Disinfection in silkworm rearing

Introduction

My dear student friends, in today’s lecture let’s study the Disinfection in silkworm rearing.

Mulberry silkworm, Bombyx mori, the primary producer of commercial silk is domesticated for many
centuries. The colonization has rendered the silkworms to lose its natural abilities to withstand the
adverse climatic conditions and pathogen infection. The breed improvement programmes aimed at
evolving breeds with certain economic parameters have further eroded their ability to withstand
pathogenic infections. This has made them susceptible to many diseases, causing crop loss or low
yield, which ultimately has lead to low silk production.

The occurrence of epizootic condition in silkworm colonies have necessitated the disinfection process
in silkworm rearing. The diseases found in silkworms are either chronic or acute in nature depending
on the pathogens involved. Flacherie is caused by bacterial pathogens alone or mixed infection with
infectious flacherie virus or densonucleosis virus. Grasserie is caused by a viral pathogen BmNPV.
Muscardine and Aspergillosis are fungal diseases, which are common during winter and rainy
seasons. Muscardine infects all stages of silkworms, whereas Aspergillosis occurs during early instars
of the larval stage. The occurrence of pebrine disease is rare and may breakout at any period of the
year. The vomit, feces and corpses of the diseased silkworm contains the pathogen and are released
into the environment. Further, cross contamination from other insect pests also increases pathogen
load in the rearing environment.

As there are no foolproof curative measures, these diseases are best prevented through proper
disinfection of rearing houses, surroundings, rearing appliances and sterilization of larval surface and
rearing bed, by application of various bed disinfectants. Disinfection is an integral activity in
silkworm rearing aimed at destruction of disease causing pathogens in silkworm rearing environment.
It involves the spraying of chemical disinfectants before the commencement of rearing and dusting of
the bed disinfectants during the course of silkworm rearing. Different disinfectants are used for
disinfection in different concentrations. The quantity to be sprayed differs depending on the floor
area, height and type of rearing house and the method of rearing, etc.

For better understanding, the topic is being explained into different sub-headings and they are:

1. Disinfection
2. Physical methods
3. Chemical methods
4. Estimate required quantum of disinfectant solution
5. Schedule of disinfection activities
6. Preparation of different disinfectant solutions
7. Bed disinfectants

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Let’s study these aspects one by one in detail. The first one is -

Disinfection

As a preventive measure, disinfection is carried out before taking up the rearing. Disinfection means
the destruction of pathogens. The use of disinfectants is recommended not only to get rid of source of
infection present in the rearing environment and appliances, but also as a prophylactic measure.
Disinfection could be achieved by physical or and chemical means. The microorganisms differ in their
resistance to destruction by physical and chemical methods. Certain factors are to be considered,
while selecting a disinfection method for the rearing room and appliances.

 The method should be effective

 The method should be simple and easy for application
 The method should be harmless to humans and domestic animals
 The method should be harmless to building and equipments
 The method should not be corrosive to the building or equipments

The second aspect of our study is Physical method of disinfection.

Certain physical disinfectants or destroyers are used for disinfection. However the physical
disinfectants do not offer perfect disinfection of rearing house appliances. The physical methods
involve the use of sunlight, ultraviolet radiation, flame, heat and steams and boiling water.

Sunlight: Sunlight acts as a disinfectant due to the heat it generates on the surface to be disinfected as
well as due to the ultraviolet radiation emitted. The rearing equipments like stands, rearing trays,
mountages, etc., are disinfected by exposing them to hot sun. Attention must be paid to expose both
the surfaces of the appliances. Many pathogens lose its infectivity when the contaminated rearing
trays are exposed to hot sun for 20-30 h.

Ultraviolet radiation: Ultraviolet light are the electromagnetic radiation emitted by the sun having a
wide range of wavelengths. Ultraviolet radiation of approximately 257 nm wavelength emitted by
ultraviolet lamps is germicidal and can be used effectively to eliminate pathogenic microorganisms on
the exposed surfaces and in air. The UV radiation disrupts DNA and RNA in living organisms.

Flame or fire: Oxidizes the microorganism into ashes. Though it is rapid, the threat of accidental fire
and damage to the equipments and tools used for silkworm rearing is a limiting factor for the use of
fire.

Heat: It is an effective destroyer of pathogens. Dry heat destroys the microorganisms by oxidation
and is non-corrosive and less effective than moist heat. Dry heat requires longer exposure time and
high temperature. Moist heat or steam causes irreversible coagulation of microbial proteins. The
action of moist heat is faster and more effective.

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Boiling water: Use of boiling water is an effective disinfection method. The maximum temperature
obtainable is 100°C and the exposure of 10-30 min, is effective. But this method is highly
cumbersome and is not a reliable sporicidal.

The third aspect of our study is Chemical method of disinfection.

Disinfection is best performed by use of chemical disinfectants. An ideal chemical

disinfectant is one having broad-spectrum of activity with the potential to kill a wide range of
microorganisms, particularly the pathogens and it is preferably harmless to humans and animals,
freedom from irritation and toxicity. Chemical means of disinfection adopts specific chemicals
having the germicidal activity against targeted microbes for an assured and effective disinfection. The
use of chemicals for disinfection gained momentum after Lister has showed in 1867 that phenol
would kill microorganisms. An ideal disinfectant should be safe to use, non-irritating, free from
toxicity and should not have unpleasant odour. Choice of chemical disinfectant depends on a number
of factors.

a. Number and nature of microbes to be destroyed.

b. Type of surface of rearing house and appliances
c. Interaction with organic matter present in the surface to be disinfected
d. Contact time and temperature conditions
e. Toxicity to individuals, residual toxicity and effect on items such as fabric and metal
f. pH, temperature, hardness of water being used and
g. Cost

For killing or inactivating pathogens causing disease in silkworms various disinfectants are
recommended. These are used to disinfect the rearing rooms, surroundings and rearing appliances,
apart from maintaining personal hygiene. Some of the disinfectants being used in silkworm rearing
environment are – formalin, sodium hypochlorite, quaternary ammonium compounds, iodine
compounds, lime, phenolic compounds, calcium hydroxide, chlorinated lime, chlorine dioxide,
organic chloric compound, sodium dichloroisocyanuric acid, trichloro iso cyanuric acid, etc. The
following are some major general disinfectants used in sericulture.

Formalin: Formaldehyde in water is formalin and is known as formic aldehyde, methyl aldehyde,
ethylene oxide, oxomethane and oxymethylene. It is a colourless gas and usually available as a 36%
solution in water and methanol. It is clear and colourless solution with irritating odour, pungent and
suffocating. Formaldehyde, the active ingredient in formalin solution is a strong reducing agent. It
forms formic acid utilizing the oxygen from the cells leading to the death of the germ. Formalin is
effective only in rearing houses which can be made air tight, at high humidity (70%) and temperature
of above 20˚C. The gaseous nature of the disinfectant demands a closed condition for a longer period.
It was till recently most used disinfectant in sericulture but its use is limited as it is reported to be a
carcinogen and is classified as hazardous chemical. Formaldehyde is highly pungent and cause

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irritation to eye, nose, throat and creates breathing discomfort. The germicidal activity of
formaldehyde is due to its strong reducing action on the germs. It takes oxygen from germ cells and
kills them. Disinfection mask, disinfection coat, Gum shoes and hand gloves are essential for
disinfection

Chlorinated lime: Commonly known as bleaching powder is a mixture of calcium hypochlorite, basic
chloride, calcium hydroxide and free slaked lime. The active ingredient is hypochlorite. It is a white
amorphous powder with a pungent smell of chlorine. For effective disinfection, an active chlorine
content of 30% must be used. Hypochlorite in combination with weak acid (CO2 in air) and water
produces hypochlorous acid and hydrochloric acid. Hypochlorous acid is unstable and resolves into
hydrochloric acid and nascent oxygen. The oxidation reaction by nascent oxygen is germicidal.
Hydrochloric acid also aids in the release of chlorine, which is also germicidal. Chlorine alters the
membrane permeability of cells allowing the macromolecules such as proteinsand nucleic acids to
leak out. The main component of chlorinated lime, Calcium hydroxide is germicidal which dissolves
the polyhedron protein to release and disintegrate virions. The action of bleaching powder becomes
efficient under wet conditions and therefore the surface of walls and equipments should be drenched
with this solution.

Chlorine dioxide: Chlorine dioxide as gas is well known as most powerful and anti-microbial agent
and its advantage as a hard surface disinfectant is widely recognised. Chlorine dioxide is a yellow to
green coloured gas with a distinct odour similar to that of chlorine. Chlorine dioxide gas is highly
toxic and vary unstable. It has been stabilized into solution form which is very effective disinfectant
against pathogens causing diseases in silkworms. It is soluble in water and like oxygen it exists as a
gas in solution. Like Ozone, chlorine dioxide reacts by chemical oxidation and do not form
chlorinated organic by-products. Stabilized chlorine dioxide is effective even at broad pH ranges. This
stabilized chlorine dioxide is superior disinfectant that chlorine in many respects. It is stable, 2.5 times
more effective than chlorine and 50 times than hypochlorites. It is a biotoxic agent causing the
membrane damage. It reacts with nucleotide and amino acid moieties of the germs.

Calcium hydroxide (Slaked lime): It is an antiviral agent and commonly used as a silkworm body and
rearing seat disinfectant. Burnt limestone (Calcium oxide) is hydrated with water to give calcium
hydroxide and slaked lime powder. The slaked lime being alkaline destroys the viruses and is most
effective against BmNPV, BmIFV and BmDNV.

Asthra: Asthra is a commercial product available in the market. 0.05% solution of Asthra is effective
against all pathogens, causing diseases in silkworms. 50 g of Asthra powder is dissolved in water and
after 2 hours, the solution could be used for disinfection.

The fourth aspect of our study is the estimation of required quantum of disinfectant solution

This process involves two steps. Step one is to Calculate the floor area of the rearing house. For each
type of rearing house there is slight variation in the calculation procedure. Different types of rearing

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houses viz., independent rearing house and rearing cum dwelling houses are being used in silkworm
rearing. Similarly rearing houses are with different types of roofs viz., RCC, thatched, tiled and
Asbestos. To calculate the quantity of disinfectant for a rearing house, the area and height should be
determined. Measure the length (L), Breadth (W) and height (H) of the rearing house with the help of
a measuring tape. Calculate the floor area of the rearing house by multiplying length × breadth of
rearing house. That is, Floor area = Floor length × Floor breadth. For example: Area of a rearing
house with length 15 meters and width 10 meters = 15 meters × 10 meters = 150 sq. meters.

Step two is to work out the height of rearing house. If the roof is flat, the height of the rearing house
is the measure between the floor and roof. If the roof is slanting at one side, then the height is
measurement between floor and flat end of room + half of the measurement between flat end and tip
of the roof. If the roof is slanting at either side, then the height is measurement between floor and flat
end of room + the measurement between flat end and tip of the roof.

Estimation of the requirement of disinfectant is done by multiplying the floor area in Sq. meter with
1.5 litres. Add 500 ml for every addition of 1 meter height of the rearing house beyond 3 meters. Add
10% extra to estimated quantity of disinfectant for shoot rearing. Add 35% extra to the estimated
quantity of disinfectant for tray rearing.

The fifth aspect of our study is schedule of disinfection activities

The effective disinfection is achieved by following the recommended disinfection schedule. By

meticulously following the schedule of activities, pathogenic infection in the silkworm crops can be
limited.

a. Collect the left out mulberry and other bed refuse immediately after marketing of cocoons and put
them into compost pit

b. Collect diseased and dead larvae/pupae/floss etc., and disinfect by sprinkling 5% bleaching
powder in slaked lime powder and dispose suitably

c. Burn the floss on the mountages using flame gun

d. Conduct first disinfection with any recommended disinfectant using power sprayer

e. Five days before brushing, clean the rearing house and appliances and wash in water. Dry the
appliances in bright sunlight for a minimum period of 10-12 hours

f. Four days before brushing, continue the sun drying of the appliances. If the prevalence of viral
diseases were high during the previous crop, disinfect the rearing house and appliances with 0.3%
slaked lime solution

g. Three days before brushing, conduct second disinfection of rearing house with suitable chemical
disinfectant

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h. Two days before brushing, dust 5% bleaching powder in slaked lime powder @ 200g/sq. m. at the
passage and rearing house surroundings

i. One day before brushing, arrange appliances for rearing

The sixth aspect of our study is the preparation of different disinfectant solutions

a. Preparation of 2% formalin solution

To prepare a known volume of 2% Formalin solution, divide the total quantity of the solution
required by 18 to get the quantity of formalin required. To this formalin, add water, 17 times to make
2% formalin solution. Or alternatively, to one part of formalin add 17 parts of water.

b. Preparation of 2% bleaching powder in 0.3% slaked lime solution

For preparing 100 litres of 2% bleaching powder solution in 0.3% slaked lime solution, Add
little water to 2 kg of bleaching powder and 0.3 kg of slaked lime and make a paste. Add this paste to
the rest of water and stir it well. Keep for 10 minutes and use the supernatant for disinfection.

c. Preparation of 2.5% chlorine di oxide solution in 0.5% slaked lime solution

For preparing 2.5% chlorine dioxide solution, use the commercially available Sanitech or
Serichlor solutions. Use 50 g of activator of every 500 ml of solution. For preparing 100 litres of
chlorine di oxide solution, take 250 g activator crystals into a basin/bucket and add 2.5 litres of
Sanitech or Serichlor solution. Stir and keep for 10 min. Add activated solution to 97.5 litres of water
and add 500 g of slaked lime. Mix thoroughly and use for disinfection.

d. Preparation of 0.05% Asthra solution

For disinfection of rearing house, its surroundings and appliances, 0.05% Asthra solution is
recommended. To prepare 0.05% Asthra solution, Add 50 g Asthra powder in 100 liters of water. Stir
thoroughly with the help of a stick. Keep for 2 hours for dissolution of the disinfectant. Spray Asthra
solution in the rearing house.

e. Preparation of 0.3% slaked lime solution

For preparing 100 litres of 0.3% slaked lime solution, take 300 g of slaked lime powder and
make a paste. Add this paste to the rest of water and stir well. Keep for 10 minutes and use the
supernatant for disinfection.

The seventh aspect of our study is use of bed disinfectants

For managing silkworm diseases in a silkworm colony, it is essential to follow hygiene practices.
These are aimed at limiting disease spread in the silkworm colony and prevention of secondary

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contamination and infection. It involves application of bed disinfectants, to prevent the spread of
infection within the colony of silkworms.

To avoid secondary contamination and spread of disease in the colony, silkworm body and rearing
seat disinfectants are to be applied. Periodic application of the bed disinfectants as per the
recommended schedule significantly prevents the spread of diseases during rearing. Application of
slaked lime powder at the beginning of each moult helps in absorbing moisture and regulates bed
humidity. Slaked lime being alkaline is an effective viricide and highly helpful in managing the viral
infections in the silkworm colony. Application of bed disinfectants after each moult is most effective
disease management practice. General silkworm body and rearing seat disinfectants are antifungal,
antibacterial, antiviral and antimicrosporidian in nature. Application of slaked lime dust in
combination with bleaching powder in and around rearing house and premises improves hygiene in
the rearing environment.

It has always been a practice to use silkworm bed disinfectants to contain the spread of diseases
during the progress of a silkworm crop. Several potential bed disinfectants have been developed in
India for disease management during silkworm rearing. Many popular bed disinfectants are in the
field for controlling various diseases of silkworms. To prevent secondary infection, occurrence and
spread of diseases in silkworm colony, silkworm body and rearing seat disinfectants are dusted over
the silkworm body and rearing seat following a schedule. Slaked lime powder is dusted every time the
larvae settle for moult. Other commercially available bed disinfectants are dusted while the larvae
come out of moult and mulberry is fed after half an hour of dusting the bed disinfectant. The
disinfectants are dusted at 3-5g/sq. ft. bed area. It is important to note that the bed disinfectants do not
cure silkworms from diseases and it only prevents the spread of diseases. Hence it should be used as a
preventive measure by dusting the powder formulation uniformly as per recommended quantity and
schedule.

The chosen bed disinfectant must be effective against all silkworm pathogens to prevent the spread of
all diseases. Certain precautions are essential while using bed disinfectant formulations. It should not
be dusted on the silkworms settled for moult or under moult and on eatable mulberry leaves available
in bed. While dusting eyes, nose, ear and mouth are to be protected. Hands should be washed
thoroughly after dusting.

Thus the topic can be summarized that, the disinfection is an essential part of silkworm rearing
for successful cocoon crop. Disinfection can be achieved by physical and chemical means or by
employing both. Selection of suitable disinfectant and disinfection method depends on the
rearing conditions or infrastructural facilities available with the rearer. Scheduling of
disinfection activity is essential for meticulous destruction of pathogens from the rearing
environment. Similarly, sanitation of the silkworm body and rearing seat is very vital for
controlling the spread and intensity of silkworm pathogens, thus saving the crops.

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