Chapter 5

Evaluation of Toxicity

5.3 TESTS WITH INSECTS

5.3.1 Topical Application

Topical application is the most commonly employed method of exposure for insects. The
insecticide is first dissolved in a relatively nontoxic and volatile solvent, such as acetone, and
then a known volume of the test solution is applied to the body surface using a microsyringe
accurate to 1 μL. Although topical application is one of the most accurate ways of treating the
insect, it is tedious, requiring too much time for some situations. Furthermore, some species,
due to size or behavior, cannot be treated successfully in this way.

5.3.2 Injection Method

Although the results obtained with the topical application procedure can be a very reliable
indication of the contact toxicity of any insecticide to an insect, the method does not indicate the
actual amount of insecticide entering the insect’s body. When this knowledge is required, the
injection method is usually employed. The insecticide is commonly dissolved in carrier solvent
such as peanut oil, propylene glycol, or acetone and injected into the body cavity. Injection is
usually made at the abdominal sterna or the intersegmental regions, avoiding the longitudinal
center line so that it does not injure the abdominal nerve cord. The needle is held in position for
a while and then pulled away gradually in order to avoid bleeding due to internal pressure
(Matsumura, 1985). Some limitations of this method include (1) difficulty finding a solvent that is
not toxic or that will dissolve sufficient insecticide to permit the injection of small volumes and (2)
the fact that some species cannot be injected (because of size or behavior).

5.3.3 Dipping Method

In the dipping method, the insects with a pair of forceps are picked up and dipped into the
insecticide preparation, which is either a suspension or an emulsion. A dipping net or a
screened container can also be devised for exposure of the insects for a few seconds. For
example, a ball-type metal mesh tea strainer (mesh size, 0.5 mm) was adopted for dipping
aphids in liquid solution for 10 s (Chandrasena et al., 2011). The dose is varied by increasing
the concentrations in a series of beakers. After treatment, the insects are placed in clean
containers, and mortality counts are made after a suitable interval.

5.3.4 Contact Method (Residual Exposure Method)

In this method, a known amount of the insecticide dissolved in a volatile solvent such as
acetone is added to a glass jar and then rotated, allowing the insecticide to coat the entire
surface while the solvent evaporates. The dose is varied by the amount of solution added to the
jar. Another method for testing deposits of insecticide is to apply a spray (solution or
suspension) to a Petri dish using a precision sprayer such as the Potter precision laboratory
spray tower (Figures 5.1 and 5.2). This device delivers known volume of spray in a fine mist that
settles on the surface to be treated. In tests with aphids or spider mites, a leaf substrate
contained in a Petri dish can be sprayed by the spraying apparatus.

Figure 5.1 Potter precision laboratory spray tower.

Figure 5.2 Sectional diagram of Potter precision laboratory spray tower. (From Potter, C. and Way, M.J., Precision spraying, in
Methods of Testing Chemicals on Insects, Shepard, H.H., ed., Vol. 1, Burgess Publishing Company, Minneapolis, MN, 1958, p.
154.)

5.3.5 Fumigation Method

This method is suitable for stored product pests. The evaluation of toxic vapor is a somewhat
similar operation. The insecticide is introduced into a sealed container along with insects, and
mortality is recorded at different intervals. Various fumigation techniques were described by
Busvine (1971), including (1) small-scale techniques for measuring the susceptibility of exposed
insects and (2) semipractical tests using large fumigation chambers.
5.3.6 Feeding Method
This is used for testing insecticides against larvae. For phytophagous insects, larvae are placed
individually in a container and starved for a few hours. They are then fed individually a small leaf
disk that had been topically treated with 1 μL of acetone containing various concentrations of an
insecticide. Mortality counts are made after 24 h.
Most of the previously mentioned methods are described in detail by Busvine (1971). These
methods are often used by pesticide companies in their screening of candidate insecticides or in
the comparison of formulations.

Extracted from:
Yu, S. J. (2015). The Toxicology and Biochemistry of Insecticides (2nd ed.). Boca Raton,
USA: CRC Press. pp. 104-106.

References of some procedures mentioned:

Chandrasena, D., Difonzo, C., and Byrne, A., An aphid-dip bioassay to evaluate susceptibility of
soybean aphid (Hemiptera: Aphididae) to pyrethroid, organophosphate, and neonicotinoid
insecticides, J. Econ. Entomol. 104, 1357, 2011.
Busvine, J.R., Critical Review on the Techniques for Testing Insecticides, Commonwealth
Agricultural Bureaux, London, U.K., 1971.