2012

A Practical Manual on,

Sericulture
Submitted To:
Mr. Minraj Pokhrel Lecturer and Course In Charge Department of Entomology IAAS, Rampur, Chitwan

Submitted By:
Babu Ram Panthi B.Sc.Ag. 8th sem (Entomology as elective) Class Roll No: 58 Exam Roll No: 69 IAAS, Rampur, Chitwan

Practical Index of Sericulture

SN 1 2 3 4 TITLE OF PRACTICAL IDENTIFICATION OF REARING TOOLS AND EQUIPMENTS USED IN SILKWORM REARING PRACTICES OF MULBERRY RAISING, PROPAGATION, TRAINING AND PRUNINIG HARVESTING OF MULBERRY LEAVES, SHOOTS AND YIELD ESTIMATION STUDY OF DIFFERENT INSECT PESTS OF MULBERRY PLANT STUDY OF DIFFERENT DISEASES OF MULBERRY PLANTS STORAGE, INCUBATION AND TRANSPORTATION OF SILKWORM SEEDS PROCESS AND PROCEDURE OF SILKWORM SEED PRODUCTION STUDY OF LIFE CYCLE OF MULBERRY SILKWORM OBSERVATION OF MOULTING AND APPLICATION OF RKO STUDY OF INSECT PEST AND DISEASES OF MULBERRY SILKWORM REARING TECHNIQUE OF YOUNG AND GROWN UP AGE OF LARVAE CLEANING AND DISINFECTION OF REARING ROOM PREPARATION OF SILKWORM FEED BED AND BED CLEANING LARVAL MOUNTING AND CARE DURING COCOON SPINNING COCOON HARVESTING, GRADING AND PRICING POST HARVEST PROCESSING OF COCOON Page No. 1 3 5 6

5 6 7 8 9 10 11 12 13 14 15 16

8 10 11 13 14 15 19 21 22 24 25 27

17 18 19 20 21

SILKWORM RECORD KEEPING STUDY OF MORPHOLOGY OF DIFFERENT STAGES OBSERVATION OF SEX DIFFERENTIATION OF LARVAE, PUPA AND ADULTS OF SILKWORMS ANATOMY OF SILKWORM (BOMBYX MORI) LARVAE STUDY OF DIFFERENT GLANDS PRESENT IN BOMBYX MORI

28 30 32 33 34

A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical No: 1 IDENTIFICATION OF REARING TOOLS AND EQUIPMENTS USED IN SILKWORM REARING. Objectives To be familiar with tools and equipments used in silkworm rearing. To be acquainted with the use and application of the tools and equipments.

Introduction Silkworm are very sensitive to weather and other conditions, therefore special care is to be taken during handling of worms. Different equipments are needed for proper rearing of silkworms. The following are the major: 1. 2. 3. 4. 5. 6. 7. 8. Rearing stands Ant wells Rearing trays Paraffin paper Foam rubber strips Chopsticks Feathers Leaf chambers 9. 10. 11. 12. 13. 14. 14. 15. Chopping boards Cleaning nets Mountages Hygrometer Thermometer Sprayer Wooden stands Leaf baskets

Methodology Observation of different tools and equipments used at Bhandara Sericulture Farm and Khopasi Sericulture Development Farm. Note taking, drawing of the tools and equipments. Referencing for the details of the tools and equipments.

Description of the equipments I. Rearing stands These are made of wood or bamboo, portable and are 2.5 m high, 1.5 m long and 1 m wide with 10 shelves with a space of 20 cm between each shelf. The rearing trays are arranged in the shelves. In whole shoot feeding method,Rearing stand width should be 5 -5.5 ft.With One ft above the ground level.and with 2 ft.Distance between rearing stand & wall. Ant wells These are provided around the legs of stands to stop ants crawling on to the trays and attacking the worms. These are made of concrete stone base 20 cm2 and 7.5 cm high with groove of 2.5 cm depth to hold water. The legs of the stands rest in the center of the block. Rearing trays

II.

III.

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

These are used to rear silkworms and are made of bamboo. They are round (1.2 to 1.4 m dia) or rectangular (0.9 to 1.2 m x 0.7 to 0.9 m) with a depth of 7.5 cm. IV. Paraffin paper It is a thick craft paper coated with paraffin. It is used for rearing early age worms, on the bottom and as a cover of the rearing bed to prevent withering of chopped leaves and to maintain humidity in the rearing bed. Foam rubber strips Long foam rubber strips of 2.5 cm wide and 2.5 cm thick dipped in water are used to keep all around silkworm rearing bed during first two instars to maintain humidity. Chopsticks These are made of bamboo, 17.5 - 20.0 cm long and thin and tapering to one end. A pair of chop sticks are used to pick early age larvae and also for preparing the bed. Feathers Bird feathers, preferably white ones are used for brushing the delicate newly hatched worms onto the rearing bed. Leaf chambers These are maintained in rearing house for storing the harvested mulberry leaf ready for feeding the worms at set intervals.

V.

VI.

VII.

VIII.

Summary and conclusions: Thus we were able to identify different tools and equipments used in insect rearing at Bhandara and Khopasi Farm.

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical no: 2 PRACTICES OF MULBERRY RAISING, PROPAGATION, TRAINING AND PRUNING. Objectives: To know about the practices of mulberry raising, propagation, training and pruning. Introduction: Propagation of mulberry is exhibited by sexual and asexual means. In sexual means of propagation seedlings are raised by sowing viable seeds. Asexual or vegetative means of propagation includes grafting, layering, cutting and micro propagation through tissue culture. Sexual propagation: this is the most conventional method of propagation. Sowing of viable seeds in soil for germination under adequate condition and raising of new mulberry plants for direct field utilization is done. In this propagation system, time for propagation is short, the co-efficient of propagation is large, the root system grows vigorously and the predominance of useful mutations is numerous in the seedling populations. But in some species of mulberry cant be propagated by means of sexual propagation. Such can be propagated by means of asexual propagation. Asexual propagation: This method implies the utilization of a certain part of the vegetative organs in a favorable condition for culturing another living mulberry plants. Since mulberry is highly heterozygous plant, by means of asexual propagation the inherited characteristics of a parent stock are retained. In mulberries the asexual/vegetative means of propagation can be accomplished through grafting, layering and micro propagation through tissue culture. 1. Grafting: in a grafting a part of mulberry plant is called scion is artificially united to the root of bark of another plant called stock which keeps on growing. Grafting facilitates the propagation of a variety having desirable qualities and which cannot be propagated by other means, however the success depends on the quality of both stock and scion. It is very easy in mulberry. Normally one year old seedlings are used as stock. Depending upon the part use as the stock the grafting can be classified in to following, a. Shoot grafting i. Wedge grafting ii. Crown grafting iii. Whip grafting b. Root grafting i. Bag grafting ii. Inverted bag grafting c. Bud grafting i. Patch budding ii. T- budding iii. Flute budding 2. Layering: it means the development of roots on a steam while attach to the parent plant and the root is then detached to become a new plant growing on its own roots. There are three types of layering : a. Simple layering b. Air layering c. Trench layering

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

3. Cutting: It is simple and common form of asexual propagation and is meant by inserting a fragment of plants vegetative organ in the soil when roots and shoots grow out to form a new plant. Both hard and soft wood cuttings from a stem could be utilized as planting material. 4. Micro propagation: the higher multiplication rates, round the year propagation, control over chemical and physical environment factors and possibilities of micro propagation. The use of callus, cell , organs, meristem and protoplast cultures in mulberry can well fitted in this direction. Summary and conclusion: Thus we are able to done propagation on mulberry plant and also the training & pruning operation in mulberry plant.

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical No: 3 HARVESTING OF MULBERRY LEAVES, SHOOTS AND YIELD ESTIMATION Objective To know the methods of harvesting of mulberry leaves and shoots To calculate the tentative yield of the mulberry crop Methodology Observation of orchard at Bhandara Note taking Referencing for the detail Introduction The method of harvesting leaves varies in different areas according to the rearing methods used and also to suit the intensity of rearing and availability of labor at the given time. Three methods of harvesting are prevalent for mulberry: a) Leaf picking: Leaves are picked individually from main stem with petioles. At the same time, terminal buds are nipped off so that lateral shoots develop rapidly. It requires more labor and leaves Wither quickly. b) Branch cutting: The entire branch is harvested and used to feed worms after third moult directly. It requires less labour and leaves retain succulence for longer period. c) Whole shoot harvest: The branches are cut to ground level by bottom pruning. The entire shoot is fed to the larva. Shoots are harvested at 10-12 weeks interval and 5-6 harvests are made per year. Leaf harvested during afternoon contain less water and more of carbohydrates due to active photosynthesis and transpiration taking place in day time and such leaves wither very quickly. Hence, leaf harvesting in early morning hours is recommended. Yield estimation Under normal condition a hectare of irrigated mulberry yields about 30,000 kg of leaf a year in Chitwan condition. For raising 100 DFL S of multivoltine bivoltine hybrids 700 kg of leaves will be required per rearing. Summary and conclusion Thus we were able to estimate the tentative yield of mulberry and knew about different harvesting methods of mulberry leaves and shoots.

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical No: 4 STUDY OF DIFFERENT INSECT PESTS OF MULBERRY PLANT Objectives To be familiar with different insects pests of silkworm To know about the lifecycle, damage symptoms and management of mulberry insect pests. Introduction: Many species of insects pest, diseases and deficiency symptoms are observed in mulberry. However a few of them are economically important. Due to the pests the economy of sericulture is severely affected since both the quantity and quality of the leaf produced are affected. 1. Hairy caterpillar : Spilosoma (Diacrisia) obliqua Arctidae; Lepidoptera It is voracious feeder causing extensive damage to mulberry. Life history: Adults are light brown in color with brick red abdomen, having dark rows of spots laterally and dorsally. Female lays about 1000-2000 eggs in batches on ventral surface of leaf. The eggs hatch in 5-7 days. The young larvae are gregarious in nature. The grown up larvae possess prominent setae (hairs) dorsally and laterally. The anterior and posterior regions are black in color while rest of the body is reddish brown. The caterpillar moults 6 times and the larval duration lasts for 28-30 days. The pupation occurs in soil in a loose cocoon like structure. The pupa is brown in color and its period lasts for 12-14 days. The life cycle is completed in about 48 days. Symptoms: The young caterpillars feed on the chlorophyll layer of the leaf exposing the veins which impart dried/dead appearance to the leaves (Skeletonization). The grown up larva feed on the entire leaf rendering the branches without leaves. Management: Monitor the garden regularly for collection and destruction of egg masses and gregarious young caterpillars. Ploughing the caterpillar infested garden to a depth of about 15cm to expose pupae to scorching sun on to the sight of its natural enemies like birds.

Flood irrigation will help to kill the pupae. Digging trenches (30 cm width and 30 cm depth) around the garden and dusting with Methyl Parathion 1.5 % dust helps to check migration of caterpillars from other gardens. Spray DDVP @ 0.2% or dimethoate @ 0.2% prepared in 0.5% soap solution. Safe period is 12 and 13 days respectively. 2. Jassid: Emposa sp.

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Both adult and nymph suck sap from buds and leaves. The insect over winters as eggs inside the bark. Produce 3-4 generations a year, very active Oct-May. The little greenish hoppers suck the sap causing the characteristics symptoms hopper burn, the first symptom is the appearance of a triangular brown spot at the top of leaf. Entire margin may roll up and form cup shape as vector of virus. Secrete honey-dew and shooty mould develops. Life Cycle: Female lays pale yellow elongated eggs in epidermis. Egg hatches after 6-10 days. Nymph is very small pale green colored, feed from underside of leaf. They run sideways over the edge of leaf. Adult is Light green 2.5-4.7 mm. Control: Light trap attracts the adult hoppers; removal of diseased plants (virus) when rigor sprayed 10 days, spraying during early spring to kill newly coming adults, sprinkler irrigation is effective. 3. Thrips : Bathrips melenicornis, Scirtothrips dorsalis Thripidae: Thysanoptera Biology : The females are dark brown, measuring 0.9 mm in length. They reproduce both sexually and parthenogenitically. Female lays eggs on ventral side of tender leaves which hatch in 6-8 days. The nymphs are pale yellow and lasts about 15-18 days. The pest completes 7-8 generations a year. Nature and extent of damage : Both adults and nymphs lacerate the leaf tissue and suck the oozing cell sap from young buds and leaves. The infested parts get hardened; leaves become brittle, malformed with reduced leaf area. In addition, sap extraction by the thrips results in necrosis and drying up of leaves. Due to thrips attack, the epidermal cells get punctured, leaves and buds become rudimentary resulting in premature fall. During laceration, the thrips secrete saliva which coagulate with sap resulting in the formation of white streaks in early stage Management: Removal of alternate hosts/weeds helps in regulating thrips. Periodical ploughing and digging helps in exposing the thrips pupae to hot sun and natural enemies. Spraying with DDVP @ 0.2% / Dimethoate @ 0.1% with a safe period of 3 and 15 days, respectively.

Summary and conclusions: Thus we were able to know about the different insect pest of mulberry plant.

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Practical No: 5 STUDY OF DIFFERENT DISEASES OF MULBERRY PLANTS Objectives: To be familiar with the different diseases in mulberry plants. To know the different control measures for different diseases.

Date: 2068/11/09

Introduction: Many species of insect, pest and diseases are observed in mulberry. However few of them are economically important. Due to diseases the economy of sericulture is severely affected since both the quantity and quality of leaf produced are affected. There are number of diseases which affect the normal growth of mulberry, such as fungal, bacterial, viral, microplasma and nematode disease. a. Powdery mildew Phyllactinia corylea(pers) karst - Initially white powdery powdery patched appear on the ventral surface - Later entire leaf is covered with white powder and - Then leaf turn yellowish brown brown to black in color - Affected leaf dry , leathery and fall off - Commonlly occurring sept-dec Dusting sulphur powder, diethane/sulfur suspension, lime sulphur spray, karathene spray (0.1%) b. Leaf spot Cercospora moricola( cooke) - Appearance of small whitish spots - The affected leaves fall of - Common during rainy season Spray difolatan(0.2%) , Bavistin(0.05%) 2-3 at 10-15 days interval. Pick and burn diseased leaves, remove weed as a alternative host of this diseases. c. Mulberry dwarf caused by virus and MLOs New leaves are smaller with yellowish color, no glossy appearance Leaf crinkles and hardens Margin and tip curl up turning round shapes Affected branches are shorter then normal When started within4-5 yrs entire plant become diseased One year old plant if diseased die within 2-3 yrs Leaf hopper transmit the pathogen when fed 3 hrs or more, but require > then 7 hrs sucking from infected plants, remove affected plants, plant quarantine to import or export plant material. Planting in isolation, chemical control of leaf hoppers(0.05) monocrotophos first early spray after winter , hydrochloric terraycin ingestion in tall and median tree.

d. Leaf blight Pseudomonas mori - Appearance of small water-soaked spot on the lower surface of leaves - Vein and veinlets also are affected some time

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

e. -

Lesions are dark, irregular, shrunken and excrete yellowish bacterial pus Infected leaves curl and roll up More serious in rainy season Phloem rot a turning blakish brown Buds and leaves in affected branch wither rapidly and fall off. Root knot nematode Meloidogyne sps Beginning shows like deficiency of water and nutrient Leaves turn yellowish and plants are stunted In severe cases affected plants are wilt and die Roots show knotty galls, yellowish white initially, later turn brown finally black and not rotten In sandy soils infected plants die within 4-5 years. Planting disease free saplings, soil disinfection by formaldehyde, nemagan(80%) @ 45 kg/ha( 1 lit in 50-100 lit water) use in ditch cover soil, plant 8-10 after , follow rotation with rice, corn, sugarcane.

Conclusion: Thus we were able to identify the diseases in mulberry and their control measures.

Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical No: 6 STORAGE, INCUBATION AND TRANSPORTATION OF SILKWORM SEEDS Objectives To know about the procedures of storage, incubation and transportation of silkworm seeds. To know the precautions to follow while doing this. Introduction Silkworm egg is the basic seed for sericulture industry. The ultimate success of of silkworm crop depends much on the efficiency of silkworm seeds. Grainage is the place where silkworm seeds (eggs) are prepared. The basic function of grainage is to produce quality, disease free silkworms seeds. Storage of seed cocoons 1. Building should be well ventilated. 2. Avoid direct sunlight and heavy wind on cocoons. 3. Remove floss from the cocoon. 4. Spread cocoons in thin layers in tray. 5. Cut the cocoons for easy emergence. 6. Provide separate room for different races. 7. Maintain 25C & 80% RH. 8. Preserve cocoons in normal light during day and dark at night. 9. Pupae are preserved in corrugated papers or paddy husk or saw dust. 10. Perforated absorbent paper or newspaper is spread on the cocoons for easy picking of moths during emergence. 11. Sex separation is done in the pupal stage for hybrid egg preparation. 12. Incubation of silkworm seed: Heating of the rearing room is done through a heater by indicating the actual reading of the thermometer. The egg box is opened after washing the hand and sterilization and spreaded over the tray for incubation. Transport of cocoons 1. Dont expose seed cocoons to hot sun or rain. 2. Avoid long distance transport. 3. Transport on the cooler part of the day or at night. 4. Keep cocoons in thin layers. 5. Avoid severe shock. 6. Pupae inside the cocoon shouldnt get damaged. 7. Dont keep cocoons tightly. Summary and conclusion Thus we became familiar with the different storage, incubation and transportation techniques of silkworm seeds along with the precautions to follow while doing this.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical No: 7 PROCESS AND PROCEDURE OF SILKWORM SEED PRODUCTION Objectives To be familiar with process and procedure of silkworm seed production Introduction Production of superior and diseases free silkworm eggs is one of the major requirements of sericulture industry. High technical knowledge and good facilities are required to produce the quality silkworm seeds. For an individual farmer it will be hard to produce the required quality eggs. Hence such eggs can be produced only by government agencies or trained persons. For such production they must have a model grainage having seed cocoon preservation, pairing, depairing, egg laying, egg processing, egg examination, acid treatment and incubation room. Major grainage equipments are foot cleaning trey, ant well, grainage stand, grainage tray, working stand , cellule, bamboo trey , moth crushing set , moth testing table , moth examination stool, thermometer and hydrometer. Selection of seed cocoon is done by mid gut examination and forced eclosion test. Cocoon should be uniform in size, shape and hardness and should have more silk percentage. Multivoltine females are crossed with bivoltine males to get the hybrid eggs required for the farmers. Bivoltine eggs will always go for hibernation which will not hatch immediately if these eggs are to be hatch , it can be done artificially by subjecting these egg to low temperature and HCL. Grainage: Grainage is the place where silkworm seeds are prepared. The basic function of a grainage is to produce quality, disease free silkworm seeds. Selection of good cocoons: Cocoons of uniform size, shape and hardness with less percentage of double cocoons and more silk percentage are selected. Cocoon should have more sound pupae and free from diseases. Sex separation: Sex separation is to avoid selfing. This is essential for hybrid seed production. In pure races it is done for specific mating to get better economic characters. Usually sex separation is done at the pupal stage. Female are identified by their large size with broader abdomen and an X mark on the ventral side in 8th abdominal segment. Males are smaller in size with tapering abdomen and a dot in the 8th abdominal segment. Pairing: The copulation between male and female moth is called pairing. Sufficient number of females is collected in a tray and equal no. of males sprinkled over the female. In a few minutes males will pair with female. The mated moth are removed and kept in separate trays covered with cellules. Depairing: The removal of male from the female moth is known as depairing. It is done after 4 hours of mating carefully. it is done gently holding the female with two fingers by the sides and with one finger male is removed. Oviposition:

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Egg laying is otherwise known as Oviposition. Female moth starts laying eggs when they are detached from male moths. After depairing females are kept on the old newspaper and paper is tapped gently. This will induce moth to pass urine. After this moth are arranged on egg sheets covered with cellules kept undisturbed. Processing of silkworm eggs: The disease free multivoltine silkworm eggs are washed in 2% formalin or 2% bleaching powder solution for 5 minute and aging washed in water and dried in shade. This enables surface disinfection of the eggs. Preparation of loose eggs: In this method the female moths are allowed to lay eggs on egg sheets smeared with starch paste and dried. The female moth after pairing kept for egg laying. The sheets are kept in water for 1520 minute. The eggs will be loosened and if the egg sheet is brushed with hand all the egg will come off from the egg sheet. The loose eggs are collected and the unfertilized and dead eggs are removed by subjecting these eggs to 1.06 and 1.1 specific gravity Nacl solution. Summary and conclusions: Thus we were able to know about insects pest and diseases of silkworm.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Practical No: 8 STUDY OF LIFE CYCLE OF MULBERRY SILKWORM Objectives: To know the different stages of mulberry silkworm.

Date: 2068/11/09

Introduction: Mulberry silkworm passes through complete metamorphosis and thus has distinct four developmental stage- eggs , larva , pupae and adult. Silkworm races are classified according to the difference in their physiological and ecological characteristics. Each race possesses it own characters , which provide information required for care in their rearing and other handling. It is necessary to understand social characters and their biology in order to successfully rear the worms. Eggs : silkworm egg are tiny , round or oval, with a micropyle at the anterior pole. The eggs weigh about 0.55 to 0.66 mg so that there are about 2000 eggs/gram. The eggs are first laid are heavier then those laid later by the same moth. The color of eggs are different in different races. The white cocoon races lay pale eggs compared to yellow cocoon races. Diapusig race egg color changes to deep brown white enterning in to diapause. But in non-diapausing races,color does not change till development is complete and one day before hatching eggs enters into blue stage(E5). The egg measures 1 to 1.3mm in length and 0.9-1.2mm width. The eggs are attached to substratum through glue secretion from the accessary glands and is removed by washing with water in preparation of loose eggs. Larvae: the larvae is of eruciform or polypod type with abdominalprolegs. Its body is covered with the chitinous skin. The bristles or sitae are situated on four pairs of tubercles in each segment. Body is divided into head, thorax and abdomen with three pairs of prolegs, four pairs of abdominal legs and pair of caudal legs with caudal horn on the eight segment. The newly hatched larvae is about 3mm long, black in color and its body is covered with bristles. It is called ant. As the larvae grows, its color become lighter and its cuticle become smoother. The last instar is about 10cm long. Pupae: pupae are soft and white after the moult that occurs within the cocoon spun by the final instar larvae. It takes 48-72 hours to spin its cocoon and transforms it in to pupae in 2-3 days and after another ten days emerges as moth. The pupae is a non-motile and non-feeding stage. This stage is devoted for the development of cuticle, compound eyes and wing pad and it is useful for gross examination of the age of the pupae. Adults: from the cocoon the moth emerges and emergence takes place in morning. Copulation also takes place just after the emergence. The moth have prominent bipectinate antennae and compound eyes on head . the coiled proboscis is non functional. There are three pair of thoracic legs and two pair of wings on the thorax. Moth cant fly but male are more active in movement then females. Conclusion: Thus we are able to identify the different stages of mulberry silkworm.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical No: 9 OBSERVATION OF MOULTING AND APPLICATION OF RKO Objectives To know the morphological changes in larva during moulting To know the handling procedures during rearing To know the use and application of RKO Methodology Note taking Referencing for the detail Introduction Moulting The insects do not grow by regular, gradual and imperceptive degrees as the sclerotized exoskeleton will not permit like those of mammalian skin. The growth of insects is limited by the cuticle. Cuticular growth and change in form only occur when an insect moults. So for any marked in size, the cuticle must be shed and replaced. Casting or shedding off cuticle is commonly known as moulting. The moulting involves the sequence of two events. The beginning is with the separation of old cuticle from the underlying epidermal cells (apolysis) and ending with the casting of the remnants of old cuticle after the production of new one (ecdysis). The discarded skin is known as exuviae. The interval between apolysis and ecdysis is said to be pharate. Care during moulting Silkworms moult four times during their larval period. Handling of worms during this period is of particular importance. During moulting worms stop feeding, hold their head & thorax vertically and wriggle out of their old skin. Temperature and RH should be about 23C and 65% Spreading of rearing bed should be done at the initial stage to make good moulting conditions. A very thin layer of lime powder or dried rice bran is dusted over the bed to keep the bed dry. Late or early moulted larva are segregated and reared as separate batch. Application of resham kit okhati(RKO) Due to centuries of domestication the silkworm has no natural resistance and shows neither morphological nor behavioral adaptation to escape parasites and pests. Development of disease resistance species has been partially successful and no race is completely immune to diseases. Silkworms are infected by protozoa, bacteria, viruses and fungi. The occurrence of fatal fungus disease is prevalent in dampy condition. To protect the larvae from fungal infection an anti muscardine bed disinfectant powder known as Resham Kit Okhati(RKO) is dusted on the worms after each moult 30 minutes before they resume feeding. Summary and conclusion Thus we knew about the moulting, changes during moulting, handling during moulting and the application of of Resham Kit Okhati.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical No: 10 STUDY OF INSECT PEST AND DISEASES OF MULBERRY SILKWORM Objectives To be familiar with insects pest and diseases of silkworm. To know about different insect pest and diseases and their control measures. Introduction The pest problem of mulberry silkworm during rearing and cocoon storage is very less than the occurrence of various diseases of silkworms. Among the insects pests, various species of tachinid Uzi fly, dermestid beetle, earwigs and ants are the major. Mites, lizards, birds, rats and squirrels are the other pests. A. Uzi fly: Indian Uzi fly (Exorista sorbillans Syn Tricholyga bombycis) of Tachinidae, Diptera is the most serious. The other important are Japanese, Black and Tasar Uzi flies. These insect are parasites and the female fly lays more than 300 eggs on as many as 300 fairly grown up silkworm larvae usually from the third instars onwards. The eggs hatch in 2-5 days and the maggots penetrate through the skin of the silkworm and enter the body to feed on internal body contents. In cases, the mature maggots escape by piercing the cocoon and rendering the cocoon unfit for mass reeling. Control measures : Use of nylon net (40-70 mesh) to prevent entry of uzi fly into rearing house. Collection and destruction of uzi fly affected silkworm larvae and cocoons. Destruction of uzi fly maggots and pupae collected from rearing trays, mountages, cracks and crevices in the floor of rearing house. Application of diflubenzuron (Dimilin 25 WP) with levigated China clay as diluent (1:9) on third instar maggots. B. Dermestid beetles : Dermestes cadverinus Dermestidae: Coleoptera A number of species of dermestid beetle have been reported as pests of coccon. Adults and grubs attracted to the smell of cocoons in storage. The stored cocoons after stiffling are mostly infested by the grubs of dermestid beetles. They make hole and feed on the pupae inside. Control : Cleaning of rearing house and cocoon store room. Do not store rejected cocoons and perished eggs for long time.

Fumigate the dried cocoon storage rooms with Methyl bromide @ 0.5 kg/283 m2 for a day or with chloropicrin @ 0.5 kg/283m2 for 3 days.

C. Mites, Pediculoides ventricosus : The female mites attack silkworm larvae, pupae and adults causing death. The body surface of silkworms struck with mites develops black specks. The infested ones loose appetite, become

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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inactive and have difficulty in excreting and the excreta is attached bead like to the anus. If severely infested, the worms vomit yellowish green fluid. The mite takes nutrition from silk worm and gives out a toxic substance which kills the silkworm. Control : Avoid the storage of wheat/rice straw near rearing house, Treat the building and thatched materials with acaricide or fumigant before use. D. Ants : They attack silkworms in rearing trays. They can be prevented by placing ant wells with water below the rearing shelves. E. Nematode, Hexamermis microamphidis : It is mostly found in silkworms of late autumn rearing. The nematode attacks the young stage l a r v a e a n d p e n e t r a t e s i n t o t h e i r b o d i e s . The h e a d o f a f f e c t e d s i l k w o r m b e c o m e s transparent and the body milky white. Diseases of mulberry silkworm: The silkworm Bombyx mori is prone to the attack of a number of diseases. Among the diseases, pebrine, grasserie, flacherie and muscardine are important. 1. Pebrine: It is caused by a protozoan parasite, Nosema bombycis. Pebrine disease is also known as Pepper disease or corpuscle disease. Infected seeds are the major factors responsible for disease spray and infect all stages from eggs to adults. Mode of Transmission: By contact with the diseased silkworms, ingestion of contaminated food, transovarial transmission Symptoms of infection: Egg stage :Few number of eggs on egg cards, overlapping of eggs one over the other, instead of closely side by side, easy to detach eggs from egg card due to lack of adhesiveness, poor/less egg hatching. Larval Stage: With advance in disease, larvae become sluggish and dull, poor apatite, retarded growth resulting in irregular moulting, presence of larvae in unequal size in the rearing bed, appearance of irregular dark brown spots or black spots on the body of larvae, hanging down of the head, instead of holding up, Infected larvae may die before spinning or it may spin only a poor and flimsy cocoon, affected larvae lack luster and in later instars turn rusty brown. Pupal stage: The abdominal area is soft, swollen and dark in color, Black spots present on sides of the abdomen; Pupa loses its luster and become dull in its movements. Adult Stage: Discolouration of scales on abdominal area, Black spots may be seen on abdomen, deformed wings, distorted antennae, and Low fecundity. Disease Management: The disease can be managed by using diseased free females and eggs, sterilization of eggs, equipments room and workers with 2% formalin, destruction of infected seeds and laying females.

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2. Muscardine Disease: The disease is fungal and the characteristic is mummification of the infected larvae by deposition of calcium oxalate salts and sometimes called calcino. The different fungus has variety of colored spores but mode of infection, the symptoms and control measures are similar. Causative organism: Caused by Beuvaria bassiana (white muscardine); Spicaria prasina (Green muscardine) or Iscaria farinose (yellow muscardine). Mode of transmission: Dominantly by penetration and very few through mouth or spiracles. Symptoms: Initially moist or oily speaks appears on the body;the larvae vomit or purge violently, the infection leads hyphae come out from intersegmental membrane, skin shrinks and skin becomes inelastic. Affected larvae die before spinning. The dead larvae are covered with white mycelia at first, which later changes to yellow, black or green due to formation of spores. The dead larvae become mummified into chalky white structure. Late instars spin flimsy cocoon and pupae die within calcified cocoon. Predisposing factors: Diseased worms, fecal matter. Prevention and control: Disinfection of room/equipment with 2% formalin / 5% bleaching powder. Reduce humidity in bed with lime powder. Application of formalin chaff @ 0.4, against I and II instars and 0.5, 0.6 and 0.8%, against III, IV and V instars, respectively. 3. Grasserie The viral disease complex is called jaundice as the infected larvae appear yellow in color and excrete white faeces filled with viral polyhedral. Nuclear polyhedrosis virus: (Grasserie, Jaundice, Hanging disease) Caused by borrelina virus. Polyhedrons are hexagonal or octagonal. Measure 0.5 to 8.0 in size. Virion measures 400 x 80 nm. The polyhedra engulfed along with food will get dissolved in midgut and virions infect the midgut cells. Later, they enter haemocoel and invade fat bodies, silk glands, trachea, haemocytes and the polyhedral formation occurs inside the nucleus of the target cells. Fourth and fifth instar larvae are more susceptible. Symptoms: No external symptoms in early stage of disease Larvae become sluggish and lose appetite Swelling of inter segmental region Shining yellowish, fragile skin without elasticity Larvae become restless Crawling out of larvae from rearing tray and falling down, thus resulting in rupturing of fragile skin, there by releasing turbid dirty whitish haemolymph containing numerous polyhedral The diseased larvae lose the holding power of legs except the last pair with which they hang head downwards. Predisposing factors : High temperature, high humidity and their fluctuation in rearing room, Excess moisture in leaf and rearing bed, Insufficient ventilation, Overcrowding during rearing, Inferior quality mulberry leaf, Feeding mature leaf followed by tender leaf, Feeding leaf with accumulated dew drops and rain drops. Prevention and control : 1. Strictly avoiding the predisposing factors 2. Surface sterilization of eggs with 2% formalin 3. Use of bed disinfectants like dusting reshamkeetoushad @ 2-2.5 g/sq. ft during chawki rearing and @ 3.5-4.5 g/sq. ft during late age rearing once after each moult.

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Midgut cytoplasmic polyhydrosis: the disease is characterized by the multiplication of the virus and formation of polyhedral in the host cytoplasm of midgut cells. Causative agent: Smithia is a RNA virus that forms hexa or tetra-gonal polyhedral. Mode of transmission: oral and induction due to bad environment. Symptoms: Loss of appetite, lag in development, shrinkage of the body and leads to transculent cephalothorax. Supernumerary molts and white faeces excretion, midgut becomes opaque. Pre-disposing factors and control measures: Same to NPV 4. Flacherie: the disease is caused by multiplication of bacteria in the alimentary canal and manifested symptoms of diarrhea and vomiting. Causative agent: caused by bacteria Bacillus thurungiensis and other bacteria. Mode of transmission: the larvae are most susceptible and infection occurs by ingestion of spores and induction by bad rearing condition. Symptoms: Symptoms are manifested as loss of appetite, slow growth, diarrhea, vomit and in most case body purifies and become black or even green. Pre-disposing factors: Similar to muscardine, bad rearing conditions like high temperature and humidity, poor ventilation, overcrowding, bad leaves and over feeding are the pre disposing factors for disease spread. Control measures: Selecting sturdy race, proper rearing and incubation of eggs are preventive measures. 5. Septicemia: the infection of haemolymph causes this disease. Causative agent: Bacillus, Streptococcus and Staphylococcus lowers gut pH and gut wall degenarates allowing the bacteria to enter the haemolymph and causes septicemia. Mode of transmission: Contact with wounds and oral. Symptoms: Softening of the body, change in the body coloe to brown and production of liquid excreta, loss of clasping power of prolegs and death of larvae. Control measures: Provision of good sanitation, removal of infected larvae and careful handling of the larvae to avoid skin wound are the preventive measures. Summary and conclusions: Thus we were able to know about insects pest and diseases of silkworm.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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Date: 2068/11/09 Practical No: 11 REARING TECHNIQUE OF YOUNG AND GROWN UP AGE OF LARVAE Objectives: To know the rearing technique of young and grownup age of silkworm Introduction: Silkworm rearing is mainly for cocoon production either for reeling or for seed production. Mulberry silkworm is a delicate , domesticated animal and hence is rearing is also highly skillful job, silkworm rearing needs lot of care and patience. Being a delicate creature, silkworm responds greatly to the environmental condition , especially temperature and humidity. It is also susceptible to a number of diseases which sometime take a very heavy toll of worms. This all clearly indicate that silkworm rearing is a skilled job and required highly skilled hands in order to ensure stable yield of cocoons. Healthy development of silkworm in all stages is of primary concern for which all the condition such as temperature , humidity , timely feeding with quality leaves under a well ventilated and protected environment are required. Therefore , the silkworm rearing requirements can be categorized in to good rearing environment , rearing facilities and continuous supply of quality leaves. All these together exerts a combined effect upon the physiological activities of the worms thus affecting their life. Silkworm rearing requires many pre information such as leaf, labour,variety, consideration like temperature, humidity, air, light, leaf quality and involves many operation i.e. disinfection, brushing, maintenance of optimum temperature and humidity in rearing room, feeding , bed cleaning, spacing, care during mounting of mature worms for spinning. The ripe worm generally requires an area about a square of its body length for spinning its cocoon. About 24-260c temperature and 60-70% relative humidity are optimum codition fro spinning. Too low temperature causes delay in spinning and also affects color, lusture and texture of cocoon, while to high temperature hasten spinning and causes waste of silk in preliminary web. Proper ventilation is needed to dry the wet silk into a firm cocoon. It is particularly important that during the first 50 hours after mounting ideal conditions are maintained for spinning cocoon. The worm anchors itself first to the mountage and then by continuous movement of head silk fluid is excreated in a minute quantity forming a continuous thread. First it spins a loose hammock forming the framework of the cocoon. This layer is called floors which is about 2% by weight of a cocoon in bivoltine and 50% in multivoltine. The larvae continue webbing in compact form inside and finally shrinks and covers itself with agrossmar or thin layer ofsilk. The last inner layer of thin silk is called pelade layer and is unreelable. Larvae spins cocoon approximately @ of 6 inches thread\minute continuously for 2-3 days. If the worms are disturbed during this process it causes suspension of spinning and breaking of thread.

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The basic requirements for growth and development of mulberry silkworm Descroiption/Instars Room temperature(oc) Relative humidity(%) Feeding duration(days) Feeding frequency(times) Amount of food(kg) Bed area(sq.foot) No of bed cleanings(times) Mulberry feeding Percent of total food(%) I 26-28 85 3-3.5 10-12 5 5-10 1 Twigs with 5 leaves 0.3 II 26-28 85 2-2.5 7-8 15 15-45 2 Twigs with 5 leaves 1 III 24-26 80 3-3.5 10-12 50 45-90 2 Twigs with 5 leaves 5 IV 24-25 75 4-5 12-15 230 90-180 2 V 23-24 70 8-10 21-27 800 180-360 4-5

13.7

80

Conclusion: Thus we are able to rear the young and grownup stages of silkworm.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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Date: 2068/11/09 Practical No: 12 CLEANING AND DISINFECTION OF REARING ROOM Objectives To know the cleaning and disinfection techniques To know the tools and reagents used during cleaning and disinfection Introduction Due to centuries of domestication the silkworm has no natural resistance and shows neither morphological nor behavioral adaptation to escape parasites and pests. Development of disease resistance species has been partially successful and no race is completely immune to diseases. Silkworms are infected by protozoa, bacteria, viruses and fungi. In order to prevent and control silkworm diseases, disinfection and thorough cleaning of rearing room and appliances must be carried out prior to the commencement of rearing. The commonly used disinfection methods are: 1. Sun drying It is the cheapest means of controlling pathogens. UV-rays denature the protein of pathogen & infra-red rays make the pathogen heated and dried. The rearing appliances must be washed and exposed to the sunlight. It can only be carried out in summer and only applicable for surface sterilization. 2. Hot water treatment This is simple, feasible and effective disinfection method. Small rearing tools like nets, chopsticks etc. are boiled in hot water for 30 minutes and taken out. 3. Steaming Rearing room and appliances are steamed under high pressure in specially designed cabinets for hrs. in addition to initial investment hot steam is harmful to the appliances made up of bamboo or sticks. 4. Hot air treatment It is very effective method but laying pipes and installing facilities are very expensive. 5. Chemical treatment This is most commonly used method. The rearing house as well as the appliances used in rearing should be disinfected with2% formalin prior to commencement of every rearing. For effective disinfection, the rearing house should be made air tight as far as possible and with the rearing appliances kept inside, the walls, windows, doors and the appliances should be sprayed with 2% formalin @ 7-8 lit for 100 sq. meter and the doors closed immediately. After 24 hours of disinfection the doors and windows should be opened and the rearing house should be completely aerated at least 24 hours before the commencement of brushing. Summary and conclusion Thus we became familiar with the cleaning and disinfection procedures, tools and chemicals used during this and precautions to follow.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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Date: 2068/11/09 Practical No: 13 PREPARATION OF SILKWORM FEED BED AND BED CLEANING Objectives To be familiar with silkworm feed bed and bed cleaning. To know about the spacing required for the different stages of larvae for feeding. Introduction Silkworms grow and develop very fast. As the size of worm increases, the size of rearing bed should be increased accordingly. They grow approx. 15 times from hatching to 1st instars, 4-5 times each at second and third instars, 25 times each at fourth and fifth instars. Accumulation of wet leaves and faecal matters favor development of certain diseases which often take a heavy toll of silkworms. Removal of such material including dead and unhealthy worms from the rearing bed is called bed cleaning. Bed area To attain full larval growth, good survival and successful cocoon crop, maintenance of optimum bed area is important. For multivoltine x bivoltine hybrid 360 ft2 and for bivoltine x bivoltine 480 ft2 / 100 dfls are found optimum by the end of V instar. Rearing bed area has to be increased daily to match the growth of silkworms. The space requirement for larvae during chawki rearing is given below. ------------------------------------------------------------------------------------------------------------------------------Instar Uni/Bivoltine Multivoltine Beginning Beginning End End I 0.2 0.8 0.2 0.5 II 1.0 2.0 0.5 1.5 III 2.0 4.5 1.5 3.0 Bed spacing The successes of silkworm crop and cocoon quality also depend on spacing. Over crowded rearing leads to insufficient consumption of feed, poor growth, susceptibility to diseases and low cocoon yield of inferior quality. The spacing should be increased daily in proportion to the growth of silkworms. The rearing bed spacing recommended for IV and V instars is as follows. Instar Space in mm2/larva Uni/Bivoltine Beginning IV V 5.0 10.0 End 10.0 20.0 Multivoltine Beginning End 3.0 9.0 9.0 18.0

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Bed Cleaning Leftover leaf and litter accumulate in rearing bed leads to increase in humidity and temperature and multiplication of various pathogens. Hence, regular cleaning of bed is necessary. Since the young age worms are delicate, cotton / nylon nets are used for bed cleaning. The cleaning net is applied on the bed, one feed before cleaning, and the feed is given above the net. The worms crawl through the meshes and come up to feed on the leaves on the net. At the next feed the net along with the worms above is transferred to another tray and fresh feed given. The feces and left over feed are collected and put in manure pit. Cleaning is done once in I instar prior to I moult (on 3rd day), twice during II instar, first on resumption of fresh feed and the second prior to II moult, and thrice during III instar, first on resumption of fresh feed, second during the middle and the third prior to III moult. The mesh size of nets used for I & II instars is 2 mm2 and for III instar 10 mm2. During IV and V instars bed cleaning is done every day with nylon or cotton nets of appropriate mesh size (20 mm or 2 cm X 2 cm). As in chawki rearing the net is fixed over the rearing tray one feed prior to cleaning and feed is given above the net. Before next feed the net along with worms and leaves is transferred to another tray. The faeces and left over leaf are collected and put into manure pit. Paddy husk or charred husk is also used for bed cleaning in late age rearing. They are spread in a thin layer over the bed prior to feeding. The worms crawl through this layer and start feeding on fresh leaves. The worms and the leaves are removed to a fresh tray and the litter and old leaves are put in a manure pit. Summary and conclusions: Thus we were able to know about the preparation silkworm feed bed and bed cleaning.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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Date: 2068/11/09 Practical No: 14 LARVAL MOUNTING AND CARE DURING COCOON SPINNING Objective: To know about the mounting operation in silkworm larvae. To know the process of cocoon spinning. Introduction: Mounting is the final and the most busy operation in silkworm rearing. Toward the end of the 5th instar, the silkworm stops feeding, select a suitable site , exudes silk through its spinneret and spins a cocoon. For providing optimum spinning condition, the ripe worms are tranfered to mountage. Soon after mountage the worm passes out its final excreta and emptying its gut and start spinning cocoon.The worm ripe and are ready to mount after5-7 days of feeding in 5th instar for multivoltine and bivoltine rave in tropics and 7-9 days for univoltine and bivoltine in subtropical regions. The ripe worm can readily be distinguished by its translucent and yellowish body appearance shrinkage of body length and visible constriction at the 4th and 5th abdominal segment. The process of spinning cocoon takes about 12 days in multivoltine and 2-3 days in uni/bivoltines. The ripe worm generally requires an area about a square of its body length for spinning its cocoon. The optimum spacing for chandrika is 50 worms/0.1 m2. About 24-26C temperature and 60-70% relative humidity are optimum for spinning. To low temperature causes delay in spinning and aslo affects the color, lusture and texature of cocoon, while to high temperature hasten spinning and causes waste of silk in preliminary web. Proper ventilation is needed to dry the wet silk into a firm cocoon. It is particularly important that during the first 50 hours after mounting ideal conditions are maintained for spinning cocoon. The larvae anchors itself first to the mountage and then by continuous movement of head silk fluid is excreted in a minute quantity forming a continuous thread. Movement of head by larvae is about 65 times/minute. First it spines a loose hammock forming the framework of the cocoon . this layer is called floor which is about 2% by weight of a cocoon in bivoltine and about 50% in multivoltine. The larvae continues webbing in compact form inside and finally shrinks and covers itself with the agrossmar or thin layer of silk. The last inner layer of thing silk is called pelade layer and is unreelable. Larvae spins cocoon approximately @ of 6 inch thread/minute continuously for 2-3 days. If the worm are disturbed during the process it causes suspension of spinning and breaking of the thread.

Conclusion: Thus we are able to do mounting process in silkworm and also the care taken during cocoon spinning.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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Date: 2068/11/09 Practical No:15 COCOON HARVESTING, GRADING AND PRICING Objectives To know the methods of cocoon harvesting , grading and pricing To be familiar with precautions while doing these Introduction Harvesting The harvesting of silkworm cocoon is the final stage of the process of silkworm rearing. It should be done neither too early nor too late. It is done after hardening of the pupal cuticle i.e.5th day after spinning for tropical and 7th day for temperate races. Delay results pierced cocoons due to moth emergence. Early harvest results in spoiled cocoons due to injury. Harvesting is done normally by hand. Harvesting of cocoon is done on the 5th day of spinning. In winter, it may be done on 6th day in case of multivoltines for commercial use. Seed cocoons may be harvested on 8th/9th day of spinning depending on temperature. Harvesting should not be done immediately after pupation, since the soft cuticle of the Pupae may rupture due to bad handling.

Grading Cocoon grading is of paramount importance to determine the amount of quality cocoons so that a price can be determined. After examination of the cocoon characters the price of cocoon is fixed. Following are the procedures before grading: Put cocoons for separation on the table. Separate stained and defective cocoons. Weigh the cocoons. Take a sample. Transport safely. Calculate the shell ratio Determine the grade Pricing The commercial characters of cocoons for fixing price are: 1. Heritable characters This includes colour and shape of cocoon. Colour is racial character and varies from white to yellow. Similarly different shapes are- round, oval, egg shaped, peanut shaped, spindle shaped etc. 2. Environmentally induced characters This refers to weight and size of cocoons. The weight of cocoon used to produce one unit of raw silk is called renditta. The numbers of in one renditta is considered for price fixing.

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3. Reeling and silk characters Shell ratio is one of the important basis used for price fixing. Shell ratio= (weight of the shell/weight of entire cocoon) 100% Based on SR ratio price is determined as follows: Shell ratio (%) More than 21 19-21 16-18 Less than 18 All bad and defective cocoons Cocoon grade A grade B Grade C Grade D Grade E Grade Price(Rs/kg)

Summary and conclusion Thus we knew the methods of cocoon harvesting, grading and pricing of cocoons and the optimum stage of cocoon harvesting so that higher price could be achieved due to high quality. Also the handling methods were also dealt.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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Date: 2068/11/09 Practical No: 16 POST HARVEST PROCESSING OF COCOON Objectives To be familiar with post harvest processing of coccon. To know about the quality of coccon required and its preservation. Introduction As the cocoons are high valued and perishable commodity, they should be harvested and sold at the right time to get better yield and quality cocoons to secure a good price. Every persons involved in sericulture should aim at the production of food quality reeling cocoons to procure high quality international grade silk. The quality of silk depends upon the quality of cocoon which in turn depends on the silkworm race, rearing condition, harvesting and proper grading. Poor quality cocoons are formed mainly due to inappropriate treatments and adequate care during cocooning. Processing The silk yarn production begins with selecting for their quality. The cocoons will be sorted into normal and abnormal cocoons. The fresh cocoons normal are reelable to produce raw silk but the abnormal cocoons are unreelable. The abnormal cocoons are Double cocoons, perforated cocoons, inside soiled cocoons, outside soiled cocoons, thin-end cocoons, thin-middle cocoons, thin shelled cocoons, double layered cocoons, malformed cocoons, etc. After the first selection of cocoons, they have to be dried. The first goal of cocoon drying is the protection of cocoon quality, to preserve conditions for reeling cocoons and prevent damage that might be caused by long periods of storage. Drying kills the pupa and evaporates moisture that would otherwise ruin cocoons. Dried cocoon storage conditions are designed to keep the raw material for long periods without any damage from moulds and pests. Cocoon storage should preferably be built with double walls. Cloth or polyethylene bags are recommended as containers for dried cocoon storage. In most modern factories which aim at producing high-grade raw silk, the cocoons are graded on visual inspection or by mechanical tests are actually mixed as in the required proportions. This is called cocoon mixing or blending and is done to ensure speed and uniformity of reeling as well as to obtain the desired effect in raw silk. The reelable cocoons have then to undergo the reeling processing, after which they are either processed further to raw silk or to thrown silk. The unreelable silk is processed further to spun silk. Summary and conclusions: Thus we were able to know about the post harvest processing of coccon.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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Date: 2068/11/09 Practical No: 17 SILKWORM RECORD KEEPING Objective: To know about the method of silkworm record keeping. Introduction It is very important to keep the record of different activities and operation done during silkworm rearing and cocoon production. System of keeping record is helpful for planning in future and hence ensures the maximum production of silk. For record keeping following types of tables should be used. The main product of silkworm is silk cocoon so record keeping also include the grading and recording of cocoon. For keeping the record of cocoon following format is used. Cocoon grading system Cocoon grading system is very important for transaction of cocoons Pricing system for marketing cocoon has to be based on grade of cocoon Therefore cocoon grading system should be fair and proper for seller and buyer. Purchase of green cocoon depends on the contents, quality and shell percentage of the single cocoon. The basic parameters to be considered for grading purposes are the general cocoon qualities such as cocoon weight, shell weight and cocoon shell ratio(CSR) Cocoon assessment is done for cocoon shell content to fix the proper cocoon price. Weight of cocoon: The most important commercial character of cocoon is weight. The lower the number of cocoons, the greater, the greater the silk contents. Weigh separately the good cocoons and sort out cocoons (bad) of each farmers crop. For the cocoon assessment, take randomly 50g of cocoons with live pupae form sample, and count the cocoon numbers of 50g for calculation of cocoon weight. Shake the cocoons randomly to identify the live pupa inside the cocoons. Weight of the cocoon shell: More important than the weight of the whole cocoon is the weight of the silk shell, as it is the shell that yields the silk for reeling. Hence, larger the weight of the shell, greater the silk yield from it. Cut 50g of cocoon and separate the pupae and cocoon shells, then weight cocoon shells and keep record. Cocoon /Shell ratio: Shell ratio calculation is based on the 50g cocoons and weight of cocoon shells. Shell ratio = weight of cocoon shell/weight of cocoon(50g) * 100

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Cocoon grading format: Serial no: Name of farmer: Address of farmer: Rearing season: Cocoon receive d by farmer s Amount of cocoon(kg) during grading Sample cocoon (Nos/50g ms) Wt of sample cocoon(g m) Wt of cocoon shell(gm ) Shell ratio( 5/4) Cocoon grade/cla ss Price of good cocoons( good cocoon*SR*pri ce) Cocoon price Total cocoon price( price of good + bad cocoon s)

Total cocoon/ Bad cocoon 1

Good cocoon/ Bad cocoon 2

Conclusion: Thus we were able to keep the production and pricing record of cocoons.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

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Practical No: 18 STUDY OF MORPHOLOGY OF DIFFERENT STAGES Objectives To know the morphology of silkworm at different stage To be able to identify different stages

Date: 2068/11/09

Introduction Silkworm passes through complete metamorphosis and thus has four distinct developmental stagesegg, larva, pupa and adult. Egg: Larva: Eggs of European races. Eggs laid early during oviposition are larger. Indian crossbreed- multivoltine , 400 eggs per laying - very small Av. egg wt. 0.55 - 0.6 mg (2,000 eggs/g) White cocoon races lay paler eggs compared to yellow cocoon races. The color of the eggs changes to deep brown when they enter into diapause. Non-diapausing races - color does not change till one day prior to hatching - Blue egg stage. Eggs are ovoid, spherical or ellipsoid in shape and are flat on one side i.e., - egg dimple. Protective covering of the egg is chorion and its opening is micropyle The eggs are centrolecithal. Eruciform - polypod Ant, newly hatched black larva with bristles 3 mm long The bristles or setae are situated on four pairs of tubercles in each segment. The larva are10 cm long. The three body regions (Tagmata) of a larva are head, thorax and abdomen.

Head: Hypognathous formed by the fusion of cranium Apical plate is Epicranium with epicranial suture. Prominent sense organs are three pairs of ocelli. Antennae, from 2nd head segment are short, 3 or 5 jointed each joint has 30-40 Segments - they perceive odour. Thorax: Prothorax: Smaller with a pair of spiracles Meso and Meta thorax: Project slightly upwards and form dorsal hump. A dorsal eye - spot on mesothorax. Each thoracic segment has a pair of true legs. Abdomen: Formed by 11 segments, but 9 only visible (9, 10, 7, 11th abdominal segments fuse to form apparent 9th segment, the caudal plate and paired caudal prolegs) Paired prolegs are also present in 3rd, 4th, 5th and 6th abdominal segments.

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Pupa: Adult:

Projection on the dorsum of the 8th segment is Caudal Horn. First eight abdominal segments have a pair of spiracles laterally. Pupal moult occurs within the cocoon, non-motile and non-feeding. Head with a pair of compound eyes and antennae, and mouth parts vestigial. Thorax with two pairs of wing pads and three pairs of limb buds immovably pressed. Abdomen with nine visible segments and seven pairs of spiracles in the first seven segments. Last pair of spiracles non-functional. Female pupa is larger with broad abdomen, ventral line in the center of the 8th abdominal segment. Male pupa has small round spot on the ninth segment. Moth is about 4 x 2 cm, no flight, does not feed and short life span of 3-6 days. Hypognathous head, bipectinate antennae and non-functional coiled proboscis. Thorax with a pair of wings and three pairs of legs. Abdomen with eight visible segments in male and only seven in female Six pairs of spiracles in the abdomen.

Summary and conclusion Hence we were able to identify different stages of silkworm knowing their morphological differences.

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A Practical Manual on Sericulture

2012

Date: 2068/11/09 Practical No: 19 OBSERVATION OF SEX DIFFERENTIATION OF LARVAE, PUPA AND ADULTS OF SILKWORMS Objectives To be familiar with different stages of silkworm. To know about the sex differentiation in different stages of silkworm. Introduction Sex differences in the larva: Female larvae has a pair of sex marks on ventral side of 8th and 9th abdominal segments, called Ishiwata's Fore glands and Ishiwata's Hind glands, respectively. Ishiwata's fore glands later modify into bursa copulatrix and receptaculum seminalis. Ishiwata's hind glands become posterior part of oviduct and the accessory glands. Male larva has a single median opening at the junction of 8th and 9th segments called opening of Herold's gland. It becomes seminal duct and ejaculatory duct of the adult. Then openings are seen only in freshly moulted larvae. Hence, marker genes in the sex-chromosomes are used in special races for easy separation. The sex marker genes are located on the w-chromosome of the female. Pupa: Pupal moult occurs within the cocoon, non-motile and non-feeding. Head with a pair of compound eyes and antennae, and mouth parts vestigial.Thorax with two pairs of wing pads and three pairs of limb buds immovably pressed. Abdomen with nine visible segments and seven pairs of spiracles in the first seven segments. Last pair of spiracles non-functional. Female pupa is larger with broad abdomen, ventral line in the center of the 8th abdominal segment. Male pupa has small round spot on the ninth segment. Adult: Moth is about 4 x 2 cm, no flight, does not feed and short life span of 3-6 days. Hypognathous head, bipectinate antennae and non-functional coiled proboscis. Thorax with a pair of wings and three pairs of legs. Abdomen with eight visible segments in male and only seven in female. Six pairs of spiracles in the abdomen. Sexual dimorphism in adults: Character Female Male Color Paler Darker Activity Less Active More Active Antennae Small Large Body Size Large Small Abdomen Flat with 8 Segment Narrow with 7 segment External Genitalia The caudal end has a median The caudal end has a pair of knob like projection with hairs. hooks, harpens helping in copulation. Summary and conclusions: Thus we were able observe the sex differentiation of larvae, pupa and adults of silkworm.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Practical No: 20 ANATOMY OF SILKWORM (Bombyx morri) LARVAE Objectives: To know the anatomy of mulberry silkworm( Bombyx morri).

Date: 2068/11/09

Introduction: The larvae are polypodus or eruciform type. The head is hypoganthus , heavely chitinized and bright brown in color. There is spinneret as the median process, at the distal portion of the labial prementum through which silk gland open and silk exdues. It includes different system. Internal anatomy of silkworm larvae include following systems: Digestive system Circulatory system Excretory system Respiratory system Reproductive system Silk secretory system In above system, most of internal function of different system is similar to the lepidopteran moths but silk secretory system is somewhat different. Silk gland: there are a pair of silk glands, which are transformed labial glands. They are derived from invagination of labial ectoderm. They are important organ which produce liquid silk as the source of cocoon fibre, which is used for spinning the cocoon. The cocoon protects the pupa. There are situated on the ventolateral sides of mid intestine. The posterior ends are blind. The paired ducts unite anteriorly and open into spinneret, the median projection in the labium. Each silk gland is divide anterior, middle and posterior parts, of there middle parts is the largest and is twisted in s shaped. The glands are well differentiated in the fourth and fifth instar larvae. They are so large in the fully grown final instar larva that they occupy almost the entire body cavity and account for 50% of the total weight of the larva. The silk glands are tubular in nature and width of the tube varies in different regions of the gland. The anterior part is in the form of a straight thin tube of more or less uniform in diameter. There are no trachea in this part. It receives its oxygen supply through blood circulation. It has no secretory function. The middle region has three parts , anterior , middle and posterior. The middle part is widest and the endof posterior part is narrow. The right and left anterior parts of the gland unite behind the spinneret. A pair of fillips gland are situated at the junction of two anterior region. A viscous fluid is secreted by these glands which coats the silk threads. The wall of the silk gland is composed of three layers, the outer tunica propria of uniform thickness throughout the gland, the middle glandular layer and inner tunica intima enclosing the lumen of the gland. The fibroin and sericin are secreted in the fourth and fifth stages. Fibroin, the silk protein is secreted form the posterior region. The middle region acts as the reservoir for the maturation of fibroin and it secrets sericin and fibroin. Conclusion: Thus we were able to know about the anatomy of mulberry silkworm.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com

A Practical Manual on Sericulture

2012

Practical No: 21 STUDY OF DIFFERENT GLANDS PRESENT IN Bombyx mori

Date:2068/11/09

Objectives To know about different glands present in the silkworm To know in detail about silk gland Introduction Salivary Glands are the transformed labial glands, cylindrical and tubular in structure. They are situated on the ventro-lateral sides of the mid intestine. The slender duct passes through chitinous state of mandibular flexar muscles and open below the mandible in to mouth cavity. Silk glands: The silk thread of the cocoon is secreted by a pair of glands which are actually modified labial glands. These glands are well differentiated in the 4th and 5th instar. They lie below the alimentary canal. They are large and they are fully grown. In the final instar it occupies most of the body cavity ventral to the alimentary canal and accounts for 50 percent of the body weight. The silk glands are tubular in nature and the width of the tube varies in different regions of the glands. A pair of modified labial glands well differentiated in the 4th and 5th instar larvae lie below alimentary canal; occupy most of the body cavity and accounts for 50% of the weight and 4 times the length of larva. Formed of 3 layers, outer tunica propria of uniform thickness, middle glandular layer and inner tunica intima which is very thick in the anterior region and is shed at each moult and in the other regions it is thin and is not shed at each moult. The nuclei of glandular cells are round in young worms and are branched in mature worms. The three distinct regions of silk glands are posterior, middle and anterior regions. The silk filament is made up of inner fibroin (75%), a tough elastic protein and outer sericin(25%), a gelatinous protein The posterior most regions highly folded and secrete major protein precursor of the silk, fibrinogen, which gets denatured to fibroin. The widest middle region is folded into w-shape with three limbs, the posterior, the middle and the anterior limbs. It acts as reservoir for fibroin and it gets matured here. The posterior limb of the middle region secretes sericin I. Middle limb secretes sericin II around sericin I Anterior limb secretes sericin III around sericin II. The anterior region is uniform, thin, no secretions but conducts the silk fiber assembled in the middle region to the spinneret. The anterior region of the two sides open at the base of the median projection in the labium called spinneret, which draws out silk in the form of a fine filament. A pair of glands known as Filippis's or Lyonnet's glands is situated at the junction of the two anterior regions. A viscous waxy fluid is secreted by these glands serves as lubricant at the time of silk thread ejection The threads of the two sides are called the Brins, and the sericin layer of the two binds them together into a single filament or Bave. Summary and conclusion Hence we were familiar with the different glands present in silkworms and knew in detail about the silk gland.

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Prepared By: Babu Ram Panthi, B.Sc.Ag. Elective (Entomology) panthibabu@outlook.com