PHYSICAL AND CHEMICAL
METHODS OF
DETECTION OF INSECT PESTS
PHYSICAL AND CHEMICAL METHODS TO DETECT HIDDEN INSECTS
I. PHYSICAL METHODS
1. Visual examination:
Preliminary method
Damaged grains in the samples is indicated by loss in its
natural luster- Dull -lifeless.
Trained worker can very easily determine the qualities of grain
by its appearance.
[Link] damage and insect emergence holes:
Very simple and gives a good index of deterioration or damage to
the field workers
Does not indicate the damage going on inside the kernels
Per cent kernels damaged by insects including the germ eaten
kernels is assessed
100 g of grain sample is evenly spread out on a glass plate, the
damaged kernels are picked up to assess the percentage of insect
damaged kernels
3. Determination of foreign matter, insect infestation etc.
Using sieves insect debris, webbing, larvae, their cast off exuviae
etc, physically separated and hand picked
Rough idea of the damage
Foreign matter is also examined under microscope
4. Total damage: Quality test
Damage by heat, sprouted, mould and rotten wheat kernels are
physically separated.
This method is followed to assess the quality of grains–in
commercial
5. Germination tests:
Viability of the wheat and other food grains is reported in terms of
percentage of kernels developing strong sprouts under controlled
conditions
10 g seeds on the moist filter paper at optimum temperature.
Reduced viability indicates increase in deterioration
6. Loss in weight:
Estimated by determining volume weight ratio
A decrease in volume / weight ratio indicates an increase in
damaged kernels
(Und) – (DNu)
Dry mass loss (%) = -------------------- x 100
U (Nd + Nu)
Where,
Nu = Number of undamaged grains ;
Nd = Number of damaged grains
U = Dry mass of undamaged grains, and
D = Dry mass of damaged grains
7. Acoustic method
Sounds produced by the insects due to their movement or feeding
has been utilized in detecting insect infestation quantitatively
Possibility of detecting the dead insects are serious limitation
8. X-ray radiographic method
Internal insect infestation can be detected
All stages of development of the insects can be observed rapidly
Recently Polaroid radiographic media
9. Traps
TNAU Insect traps are excellent insect detection devices and more
effective in the detection of stored grain insects
TNAU Insect Probe Trap TNAU Pit fall trap Indicator device
10. Near Infrared Spectrometer (NIRS)
Detects and measures the chemical composition of biological materials.
Molecules comprising organic matter vibrate at frequencies corresponding to
wavelengths in the infrared region
Optical sensors measure this absorption and quantified
11. Development of a new loss-assessment method [MM]
This method is capable of determining the frass activity of beetles tunneling
through the dried chips
The increase in inner volume of a cassava chip is measurable by means of
vacuum equipment and can be converted into weight loss.
Vacuum equipment
B. Chemical method to detect Hidden infestation
[Link] of stains
a) Acid fuchsin
Mix 50 ml. glacial acetic acid in 950 ml of distilled water and add 0.5
g acid fuchsin
Soak grain samples in warm water for 5 minutes
Then immerse the soaked grain samples in the stain for 2 to 5 min.
Remove excess stain by washing in tap water.
Observe under microscope – egg plug stains show bright cherry red
While feeding punctures including mechanical injuries in light pink.
b) Gentian violet
Prepare 1% aqueous stock solution of gentian violet in 50 ml of 95%
ethanol.
Immerse the sample for 2 minutes in a solution containing 10 drops
Observe under microscope – egg plug stains show purple colour.
[Link] sulphate
Kernels are immersed in dilute solution of the dye (20 parts per
million) for one minute
Rinsed and examined under ultra violet light
Egg plug stains show intense greenish yellow under ultra violet light.
2. Floatation or density method
Involves the use of two solutions of different specific gravity.
Sodium silicate in water (sp. gravity 1.160 to 1.190) and
Methyl chloroform (sp. gravity 1.30) with debase oil
Grain is immersed in the fluids
A three layer separation occurs
Non-infested kernels sink to the bottom
Infested one floats and
Light weight kernels including those infested by early stages of
insect hang in the line of separation between the two fluids.
3. Gelatinization method
In this method the grain is boiled for ten minutes in 10%solution of
sodium hydroxide.
The treatment renders the kernels translucent, thereby, revealing
the presence of internal infestation
4. Cracking floatation method
Cleaned grain is coarsely ground to release the internal insects
Soaked either in a water-alcohol mixture or in boiling water
Mix with gasoline or mineral oil.
Insects floats with the oil layer in a flask
Collect it on a filter paper and count it
5. Fragment count or acid hydrolysis method
Presence of insects fragments such as elytra, head capsules,
mandibles, counted basically involving a flotation technique but
in a modified way.
A mixture of oil and aqueous phase (besides surfactants) is used in
making the fragments to float
Test involves digestion of a sample with an acid, wet sieving, or a de-
fating treatment using a detergent or solvent
Insect fragments which are olephilic, are separated from food
particles by the attraction of the oil phase (light) mineral oil in an oil-
aqueous mixture).
Floating fragments are trapped or filtered and examined under a
microscope. Infestation detection by the fragment count chocolate
and powdered spices.
6. Spectrophotometric analysis
De-hydroxyphenol occurring in insect cuticle is estimated by
spectrophotometer
Phenols produce certain dyes and are reacted with dichloroquinne
chlorimide.
7. Ninhydrin colour reaction: chemical indicator technique
Insects body fluid produces a colour with ninhydrin impregnate filter
paper (0.7% soln. in Acetone).
High sensitivity and specifically suitable to mechanization
A machine has also been manufactured in which wheat kernels are
crushed on a treated strip of paper tape and the same observed for
stains The machine is known as "Ashman Simon Hidden Infestation
Detector".
8. Carbon dioxide determination method
Quantity of carbon dioxide produced by a given sample of grain in
24 hours, is measured and the extent of internal infestation can be
estimated.
Representative grain sample free from moving insects in incubated
for 24 hours at 2.5°C.
The concentration of CO2 is measured with the help of gasometric
method accurate to ± 0.2 %.