in performance shown the
F1 progeny over either of its parents or mid parent value. Hybrid vigour is the manifestation of interaction
between the genes or alleles of two pure lines. However the phenotypic expression is always dependent
on the interaction between genotype and environment. Even if the organism posses a very good genetic
makeup the phenotype is the result of its interaction with the environment. Thus, for optimum expression
of the genotype congenial environment is necessary. The phenotypic expression is also dependent upon
the genetic plasticity or buffering capacity of the organism. The extra vigour shown by the out cross
between two inbred lines of wilkworm could be due to the extra buffering capacity of resultant organism.
During the different stages of silkworm development many characters have been found to have a
relationship with the qualitative and quantitative aspects of silk yield (Ohi el al., 1970). However, the
characters which reveal intense manifestation of heterosis are as follows:
i. the duration of feeding in hybrids becomes sh0l1er than that of the parents or the mid parental
values
26
ii. (MPV),
iii. the mortality rate is lower than that other parents,
iv. the double cocoon rate is higher than that of the parents,
v. the cocoon weight is higher than that of MPV,
vi. the cocoon shell layers are heavier than that of MPV,
vii. the length of silk fibre is longer than that of MPV, and
viii. the cocoon fibre weight is heavier than that of MPV.
The earlier results of Osawa and lIarada (1944) show the following values of hybrid vigour for different
characters considering the MPV as 100.
Fig.3.1 Silkworm larvae and cocoons, hybrid combinations of APMW1xAPS8 and APMG1xAPS8
Table 2.1 Heterosis for different characters in the silkworm (Index value of 100 is taken for
mid parental values)
S.No. Characters Heterosis

1. Duration of feeding 97
2. Larval mortality 56
3. Double cocoon rate 146
4. Silk filament size 103
5. Cocoon shell weight 124
6. Egg number 123
Heterosis in different crossing systems
The hybrid vigour is at its best in the single cross of the genetically distinct populations, which decreases
gradually like F1 > F2 > F3 > F4 ... and the phenomenon of heterosis disappears in about F14 generations in
the silkworm (Hirobe, 1985).
Silkworm varieties could be crossed into single cross (A x B), three-way cross (A x B) x CJ and double
cross patterns (A x B) x (C x D). Generally, single cross hybrids manifest the highest rate of hybrid
vigour as compared to three-way and double cross hybrids. Studies conducted on variation in quantitative
characters in parental strains and hybrids of different crossing systems reveal that variability for
quantitative traits is smaller in single cross hybrids than in parental strains, three-way and double cross
hybrids (Watanabe, 1961; Sohn, 1983; Yokoyama, 1973b).
Check Your Progress
b) Compare your answer given at the end of the unit
1. Hybrid vigour refers to positive ________________.

2. Heterosis is defined as the _________________.
27
3.4 SUMMARY
Heterosis, hybrid vigor, or out breeding enhancement is the improved or increased function of any
biological quality in a hybrid offspring.
Heterosis is defined as the ‗extra vigour‘ or superiority shown by the F1 progeny over either of its parents
of the mid parental value.
The phenomenon of heterosis if fully exploited in silkworm and maize breeding. In the case all other
animal breeding, pure line selection method is made use of.
Hybrid vigour refers to positive heterosis. It is the extra vigour or improvement in performance shown the
F1 progeny over either of its parents or mid parent value.
1. Heterosis.
2. Extra vigour
I. Answer the following questions in about 30 lines each.

1. Write about heterosis.
2. Detail about hybrid vigour in silkworm.
II. Answer the following questions in about 10 lines each.
1. Write about heterosis in different crossing systems.

2. What are the characters which reveal intense manifestation of heterosis?
28
UNIT-4 PARENTAL RACES
Contents
4.0 Objectives
4.1 Introduction
4.2 Parental Races and distribution
4.2.1 Based on Origin of Place
4.2.2 Based on Voltinism
4.2.3 Based on Moultinism
4.3 Summary
4.0. OBJECTIVES
 learns the importance of parental races.
 understands about distribution of races.
 knows about voltinism, moultinism.
4.1. INTRODUCTION
The history of silk is as old as the history of mankind and so is the history of silkworm. According to
archaeological and bibliographical evidence, it is probable that sericulture was practiced in China about
2,500 B.C. and Bombyx mandarina L. a kind of wild silkworm were reared in North China be guessed.
Repeated selection and purification by humans for a long time has become the silkworm of today.
Different climates and areas produced several region specific silkworm races with different
characteristics, due to geographical isolation for long time. On the basis of native regions these silkworms
are four types. They are Japanese, Chinese, European and Tropical races. European races are native of
Europe and Central Asia. Tropical or Indian races are native of India and south East Asia. The univoltine,
bivoltine and multivoltine races are found in Japan, China, Europe and India. But some of these varieties
like multivoltine are popular in warmer regions and reared in India.
However, these parental races confined to a particular Geographic region are involved in certain
combination to evolve new hybrid varieties. Some of them are maintained (stock) in basic seed forms for
future purposes. In Japan alone, more than 2000 genetically identified races are maintained (JOCV,1981).
In India there are about 200 races maintained at different breeding centres (FAO,1981). At present there
are separate breeding stations in India for evolving commercial races of multivoltine and bivoltine
varieties.
4.2 PARENTAL RACES
As detailed earlier in this chapter there are different races in silkworms. Their classification is based on
native regions, the number of hatchings in a year, rearing period, body markings, cocoon colour, egg
colour etc.
29
European Univoltine China Univoltine Korean Univoltine
China Bivoltine Japanese Bivoltine
Tropical Polivoltine Japanese Univoltine
4.2.1 Based on Place of Origin
There are four types of races. They are as follows.
(a) Japanese race.

(b) Chinese race.
(c) European race and.
d) Tropical or Indian race.
Above said races can be distinguished from one another on the basis of morphological characters of Egg,
Larva, Pupa and Adult. Biological characters like duration of lifecycle, diapause, moults, environmental
factors, commercial characters like filament length, denier, reelability, mortality ,shell ratio etc..
A. Japanese Race
Generally the larvae of these races are strong and resistant to adverse environments but larval period is
comparatively long. It has univoltine and bivoltine silkworms. The eggs are in many colours and are non-
hibernating. Many white, rotten eggs are produced in which many eggs die after pigment stage. The
larvae are healthy and strong but grow slowly. The markings on the skin of the larvae are normal but
sometimes show quail markings. The body is black in colour. The larvae are yellowish during moult and
red in ripe stage. The larvae are slow in eating thus the duration continues for a long period. Worms do
not identify leaf quality. Nf (Neurofibromatosis) type virus disease is observed. Cocoons are dumbbell
shape or peanut shape or spindle shaped with white or straw colour, but a few races are green or yellow.
These races produce more. Many of these are double cocoons and slightly short and thick cocoon
filament. Cocoons are inferior with less fibre length and reeling is generally poor.
B. Chinese Race
The silkworm larvae of Chinese races eat mulberry leaves actively and the progress of larval growth is
comparatively fast and uniform. The larval marking of many races is plain and the larvae are not sensible
to high temperature and muscardine but sensible to high humidity. Chinese eggs are light yellowish in
colour. The larvae are white with round body form. They are white during moult and blue in spinning
stage. They grow rapidly by feeding actively on mulberry leaves. These are trimoulters, cannot identify
the differences in temperature, humidity and air. Cocoon are round or oval or spindle shaped and are
white, golden yellow, green, flesh colour, red or even pink in colour. Cocoon filament is long with good
reelability. The cocoon filament is thin and long and its reelability is good. Univoltine, bivoltine and
polyvoltine are comprised in these races, and a few trimoulters races are also included in this group,
though most of the races are tetramoulters. This cocoons are uni, bi, multivoltine types.
C. European Race
All races of this group are univoltine, and eggs are big and heavy and dark brown in colour compared
with the other races. The hatching is irregular. The larvae are long, big with pale normal markings. The
larvae are yellowish during moult and red in ripe stage. The larval marking is lightly normal, and
silkworm larvae eat mulberry leaves actively. The duration of larval stage, especially 5 th instar stage is
30
long and long larval body. The larvae grow fat easily but slightly difficult to rear as they are sensible to
bad environment, pebrine, muscardine and C-type virus disease. These are weak against the high
temperature and humidity. They spin large and long oval cocoons with little constriction, and double
cocoons are rare. The majority of cocoons is white or flesh colored or yellow in colour. The cocoon shell
weight of this race is high and cocoon filament is long, with much sericin and good reelability. Double
cocoons are less. The sericin counter is more, making easy reelability. These are univoltine.
Fig.4.1 Parental races
D. Tropical Race
The tropical races are multivoltine, which produce non-hibernation (non diapause) eggs and has a white
egg 3 (w-3, 10:19.6) gene. Eggs are small and are light weight with lustrous shell. The silkworm larvae
are small, long, robust and tolerant to high temperature and humidity, and the larval duration is very short
except Pure Mysore race. These races produce small cocoon with spindle shape, which are green, yellow
or white in color and flossy. The cocoons are loose and flossy with light cocoon shell weight and thin
cocoon filament. They few double cocoons but sericin percentage of these races is very high (about 30%),
so raw silk percentage is very low. However the cocoon filament is fine, thin and clean with little
lousiness. Muscardine disease is common.
Fig.4.1 Parental races

E. Korean Race
Korean races are all univoltine and tri-molters, and larval
duration is comparatively short. The cocoon of these
races are small with low cocoon shell ratio and the
cocoon color are almost yellow, yellow green, and a few
white. Short and thin cocoon silk filament.
Fig.4.3 Korean race cocoons
31
Table.4.1 General characters of regional origin races
4.2.2 Based on Votinism
Voltinism is the ability of silkworm to produce one to several generations in a year.

Based on voltinism the silkworms are broadly classified into three types.
(a) Uni-Voltine
(b) Bi-Voltine
(c) Multi or poly-Voltine
Here are the details of different voltinisms in silkworms.
(a) Uni-Voltine
Silkworms producing one generation in a year are called uni-voltine. The univoltine races are suitable for
the cold regions. These lay diapause eggs due to absence of juvenile hormone. Larval duration of these
races is long and the larval body is large in size. Cocoons are large, cocoon filament is of good quality,
but they are not suitable for the hot season rearing because the larvae are not resistant to the bad
environmental condition such as high temp and humidity. They are unsuitable for summer and autumn.
The diapause eggs are reared by artificial breaking. The cocoon filament is of good quality. The shape of
cocoon is round and oval which is white and pale yellow in colour.
(b) Bi-Voltine
They produce two generations in a year. The first generation adult lays non–diapausing egg. The second
generation adult developing from non-diapausing egg laid during first generation lays diapause eggs due
to absence of juvenile hormone, which is dormant till next spring. These races are suitable for the
temperate zone. The larval duration is short compared to univoltine and the larvae are robust and tolerate
environmental conditions. The cocoon quality of this race is inferior to that of univoltine races but most of
present commercial varieties are bivoltine to which the characters of good cocoon filament of univoltine
races were introduced. They can be reared summer and autumn. The leaf cocoon ratio is less. Cocoon
weight, shell weight, silk percentage and filament length are lesser than uni-voltines. Cocoons are
dumbbell or oval in shape, white or pale yellow in colour.
Example: NB4D2, NB18, KA, NB7 etc.
32
(c) Multi-Voltine
They produce more than 5-6 generations in a year and suitable for the tropical region. The duration of
larval stage of these races is very short, and the larvae are very robust and tolerant of high temp climate.
The leaf cocoon ratio is high. Cocoons are small in size the percentage and the yield of raw silk is poor.
However, the cocoon filament is thin in size and clean with little lousiness. The cocoons produce inferior
quality silk than uni and bivoltine races. The shell ratio is less. The filament length is short. The filament
is fine and clean with little lousiness, but with more lustrous. The larvae are robust and can tolerate
fluctuating environmental conditions and hence best suited for tropical climates. They lay only non
diapausing eggs due to presence of juvenile hormone.
Example: Pure Mysore, C.nichi, Hosa Mysore.
4.2.3 Moultinism
Moltinism is one of the most important economic characters along with voltinism. There are at least 5
different types of molting in the silkworm, namely di-, tri-, tetra-, penta-, and hexamolters. The most
common type in the primitive domestic silkworm varieties and in the wild moths are trimolters, where as
almost all present day commercial strains are tetramolters. Tetramolting character has been considered as
the standard (normal) type in silkworm genetics. The expression of molting character is also affected
partly by the sex-linked maturity genes (Lm).
The dominance relations are M3> +M> M5
The manifestation of moulting characters is also controlled by the balance of juvenile hormone of corpus
allata and ecdysone of prothoracic gland.
Silkworm larvae cast off its old skin and develop new skin and this process is known as Moultinism. Each
moulting period lasts for 15-30 hrs. It is a hereditary character. The moulting is a physiological process
under the control of ecdysone hormone. Based on number of moults the Bombyx mori can be of
trimoulter, tetramoulter and pentamoulters. In tri-moulters the length of the larval stage is short. The
larval body and cocoons are small and the cocoon filament is fine. The tetramoulters larval duration is
medium and produces thin fibre. Most of the commercially reared worms are tetramoulters with five
instars. These worms are reared most widely. In Pentamoulters larval length is long, the body, cocoon and
the filament length is large in size.
Check Your Progress

1. Name races based on origin________________________________________________________

2. Mention races based on voltinism_____________________________________________________
3. How many moulters are there in Bombyx?___________________________________________
4.3 SUMMARY
These parental races are distributed in different geographical parts. There are Japanese, Chinese,
European and Indian races. Races are classified on the basis of native origin, moultinism, voltinism,
cocoon colour, larval marking etc.
33
The race can be differentiated from one another on the basis of morphological, biological and commercial
characters.
Japanese race lay non-hibernating eggs, larvae grow slowly, black in colour, slow eaters, inferior
cocoons. Chinese races are robust to high temperature, rapid growth, tri moulters, good reelability,
filament is long. European race eggs are big, heavy, and irregular in hatching, larvae are long, long life,
fast growth, and weak against high temperature. Cocoons are long with more shell weight with long fibre
and good reelability.
Indian race eggs and larvae are small, robust against temperature and humidity. Cocoons are flossy with
less shell weight, filament is fine.
Silkworms are of three types on the basis of the number of generations produced per year. They are uni-
voltine, bi-voltine and multi-voltine.
Moultinism is a racial character based on which the silkworms are classified into three groups. They are
Tri-moulters, tetra-moulters and penta-moulters.
1. Japanese race, Chinese race, European race, Tropical or Indian race.

2. Uni-Voltine, Bi-Voltine, Multi or poly-Voltine
3. Tri, tetra, penta moulters.
1. Detail about parental races based on place of origin.

2. Write about races based on voltinism.
1. Write about China races.

2. Write about Japan races.
3. Write about European races.
4. Write about Temperate region races.
5. What is moultinism?
34
UNIT-5 BREEDING STATIONS
Contents
5.0 Objectives
5.1 Introduction
5.2 Breeding Stations
5.2.1 Silkworm Seed Organization
5.2.2 Basic seed multiplication centres
5.2.3 Seed Cocoon Rearer and Seed crops
5.2.4 Seed Cocoon Markets
5.2.5 Seed Legislation Act
5.3 Summary
5.0 OBJECTIVES

 learns the importance of seed grainages and silkworm seed organization.
 understands about seed multiplication centers.
 Knows about seed crops and seed markets.
 appreciates seed legislation act.
5.1 INTRODUCTION
Silkworm eggs are very important basic material for the production of silk. ‗Good quality seed‘ can be
defined as that which is free from diseases, has more number of good eggs, gives uniform hatching and
assures a stable cocoon crop.
5.2 BREEDING STATIONS

For maintaining racial purity, quality and disease freeness etc., reproductive seeds are produced as well as
multiplied in number in a series of breeding centre called breeding stations. The State Departments of
Sericulture (DOS), National Silkworm Seed Organization (NSSO) of Central Silk Board and private
Licensed Seed Producers (LSP) are the agencies involved in the commercial silkworm seed production. In
India, about 95% of silk production is multi, bivoltine in nature. Requirement of silkworm eggs indicates
a shortage in the country by 6.43%. Hence, the seed production sector provides an opportunity for profit
making enterprise besides generation of employment.
These are the stations where reprodutive seeds are produced. The production of reproductive seeds is
different fro the production of commercial seed. Commercial seeds are mostly F1 hybrids produced by
corssing a female of hardy pure line local race with a pure line male of high yielding, generally, a
bivoltine race. Reproductive seeds are generally pure lines of both the local and high yielding races or
rarely, F1 hybrids used for the production of a double cross commercial seed.
The parents to be used for commercial seed production are reared in the breeding stations. The number of
pure breed parents required for a grainage are too large to be obtained by a single multiplication step from
the stock in the germplasm. So, generally a three (for bivoltines, a four) stage multiplication is carried out.
35
In order to preserve the racial characters and avoid mixing up of the races, each of these multiplications
are carried out in different stations rather than in the same place.
5.2.1 Silkworm Seed Organization
"Seed organisation" comprises the maintenance of breeder's stock and its multiplication for the ultimate
production of large quantity of commercial hybrid seed. Therefore, the maintenance of breed
characteristics (purity, vigour etc.) is of utmost importance. The breeder's stock maintenance should be
the responsibility of breeders of Research Institutes, which in turn (now and then) should supply the basic
seed for further multiplication. The breeder's stocks will be multiplied 3-4 times in a year (favourable
months) and the different multiplication levels are designated as P3, P2 and Pl. However, a three tier
system is considered more ideal and efficient which is followed in all sericulture advanced countries. The
breeder's stock and multiplication centres should be under the control of Government agencies and well
trained persons.
Multiplication cycles
 Reduce the cycles of basic seed multiplication from the present 4 cycles to 3 cycles.
 Produce only required quantity of quality cocoons and layings for further supply.
 Stop the present tendency of producing large quantity of cocoons and layings which are mostly
not being used.
P4 and P3 stations
Normally, the seeds of multivoltine mulberry silkworm races (whether cross breed or pure breed) are
multiplied in a three tier system starting from the P3 stations. But exotic pure breed races of bivoltines, as
well as, rare multivoltine races have a four tier multiplication system beginning with P4 station. These
statiotions are also called as the Race Breeding Stations. They are also major germplasm banks of the
races. They are under Government control and the places where the great great grandparents or the great
grant parents of commercial grainage seeds are multiplied. They also undertake research experiments on
the breeding techniques for evolving new races. The aim of the breeding stations is to maintain the purity
of the races. They rear only pure breeds nd no hybridisation is done except for research purposes.
Different pure breeds differ in their resistance to disease, tolerance to environmental conditions, response
to the variety of mulberry and other factors. Rearing of each race is, therefore, done separately by what is
called the cellular methods of rearing. Each laying of each race is reared separately in different trays, each
is fed with the mulberry variety suitable for it and mounted and harvestd separately. There is no mass
rearing of even the same race. As this requires knowledge of the physiology of each race, rearing is done
only by technically qualified persons. 25% of the best cocoons are selected and sent to the station at the
next lower level for multiplication. The remaining cocoons are used in station itself for preparing eggs
called Breeder‘s Stock Egg.
The P4 and P3 stations are located at the cooler parts of the country which are ideally suited for bivoltine
rearing and where multivoltines can also be reared.
Procedure for multiplication of basic stocks at P4 and P3
 Plan and programme the basic seed production based on the requirement of eggs for next level of
multiplication. There should be an integrated programme to supply the eggs to various
multiplication centres.
36
 Prepare a flow chart and supply programme for one year in advance for effective monitoring of
the production and supply of basic seed.
 Brush 10-30 dfls of each breed in cellular batches (individual laying) depending upon the demand
and supply of layings.
 Count all the larvae after third moult and retain till spinning.
 Sort out dead pupae-cocoons, malformed cocoons, stained cocoons, thin shelled cocoons etc.,
after the cocoon harvest. Live pupae cocoons only are taken for calculation.
 Calculate pupation rate on the basis of live pupae. Live pupa can be known by the sound when
the cocoon is gently shaked. Extreme low pupation percentage batches are to be rejected
(e.g.:1,0Vo below the average).
 Calculate the pupation rate for the number of larvae brushed/third moult larvae. Deduct the
number of uzi infested larvae and muscardine affected if any, from the original number and
calculate the pupation rate only for the remaining basic number of larvae.
 Take 50-60 cocoons at random in each batch and sex them. After sex separation, record mass
cocoon weight, shell weight and cocoon shell ratio separately for both male and female cocoons.
Then mean of both the sexes in each batch.
 Calculate the mean of cocoon weight, shell weight and shell ratio of all the batches of the breed.
 Select only the batches above average for pupation (in each breed).
 Eliminate the batches which have produced cocoons with undesirable shapes, thin shell cocoons
and poor growth performance.
 In case of P4, select the batches scoring above the average for all the three traits viz., cocoon
weight, shell weight and shell ratio (minimum 3 batches are to be selected).
 In case of P4, select individual cocoons. This should be done as per the guidance of the breeder.
 Whereas in P3, select the batches above average or nearer to average for the three traits like in P4
(minimum 3 batches are to be selected).
 Do not resort to cocoon selection for P3 batches. Select only the batches based on their merit.
 For every rearing the P3 layings should be obtained from P4 stock.
 The selected batches of each breed both in F4 and F3 are to be interbred/inter-batch crossing for
raising next generation (e.g. 1x3, 2x 3, 3 x 1).
 Depending on the required number of dfls, keep equal number of male and female pupae for egg
production (e.g. to prepare 100 dfls 300 female and 300 male pupae are to be kept). If there is
shortage (less) of males, use the same males two times for egg production.
 After egg laying, all the mother moths should be subjected for pebrine test individually.
37
 Make provision to preserve (P4 and P3) eggs under different hibernation schedules.
 Replace the breeder's stock from breeders once or twice in a year.
P2 stations
These produce the Foundation Stock seeds and serve also as thee germplasm banks. They receive he seed
cocoons for rearing from breeding station above them ie., P3 station. Here also multiplication of the pure
breed races are carried and no hybridixation. The layings of each mother moth are reared individually and
mounted separately by the cellular method of rearing so that there is no change of mingling of racial
characters. The P2 stations select 25% of their best cocoons of each layings and send to the nest station
lower down for multiplication. The remaining are used either for replenishing the stock or for reeling.
Since the commercial grainages are two levels below these stations, they are the place where grandparents
of the commercial laings are reared.
Multiplication of silkworm breeds at P2stage

 For every rearing, the layings should be obtained from P3 stock.
 Depending upon the requirement of P1 dfls, the P2 dfls rearing should be prepared. To produce
1000 P1 dfls, 6000 pupae are required i.e., 3000 males and 3000 females. The approximate
survival rate is 50 %o of the total larvae brushed i.e., 12,000larvae.
 5-10 dfls are brushed in mass
 After cocoon harvest, sort out dead pupae-cocoons, malformed cocoons stained cocoons, thin
shelled cocoons etc. Live pupae-cocoons are only taken for calculation of pupation rate.
 Adopt mass cocoon assessment of 50 males and 50 females in each batch to record cocoon
weight, shell weight and shell percentage.
 Record data systematically.
 Cull out undesirable cocoons. Do not resort to any cocoon selection.
 Utilize all good cocoons for P1 laying production.
 After egg laying, all the mother moths should be subjected for pebrine test individually.
 Create data base systematically in Basic Seed Farms (BSF).
P1 stations
These are the stations where the reproductive seed cocoons required by the grainages are reared ie., the
parents of the commercial layings. They receive seeds from P2 stations. The seed cocoons are mass reared
and the first hybridisation of double crosses is done here and the hybrids produced are called Foundation
Hybrids. They send 25% of the cocoons produced to the grainages or to seed cocoon rearers for rearing
the seed cocoons which act as parents for the commercial layings.
The purpose of the three or four tier system of multiplication is to build up the population of the pure
breed races without any deterioration in their characters. For example, if we assume that an average
laying of a couple of moths is 400 (this varies from 600 in bivoltine to 250 in local multivoltines), and
that 400 lakh layings are the requirement of a particular region, the grainage requires one lakh or 5000
38
pure breed moths of the local race and an equal number of bivoltine pure breed moths for producing the
required amount of layings. This large number of the parent moths cannot be obtained from the available
germplasm banks in a single multiplication. From 10 moth germplasm banks (5 pairs) the P3 station will
produce 2000 seed cocoons of which only 25%, ie., 500 or 250 pairs re sent to the next station and thus
the population is built up fifty fold t each level of multiplication. This system ensures that during the
multiplication process, the racial characters are not diluted and sufficient stocks are maintained at each
level of breeding.
Flow chart for multiplication of P4, P3 and P2

P4 and P3level
Individual laying in cellular brushing
Batch - (bed) wise cocoon harvest
Calculation of Pupation rate
Assessment of 50-60 cocoons (male and female) bed wise for cocoon weight,
shell weight and shell ratio
Calculation of mean values of all the batches of the breed

for cocoon weight, shell weight and shell ratio
Selection of batches above average in F4 nearer to average in P3
Culling undesirable cocoons
Cocoon cutting and sexing
Inter batch (bed) crossing
Preparation of P3 layings from P4 and P2 layings from P3
P2 level
Mass rea-ring of 5-10 dfls
39
Batch-wise cocoon harvest and assessment
Mixing of good cocoons from all the batches
Preparation of P1dfls
5.2.3 Basic seed multiplication centres
Systematic collection of data in basic seed farms
The collection of systematic data on rearing performance of various breeds is very important in Basic
Seed Farms. This helps in evaluating the merits of various silkworm breed characters to facilitate
selection of batches on merit to provide quality seed for further multiplication.
Production of Pl cocoon
The production of parent seed cocoons (P1) needed for industrial hybrid seed production is organised
under private sector. The seed areas are to be distinct for bivoltine and multivoltine breeds.
Parent seed cocoon production can also be organised through selected seed rearers by the respective egg
production centres. In such areas the agency which is responsible for hybrid seed production should
involve directly to produce the seed cocoons by adopting a requisite number of seed rearers.
Points for attention in basic seed farms
 Wash hands and feet by running water (tap water) before entering the rearing house.
 Keep separate set of slippers in young age and late age rearing houses and leaf storage room.
 Entry to the rearing house should be restricted to the workers only. Do not allow any visitors
inside.
 Do not allow leaf supplier to enter the rearing house. Leaf baskets should be collected by the
rearing staff at the door step.
 Do not touch the worms with hand. Use chop sticks or forceps for picking the worms and
spreading of the rearing bed.
 Collect diseased, dead and under sized worms using un-used leaves or old news paper and put
into the basin containing formalin.
 Do not throw litter on the floor. Start bed cleaning after spreading the rexin sheet or gunny cloth
on the floor. Disinfect the rexin sheet or gunny cloth every day after use.
 Use disinfected cleaning nets for bed cleaning. Keep two sets of nets.
 Apply lime and bleaching powder mixture around the rearing house regularly.
5.2.4 Seed Cocoon Rearer and Seed crops
Since the seed cocoons must be healthy, hygienic and preserve the racial characters, its rearing requires
technical skill. Seed cocoons are generally rained in P1 stations but if they are unable to cope with
required number of cocoons to be raised, a part of the rearing of seed cocoons is entrusted to highly
skilled farmer who are given special license for carrying this worm and who will be give a premium price
id 50% higher than the commercial cocoon price. The choice of the rearer is made after taking into
consideration of the following.
40
1. The rearer should have a scientific knowledge of grainage operation and silkworm rearing and
have an interest in sericulture and should cooperate with the grainage personnel.
2. The mulberry gardens should be cultivated following the new package of practices and should
supply good quality leaves.
3. The rearing house should be located in an area suitable for pure breed rearing, with facilities for
providing optimum rearing conditions. It should be free from germs of silkworm diseases and
have an easy access to the grainage for technical support and advice.
5.2.4 Seed Cocoon Markets
The basic criteria for procurement of seed cocoons are that it should be pebrine free and confirm to the
norms, especially regarding the pupation rate. The norms are fixed according to the season for the
characters such as cocoon weight, pupation rate and racial characters. The cocoons must be uniform with
regard to size, shape and colour. The grainage authorities must enquire about the nature of rearing,
environmental conditions during the rearing and healthiness of the cocoons before the selection of
cocoons have been made. In the cocoon market, the grainage authorities will purchase the required
quantity of multivoltine cocoons in an open auction with the presence of govt. market officer as per law.
Half quantity of the bivoltine cocoons should also purchased from bivoltine cocoon markets for the
preparation of hybrid seeds. The seed cocoons are packed loosely in perforated boxes or bamboo baskets
in small quantities and are transported during cooler hours of the day. If the live cocoons are tightly
packed, they get heated up due to respiration. Exposure of hot sun results in the dead cocoons and poor
emergence.
5.2.5 Seed Legislation Act
The five traditional sericulture states namely, Karnataka, Andhra Pradesh, Tamil Nadu, West Bengal and
Jammu & Kashmir have passed Silkworm Seed Legislation Act. The respective State Departments of
Sericulture controls:
a. Organizing generation of parental seed cocoons and their transaction in the market to the different
agencies.
b. Issue of license to take up silkworm seed production, inspection, renewal or cancellation of
licenses, etc.
c. Quality certification of the silkworm seed produced.
d. Disease monitoring / control measures in seed areas.
e. Imparting training in seed production technology
Check Your Progress

1. Expand NSSO_______________________________________________________________
2. ______________________are the breeding stations.
3. Expand LSP__________________________________________________________________
5.3 SUMMARY
These are the stations where reprodutive seeds are produced. The production of reproductive seeds is
different fro the production of commercial seed. Commercial seeds are mostly F1 hybrids produced by
41
corssing a female of hardy pure line local race with a pure line male of high yielding, generally, a
bivoltine race.
Reproductive seeds are generally pure lines of both the local and high yielding races or rarely, F1 hybrids
used for the production of a double cross commercial seed.
Seed organisation comprises the maintenance of breeder's stock and its multiplication for the ultimate
production of large quantity of commercial hybrid seed.
The seeds of multivoltine mulberry silkworm races (whether cross breed or pure breed) are multiplied in a
three tier system starting from the P3 stations.
The P4 and P3 stations are located at the cooler parts of the country which are ideally suited for bivoltine
rearing and where multivoltines can also be reared.
P2 stations produce the Foundation Stock seeds and serve also as thee germplasm banks.
P1 stations are the stations where the reproductive seed cocoons required by the grainages are reared ie.,
the parents of the commercial layings.
The production of parent seed cocoons (P1) needed for industrial hybrid seed production is organised
under private sector.
1. National Silkworm Seed Organization Transcription, translation.

2. P4, P3, P2, P1.
3. Licensed Seed Producers.
1. Detail about P4 and P3 breeding stations.

2. Detail about P2 breeding station.
3. Write about basic seed multiplication centres.
1. Write about seed cocoon rearer and seed crop.

2. Write about Seed Legislation Act.
3. Write about flow chart in P1 station.
4. Write about seed cocoon markets.
42
UNIT-6 SELECTION OF SEED COCOONS
Contents
6.0 Objectives
6.1 Introduction
6.2 Selection of Seed Cocoons
6.2.1 Cocoon Selection
6.2.2 Pupal Examination
6.2.3 Processing of Cocoons
6.3 Summary
6.4 Check Your Progress Answers
6.0 OBJECTIVES

 learns the importance of pupal examination in seed cocoon selection.
 identifies bad cocoons.
 knows the procedure for fixing the price.
6.1 INTRODUCTION
Selection and preservation of silkworm seed cocoons is very important as it decides the quality and
quantity of cocoons and its characters. Further it favours the grainage for the production of quality seed.
However identification and removal of bad cocoons is to be carried and if the percentage of bad cocoons
is more that lot should be rejected before further processing.
6.2 SELECTION OF SEED COCOONS
The basic criteria for procurement of seed cocoons are that it should be pebrine free and conform to the
norms especially regarding the pupation rate. The norms are fixed according to the season for characters
such as cocoon weight, pupation rate and racial characters.
6.2.1 Cocoon Selection
The seed cocoons should be harvested a day later than the industrial cocoons. They can be harvested six
days after completion of spinning in hotter areas and seven days in cooler zones. After harvesting, a
preliminary sorting is necessary to eliminate flimsy and poor cocoons. Double cocoons, if any, can be
retained for seed production.
The seed cocoons are packed loosely in perforated boxes or bamboo baskets in small quantities and are
transported during cooler hours of the day. lf the live cocoons are tightly packed, they get heated up due
to respiration. Exposure to hot sun results in dead pupae and poor emergence.
6.2.2 Pupal Examination
In order to produce good quality and healthy eggs the seed cocoons used for the purpose must be of high
quality and in good health, and therefore the seed cocoons arriving at the grainages are subjected to rigid
selection. In selection only sound and uniform cocoons conforming to the characteristics of the race of the
43
parental stock are selected and defective and deformed, under and over sized cocoons, double, stained and
dead, uzi infested, thin end, open end, melted cocoons etc. are rejected.
Advance detection of pebrine disease, if any before the commencement of operation of each batch helps
in averting great loss to the grainages. This is facilitated by investigations at three stages
1. Pupa test
2. Forced eclosion test
3. First day moth examination.
Pupa Test
In this test, the gut of pupa is a more reliable test for pebrine detection than the entire pupa. A sample of
pupae is tested from each batch of cocoons. For this purpose, the pupa is cut ventrally just below the wing
bud by a scissor by holding the pupa between thumb and for finger in left hand. After cutting the pupa is
pressed gently. The midgut oozes out as a brown body from the cut portion. This midgut is collected and
crushed with few drops of potassium hydroxide in a moth crushing set. The fluid is taken on the slide and
examine under the microscope with 600 X magnification. If the stock is suffering from pebrine, the entire
batch of cocoons is rejected and sent to market. Such cocoons should never be used for preparation of
silkworm seed under any circumstances.
6.2.3 Processing of Cocoons
It is also called as cocoon sorting where good and bad cocoons are separated from the cocoon lots. The
seed cocoons declared as free of pebrine disease are preserved for egg production. These cocoons are
sorted to separate deformed, flimsy, stained, double, flossy, thin, pointed, malformed, dead cocoons and
then rejected. Batches of cocoons showing higher percentage of melting are sent to reeling centres. Only
good cocoons of quality conforming to the breeds are selected and preserved. Deflossing of cocoons is
done to facilitate easy eclosion of moths. Bad cocoons are either stifled or sent to cocoon market. It is
unhealthy to keep such cocoons in the grainage.
Fig.6.2 Identification of
bad cocoons
44
Calculate good and bad cocoon percentage based on number and weight.
Weight of bad cocoons
A. Percentage of bad cocoons  X 100
Weight of total cocoons
Total number of bad cocoons

or  X 100
Total number of cocoons
Weight of good cocoons
B. Percentage of good cocoons  X 100
weight of total cocoons
Total number of good cocoons

or  X 100
Total number of cocoons
Total cocoons = good cocoons + bad cocoons

Model Problem: Calculate good and bad cocoon percentage on the basis of weight and number using the
following values.
Number Weight (gr)

Dead cocoons 100 120
Double cocoons 20 85
Pierced cocoons 15 18
Malformed cocoons 20 30
Stained cocoons 140 190
Thin cocoons 20 50
Good cocoons 1150 1870
Total cocoons 1475 2363
Solution:
Total number of good cocoons = 1150
Total weight of good cocoons = 1870gr.
Total number of bad cocoons = 325
Total weight of bad cocoons = 493gr
Total number of cocoons = 1475
Total weight of cocoons = 2362
325
Percentage of bad cocoons by number  x 100  22%
1475
493
Percentage of bad cocoons by weight  x 100  20.86%
2363
1150
Percentage of good cocoons by number  x 100  77.96%
1475
1870
Percentage of good cocoons by weight  x 100  79.13%
2363
45
Percentages of individual bad cocoons
By number By weight
110 120
1. Dead cocoons x 100  7.45% x 100  5.07%
1475 2363
20 85
2. Double cocoons x 100  1.35% x 100  3.59%
1475 2363
15 18
3. Pierced cocoons x 100  0.01% x 100  0.76%
1475 2363
20 30
4. Malformed cocoons x 100  1.35% x 100  1.26%
1475 2363
140 190
5. Stained cocoons x 100  9.49 % x 100  8.04%
1475 2363
20 50
6. Thin cocoons x 100  1.35% x 100  2.11%
1475 2363
Total = 22% = 20.83%
Check Your Progress
1. ________________________________are bad cocoons.
2. Bad cocoons are not suitable for ______________________________
6.3 SUMMARY
Selection and preservation of silkworm seed cocoons is very important as it decides the quality and
quantity of cocoons and its characters.
The basic criteria for procurement of seed cocoons are that it should be pebrine free and conform to the
norms especially regarding the pupation rate.
The seed cocoons should be harvested a day later than the industrial cocoons.
In selection only sound and uniform cocoons conforming to the characteristics of the race of the parental
stock are selected and defective and deformed, under and over sized cocoons, double, stained and dead,
uzi infested, thin end, open end, melted cocoons etc. are rejected.
In pupa test the gut of pupa is a more reliable test for pebrine detection than the entire pupa.
It is also called as cocoon sorting where good and bad cocoons are separated from the cocoon lots.
1. Double, thin, flimsy etc.

2. Seed production.
46
I. Answer the following in about 30 lines each.
1. Discuss about selection of seed cocoons.
II. Answer the following in about 10 lines each.
1. Write about cocoon selection.

2. Write about pupal test.
3. Write about processing of cocoons.
47
UNIT-7 GRAINAGE
Contents
7.0 Objectives
7.1 Introduction
7.2 Grainage
7.2.1 Model Grainage
7.2.2 Equipment and Uses
7.2.3 Disinfection
7.3 Seed Production Process
7.3.1 Procurement of seed cocoons
7.3.2 Transport of seed cocoons
7.3.3 Preliminary Examination
7.4 Processing of Seed Cocoons
7.4.1 Preservation of Seed Cocoons/Pupae
7.4.2 Sex Separation
7.5 Summary
7.0. OBJECTIVES
 understands about seed production centres.
 learns about types of grainages.
 knows the grainage plan and equipments .
 understands about transportation and preservation of seed cocoons.
7.1. INTRODUCTION
Grainages are the centers for production of large scale quality and quantities of disease free layings of
silkworm. It is for the production of pure and hybrid seed. These centers are more popular as commercial
egg production centers because they have a direct link with seed rearers. These centers encourage
progressive farmers and seed rearers to produce seeds commercially. Farmers always intense to produce
good quality cocoons, hence the farmer look forward to the Grainage for the supply of high vigor disease
free commercial seeds. These seeds produce cocoons with rich silk content and high yield.
7.2 GRAINAGE
The Grainage should have proper facilities, good environmental conditions, and well spacious rooms,
without water stagnation around the Grainage building and should be away from factories and pesticide
industries. A Grainage must be established where sericulture is popular among the villages. It can also
help the farmers technically .These Grainage centers conduct training program for the unemployed youth
to create awareness on commercial rearing
7.2.1 Model Grainage
There are certain prerequisites for grainage operation among which building location, grainage building,
equipment, technical staff are main.
43
a. Location: The location of the grainage should be in commercial cocoon producing areas to cater
the needs of commercial rearers. If the grainages are located away from seed areas, it is
disadvantage in transportation of seed cocoons and eggs. The transport of seed cocoons and eggs
is hazardous especially in summer. The high temperature leads to pupal death, melting of
cocoons, irregular hatching, more number of dead eggs which leads to hamper the complete
rearing activity. On the other hand, seed cocoons transported in high temperature leads to poor
moth emergence which increases cost and reduces the number of layings. Thus grainages are to
be located in commercial cocoon production areas for easy transport of seed cocoons for laying
preparation and also to transport layings to rearing centers. Further the surrounding must be free
from polluted air which is clinically unsuitable for egg production.
b. Labour: The life span of seed cocoons is very short thus the large number of cocoons is utilized
for egg production. Processing of egg production is highly labor intensive and requires large
number of laborers. Thus besides technical staff availability of labour is one of the prerequisite
otherwise grainage becomes uneconomical.
c. Seed rearers: Grainage should have a proper number of seed rearers whose rearing centers are
nearer to grainage for technical support and supervision. These seed rearers are identified by the
technical staff and all the rearing activities are under the supervision of technical staff.
Model Grainage
Egg production and processing is carried on with lot of care and technique. It requires a specific
environment for each and every grainage activity. Thus a separate, convenient building is required, where
each activity is confined to a particular room with suitable environment for successful completion of
processing. Light, temperature and humidity are the factors to be controlled for various activities of the
grainage. Some rooms require good ventilation, some are to kept dark and a long and spacious room for
preservation of seed cocoons is required. The building should be well planned for all the activities. The
following are essential in a grainage building.
Fig.2.1 Grainage building ground plan
44
a. Components of grainage building:
1. Seed cocoon reception and processing rooms
2. Seed cocoon/pupae preservation rooms.
3. Coupling and decoupling room.
4. Egg laying chambers.
5. Moth examination hall.
6. Egg processing lab.
7. Incubation chambers.
8. Cold storage rooms.
9. Office and dormitory.
10. General and pierced cocoon store.
The size of grainage building varies with the target of egg production. A model grainage for industrial
seed production of a capacity 25lakh dfls per annum is sketched below.
b. Staff: The grainage activity has three components ie., supply of parent seed cocoons; processing
of cocoons; moth examination and disposal of seed. All these activities are carried by a group of
technical staff. A grainage of 15lakh capacity should have 16 technical staff to carry on all the
process of egg production.
7.2.2 Equipment and Uses
The grainage equipments are designed for a specific function. These equipments are made in such a way
to transport and handle easily. Equipment is used to control disease incidence, to sterilize, to check attack
of ants, to check light, to keep moths undisturbed. The grainage equipment and their uses are detailed.
1) Cocoon preservation rack

It is for keeping the trays containing cocoons, pupae and moths. Each rack accommodates ten
trays. This rack measures 228.6cms height; 144.8cm length and 61cm breadth.
2) Cocoon preservation trays

These are used to spread pupae and cocoons. These are in different sizes and shapes.
i. Bamboo trays: These are used to spread cocoons. These are very light, easy for transport
and cleaning. These can be made in every village and available at a low cost. Each tray
has 137.2 dia. in size.
ii. Wooden trays: These are of two types. The bottom of tray is made of plywood or wire
mesh. These trays are mainly used for copulation, collection of moths. The plywood
bottom trays is used for oviposition. It measures 91.5 x 61cm.
3) Ant wells
Ants cause lot of damage to cocoons and moths. The ant wells are used to control the attack of
ants. These are kept under the legs of rack. The ant well has 4cm deep furrow which is filled with
water to prevent the ant crawling on to the stand. These ant wells are made of cement with 21 x
21 cm size.
Ant crawling can also be controlled using enamel plate, plastic plate/dish. A cloth dipped in
kerosene, gamaxine powder spread around the rack legs also serves the purpose.
45
Fig. 7.2 Grainage Equipment
46
4) Basin stand (Iron)
It is a tripod stand with a height of 86.5cms to hold a basin (30.5cm dia). The basin is filled with
2% formalin. This liquid is to wash and sterilize hands while entering into room. It is kept nearer
to cocoon preservation room, moth preservation room, egg laying room.
5) Cellule
It is plastic, black color equipment with 3.2cm height, 5.1cm dia size. It is used in pairing
oviposition to provide partial darkness which favours maximum fertilization of eggs.
6) Microscope
Moth examination is essential immediately after egg laying to identify pebrine disease. This
microscope having 600 magnifications helps to examine the moth and to produce disease free
layings (DFLs).
7) Moth crushing set

This enamel set has ten mortars and pestles. It is used to grind the moths after oviposition. The
liquid is taken to examine under the microscope for identification of pebrine spores. Diseased
moth eggs are discarded before processing. The same work can also be done by a machine and
pebrine separator.
8) Moth examination table, stool (wooden)

These are used to keep microscope during moth examination. The table (182.9 x 76.2 cm size) is
to keep moth crushing set, slide, cover glass packet, water or KOH, microscope and observation
note book. The stool (61 x 50.8cm size) for sitting while examination.
9) Dry and wet thermometer

It is to measure room temperature and relative humidity.
10) Hygrometer
It is used to measure the room humidity directly.
11) Acid treatment bath

It is a chamber with a proper lid used for acid treatment of univoltine and bivoltine eggs to stop
hibernation which favours hatching.
12) Hydrometer
It is used in acid treatment, loose egg preparation to measure specific gravity of salt water.
13) Feeding stand (wooden)

The stand is used to keep trays containing cocoons moths during grainage activity. It is also used
to keep the rearing tray to examine the worms. The grainage activities are continuous and to be
performed very actively by standing thus a stand is used.
14) Deflossing machine

It is used to defloss the cocoon before preservation for moth emergence. This mechanical
equipment is operated by pressing the pedal. The deflossed cocoons are collected to preserve.
15) Cocoon cutting machine

It is used to cut the cocoons and separate the pupae which are one of the important grainage
activities. It is mechanized equipment.
47
Fig. 7.3 Grainage Equipment
16) Sprayer
It is for disinfection of grainage rooms and equipment. It is also used to wash the equipments.
17) Crates
It is for storing male moths in refrigerator to use them for pouring (first and second).
18) Refrigerator
It is used synchronization of moths, to preserve male moths for using second pairing.
19) Incubator
To incubate eggs at 23o-25oC temperature and 80-85% humidity for even development of
embryo.
20) Formalin mat

A gunny bag or cloth piece is spread in an iron sheet tray having 2% formalin or any other
disinfectant and kept in front of the door. While entering into the room one should keep their foot
in the tray for disinfection.
48
21) Mask
The lepidopteran silk moths have plenty of scales on the body including wings. These scales are
spread in the moth emergence room during their emergence. Thus mask is used to prevent the
entry of scales, dust, formalin fumes into nose/mouth.
22) Others
Equipments like air conditioner or air cooler, slide box, cover glasses, electric stove, egg sheets,
loose egg boxes, clock are also required.
Maintenance of equipment is highly essential for its long lasting and proper functioning during
the preparation of eggs. Cleaning/washing (disinfection) are to be carried out regularly. Oiling of
machines at regular intervals is also necessary.
23) Disinfectants
The basic chemicals used for disinfection as spray, dusting, fumigation are
i. Chlorine as chlorine compounds like chloramines and hypo.
ii. Iodine as iodophors.
iii. Phenol derivatives like cresol, hexachlorophene.
iv. Cetylyridinium chloride and Benzyl alkonium chloride.
v. Formaldehyde or paraformaldehyde.
vi. Bleaching powder.
vii. Sodium hypochloride.
viii. Lime powder.
7.2.3 Disinfection
It is called as extermination of disease causing germs. The most effective and simple method is carried
using chemical and disinfectants. While selecting the disinfectants certain aspects ie., effectiveness
against different pathogens, simplicity, ease and swiftness of its application, harmless nature to human
beings, domestic animals and also to building and equipments, easy availability, cheap cost are
considered. The effectiveness depends on concentration, temperature, sprayed surface, humidity. The
grainage and rearing room including equipment are first cleaned to remove and drive out dirt, dust
particles. Then washed with fresh water after closing the crevices using clay-lime, cement etc. Then
disinfection process is started.
Fig.7.4 Cleaning process
a. Spraying: The
grainage and rearing
equipments are dipped
in a tank containing
0.5%, sodium
hypochlorite or 5%
bleaching powder
solution for about 10
minutes followed by
sun drying. Nets, foam
49
pads are soaked in 2% formalin solution. The bigger equipment are sprayed with 2% formalin.
Spraying of formalin solution makes metallic equipments rusted thus care is to be taken. The
grainage and rearing rooms are also sprayed with 2% formalin. Ideal room temperature should be
maintained at 25oC during disinfection for quick diffusion of formaldehyde gas. When the
temperature falls after disinfection, the sterilization power falls. The proper time for disinfection
is 11am. After spraying the room should be closed at least for one day. Formalin causes irritation
in nasal mucosa and eyes, further skin hardens. Thus care should be taken during disinfection by
keeping face mask and gloves.
Fig.7.5 Disinfection of rearing house and equipment Fig.7.6 Ideal entrance
b. Fumigation: Disinfection in gaseous form is called fumigation. This process is effective in only
air tight rearing houses. It is convenient and easy to disinfect room and equipment
simultaneously. The quantity of original formalin solution required for disinfection is calculated
according to room size. Then it is diluted 4-5 times. The solution is allowed to evaporate in a pan,
by heating from beneath using electric or charcoal stove. Slowly the formalin gas comes out, care
should be taken that the chemical should not catch fire, which nullified the disinfecting effect. In
any case it is necessary to fumigate for 4-5 times. The temperature of 20oC or higher and 70% or
above humidity is preferable for fumigation. Metallic equipments can be disinfected by this
method.
For preparing required strength of formalin the following methods is used.
Strength of Original formalin - strength of formalin required

Required strength of formalin 
Strength of formalin required
Or
Required concentration X Required quantity of solution
Quantity of formalin required 
available concentration of commercial formaldehyde
7.3 SEED PRODUCTION PROCESS
The local races of silkworms are adapted to prevailing climatic condition of our country which yields
poor silk cocoons. Whereas Chinese, Japanese exotic varieties produce superior quality of cocoons but
50
intolerant to local conditions of our country thus cannot be reared for commercial production. But these
exotic varieties with superior yield and quality are combined with strong, resistant local variety to
produce hybrid silkworms. The bivoltines and multivoltines pure races are reared systematically to cross
breed. However certification of disease free laying is a must.
a. Stock maintenance: To produce good quality seed there must be a sound seed organization.
Considering the importance of quality seed, the sericulture countries have established a network of
institutions for egg production and also to impart training to staff working in grainage. Silkworm
seeds are two types ie., reproductive and industrial seeds.
b. Reproductive seed: These are used for producing seed cocoons (parents of seed cocoons) which
are required in large numbers for producing commercial seed. The main purpose is to maintain
racial purity. The selling price of these is 30-50% more than reeling cocoons. These seeds are
multiplied in number in a series of breeding centers called breeding stations (in three or four
stages) in order to ensure that the racial characters are not diluted during multiplication.
c. Industrial or Commercial seed: These re specific hybrids between two or more pure lines of
races of silkworm and are reared by normal rearer to produce commercial cocoons for reeling
purpose. These seeds are produced in grainages. Seed rearers are selected by sericulture dept that
carryon grainages. Seed cocoons must be healthy, hygienic and preserve racial characters.
Procurement of seed is based on certain standards fixed in respect of different races and seasons.
These are produced under ideal climatic condition, free from diseases, exclusively for purpose of
reproduction.
Table 7.1 Norms for production of seed cocoons.

S.No. Character Bivoltine Multivoltine
1. Egg/laying 400-450 300-350
2. Percentage of hatching 85-90% 90%
3. ERR 20000 cocoons/100dfls 18000 cocoons/100dfls
(30kg) (18kg)
4. Pupation rate 90% 90%
5. No. of cocoons/Kg Not above 700 Not above 1000
6. Shell ratio 18% 12%
After confirming all the above factors certain commercial characters such as ERR, pupal weight, shell
weight, shell ratio, good and bad cocoon percentage, filament length, floss and denier, raw silk
percentage, renditta are calculated for fixing the price.
7.3.1 Procurement of seed cocoons
Seed cocoons are those which are produced under ideal climatic condition, free from disease, exclusively
for the purpose of reproduction. Seed cocoons should confirm to the racial characters. In practice seed
cocoons are raised in seed areas. The quality of seed cocoons determines the quality and productivity of
seeds, it is very important that the cocoons procured for seed preparation should be of high standard.
Keeping all these facts, standards are fixed in respect of different races and season which are as follows.
a) Purchase of cocoons which have been closely watched by the extension staff and health
certificate affixed on inspection card.
b) Gut examination of pupae must be conducted before purchase.
c) The seed crop should be free from diseases especially pebrine.
51
d) Seed crop should have been reared under ideal conditions and fed with nutritious mulberry leaf.
e) Seed crop should have a good survival rate.
f) Seed crop should have a high pupation rate. But do not purchase cocoons which are with very
heavy pupal eight as that might lead to melting.
g) Cocoons should have good cocoon weight.
h) Cocoons not confirming to the characters of the race should not be purchased.
i) Crops showing an average yield of cocoons and above as fixed by the norms only should be
purchased.
j) Rates are fixed as per the standards.
k) Purchase officer must certify for the quality of cocoons and its disease freeness.
l) Details of rearer, quality and quantity of cocoons, race, spinning date, cost, total amount paid,
name of the market are recorded and sent to the grainage along with cocoons.
m) The following are the norms for production of cocoons for multivoltine and bivoltine breeds.
S.No. Characters Bivoltine Multivoltine

1. Egg/laying 400-450 300-350
2. % of hatching 85-90% 90%
3. ERR 20000 cocoons/dfls (30kg) 18000 cocoons/100dfls (18kg)
4. Pupation rate 90% 90%
5. No. of cocoons/kg Not above 700 Not above 1000
6. Shell ratio 18% 12%
After confirming all the above factors certain commercial characters such ERR, pupal weight, shell
weight, shell ratio, good and bad cocoon percentages, filament length, floss are calculated. The cocoon
rate is to be fixed on procurement of seed cocoons.
a. Price fixation:
There are certain norms for fixing the price of cocoons which are periodically revised by the government
in favor of seed cocoon producers. The yield of cocoons must not be less than 30kg/100dfls on the day of
marketing to the grainage in bivoltines and 20kd/100dfls in multivoltines. And number of cocoons/kg of
bivolting must not be less than 550-700; 800-1100 in multivoltine.
b. Principles for price fixation:
i. Standard Cocoons: bivoltine 650/kg ; multivoltine 1000/kg

ii. Standard Rate: It is fixed by the government from time to time
iii. Rate fixed/kg of cocoons brought by the farmer
Standard rate X No. of standard cocoons/kg

Cost of kg cocoons 
No. of cocoons per kg of the farmer
c. Model Problem-I
Venkaiah brought 40kg of bivoltine cocoons which weigh 620 number per kg. the standard rate is Rs.
450/kg. Calculate the cost cocoons per kg and total amount due to the farmer.
Standard cocoons = 650/kg

Standard rate/kg = Rs. 450/-
No. of cocoons/kg of the farmer = 620
52
450 X 650 292500

  471.77
620 620
Cost of one kg cocoons is Rs. 471.77
Total number of cocoons of the farmer = 40kg
471.77 X 40 = Rs. 18,870.80
Total amount due to Venkaiah is Rs. 18,870.80
d. Model Problem-II
Venkaiah brought 40kg of bivoltine cocoons which weigh 620 number per kg. the standard rate is Rs.
450/kg. Calculate the cost cocoons per kg and total amount due to the farmer.
Standard cocoons = 1000/kg

Standard rate/kg = Rs. 350/-
No. of cocoons/kg of the farmer = 900

350 X 1000 350000

  388.88
900 900
Cost of one kg cocoons is Rs. 388.88
Total number of cocoons of the farmer = 40kg
388.88 X 40 = Rs. 15,555.20
Total amount due to Venkaiah is Rs. 15,555.20
7.3.2 Transport of seed cocoons
Carrying seed cocoons safely from production centers to egg processing centers. Safe transportation is
necessary not to affect pupae ad cocoons which hamper the mother emergence. The seed cocoons are
harvested on the 5th or 6th day after spinning. This stage is suitable for transportation. If the pupa has
turned dark brown color and is hard to touch then the seed cocoons are fit to be transported.
Transportation at the late pupal stage causes damage.
The best time for transportation of seed cocoons is the cooler hours of the day ie., early morning or late
evening. Hot days damage the pupae due to heating up of cocoons such cocoons emerge weak moths
either die or lay poor egg number.
53
Seed cocoons should be loosely packed in the containers (cloth bags, bamboo conical baskets, plastic
perforated bins) so as to allow sufficient space for aeration. Each container is perforated and containers
are placed horizontally one upon the other in rows.
7.3.3 Preliminary examination
It has two methods – pupal gut examination and forced eclosion. As the pebrine spores tend to
concentrate in the gut region, pupal gut is extracted and examined. At the beginning of infection
identification is difficult. To overcome this, cocoons are subjected to high temperature (30 o -32oC) for
early eclosion. The emerged moths (after 2-3days) are examined for pebrine spores. However both types
of tests are advised.
In each test examine 2 to 3 smears and 8-10 fields carefully. The spores if present appear as oval shining
bodies under 600 magnification of the microscope. If pebrine is noticed, further processing of cocoons is
stopped and they are immediately disposed for reeling. This saves cost of seed cocoons, labor and
material.
Later about 10 male and 10 female cocoons are weighed to calculate cocoon weight, shell weight and silk
ratio and recorded to confirm the quality.
shell wright
a. Shell ratio   100
cocoon weight
b. Filament length = one revolution of epprouvette = 1.125mts.

or 400 revolutions on epprouvette = 450mts.
reeled silk weigh t (g)

c. Denier  X 9000
filament length
7.4 PROCESSING OF SEED COCOONS

seed cocoons declared as free of pebrine disease are preserved for egg production. Cocoons are sorted to
separate the following. Only good cocoons of quality confirming to the breeds are selected and preserved.
Deflossing of cocoons is done to facilitate easy eclosion of moths. Bad cocoons are either stifled or sent
to cocoon market. It is unhealthy to keep such cocoons in the grainage.
weight of bad cocoons

a. Percentage of bad cocoons  x 100
Total no. of bad cocoons
Or  x 100
Total no. of cocoons
weight of bad cocoons
b. Percentage of good cocoons  x 100
54
Total no. of good cocoons
or  x 100
Total no. of cocoons
Total cocoons = good cocoons + bad cocoons
7.4.1 Preservation of seed cocoons/pupae
Good cocoons are arranged in a single layer in bamboo trays. Each tray can hold 1000-1200 multivoltine
cocoons or 800-900 bivolting cocoons by arranging in stands. Pupae are stored in wooden rectangular
trays. Over crowing is avoided which leads to pupal mortality.
Temperature and humidity play vital in cocoons and

pupa preservation. The proper temperature range of
23o-26oC and 70-80% humidity are required. The
uniform eclosion of moths depends on the intensity
and duration of light. Cocoons should be exposed to
diffused light during the day and darkness during the
night. The higher the temperature the lower will be
the eclosion rate. At a temperature of 30oC and
above, the eclosion rate get reduced to minimum and
male sterility sets in. moths become weak and not be
able to copulate and eggs laid if any do not hatch. At
lower temperature of 20oC the eclosion is delayed
and egg laying period is extended, becomes irregular,
fecundity is affected, the size of the egg is reduced
and percentage of unfertilized eggs increase.
The trays are covered with perforated paper through

which emerged moths make their way out, this
facilitates easy picking of moths for coupling. Fig.7.7 Preservation of Seed Cocoons
7.5 SEX SEPARATION OF SEED COCOONS
Industrial seeds are hybrids of two or more races, thus separated sexually facilitating mating desired
variety. If they are not separated before emergence (eclosion) pairing occurs in the same race. Sexing
helps in true hybrid preparation. The sexes can be separated either at the pupal stage or at the moth stage.
Separating the sex at moth stage is laborious as the worker needs to be vigilant throughout the emergence
period. Sexing at the pupal stage is better and more efficient. The best period for sex separation is when
the pupae are 2-3 days old. The pupae are dark brown in color, hard to touch and sex markings are clearly
visible. A skilled worker can separate 10000-12000 pupae in eight hrs. with less than 5% error. It can be
done mechanically based on body width and weight but the error would be 20-30%.
Fig.7.8 Bombyx pupa
55
Male Female
Small in size with a narrow and Larger in size with a broader
tapering abdomen. abdomen.
A dot like structure on 8th An ‗X‘ mark on the 8th abdominal
abdominal segment segment
Check Your Progress

1. _________________is used to prevent crawling of ants.

2. __________________________________equipments are used for moth examination.
3. Bombyx female pupa is________________while male is____________________
4. __________________________is used for disinfection.
7.5 SUMMARY
Grainages are the centers for production of large scale quality and quantities of disease free layings of
silkworm. There are certain prerequisites for grainage operation among which building location, grainage
building, equipment, technical staff are main.
Egg production and processing is carried on with lot of care and technique. It requires a specific
environment for each and every grainage activity.
The grainage equipments are designed for a specific function. These equipments are made in such a way
to transport and handle easily.
Disinfection is called as extermination of disease causing germs. The most effective and simple method is
carried using chemical and disinfectants. Disinfection in gaseous form is called fumigation. This process
is effective in only air tight rearing houses.
Seed cocoons are those which are produced under ideal climatic condition, free from disease, exclusively
for the purpose of reproduction. There are certain norms for fixing the price of cocoons which are
periodically revised by the government in favor of seed cocoon producers.
Carrying seed cocoons safely from production centers to egg processing centers. Safe transportation is
necessary not to affect pupae ad cocoons which hamper the mother emergence.
Preliminary examination has two methods – pupal gut examination and forced eclosion. As the pebrine
spores tend to concentrate in the gut region, pupal gut is extracted and examined.
seed cocoons declared as free of pebrine disease are preserved for egg production.
Good cocoons are arranged in a single layer in bamboo trays. Each tray can hold 1000-1200 multivoltine
cocoons or 800-900 bivolting cocoons by arranging in stands.
56
Industrial seeds are hybrids of two or more races, thus separated sexually facilitating mating desired
variety. If they are not separated before emergence (eclosion) pairing occurs in the same race. Sexing
helps in true hybrid preparation.
1. Ant well.
2. Moth crushing set- mortor and pestle.
3. Big abdomen, narrow abdomen
4. Formalin
1. Detail about grainage equipment.

2. Write about prerequisites of grainage .
3. Detail about disinfection of rearing room and equipments.
4. Write about seed production process.
1. Write about preliminary examination of seed cocoons.

2. Write about transport of seed cocoons.
3. Write about preservation of seed cocoons.
4. Write about pupal sex separation.
5. Mention the importance of pupal examination.
6. Write about moth crushing set.
7. What is the importance of thermometer, hygrometer, humidifier?
8. What is the use of formalin in grainage?
9. Write about processing of cocoons.
57
UNIT-8 SEED PRODUCTION
Contents
8.0 Objectives
8.1 Introduction
8.2 Seed Production
8.2.1 Synchronization of Moths
8.2.2 Moth Emergence
8.2.3 Selection of Moths Coupling and Decoupling
8.2.4 Oviposition
8.3 Egg Production
8.3.1 Oviposition
8.3.2 Flat Card Method
8.3.3 Loose Egg Preparation
8.3.4 Effect of Environmental conditions on egg laying
8.4 Moth Examination
8.4.1 Kinds of Examinations
8.4.2 Surface sterilization
8.4.3 Assessment of layings
8.4.4 Characters of layings
8.5 Summary
8.0. OBJECTIVES
 knows about moth synchronization.
 understands to select moths for coupling.
 understands about oviposition.
 understands about types of egg production.
 knows about moth examination.
8.1. INTRODUCTION
Grainage produces pure and hybrid seed. These centres are more popular as commercial egg production
centres because they have a direct link with seed rearers. These centres encourage progressive farmers
and seed rearers to produce seeds commercially. Farmers always intense to produce good quality cocoons,
hence the farmer look forward to the Grainage for the supply of high vigour disease free commercial
seeds. These seeds produce cocoons with rich silk content and high yield.
8.2 SEED PRODUCTION
The aim of a seed production is to produce quality seeds. The centre should have proper facilities, good
environmental conditions, and well spacious rooms, without water stagnation around the Grainage
building and should be away from factories and pesticide industries. A seed production centre must be
established where sericulture is popular among the villages. It can also help the farmers technically. These
Grainage centres conduct training program for the unemployed youth to create awareness on commercial
rearing.
58
8.2.1 Synchronization of Moths Moth Emergence
While preparing hybrid seed, moths of different races are made to emerge (eclosion) simultaneously on
the same day, so that male and female moths are readily available for hybridization. This phenomenon is
called as ―moth synchronization‖ or ―adjustment of emergence of moths‖. Brushing time can be adjusted
to synchronize moth emergence. If one race takes two days more from incubation to eclosion, it must be
incubated two days earlier. The emergence can be adjusted by refrigerating the cocoon or pupae, thus
delays moth emergence. On the similar lines moths can also be refrigerated. Pupae of 2-3 day old or just
at the time of emergence are ideal for refrigeration. The safe period of refrigeration varies with stages and
sexes. The male pupae and moths can be refrigerated for 7 days. The female pupae and moths can be
refrigerated for 2-3 days. The ideal temperature for refrigeration of cocoons/pupae/moth is 5oC with a
humidity of 65% for cocoons and pupae and 75-8-% for moths.
8.2.2 Moth Emergence
The silkworm after spinning the cocoon, lodges inside the cocoon and transforms into a pupa. The pupa
later metamorphoses into moth, which comes out of the cocoon. This process of moth coming out of
cocoon is called ―moth emergence‖.
Generally moths emerge after 9-14 days of spinning. But the pupal age varies according to races and
seasons. Multivoltine races emerge earlier than other races, followed by China, Japan bivolting races. The
expected time of moth emergence can be predicted by observing the pupal and cocoon characters. A day
before becoming moths, the pupae appear soft, dark in color with loose pupal skin, prominent eye
formation, wing parts, appendages appear. A day before emergence a distinct wrapping sound can be
heard in the seed cocoon room. This is due to transformation of pupae into moths, which are trying to
rupture pupal skin to come out.
It requires a specific environmental condition for cocoon or pupae before emergence of moths. These are
to be kept in darkness. On the expected day of emergence bright lights are illuminated on early in the
morning at 4-6pm. Temperature of 25oC with 75-80% humidity has to be maintained during emergence.
Moths start emerging 6-8am, which are picked and kept in a try for urination. The healthy moths are taken
for coupling or for preservation. Emergence decreases as the day light intensity increases. And after
8.30am moths do not emerge. Then the room must be closed and dark light is provided only in the
subsequent days of emergence. Moths emerge only for 3-4 days only.
Sometimes due to mistakes in sex separation moths of other sex mix and emerged moths mate. Such pairs
are rejected. While
emergence the moth ruptures
the pupal skin and secretes an
alkaline juice on one end of
the cocoon and makes it wet.
Then moth uses its legs and
head, wriggles out of the
cocoon. After coming out it
stretches its wings in about 5
minutes. After passing urine
especially male moths are
seen moving actively in the
tray.
Fig.8.1 Moth emergence
59
8.2.3 Selection of Moths, Coupling and Decoupling
The moths which are unhealthy, injured with crumpled wings and exposed body is rejected. Only healthy
moths are selected for mating. The female moths (300-400) are collected into a fresh tray into which
desired male moths are broadcast over the females gently. The moths pair within 5-10 minutes. The moths
pair within 5-10 minutes. The left over moths (unpaired) are collected in to another tray. If the males
available are in excess, they should be preserved at 5oC for later use.
Each paired moths are kept in cellules arranged in a tray (90 x 60 cm size). All the trays are kept in dark
room for about 3-4hrs. temperature of 25oC with 75-80% humidity and semi dark conditions are provided.
In a span of 3-4hrs of mating, male moth ejaculates twice. The first ejaculation occurs just after half-an-
hour and second after 90 minutes. Therefore, pairing for 4-6hrs favors complete coupling to fertilize
maximum number of eggs. Further, it favors uniform age of eggs. It is also helps to carry on acid
treatment. The pairing process should be completed before 8am and cannot be carried for 8-12pm.
Fig.8.2 Coupling, Oviposition, healthy and deformed moths
After six hours of mating the moths are separated and it is called ―depairing‖. The males are separated by
holding the female twisting male gently without injuring the external genitalia of the female.
Male moths second pairing
Properly preserved male moths pairs for eight times. But the number of pairings decreases fertilization
rates. Thus male moths can be conveniently used for three times. The male moths are preserved at 5-
60
7.5oC temperature for about 4-5 days. Moths are preserved in a single layer. Prior to second pairing the
male moths preserved are released at room temperature for 5-10minutes. Second pairing duration for
about 4 hours with a temperature of 25oC and 80 percent humidity.
8.3 EGG PRODUCTION
Egg production is one of the critical phases in grainage. Complete activity of the sericulture industry is
based on grainage activity. Production of good and disease free layings definitely improves the industry.
Silkworm eggs are divided into hibernating and non-hibernating eggs. In hibernating eggs the embryo
develops only halfway and undergoes a stage of dormancy called diapauses and hatches out in the
following spring. In non-hibernating eggs the embryo develops without undergoing diapauses and hatches
out in normal way.
Generally univoltines lay only hibernating eggs and multivoltines lay non-hibernating eggs while
behavior of bivoltine eggs is intermediate. This trait can change under different environmental conditions
but silkworms are broadly divided into univoltine, bivolting and multivoltine.
8.3.1 Oviposition
After decoupling females are first place on a paper and tapped to induce them to pass urine. Later moths
are placed on egg cards, covered individually with a cellule or moth funnel. This provides darkness and is
not disturbed. In case of loose egg preparation 50-100 moths are put on glue coated craft paper and placed
on a tray which is closed on all sides. The moths normally lay eggs from afternoon onwards and reach
peak stage by early night hours. By the next morning the moths complete the laying process. On an
average multivoltines lay 300-400, univoltine and bivolting 400-500 eggs in 24hrs. the oviposition room
is kept dark by closing the doors and windows. The temperature of 24±1oC and humidity of 80% are
maintained. Low humidity dries up the gummy substance of ovipositor secreted by female and obstructs
egg laying. As a result we get less number of eggs.
There are two types in preparation of laying.

a. Segregated egg laying has two methods.
i. Pasteur‘s method (followed in China, Japan)
ii. Cellular bag method (followed in European countries)
b. Mixed egg laying also has two methods.
i. Flat card method
ii. Loose egg method.
8.3.2 Flat card Method
It is used for industrial egg production on a very large scale. Egg sheets contain 20-42 squares are
arranged in 60 x 90cm tray. Each female moth is kept in a square and covered with cellule. Such trays are
arranged in one tier in a wooden rack kept in oviposition room. The room is maintained dark and
optimum temperature and humidity are provided, for better laying of eggs. The egg sheets along with
moths are taken to moth examination hall after 24hrs of egg laying. The moth samples are taken for
crushing and examined for pebrine disease. If a certain percentage of samples are found to have
pebrine, then entire lot is disposed. The advantages of this method are that pebrine inspection is perfect
and eggs laid by pebrine infected moths are eliminated/scraped. Hence reproductive seeds are prepared on
cards.
61
Fig.8.3 Female moth
laying eggs
and Egg card
8.3.3 Loose Egg Preparation
It is similar to flat card method except that the eggs are laid on smooth side of craft paper or on starched
paper. This method has large advantages. A large number of moths are allowed to lay eggs and only
sample moths are drawn for examination. This method is adopted for commercial seed production.
About 100-200 gms of brushed to remove the eggs from the cards. Arron root or Maranta starch is added
to one litre of water ad boiled to prepare a paste. After cooling the paste is smeared uniformly on craft
sheet (90 x 60cm) or cloth in thin layer. These sheets are spread in a wooden tray. Female moths (30-200)
after urination are transferred in the oviposition room and moths are allowed to lay eggs for 1-2 days. On
the next day moths are removed for examination. After examination the egg sheets/cloth are soaked in
water for 15minutes. It is gently brushed to remove the eggs from cards. Then eggs are collected by
removing the sheet and filtering through a muslin cloth. These eggs are soaked in 0.5% bleaching powder
for 5-10minutes to remove the gum and to avoid formation of clumps of eggs. Eggs are washed in water
and transferred to salt solution with a specific gravity of 1.06 – 1.09 at room temperature. The fertilized
eggs having higher specific gravity sink in the solution. The floating eggs with low specific gravity are
rejected. The good eggs are washed again in water then with 2% formalin solution of 20minutes. Again
eggs are washed in water and dried in shade. These eggs are packed in box (unit) consisting of 20,000
eggs. The boxes are sealed and labeled. The label should have the details of race, date of egg laying,
quantity of eggs, name of the grainage, technician signature. In multivoltine a gram of eggs has about
2000 eggs which 1800 in bivoltines. One kg of bivoltine cocoons yield about 55gms of eggs. In cross
bred 1kg of multivoltine and 0.7kg of bivoltine cocoons yield about 55gms of hybrids. This is due to
rejection of bivoltine females and multivoltine males. In reciprocal cross 20gms of seed recovery is
expected.
Fig.8.4 Loose egg preparation
62
Advantages of loose eggs:
1. Easy for mass egg production, handling, transportation and incubation. Facilitates elimination
of poor quality and unfertilized eggs before supply.
2. Surface sterilization and hydrochloric acid treatment is more effective and uniform.
3. More accurate than sheet eggs for comparison of scientific data like individual yields, race-
wise, yields, region and season-wise yields.
4. Different between sheet egg and loose egg production Sheet egg production
5. Ordinary craft sheets without starch coating are used for egg laying.
6. Kraft sheet used for egg laying is of 22.5 cm x 28 cm in size.
7. On each sheet 20 moths are left to lay eggs.
8. Cellules are placed over the moths to restrict the egg laying area.
9. Eggs stuck to sheets are supplied after surface sterilization and acid treatment.
10. Followed for P3, P2 and P1 production.
11. Loose egg production
12. Ordinary craft sheets coated with starch are used for egg laying. Starch coated craft sheet used
for egg layings is of 65 cm x 95 cm in size.
13. On each sheet 250 to 300 moths are left to lay eggs.
14. Cellules are not used, but the moths are spread free on the sheet.
15. Eggs laid on the sheet are loosened, surface sterilized, acid treated, measured, packed in loose
egg boxes and supplied.
8.3.4 Effect of Environmental conditions on egg laying
Oviposition room should have 24oC±1oC temperature and 80% humidity. If the humidity is less the glue
secretions released by accessory glands of female from oviduct during oviposition dries and eggs cannot
descent down. Thus oviposition is is stopped. This causes very less number of eggs. Thus proper humidity
is to be maintained for which electrical humidifiers, wet gunny cloths, soil beds are the best options.
When the temperture rises the glue released from oviduct glands will dry and again causes a hirdle in egg
laying. During low temperature the glue will not dry soon. Thus both the factors ie. Temperature and
humidity are to be maintained properly. This favors increment in egg number.
8.4 MOTH EXAMINATION
Pebrine is Transovarially transmitted disease. Infected moths transfer the disease through eggs to next
generation. The elimination of eggs laid by a disease moth is important. Hence it is necessary to examine
mother moth.
There are two methods.

a. Fresh moth or green moth examination: In this moths are examined soon after egg laying when
the seeds are required for immediate use.
b. Dry moth examination: When the eggs are to be hibernated, the moths can be dried and tested at
leisure. A sample of 30 moths is oven dried at 70±5oC for 6hrs.
8.4.1 Kinds of Examinations
Kinds of examinations are of two types depending on the purpose for which the seed is used ie.,
reproductive seed or commercial seed.
63
a. Individual moth examination
It is conducted for producing reproductive seeds. Therefore all moths are examined individually
to make sure that they are completely free from pebrine. After oviposition moths are transferred
to mortar and crushed with pestle by adding 2-3 drops of 2% KOH solution. A smear is taken to
slide and covered with cover glass and observed under microscope at 600 magnifications.
Infected moths are marked to scrap the eggs laid. This process is laborious but perfect for
identifying the pebrine.
Fig.8.5 Moth examination
b. Mass moth examination

It is followed for commercial/industrial seed production where only samples of moths are
examined in groups of 10-30. After oviposition moths are taken in groups for crushing by adding
90-100ml of 0.5 potassium carbonate solution. Moths are crushed at 10000 rpm to separate
pebrine spores from tissue. After that material is filtered through a coarse filter paper. And filtrate
is centrifuged at 200rpm for 3min. and ppt is dissolved in 2-3 drops of KOH. The smears are
taken for observing under microscope.
c. Identification of pebrine spores
These spores appear as shining oval bodies under microscope. Though the spores are colorless with a
luster, decreased intensity of light gives a satisfactory contrast of light and shade making the observation
clear.
8.4.2 Surface sterilization
The eggs are processed after moth examination. While egg laying the surface is stained with moth urine
and covered with moth scales and disease causing organisms. Thus egg surface has to be washed and
surface is disinfected to remove the stains and surface contamination which is called egg processing. Egg
sheets/cards with standard laying after removing the poor layings and dead layings soaked in 2% formalin
for 5-10min. These eggs are washed in water and allowed to dry. These egg sheets carry the name of the
grainage, hybrid combination, date of egg laying, signature of moth examiner, number of good layings.
Such eggs are now ready for sale. Loose eggs are sterilized with 0.5% bleaching powder.
8.4.3 Assessment of layings
The egg sheets sometimes contain deformed, poor, unfertilized, diseased eggs which are identified and
marked during moth examination. These eggs are removed or scraped from egg card/loose eggs. The egg
64
processing refers to sort good and bad/defective eggs and removal of surface stain and infections by
adopting suitable methods.
8.4.4 Characters of different layings
Total number of eggs laid by a single moth is called as one laying. Only good layings are supplied to the
rearer.
Good layings
1. Each laying should have minimum of 300 eggs.
2. Layings with less fertilized eggs are not considered.
3. There should not be any piled eggs.
4. The eggs are laid in a single layer, side by side.
5. Good laying has maximum number of eggs.
6. The disease free layings (DFLs) are called as good layings.
Multivoltine hybrid seed

1. Eggs are non-pigmented and non-hibernating.
2. Eggs hatch after 10 days of egg laying.
3. Acid treatment is not required to make eggs hatch.
4. Sheet egg method is applied for egg production.
5. Eggs cannot be stored for more than 20 days in 5 °C.
6. Bivoltine male moths and multivoltine female moths are used for production of hybrids.
7. Sexes are separated at moth level for production of hybrids.
Multivoltine hybrid seed (CSR)

1. Eggs are pigmented and hibernating.
2. Eggs do not hatch after 10 days of egg laying naturally.
3. Acid treatment is required to make the eggs hatch after 10 ~ 11 days.
4. Eggs are produced by loose egg production method.
5. Eggs can be stored up to 6 months under 2.5 °C and 5 °C.
6. Male and female moths of bivoltine are used for crossing reciprocally.
7. Sexes are separated at pupa level for production of hybrids.
Check Your Progress
1. ____________________________moths are not suitable for egg preparation.

2. Paired moths are kept in________________for copulation.
3. Sheet eggs are prepared on _____________________
4. ____________________examination is compulsory before finalizing eggs for packing.
8.5 SUMMARY
Grainage produces pure and hybrid seed. These centres are more popular as commercial egg production
centres because they have a direct link with seed rearers. The aim of a seed production is to produce
quality seeds.
While preparing hybrid seed, moths of different races are made to emerge (eclosion) simultaneously on
the same day, so that male and female moths are readily available for hybridization. This phenomenon is
called as ―moth synchronization‖ or ―adjustment of emergence of moths‖.
65
Generally moths emerge after 9-14 days of spinning. But the pupal age varies according to races and
seasons. The moths which are unhealthy, injured with crumpled wings and exposed body is rejected. Only
healthy moths are selected for mating. Properly preserved male moths pairs for eight times.
Egg production is one of the critical phases in grainage.
After decoupling females are first place on a paper and tapped to induce them to pass urine. Later moths
are placed on egg cards, covered individually with a cellule or moth funnel.
Loose egg preparation is used for industrial egg production on a very large scale. Egg sheets contain 20-
42 squares are arranged in 60 x 90cm tray. Each female moth is kept in a square and covered with cellule.
It is similar to flat card method except that the eggs are laid on smooth side of craft paper or on starched
paper. This method has large advantages. A large number of moths are allowed to lay eggs and only
sample moths are drawn for examination.
Oviposition room should have 24oC±1oC temperature and 80% humidity.
Pebrine is Transovarially transmitted disease. Infected moths transfer the disease through eggs to next
generation. The elimination of eggs laid by a disease moth is important.
egg surface has to be washed and surface is disinfected to remove the stains and surface contamination
Total number of eggs laid by a single moth is called as one laying.
1. Deformed moths
2. Cellule
3. Egg cards
4. Mother moth examination
1. Write about loose egg preparation.

2. Write about moth examination procedure.
3. Write about selection of Moths, Coupling and Decoupling.
4. Write about moth emergence.
1. Write about surface sterilization.

2. Write characters of good eggs.
3. Write about assessment of layings.
4. Write about environmental effects on egg laying.
5. Write about oviposition.
6. Write about male moth second pairing.
7. Write about synchronization of moths.
66
UNIT-9 EGG PRESERVATION AND HIBERNATION
Contents
9.0 Objectives
9.1 Introduction
9.2 Egg Preservation and Hibernation
9.2.1 Preservation of eggs produced in spring
9.2.1.1 Preservation in summer after egg laid
9.2.1.2 Preservation in autumn and winter
9.2.2 Preservation of eggs produced in autumn
9.3 Summary
9.0. OBJECTIVES

 knows about egg preservation and its importance.
 understands storage of spring and autumn eggs.
 understands hibernation schedules.
9.1. INTRODUCTION
After preparation of silkworm eggs they are to be preserved well. These eggs are of diapausing and
non-diapausing type needs lot of care in handling so that they are well preserved and utilized for the
particular season.
9.2 EGG PRESERVATION AND HIBERNATION
In silkworm rearing it is important not only to produce silkworm eggs of high quality, but also to
carefully protect and preserve them to ensure good and uniform hatching. Univoltine eggs are left
after oviposition without any treatment, undergo diapauses without hatching. But the bivoltine eggs if
incubated at high temperature become hibernating or black eggs and do not hatch for the second time
during the year.
a. Types of eggs: The eggs are of two types ie. diapausing and non-diapausing eggs. After
oviposition further processing of eggs depends upon their diapausing or non-diapausing
nature. Generally univoltine are diapausing eggs. Multivoltine eggs are non-diapausing type.
The industrial or commercial eggs are hybrids of these uni and bivoltine are non-diapausing
type.
b. Handling of eggs: In diapausing eggs a hormone is responsible for inhibition of embryo

development, and its effect is neutralized by the effect of cold temperature. Thus the eggs are
activated. The duration of aestivation period ie. the duration of higher temperature at which the
eggs are kept and the related duration of cold temperature treatment required to break the
diapauses, eggs laid in any season can be properly preserved and hatched as required during
the following spring. This handling of eggs refers to processing of eggs under optimum
conditions to obtain hatching whenever desired. It depends on the nature of breeds involved in
handling. The important factors in egg handling are as follows.
i. Eggs are handled depending on the nature of eggs.

ii. Hatching of eggs has to be obtained in desired time.
iii. Conditions required to be maintained for various purposes.
67
c. Methods of handling of eggs:
i. Handling of multivoltine eggs: These breeds produce non-hibernating eggs. They hatch
in 10-11 days after laying. If hatching has to be delayed in these eggs, the eggs on the
second day of laying should be placed for preservation at 5oC with 75-80% humidity for
20 days. During this period eggs can be released for incubation on any day.
ii. Handling of bivoltine eggs: These breeds lay hibernating eggs which do not hatch in 10-
11 days after laying because these eggs undergo diapauses. However they can be made to
hatch by following artificial treatments. Depending on the requirement of their hatching,
they are processed by different methods.
9.2.1 Preservation of eggs produced in spring
9.2.1.1 Preservation in summer after egg laid
In about a week, the embryo enters a state of diapause. The egg color is lightly yellow when first laid
and after 36-48 hours it gradually changes into reddish brown and becomes darker. On the 4-5th day,
the egg acquires its inherent color of the variety. During this period, respiration is brisk because of the
embryonic development.
Care should be taken not to shock, crush or rub the eggs. So the eggs should be kept in well ventilated
clean rooms at a constant temperature of 24～25oC and a relative humidity of 75%.
High temperature in summer is a requisite for stabilizing the diapause of the hibernation eggs. The
proper temperature for completion diapause is 25oC. Temperature of 30oC is harmful to the eggs, and
higher than 30oC is the more harmful it will be.
Lower below 20oC may disturb the diapause completion, making the hibernating eggs unable to
withstand cold storage, and disturbs uniformity in embryo development. For the purpose of safe
production of the eggs, it is advisable to preserve the eggs around 50-60 days at 23-25oC in summer.
Chilling days needed to obtain more than 80% hatchability from the eggs preserved at 25 oC for 10 to
210 days.
The optimum humidity for the preservation in summer is 75-80%. If it is too dry, the eggs will be lost
too much water and too wet, mould is apt to grow on the eggs.
Fig.9.1 Preservation of silkworm eggs

deposited in June, 1.Oviposition, 2. Inn warm
regions, a decrease of temperature may be
necessary, 3.Exposure to low temperature, 4.
Cold storage, 5.intermediate care, 6. Removal
from cold storage for incubation
9.2.1.2 Preservation in autumn and

winter
In September through October (during

autumn), the eggs are still in diapause, so a slight variation in temperature does no harm, but in Nov.
through Dec. (early winter), the eggs enter the pre-termination of diapause, and high temperature
should be avoided.
68
In order to secure simultaneous hatching, it is necessary to expose the eggs to a low temperature of
7.5 to 5oC or lower for more than 50-60 days, as shown in step 3 in figure.
At the end of Jan. or beginning of Feb., diapause will be almost terminated at 5 to 7.5 oC. At this time
the embryos increase in length and the head lobe is wide open. If kept at 5oC for a long time at this
stage, the embryos that have terminated diapause become unviable. Therefore, within about 60 days
they must be transferred to 2.5oC. Thereafter, the eggs can safely be stored for 40-60 days at 2.5oC.
Intermidiated care: To ensure survival, eggs are allowed to develop one step by conditioning at
15oC for a few days (step 5). Thus the embryos reach the stage of the 'the longest embryo', just one
day before appearance of the neutral furrow. Then, the eggs are kept at 2.5oC until the time of
incubation.
(In sub-tropical or tropical areas, the natural atmospheric temperature is too high to break the diapause
even in winter.
It is, therefore, necessary to transfer the eggs into cold storage earlier to render the eggs to go through
an artificial hibernation for the purpose of awakening the embryo from diapause)
9.2.2. Preservation of eggs produced in autumn
Eggs produced in autumn for the rearing in the next spring are called autumn eggs and they are to be
hatched in the following spring.
As the duration of preservation of the eggs is shorter, the nutrient in the eggs is less consumed and it
is easy to maintain the quality of the eggs.
The time from egg production to incubation is 6 months from Oct. to Apr.-Mar. next year, and the
time for low temperature preservation needed to terminate the diapause takes 2-3 months.
During this period, the eggs can be treated as follows:
i. Preserved at 25oC within 20 days after egg laying.
ii. Lower the temperature step by step in following 10 days. Kept at 20 oC for the first day and at
15oC for the following 5 days.
iii. After being preserved in this way for a month, the eggs are kept in cold storage for the
hibernation
9.2.3. Hibernation Schedules
a. Physical and chemical stimulants

These are stimulants which are very useful for artificial hatching of eggs. The acid treated hibernating
eggs can be utilized after 10 days up to one year at any given time.
Table 9.1 List of stimulants
S.No. Physical stimulants S.No. Chemical stimulants

1. Lowest temperature 1. Hydrochloric acid (HCl)
2 Dipping in hot water 2 Nitric acid (HNO3)
3. High electric stimulation 3. Sulphuric acid (H2SO4)
4. Rubbing with brush or feather 4. Aqua regia
5. High atmospheric pressure 5. Acetic acid
69
6. Ultra hi-frequency vibrations 6. Sodium chloride (NaCl2)
7. Exposing to sunlight 7. Hydrogen peroxide
8. UV rays/ ultra short waves 8. Enzyme treatment
9. Exposing to oxygen 9. Ozone treatment
The acid is used to treat the eggs for breaking diapauses. The dilute HCl of specific gravity is
prepared for treating eggs. The strength of HCl varies according to temperature at which eggs are
treated.
In acid treatment strong inorganic acids give good results than organic acids. The nitric acid, sulphuric
acid re very strong acids and handled carefully. Nowadays mostly HCl is used. It is mixed with
required quantity of water to get required specific gravity needed for acid treatment of eggs. The
specific gravity is tested by hydrometer and corrected accordingly.
Table 9.2 Details for preparation of one liter acid

Available specific 1.075 HCl 1.100 HCl 1.110 HCl
gravity of HCl Water Acid Water Acid Water Acid
1.150 500 500 333 667 267 733
1.155 516 484 355 645 290 710
1.160 531 469 375 625 312 688
1.165 545 455 394 606 333 607
1.170 559 441 412 588 353 647
1.175 571 429 429 571 371 629
1.180 583 417 444 566 389 611
(Required specific gravity - 1.00) x (Required HCl in ml)
Required concentration of HCl 
(Available specific gravity - 1.00)
1.0 is the specific gravity of water. The value gives the amount of water to be added to get required
specific gravity of acid.
Formalin treatment is a prerequisite for acid treatment. The eggs are surface sterilized in 2% formalin
for 15 minutes. This helps to fix the eggs to the card otherwise are released into the acid during
treatment. After the time laps eggs are washed in water.
Commercial formalin - required strength

Required % 
required strength
Check Your Progress
1. Silkworm eggs are of__________________________________________________types

2. ______________acid is used for breaking diapause.
9.4 SUMMARY
In silkworm rearing it is important not only to produce silkworm eggs of high quality, but also to
carefully protect and preserve them to ensure good and uniform hatching. Univoltine eggs are left
after oviposition without any treatment, undergo diapauses without hatching.
The eggs are of two types ie. diapausing and non-diapausing eggs. In diapausing eggs a hormone is
responsible for inhibition of embryo development, and its effect is neutralized by the effect of cold
temperature.
70
In about a week, the embryo enters a state of diapause. The egg color is lightly yellow when first laid
and after 36-48 hours it gradually changes into reddish brown and becomes darker.
In September through October (during autumn), the eggs are still in diapause, so a slight variation in
temperature does no harm, but in Nov. through Dec. (early winter), the eggs enter the pre-termination
of diapause, and high temperature should be avoided.
Eggs produced in autumn for the rearing in the next spring are called autumn eggs and they are to be
hatched in the following spring.
The acid is used to treat the eggs for breaking diapauses. The dilute HCl of specific gravity is
prepared for treating eggs.

1. Hibernating and non-hibernating.
2. HCl.
1. Detail about preservation of eggs produced in spring.
II. Answer the following in about 10 lines.
1. Mention physical stimulants used in grainage.

2. Mention chemical stimulants used in grainage.
3. Write about methods of handling of eggs.
4. Write about types of eggs.
5. Write about handling of eggs.
71
UNIT-10 ACID TREATMENT OF SILKWORM EGGS
Contents
10.0 Objectives
10.1 Introduction
10.2 Acid Treatment
10.2.1 Artificial Treatment
10.2.2 Formalin treatment of eggs to enhance adhering efficiency
10.2.3 Common methods of acid treatment
10.2.4 Cold Storage Eggs
10.2.5 Transportation of Eggs
10.3 Incubation
10.5 Summary
10.0. OBJECTIVES
 know the importance of acid treatment.
 understands what is artificial treatment.
 understands formalin treatment before acid treatment.
 Knows about incubation is important.
10.1. INTRODUCTION
The diapause, in insects, is a method of overcoming unfavourable period caused either by

physiologically
unfavourable conditions or non-availability of food. For the silkworms of the temperate region, both
these conditions will prevail during winter and thus they have developed the diapausing (hibernating)
character lo overcome this period.
10.2. ACID TREATMENT
The phenomenon of diapause is environmentally, physiologically and genetically controlled. Acid

treatment is just a method to change the physiology by blocking certain activities and inducing several
new biochemical reactions for the continuous development of the eggs.
Diapause eggs are more customarily referred to as hibernating or bivoltine eggs, while the non-
diapause
ones as non-hibernating or multivoltine eggs.
10.2.1 Artificial Hatching
When the eggs left without any treatment the diapausing eggs do not hatch on incubation. However, it
is possible to make them hatch artificially by providing various physical or chemical stimuli. The
various physical and chemical agents/methods by which it is possible to block or terminate the
diapause are given in the previous unit. Though several artificial methods have been devised, the
immersion of diapausing eggs in Hydrochloric acid (HCl) has been the best choice and most widely
adopted technique for immediate hatching, from the technical as well as practical points of view, both
at the laboratory as well as commercial levels of bivoltine seed production.
72
To bring a shift from diapause to non-diapause type, changes in the incubation schedule like shorter
exposure to light and low temperature during incubation, changes in the rearing conditions like
continuous high temperature rearing, low temperature during early stages and high temperature during
the late stages, cocoon preservation at high temperature, coupled with selection, are essential.
S.No. Name of the acid Concentration of acid Density at

at saturation (%) 20oC
1. Hydrochloric acid (HCl) 40 1.1990
2. Nitric acid (HNO3) 100 1.5129
3. Sulphuric acid (H2SO4) 100 1.8305
Selection and preparation of Acid (HCL)
In acid treatment, strong and inorganic acids give good results than organic acids. The Nitric acid and
Sulphuric acid are very strong acids and should be handled carefully. Now a days, Hydrochloric acid
(HCl) is used for acid treatment of eggs. The HCl is mixed with required quantity of water to get
required specific gravity needed for acid treatment .The Specific gravity is tested by hydrometer and
corrected accordingly.
Table.10.1 Details for the preparation of acid
Available Specific 1.075 HCl 1.100 HCl 1.110 HCl

Gravity of HCl
Water HCl Water HCl Water HCl
1.150 500 500 333 677 267 733
1.155 516 484 355 645 290 710
1.160 531 469 375 625 312 688
1.165 545 455 394 606 333 607
1.170 559 441 412 588 353 647
1.175 571 429 429 571 371 629
1.180 583 417 444 556 389 611
(Required specific gravity-1.00) x (Required HCl in ml)

Required Concentration HCl=
(Available specific gravity – 1.00)
1.00 = Specific gravity of water.
The above value gives the amount of water to be mixed to get required specific gravity of acid.
Model Problem
Prepare 15 ltrs of 1.075 Specific gravity HCl with commercially available HCl (1.160 Specific
Gravity).
73
Solution
Substitute the values in the principle
(1.075 – 1.00) x (15,000) 0.075 x 15,000 1125
= ————————— = ——————— = ——— = 7031 ml of Acid
(1.160 – 1.00) 0.160 0.160
To get 15 ltrs.acid, add 7969 ml of water to 7031ml of acid. This acid contains. 1.075 Specific
gravity.
10.2.2 Formalin treatment of eggs to enhance adhering efficiency
The mother moth while oviposition provides a thin film of gluey substance, secreted from accessory
glands on the under surface of the eggs which enables them to stick to the egg card. lf these egg cards
are dipped as such into the acid, it tends to dissolve the gluey substance. As a result, large quantities
of the eggs get detached during acid treatment and subsequent washing in water.
To overcome this problem, the egg cards, prior to acid treatment, are soaked in 2% formalin solution
for 15 minutes. Formalin acts as a fixative agent which increases the adhering capacity of the eggs to
the cards. Besides it helps in surface sterilization of eggs against different disease causing pathogens
and in removing the wastes deposited on the cards.
The egg cards thus dipped in formalin solution are either directly taken for drying or subjected to
washing in water. Washing in water helps in eliminating the irritating smell of formalin. In either
case, the egg cards are dried in shade.
It is also a practical feasibility that formalin solution is used for dilution instead of water, at the time
of acid preparation. In such a case egg sheets are directly dipped in the acid for the purpose of
treatment. This is a simple method which is already in vogue in other countries. This method saves
time, as separate soaking in formalin solution and subsequent drying process, are curtailed.
A note of precaution is that since formalin is very volatile in nature, a certain amount of formalin may
be required to be supplemented after a few treatments judging from the condition of egg dropping.
10.2.3 Common methods of acid treatment
There are two most popular methods of treating the diapausing type of silkworm eggs, to block their
diapause and eventually make them to hatch like their non-diapausing counterparts. Such acid treated
eggs are termed as artificial non-diapause eggs.
The two methods are i.e., Hot Acid Treatment, Cold Acid Treatment.
10.2.3.1 Hot Acid Treatment
In hot acid treatment eggs are dipped in HCL having specific gravity of 1.075 at 46°C for 5-6
minutes. It is always necessary to maintain proper specific gravity of HCl. Before heating the specific
gravity of the acid is first adjusted to room temperature.HCL is not heated directly. Hot acid treatment
bath is used for treatment, which favours to heat HCl indirectly. Acid is taken into a glass tube and
placed in treatment bath. When the temperature reaches to 46°C the eggs are dipped. The dipping time
is different for different types of eggs.
74
After treatment eggs are washed in water. Hot acid is treated eggs after 24 hours, otherwise eggs
cannot withstand.
Fig.10.1 Acid Treatment of eggs
10.2.3 Cold Acid Treatment
It is also called as room temperature acid treatment. There is

no heating of acid; treatment is carried at 23-30°C
temperature to eggs after 24 hours of oviposition. The
treatment is conducted on first embryo stage. Further the
eggs are likely to get separated from the egg card and the
unfertilized eggs crumble, which help in their removal. In this method HCl 1.110 specific gravity at
15°C at room temperature is used to treat the eggs. Eggs are first treated with 2% formalin before acid
treatment for egg aglutinization and washed in water to remove acid traces.
Acid Treatment of Loose Eggs
The loose eggs after sterilization are kept in porous plastic container for acid treatment. The process
of treatment is similar as explained earlier. After washing in water eggs are dried on a smooth cloth.
Stopping of Acid Treatment
Generally it is not advisable, if inevitable eggs are kept at 5°C for 5 days, up to 7 days at 2.5°C.This
process is to be carried out later they are processed as detailed earlier. The eggs are kept at 15-25°C
for 2 hrs and slowly brought to 2.5oC. This method saves spoiling of eggs. During egg preservation
70-80% humidity is to be maintained.
The hibernated eggs are acid treated to prevent diapause. These eggs are cold stored at 5°C not
beyond 3 days.
10.2.4 Cold Storage of Eggs
There are two types i.e., Short term and Long term chilling.
Short Term Chilling
Temperature of 25°C and 70- 80 % humidity is maintained during oviposition. These eggs are left for
30-35 hrs, at 25°C to reach spoon head stage and then preserved at 5°C with 75-80 % humidity for 45-
50 days. If chilling has to be prolonged beyond 60 days, it is first carried out at 5°C for about 40 days
and then at the lower temperature of 2.5°C.The eggs can be released after 35 days and up to 50 days.
While releasing, the eggs are kept at 15°C for 6-12 hrs and then at 25°C for 3-4 hrs. The eggs are
treated with HCl of 1.100 Specific gravity at 47.8 C for 5-6 minutes. Then eggs are washed in water
to remove all traces of acid. Thus eggs could be made to hatch in 45-60 days after laying.
Long Term Chilling
The eggs are kept at 25°C for 40-50 hrs and then preserved at 5°C with 75-80 % humidity for 50-70
days. Eggs turn to brown colour and reach to spoon head stage. The eggs can release on any day
during this period. While releasing they are preserved at 15°C for 6-12 hrs and then at 25°C for 3-4
hrs. Later eggs are treated with HCl having a specific gravity of 1.100 at 47.8°C for 5 min. After
75
treatment eggs are washed in water to remove all traces of acid. Thus eggs treated can be made to
hatch in 60 – 80 days after they are laid.
10.2.5 Transportation of Eggs
It means carrying the silkworm eggs from the Grainage to the rearing centres. During the period of
transportation, friction, air, light conditions, saltiness and high temperature should be avoided.
The safe transportation of eggs is highly essential in order to protect the embryo and to ensure good
hatching results, which affect the yields and quality of cocoon crop.
The silkworm eggs should be transported during the cooler hours (morning/evening). Eggs should not
be transported during the hot hours or in rainy weather.
The egg cards are loosely placed in a wooden egg carrier/perforated paper cover. For loose eggs also
same method is adopted. It is preferable to carry the eggs in specially made egg boxes, with
perforations for aeration and provision for maintaining humidity.
10.3 INCUBATION
For healthy development and uniform hatching, bivoltine eggs are to be incubated under optimum
conditions of temperature, humidity and light. Hibernated eggs are released for incubation through
intermediate temperature (15oC). After release they have to be soaked in 2% formalin for 5 minutes,
washed in water and dried before incubation. The egg cards should be spread in single layers in the
trays.
ln case of loose eggs, they must be thinly spread out in brushing frames. Incubation room and the
equipments used should be thoroughly cleaned, washed and disinfected before the start of incubation.
Incubation has a profound influence on the voltinism, larval health, as also on the yield and quality of
cocoon crop. To obtain maximum hatching, optimum incubation temperature is 25oC which is raised
by 1oC to 26oC, on the day of hatching. During incubation, humidity is maintained at 75-80% as
excessive dryness results in dead eggs, poor hatching and weak larvae. Higher humidity causes weak
larvae, although it makes hatching uniform. As the embryo grows most vigorously during incubation,
good ventilation should be provided. Care has to be taken not to overcrowd the eggs in a narrow
incubation room. Light should be provided for 18 hours a day, till head-pigmentation stage is reached.
Eggs incubated as above hatch in 10-12 days. Two days before hatching, the colour of the egg
changes to a lighter shade, with a distinct dark spot. This is the 'head pigmentation stage'. One day
before hatching, the eggs turn bluish, which is referred to as 'body pigmentation' or 'blue egg stage'.
These two stages are very sensitive to low humidity.
To obtain uniform hatching, the eggs are kept in darkness at the head pigmentation stage. Darkness
arrests hatching of the developed eggs and facilitates lagging embryos to reach the hatching stage.
After two days, when a few larvae hatch out, the eggs are exposed to light. This ensures uniform
hatching.
Refrigeration of 'blue eggs' and 'new born larvae'
Hatching of eggs under incubation can be delayed if required by refrigerating them in the 'blue egg'
stage at 5'C for 2-3 days. Care must be taken to provide 75-80% humidity during cold storage. New
born worms can also be refrigerated at 5"C for 2-3 days but this is not desirable.
76
Check Your Progress
1. __________________acid is used in acid treatment of eggs.
2. ___________________percent of formalin is used in handling of eggs.
3. Acid treatment types are___________________________________________________
4. _________________________________________are the methods of cold storage of eggs.
5. Incubated eggs hatch in________________________
10.4 SUMMARY
The diapause, in insects, is a method of overcoming unfavourable period caused either by

physiologically
unfavourable conditions or non-availability of food. The phenomenon of diapause is environmentally,
physiologically and genetically controlled.
When the eggs left without any treatment the diapausing eggs do not hatch on incubation. To bring a
shift from diapause to non-diapause type, changes in the incubation schedule like shorter exposure to
light and low temperature during incubation, changes in the rearing conditions like continuous high
temperature rearing, low temperature during early stages and high temperature during the late stages,
cocoon preservation at high temperature, coupled with selection, are essential.
In acid treatment, strong and inorganic acids give good results than organic acids.
The mother moth while oviposition provides a thin film of gluey substance, secreted from accessory
glands on the under surface of the eggs which enables them to stick to the egg card. lf these egg cards
are dipped as such into the acid, it tends to dissolve the gluey substance.
The egg cards, prior to acid treatment, are soaked in 2% formalin solution for 15 minutes. Formalin
acts as a fixative agent which increases the adhering capacity of the eggs to the cards.
There are two most popular methods of treating the diapausing type of silkworm eggs, to block their
diapause and eventually make them to hatch like their non-diapausing counterparts.
In hot acid treatment eggs are dipped in HCL having specific gravity of 1.075 at 46°C for 5-6
minutes.
It is also called as room temperature acid treatment. There is no heating of acid; treatment is carried at
23-30°C temperature to eggs after 24 hours of oviposition.
There are two types i.e., Short term and Long term chilling.
During the period of transportation, friction, air, light conditions, saltiness and high temperature
should be avoided.
For healthy development and uniform hatching, bivoltine eggs are to be incubated under optimum
conditions of temperature, humidity and light. Hibernated eggs are released for incubation through
intermediate temperature (15oC).
10.6 CHECK YOUR PROGRESS - ANSWERS
1. HCl.
2. 2%
3. Hot acid and cold acid treatments.
77
4. short term chilling and long term chilling.
5. 10-12 days.
1. Detail acid treatment methods.

2. Write about cold storage of eggs.
3. Detail about incubation of eggs.
4. Write about formalin treatment of eggs to enhance adhering efficiency
1. Write about Short term chilling of eggs.

2. Write about Long term chilling of eggs.
3. Write about Transport of eggs.
4. Write about Hot acid treatment.
5. Write about Cold cid treatment.
6. Write about stopping of acid treatment
78
UNIT-11 BYPRODUCTS
Contents
11.0 Objectives
11.1 Introduction
11.2 Byproducts and their used
11.3 Summary
11.0. OBJECTIVES
 learn about by product.
 understands that about commercial value of byproducts.
 applies knowledge to start entrepreneurship.
11.1. INTRODUCTION
In the normal course of any activity related to agriculture leaves some products which can be
conveniently used in many ways, and these products are called byproducts. These depending upon the
present circumstances a person can start an industry and can supply the product to get good income.
Sericulture is not an exception for this activity. And all the four activities mulberry cultivation,
silkworm rearing, grainage and silk reeling are left with many by products and can be used in various
industries.
Mulberry and silkworm are exploited so far only for silk production in India. If sericulture industry
needs to be kept alive in all parts of India, wastes generated during all sericultural activities can very
well be converted in to valuable products of industrial value. Thus, in the whole cycle, a substantial
quantity of residual mulberry leaves, unwanted silkworm larvae, pupae, moths and silk are available
as by-products for utilization
11.2 BYPRODUCTS AND THEIR USES
1. Waste Cocoons:
In Sericulture the cocoons are commercially produced for two purposes one is mainly for reeling the
silk which is the primary and most important commercial product of the industry and the other i.e,
grainage where in egg production activity is carried out where in we come across these cut/pierced
cocoons which mainly used as raw material for spun silk industry. The industry leaves pierced, cut
cocoons, floss, double cocoons, black or melted-flimsy, stained and urinated cocoons. A quantity of
28 tons of Bivoltine and multivoltine cut/pierced cocoons are available annually for disposal (NSSO
tender notification 2014). Apart from this bulk of waste cocoons Registered Seed Producers (RSP)
and Government Model Grainages who contribute for more than 80 % of seed production in India is
also available for use. Presently it is being observed that only 2% of these cut/pierced cocoons are
being used for making handicrafts still bulk of it is readily available for exploration. It is estimated
that for production of one lakh disease free laying a quantity of 4.25 lakh bivoltine cocoons are
required and out of which 163 kg cut cocoon is obtained.
Silk proteins from silkworm waste cocoons, mainly composed of fibroin and sericin, are candidate
biomaterials with numerous potential biomedical applications or cosmetic uses. Wild silkworm
cocoon shells are composed of approximately 70% fibroin protein (fibroin heavy chain, FibH and
fibroin light chain, FibL), and 25% sericin protein. Fibroin is a raw silk component, specifically
79
produced by the posterior silk glands, whereas sericin is a glue like protein produced mainly in the
middle silk glands and promotes cocoon cohesion by surrounding and gluing fibroin threads together.
2. Utilisation of Silk Proteins:

Genetic manipulations that alter the properties of silk glands would likely help expand the biomedical
utilities of silk proteins. Especially the modified silkworm, with loss of fibroin production, would
produce a cocoon composed solely of sericin and expand the industrial utilities of sericin. intact,
soluble, fibroin free sericin at concentrations detectable by electrophoresis could be readily prepared
from the transgenic silkworm sericin cocoons using lithium bromide. Hydrogel formation of the
intact, soluble sericin could then be induced following incubation with ethanol. The gels can also be
fabricated into creams, sheets, sponges or other 3-D shapes that have mechanical strength by air
drying or lyophilisation. Intact, soluble sericin has moisture retention properties that are superior to
collagen and can gelate in solutions of up to 96% water.
Fig.11.1 Envisaged application of sericin from the transgenic silkworms.
3. Silkworm pupae
In breeding stations and commercial grainages many pupae are rejected during the process of cocoons
selection for egg production. Such pupae can be utilized in many ways. Silk worm pupae contain 50-
60% proteins, 25-35% fats, 8- 10% sugars, few vitamins like- B1, B2 & E, minerals like calcium,
phosphorous, copper, iron etc.
i. As compost: Dried pupae can be used as compost.
ii. As food: In some parts of India and China the silkworm pupae are regarded as delicious
food because it contains water, fat, protein, Glycogen etc. Defatted pupae are
commonly used as cattle feed. Silk worm pupae are either directly used in different
purposes, like poultry feed, fish feed etc or a special oil is obtained from them which in
turn is used in different products.
iii. Pupal oil: Dried pupae consist of 28.4% oil and 51.3% protein is used for the
manufacture of pupal oil. It is used for burning lamps as well as for preparation of
homemade soaps. Used in jute industry to soften the jute fibers. Silkworm pupal oil are
now a days used in medicines having anti- inflammatory and anti tumer like effects,
besides treating sinusitis, otitis, bronchitis, asthma, tuberculosis and urinary infections.
The silkworm pupae due to their high fat content (over 30%), are used as chrysalis oil
to obtain soaps, lotions and emulsions. Varnishes and dyes used in the textile and
tannery industry are also obtained. The residue formed during the chrysalis oil's
extraction is used as natural organic fertilizer and as food for poultry birds, pigs, fish
80
etc. In some countries like China, Japan, Thailand etc, the silkworm pupae are used as
delicious human food.
iv. Defatted protein: Defatted proteins are used for making artificial fibers. It also used as
animal feed, manufacture of amino acids by chemical or microbiological methods.
4. Silkworm moth
Male silkworm moth is used in the preparation of wine. The liquid extract from the moth can be used
to treat impotence, abnormal menstruation and menopausal symptoms. They are used to prepare
pharmaceutical product for curing trauma & to strengthen the masculine function. They can be used
compost material and animal feed. Silkworm moth oil can be used for making textile dyes & soaps.
Another type of special oil (75% fatty oils) is obtained from the silkworm moths. The oil can be used
to obtain textile dyes and superior soaps. The extraction residue can be used as fodder. The moths can
also yield Cellular Cytochrome C, uric acid and few hormones for pharmaceutical use.
5. Silk worm eggs
The silkworm eggs contain albumin, fats, sugars, glycoproteins, B1 and B2 vitamins etc. The eggs are
processed into protein extract which in turn is used in the pharmaceutical industry for the preparation
of medicines having hepatoproteinic, hypolipidic and hypoglycaemic action, serving as male sexual
stimulator and are also in the food industry. This extract is sold in Romania as the Humanofort-B
product. In some countries like Bulgaria some people believe that the silkworm eggs, if eaten by
alcohol drinkers, give up drinking completely because, they start feeling alcohol disgust. But fact has
not been proved scientifically.
6. Silk worm larvae
The silkworm larvae besides spinning cocoon are used in the pharmaceutical industry for the
preparation of medicines having anti diabetic action or in the food industry as supplementary
nutraceutical. Besides this, a special type of thread is obtained from its gut which is used in surgical
purposes. A research conducted by Korean scientist Ryu etal., in 1997, proved that the silkworms
have maximum blood glucose lowering effect and the substances were found to be the four blood
glucose-lowering substances as well as the major component, DNJ (1-deoxynojirimycin), which are
nitrogen compounds.
7. Utility of Silk
Silk is actually a natural protein fibre, composed of two main proteins viz; fibroin (73.5%) and sericin
(22.28 %), besides it contains some waxes (3.02%), minerals and ash (1.11%) and negligible amount
of ether and alcohol extract. Research has found that silk proteins have moisture retention and UV ray
blocking properties, because of these very properties they are utilized in different cosmetics like:
i. Silk lotion contains rich silk proteins and silk peptides, which are good moisturizers to keep
the skin soft and moist. The lotion is meant for all types of skins.
ii. Silk Cream is a natural beauty cream. It contains all silk's eighteen amino acids which are
absorbed instantly to nourish the skin and make it moist. The cream protects the skin from
windburn, sunburn and is especially suitable for sensitive skin.
iii. Silk Night Cream contains Aloe Vera, Jojoba Oil, Silk Amino Acids, and Silk peptides. It
regulates moisture balance during the night to nourish and rejuvenate the skin besides it also
assists the flight against wrinkles.
iv. Silk Hand Cream is non-greasy cream and contains rich silk essence which gives rough
hands a silky, soft and smooth feel.
v. Silk baby cream is made from high quality nutritional elements, natural silk peptide and raw
greasy lanolin which work together to protect the baby's delicate skin.
81
8. Handicrafts
waste cocoons can easily be converted into a fulltime enterprise taking into account of capital
initiatives, expertise available, market demand and setting up of sale outlets etc. However, any
person/family from a teenage to an aged, irrespective of gender can take up this activity as a source of
secondary income even during leisure period as well. Handicrafts from unused i.e cut / pierced
cocoons are emerging as one of the rural enterprise. These cocoons are natural, cost effective, eco-
friendly. Suits well for rural youth, women and differently abled (disabled) persons. This activity
helps to develop entrepreneurship besides skill development. Provides rural employment leading to
economic stability, escalates societal status, avoids migration to urban areas
Check Your Progress

1. __________________are used to make handicrafts.

2. Male silkworm moth is used in the preparation of _________________
3. Dried pupae can be used as _____________________.
11.3 SUMMARY
In the normal course of any activity related to agriculture leaves some products which can be
conveniently used in many ways, and these products are called byproducts.
The industry leaves pierced, cut cocoons, floss, double cocoons, black or melted, flimsy, stained and
urinated cocoons. A quantity of 28 tons of Bivoltine and multivoltine cut/pierced cocoons are
available annually for disposal (NSSO tender notification, 2014).
Genetic manipulations that alter the properties of silk glands would likely help expand the biomedical
utilities of silk proteins.
In breeding stations and commercial grainages many pupae are rejected during the process of cocoons
selection for egg production. Such pupae can be utilized in many ways. Silk worm pupae contain 50-
60% proteins, 25-35% fats, 8- 10% sugars, few vitamins like- B1, B2 & E, minerals like calcium,
phosphorous, copper, iron etc.
Male silkworm moth is used in the preparation of wine.
The silkworm eggs contain albumin, fats, sugars, glycoproteins, B1 and B2 vitamins etc.
The silkworm larvae besides spinning cocoon are used in the pharmaceutical industry for the
preparation of medicines having anti diabetic action or in the food industry as supplementary
nutraceutical.
Silk is actually a natural protein fibre, composed of two main proteins viz., fibroin (73.5%) and
sericin (22.28%), besides it contains some waxes (3.02%), minerals and ash (1.11%) and negligible
amount of ether and alcohol extract.
Waste cocoons can easily be converted into a fulltime enterprise taking into account of capital
initiatives, expertise available, market demand and setting up of sale outlets etc.
82
1. Cocoons
2. Wine.
3. Compost
1. Detail about byproducts and their uses.
1. Write about uses of waste Cocoons

2. Write about uses of uutilization of Silk Proteins
3. Write about uses of silkworm pupae
4. Write about uses of pupal oil
5. Write about uses of silkworm moth
6. Write about uses of silk worm eggs
7. Write about uses of silk worm larvae
8. Write about uses of utility of Silk
9. Write about uses of handicrafts
83
UNIT-12 ECONOMICS
Contents
12.0 Objectives
12.1 Introduction
12.2 Grainage records
12.3 Economics
12.4 Summary
12.0. OBJECTIVES

 know about economics of grainage.
 understand the importance of grainage records and its maintenance.
12.1. INTRODUCTION
The production of egg is influenced by various factors. The seed cocoons are produced in seed areas
under scientific guidance and supervision, cost of seed cocoons is generally fixed by the state
government from time to time and seed cocoons are offered about 20-25 % higher rates than hybrid
cocoons. The quality of seed cocoons has direct relation in producing more number of disease free
layings. Since nearly 60 percent of the cost of production of seed goes to the cost of the cocoons, the
egg producer must concentrate on procuring good quality seed cocoons.
It is essential that accurate data of seed production are maintained in the Grainage. This will help in
efficient management, control of quality.
Generally the following are considered for working out the economics of grainages.
1. Cost of seed cocoons.
2. Establishment charges.
3. Wages.
4. Depreciation cost on equipment.
5. Interest on revolving capital.
6. Cost of chemicals, rent, electricity and water charges.
7. Total DFL‘s produced.
Utilization of seed cocoons has direct relation to the cost of seed production. If the quality of seed
cocoons is good, then recovery of DFL‘s is high.
12.2 GRAINAGE RECORDS ECONOMICS

The following are the types of records to be maintained in Grainage.
1. Seed Cocoon Register
The register should contain information about the parental race, yield, silk and its quality details, bad
cocoon percentage, seed rearer details, details of seed cocoon seller, cocoon purchase rate, weight of
cocoons purchased and their cocoon quality details.
2. Moth Emergence Register
The register should contain details of moth emergence such as number of moths obtained each day,
number of moths kept for pairing, percentage of emergence, number of moths left for egg laying
84
under each combination are recorded. This will help in forecasting the production and planning for
hibernation. These data are to be maintained separately for different batches.
3. Moth Examination Register
Maintenance of this record is important, as it provides information on the number of moths examined,
type of examination conducted, presence of pebrine, if present, percentage of infection, whether the
batch is to be rejected.
4. Egg Production Register

Information pertaining to the quality and quantity of eggs laid, unfertilized eggs rejected, quality fit
for distribution, type of egg(hibernated and nonhibernated) are maintained.
5. Hibernation and Refrigeration Register

In this register contains details of refrigeration, hibernation, date of release, temperature in cold
storage and number of moths kept in refrigerator.
12.3 ECONOMICS
In general industrial grainages are commercial in nature. In a grainage various inputs are purchased
from the market and the processed material ie., dfls are sold to rearer. The defective cocoons also get
returns. To know the cost of production of egg laying, it is necessary to work out the cost of
production of a unit quantity of layings.
The cost of establishment and operating an commercial grainage includes capital investment on land,
building, cold storage and non-recurring expenditure on equipment, furniture, fixtures and vehicles,
recurring expenditure on establishment charges, cost of chemicals, stationery, labour wages,
electricity, diesel, water charges etc. The cost of production also includes interest on capital
investment, depreciation on building and equipment and other recurring expenditure. The depreciation
is calculated based on the longevity of the building, cold storage, equipment, furniture and vehicles.
Cost of equipment Longevity Depreciation value

x
x y
y
The receipts of the grainage are sale of defective cocoons and layings.
In any case one must be sure of producing disease free layings at a lower cost utilizing all the latest
easy techniques which improve the economics of the grainage. One should not be determined to
reduce the cost of production, because quality of eggs determines the reputation, stability of a
grainage.
Utilization of seed cocoons has direct relation to the cost of seed production. If the quality of seed
cocoons is good, then recovery of Dfl‘s is high.
A. Capital Investment
S.No. Item Cost in Rs.
1. Land ..............
2. Building ..............
3. Cold storage ..............
Total Rs.
85
B. Non recurring Expenditure
1. Grainage equipment ..............

2. Furniture and fixtures ..............
3. Vehicle ..............
Total Rs.
C. Interest on items of A and B

D. Calculate depreciation on A and B
E. Recurring Expenditure
1. Establishment charges ................

2. Labour wages ................
3. Cost of seed cocoons ................
4. Cost of chemicals ................
5. Eggs sheet cost ................
6. Stationary and printing ................
7. Electricity, diesel, water ...............
8. Maintenance of vehicles ...............
9. Miscellaneous ...............
Total Rs.
F. Receipts
1. Sale of defective cocoons ...............
Total Rs.
Cost of production of one laying is calculated from the above listed tables.
1. Capital investment Table - A

2. Non recurring expenditure ― - B
3. Interest on initial investment ― - C
4. Depreciation cost ― - D
5. Recurring expenditure ― - E
6. Receipts ― - F
( C  D  E) - F
Cost of production of one laying   One DFL cost
No. of DFLs produced
Check Your Progress
1. Details related to eggs are entered in___________________________register.
86
2. Details related to seed cocoons are entered in______________________________register.
12.5 SUMMARY
The seed cocoons are produced in seed areas under scientific guidance and supervision, cost of seed
cocoons is generally fixed by the state government from time to time and seed cocoons are offered
about 20-25 % higher rates than hybrid cocoons.
Maintenance of various records pertaining to grainage activity are essential for planning and timely
activity.
Grainage aim is to produce dfls at an offer able cost. 60 % of the cost of production of seed goes to
the cost of the cocoons. Items like investment, recurring, non-recurring are considered to calculate
cost of production of eggs. One should not aim to reduce the cost of production, but to concentrate on
the quality.
1. Egg production
2. Seed cocoon
1. Write about grainage records and add a note on economics.
1. Write about grainage records.
87
Dr. B.R. Ambedkar Open University
Faculty of Science
3rd year (3 year degree course) –VI semester
MODEL QUESTION PAPER
ZOOLOGY DSC – VI
SILKWORM BREEDING AND GRAINAGE
[Time: 3 hours] [Max Marks: 80]
Section-A
[Short Answer Questions]
[Marks: 4x5=20]
Note: a) Answer any four of the following in about 10 lines each.
b) Each question carries 5 marks
1. [Block-I] Write about cross breeding.

2. [Block-I] Write about transgenic silkworm.
3. [Block-II] Detail voltinism.
4. [Block-II] Write about seed cocoon markets.
5. [Block-III] Write about pupal sex separation.
6. [Block-III] Write characters of good eggs.
7. [Block-IV] Write about grainage records.
8. [Block-IV] Write about uses of pupal oil.
Section-B
[Essay type]
[Marks: 4x10=40]
Note: a) Answer the following questions in about 30 lines each.

b) Each question carries 10 marks
9. [Block-I] Detail about inbreeding and outbreeding .

(Or)
10. [Block-I] Write about heterosis.Write about transgenic silkworm.
11. [Block-II] Detail about parental races on the basis of places of origin.
(Or)
12. [Block-II] Detail about P2 breeding station.
13. [Block-III] Detail about grainage equipment.

(Or)
14. [Block-III] Write about loose egg preparation.
15. [Block-IV] Detail about byproducts and their uses.

(Or)
16. [Block-IV] Write about cold storage of eggs.
88
Section-C
[Objective Type questions]
[Marks: 20]
Total Number of questions 20 - {15 from (Theory) and 5 from (Practicals)}
I Multiple choice questions (10 marks)
1. Which of the following lays eggs.

A) Larvaa B) Moth C) Female moth D) Pupa
2. Female pupa is
A) big in size B) small in size C) no change D) very long
3. The following is for oviposition
A) only paper B) Cup C) Cellule D) a and b
4. Acid treatments are of
A) 2 types B) 3 types C) 4 types D) one type
5. Silkworm eggs are of
6. Cold storage of eggs are of
7. The following is the parental race
A) MV B) BV C) Chinese D) a and b
8. The following chemical is used in egg preparation
A) H2SO4 B) HCl C) Formalin D) Bleaching powder
9. The following is for preventing ant crawling.
A) Tray B) Mat C) Ant well D) Board
10. Disinfection process is carried as
A) Fumigation B) Spraying C) Water spray D) a and b
II Match the following (5 marks)
1. Female moth ( ) a. Oil

2. Formalin ( ) b. Egg laying
3. HCl ( ) c. Protein synthesis
4. Moth examination ( ) d. Disinfection
5. Pupa ( ) e. Acid treatment
( ) f. Motor and pestle
III Fill in the blanks (5 marks)
1. Incubated eggs hatch in________________________.

2. Dried pupae can be used as _____________________.
3. Details related to eggs are entered in___________________________register.
4. How many moulters are there in Bombyx?______________________________________
5. Bad cocoons are not suitable for ______________________________
89

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BS619ZOO-DSE(B)-E

Dr. B. R. AMBEDKAR OPEN UNIVERSITY

Course Development Team Course Development Team (CBCS)

Associate Editor Associate Editor

Dr. B. R. Ambedkar Open University, Hyderabad.

The text forms part of Dr. B. R. Ambedkar Open University Programme.

Printed on behalf of Dr. B. R. Ambedkar Open University, Hyderabad by the Registrar.

Printed at: .................................................................................................................................

Block/Unit Title Page

BLOCK- I SILKWORM BREEDING

BLOCK -II GRAINAGE-I

BLOCK- III GRAINAGE-II

Unit –7 : Grainage 48-57

BLOCK -IV GRAINAGE-III

Unit –10 : Acid Treatment 72-78

MODEL QUESTION PAPER 88

1.2 SILKWORM BREEDING

Early history of Indian silkworm breeds

1.2.1 Inbreeding and Out breeding Concepts

It includes out-crossing, cross-breeding, and interspecific hybridization.

1.2.2 Objectives of silkworm breeding-techniques

The main objectives of silkworm breeding are

Fig.1.1 Different Breeds

1.2.3 Different types of breeding methods

Sex-limited cocoon color:

Step 3. Backcrosses for other 3 generations, but with batch rearing.

1.2.4 Line breeding, cross breeding and mutation breeding

iii. Mutation breeding:

Check Your Progress

1. Silkworms were first domesticated in _____________over 5,000 years ago.

1.5 MODEL EXAMINATION QUESTIONS

I. Answer the following in about 30 lines.

1. Detail about inbreeding and out breeding.

II. Answer the following in 10 lines.

1. Write about inbreeding.

2.2 SELECTION METHODS

 Pupation rate in %: It is calculated by the formula:

 Shell percentage: It is calculated by the formula:

 Filament size in denier: It is calculated by the formula:

 Reelability in %: It is calculated by the formula:

 Raw silk percentage: The formula used is:

2.2.2 Fixation of characters

a. Characterization and evaluation of silkworm genetic resources:

b. Molecular characterisation of silkworm germplasm:

2.2.3 Formation of new breeds

 Phenotypic evaluation of the main qualitative and quantitative breeding characters.

Segregation from foreign F1 hybrids:

Generally the method is as follows:

Separation from foreign F1 hybrids:

The method is the following:

Using populations of Japanese races:

 Initial population: J 124.

Step 3. Backcrosses for other 3 generations, but with batch rearing.

Fig.2.1 Process for generating transgenic or genome edited silkworms

2.2.4 Authorization of new breeds

Races developed by CSR&TI, Mysore

Polyvoltine breeds and hybrids of the past

Improved polyvoltine races

Productive polyvoltine x bivoltine hybrids

Productive Cross Breed (Polyvoltine hybrid) for irrigated zone

Polyvoltine hybrid for rain-fed zone

New productive bivoltine breeds and hybrids

Robust bivoltine breeds and hybrid