PHYSIOTOXI Part1

PHYSIO&TOXI
GENERAL PHYSIOLOGY & TOXICOLOGY

PHYSIO&TOXI
Disclaimer
This learning materials is used in compliance with the flexible teaching-learning approach espoused by
CHED in response to the pandemic that has globally affected educational institutions. Authors and
publishers of the contents are well acknowledged. As such the college and its faculty do not claim
ownership of all sourced information. This learning material will solely be used for instructional purposes
not for commercialization.
CatSU College of Agriculture and Fisheries
Contact Details
Catanduanes State Colleges
College of Agriculture and Fisheries
SHARMAINE C. BIÑAS
Instructor 1
09301808718
scbinas.caf.csu@gmail.com
Module Cover Image Source: https://unsplash.com/photos/J-hYMYa2jY8
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Table of Contents
Disclaimer ................................................................................................................................................... i
Table of Contents ....................................................................................................................................... ii
Definition of Key Terms..............................................................................................................................iii
PRE-TEST ................................................................................................................................................ iv
Chapter 1 - THE CONCEPT OF INSECT PHYSIOLOGY AND TOXICOLOGY ...................................... 1
Lesson 1. Understanding insects ............................................................................................................... 2
Lesson 2. External Insect Structures and Functions ................................................................................. 6
Lesson 3. Principles of Toxicology .......................................................................................................... 17
QUIZ 1 ..................................................................................................................................................... 22
LABORATORY ACTIVITY 1 .................................................................................................................... 24
Chapter 2 – INSECTICIDE USE PATTERN ........................................................................................... 25
Lesson 1. The Need for Pesticides .......................................................................................................... 26
Lesson 2. Patterns and Trends of Pesticide Use .................................................................................... 27
Lesson 3. Strategies to Reduce Pesticide Use ....................................................................................... 29
Lesson 4. Mass Rearing and Preparation of Test Insects ....................................................................... 30
QUIZ 2 ..................................................................................................................................................... 36
Chapter 3 – INSECT PHYSIOLOGY ...................................................................................................... 37
Lesson 1. Insect reproduction, growth and development ........................................................................ 38
Lesson 2. The Digestive System, Nutrition and Excretion ....................................................................... 45
Lesson 3. The Circulatory System ........................................................................................................... 48
Lesson 4. The Respiratory System .......................................................................................................... 50
Lesson 5. The Muscular System, Locomotion and Flight ........................................................................ 52
Lesson 6. Coordination and Integration ................................................................................................... 55
Lesson 7. Molting and Formation of New Cuticle .................................................................................... 59
Lesson 8. Sensory systems ..................................................................................................................... 62
QUIZ 3 ..................................................................................................................................................... 72
Chapter 4 – INSECTICIDE RESEARCH, DEVELOPMENT AND BIOASSAY ...................................... 77
Lesson 1. Research and Development Process ..................................................................................... 78
Lesson 2. Data Generation and Value Safety ......................................................................................... 79
Lesson 3. Economic and Legal Aspects of Insecticide Use .................................................................... 83
Lesson 4. Types of Bioassay and the Factors to Consider ..................................................................... 86
Lesson 5. Probit Analysis ........................................................................................................................ 89
QUIZ 4 ..................................................................................................................................................... 94
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Definition of Key Terms

Axial orientation of a bilaterally symmetrical body (refer to the figure):
Anterior – near or toward the head (or part most nearly

corresponding to a head)
Cephalic – directed toward or situated on or in or near the head
Distal –situated away from the point of origin or attachment or
central point of the body; apical
Dorsal – back; upper surface opposite the abdomen
Dorsolateral – involving or extending along the axis joining the
dorsal and ventral sides
Dorsoventral – involving the back and the sides
Lateral – relating to the side; situated on or directed toward or
coming from the side; extending from side to side;
transverse; closer to the body margin
Posterior – behind; caudal
Proximal – next to; nearest to the point of origin or attachment or central point of the body; basal
Ventral – abdominal; lower surface opposite the back
Medial – mesal; nearer to the midline,
Entomological/toxicological terms:
Apolysis - separation of old exoskeleton from epidermis.

Arthropod – any species belonging to Phylum Arthropoda such as insects, arachnids and crustaceans
having segmented body and jointed appendages, chitinous exoskeleton molted at intervals and
dorsal anterior brain connected to ventrally connected ganglia
Bioassay – the measurement of the potency of any stimulus physical, chemical, or biological, physiological
or psychological by means of the reactions which it produces in living matter.
Chitin – polysaccharide (C8H13NO5); comprises the hard, outer integument of arthropods
Communication- defined as any exchange of information between individuals
Cuticle – the external envelope of an insect secreted by the epidermis
Ecdysis – an act of molting or shedding of an outer cuticular layer
Hexapod – relating to an insect
Hormones - biologically active compounds which are produced to control certain life processes and
development.
LD50 – median lethal dose; represent the concentration (LC50) or dose (LD50) at which 50% of the
population responds
Metabolism - is the total of all the chemical and physical processes which take place within the organism.
Pesticides - any chemical substance intended for preventing, destroying, or controlling any pest, including
vectors of human or animal disease, unwanted species of plants or animals, causing harm during
or otherwise interfering with the production, processing, storage, transport, or marketing of food,
agricultural commodities, wood and wood products or animal feedstuffs, or substances that may
be administered to animals for the control of insects, arachnids, or other pests in or on their bodies.
Tanning - sclerotization of new exocuticle
Toxicology- the study of limits of the biological effects of a chemical or mixtures of chemicals.
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PRE-TEST
Let’s see how familiar you are on insects and toxicology.
I. The science of insects
Choose the BEST answer among the choices.

1. It is the branch of zoology which deals with the study of insects.
a. Entomology
b. Microbiology
c. Botany
d. Ornithology
2. Insects belong to Phylum Arthropoda. Which of the characteristics are unique to arthropods that
set them apart from any other animals?
a. Segmented bodies
b. Ventral nervous system
c. Chitinous exoskeleton
d. All of the above
3. Identify which Order do these insects belong:
Coconut rhinoceros Oriental migratory Dragonfly (Anisoptera) Black bean aphid

beetle (Oryctes locust (Locusta (Aphis fabae)
rhinoceros) migratoria manilensis)
Order ___________ Order ___________ Order ___________ Order ___________
II. External anatomy of insects
Identify the external structures of a grasshopper.

H I J
A. ________________
B. ________________
C. ________________ G
D. ________________ . F
E. ________________
F. ________________
A
G. ________________
. B
H. ________________
I. ________________
J. ________________ C
.
E
D
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III. Internal anatomy of insect
Label the parts of the internal anatomical structures of an insect. Model: grasshopper
C.
A. B.
D.
E.
F.
G.
O. H.
N.
I.
M. J.
L. K.
IV. Insect toxicology
True or False. Read each statements carefully. If the statement is true, write T otherwise write F if it is false.
_______ 1. Insecticide is used to kill or eliminate insect pests in agriculture, household and industries.
_______ 2. Insecticide toxicology differs from medical toxicology (humans) as it does not include clinical
diagnoses or treatment of human patients who are affected by insecticides.
_______ 3. DDT or dichlorodiphenyltrichloroethane is an organochlorine originally developed as pesticide
in 1874 has been widely used until today.
_______ 4. The first compounds used as pesticides include sulfur and arsenic.
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Chapter 1
THE CONCEPT OF INSECT PHYSIOLOGY

AND TOXICOLOGY
OVERVIEW
Entomology, defined as the science of insects, includes an array of biological disciplines including
evolution, ecology, behavior, anatomy, physiology, biochemistry and genetics. Entomologists work with
insects due to ease of culturing in the laboratory, rapid population turnover, and ecological abundance as
well as the minimal ethical concerns in experimental use (Gullan, 2014). There are many reasons why we
should study insects. Because of their unparalleled diversity, insects can be used to provide a better
understanding of nature and biological systems. In agriculture, knowledge about insects is important as we
manipulate ecosystems for increased food production – some entomologists believed that insect control is
imperative for human survival, as some insects are pests to agricultural crops.
Insect pests are notorious in causing production losses each year. The study of physiology is vital in
understanding toxicology of insects to pesticides. As an introduction to the course, this chapter introduces
the science of insects and toxicology - the insect’s external and internal anatomy, as well as the different
patterns of insecticide use are discussed in this chapter.
LEARNING OUTCOMES
At the end of this chapter, the students shall be able to:
1. Identify the importance of insect physiology to pesticide toxicology.

2. Identify and describe the basic structures of the external anatomy of a generalized insect.
3. Identify and discuss the functions of the basic structures, morphological adaptations of insects
(specialized mouthparts, legs, wings and head)
4. Discuss the important historical events in insect toxicology.
5. Enumerate the types of pesticides and how they are applied
6. Determine how toxicity is measured.
MOTIVATION
“A bumblebee is not designed to fly. Its body is too heavy for its light wings, wings that should not be able
to keep in the air. Bumblebees refuse to accept their limitations. It flies in spite of what it’s been told it
cannot do. It knows its strength.”
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Lesson 1. Understanding insects

Insects have lived on earth for nearly 350 million years and have evolved to develop adaptations in
almost all types of habitats and adapted morphological changes to survive. Today, insects are the most
dominant group of animals outnumbering all other terrestrial animals, occurring almost anywhere.
Entomology and entomologists
Entomology is the science of studying insects and entomologists are the people who study,
observe, collect, rear, and experiment with insects. The word “entomology” is derived from two Greek
words, “entomon” which means insect and “logia” (or “logos”) which means to study. In Greek, it literally
means “cut up or divided into segments.” Entomology is a branch of zoology, but still is a broad field
including many biological disciplines such as evolution, ecology, behavior, anatomy, physiology,
biochemistry and genetics, all unified to studying the specialized science of insects. People work with
insects due to the following reasons:
- Ease of culturing in the laboratory;
- Rapid population turnover;
- Abundance in the environment;
- Minimal ethical concerns compared to vertebrates; and
- A good model in understanding biological concepts of higher animal forms
Importance of insects in agriculture and human life
1. Insects as pests
One negative impact of insects to agriculture is their role as biotic stress on crops. There are
hundreds of insect species that cause tons of production losses. Insects are mainly controlled using
chemical pesticides which is the main source of degradation of the soil and a number of human and
animal health problems.
a. b. c. d.
Figure 1. Examples of insect pests causing damage to agriculture. (a) Fall armyworm (Helicoverpa armigera) infesting corn cob in the
field; (b) rice weevil (Sitophilus oryzae) feeding on rice grains; (c) black aphid (Pentalonia nigronervosa) transmitting Bunchy
Top Virus in abaca and (d) lice found in cattle.
2. Insects as human food (entomophagy)
Consumption of insects as food in increasingly gaining popularity. About 2000 insect species
belonging in more than 100 families are or have been used for food, especially in the central and
southern Africa, Asia, Australia and Latin America. It is said that insects are part of the traditional diet,
however due to ever increasing population and demand for food, exploitation of wild edible species
became widespread. Insects are rich in protein, vitamins and minerals and energy, contributing 5% to
10% of animal protein consumed by indigenous people. Edible insects only include those feeding on
either living or dead plant matter. Examples of these are termites, crickets, grasshoppers, locusts,
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beetles, ants, and bee brood and moth larvae. In the Philippines, June beetles (Phyllophaga sp.),
weaver ants (Oecophylla smaragdina), mole crickets (Gryllotalpa sp.), palm worms (Rhyncoporus sp.)
and locusts (Locusta migratoria L.).
3. Insects as feed for domesticated animals

Insects as alternative protein source for domesticated animals such as fish, poultry, pigs and
farm-grown minks is more acceptable. The insects involved include pupae of silkworms (Bombyx mori),
larvae and pupae of house flies (Musca domestica), and the larvae of mealworms (Tenebrio molitor).
4. Pollinators
Pollinators are flower-visiting insects that feed on nectar extracted from pollens of flowers. Bees
and butterflies forage on one flower to another thus are able to transfer the male gametes contained in
the pollen to the female gametes, resulting to pollination. Insect-mediated pollination is an important
step in the reproduction of many agricultural crops and ornamental plants. Many plants depend on
pollination for seed production and fruit set to achieve good yield. An estimated 35% of crop production
is a result of insect pollination.
5. Natural enemies
Natural enemies are insect predators and parasitoids that attack and feed on pests (insect pests
and pathogens (disease-causing microorganisms)) are important in the regulation and management of
pests (biological control). Natural enemies prevent crop pests from reaching economically damaging
levels by contributing to pest mortality.
Generally, predators are characterized as free-living, mobile, larger that the insect the prey and are
able to consume several preys throughout their life cycle. The most common insect predators are beetles,
true bugs, lacewings, wasp, dragonflies and mantis. Predators are classified as generalists or specialist.
Generalist predators feed on a wide variety of prey or even, at times,
each other while specialist predators feed on only one or few species
of prey. Unlike predators belonging to several insect orders, parasitoids
mainly belong to two orders: Hymenoptera and Diptera. Parasitoids
parasitize different life stags of their host (e.g. egg, larva, pupa and
A
adult). Adult parasitoids lay egg(s) on or within their host to complete
their development and eventually killing the host. Trichogramma egg
parasitoids are successfully used for biological control against a wide
range of Lepidopteran insects worldwide. In the Philippines,
Trichogramma evanescens (Hymenoptera) was found effective in
controlling corn borer, tomato fruitworm, sugarcane borer, eggplant
shoot and fruit borer, cacao pod borer and other lepidopterous insect B
pests. Figure 2. Preying mantis attacking another
insect (b) Female Trichogramma on
an armyworm egg.
6. Soil builders
Insects living in soils make tunnels, allowing smaller organisms, water and air to pass through.
Insects help in soil aeration and enhance nutrient cycle soil structure and tilth.
7. Production of honey, beeswax, silk and shellac

Honey and beeswax is a unique food since it can only be produce by honeybees (Apis spp.
Hymenoptera). Likewise, silk is only produced by silkworm (Fig. 3a). Shellac is produced by female
Laccifer lacca, a species of scale insect that secretes hard deposit or resin on trees (Fig. 3b&c). The
resin is processed to make liquid shellac used as brush-on colorant and wood finish.
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A B C
Figure 3. Unique commercial products produced by insects. (A) Silk worm (Bombyx mori) producing silk and (B) Lac scale insect
(Laccifer sp.) and (C) resin in trees produced by lac scale insect.
Insect nomenclature and classification
Kingdom: Animalia
Phylum: Arthropoda
Class: Insecta
Order: Hymenoptera
Family: Aphidae
Genus: Apis (Linnaeus, 1758)
Species: mellifera
Binomial nomenclature: Apis meliifera
A B
Figure 4. Arthropod design. (A) Classes of arthropods showing body segmentation and (B) theoretical evolution of arthropods from
annelid-like organism.
Arthropoda (arthros = joint; pous or poda = foot) constitutes the largest phylum of the animal
kingdom. Although differing in some structures, they agree in some fundamental characters (Fig. 4A):
- Externally segmented bodies and appendages
- Has appendages modified for feeding
- An exoskeleton with chitin
- A hemocoel instead of a coelom
- No cilia
- A ventral nerve cord and dorsal brain
No animals other than Arthropods exhibit the above combination of characters. Living in a wide
variety of habitats, developed diverse types of locomotion, have a wide range of morphological variations
and feed on a variety of food, arthropods constitute the most biologically successful groups of animals,
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evident in their great number of species. Arthropods are believed to have

evolved from annelid-like ancestor (Figure 4B). Three major lines forming the
sub-phyla are seemed to have evolved from this “ancestor”. These are:
Mandibulata or those with developed mandibles, Chelicerata or those using
chelicerae for food and Trilobitamorpha, known as the ancestor of aquatic
arthropods (only known in fossils) and had not modified their appendages for
feeding (Fig. 5).
Figure 5. A trilobite fossil.
Classes
Phylum Arthropoda
Class Onychophora Arachnida Crustacea Myriapoda Insecta
“myrios” = ten
“onychos” = claw; “insectus” =
Etymology “Arachne” = spider “crusta” = shell thousand; “podus”
“phoros”= bearing divided
= foot
--mostly terrestrial --mostly terrestrial and -mostly aquatic, -mostly are -mostly terrestrial
-small sized arthropods rarely aquatic few are terrestrial, terrestrial and rarely
-segmented body -body is divided into few are parasitic -Many aquatic (aquatic
respire through trachea cephalothorax and -cephalothorax appendages; at the early life
-excretion is through abdomen (head is fused with body is long with stages)
nephridia -four pairs of legs the thoracic numerous -body is divided
Example: Paripatus -respire through segments segments each into three
trachea, gills or book -thorax and having one or two regions: head,
lungs abdomen have a pairs of legs thorax nad
-excretion is through pair of biramous -Head is distinct abdomen
Malphigian tubules appendages in with antennae, a -wings and legs
coxal glands or both each segment pair of eyes and on thorax
Examples: spider, -Respiration is two to three pairs -abdomen has 7-
Unique
scorpion, king crab through the gills or of jaws 11segments
characters
general body -excretion is without
surface through appendages
-Excrete through Malphigian -respire through
modified tubules trachea, gills
coelomoducts -Respiration is Excrete through
(either maxillary or through trachea Malphigian
antennary glands) Examples: tubules
Examples: crab, millipede, Examples:
prawn, water flea centipede, butterfly,
spirobolus cockroach,
housefly,
mosquitoes, ants
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Insect orders
Figure 6. Phylogeny of insect orders.
Lesson 2. External Insect Structures and Functions

All species belonging to Class Insecta is
characterized as six-legged adults (Hexapoda = Greek:
hexa means six, poda means foot). The insect body is
divided into tagma or regions: the head, thorax and
abdomen, bearing appendages which may vary from one
order to another, or species to species (Fig. 7). Likewise, the
differences in the anatomy of insect mouthparts, legs, wings
and abdominal appendages are important in the
identification of each insect orders, families and genera. The
understanding of the external anatomy is necessary in the
elucidation and appreciation of the functions of various
insect forms and distinction from other hexapods. Figure 7. Morphology of a generalized insect.
The Head
The insect head (Fig. 8) is a rigid capsule formed from several sclerotized plates and has two
openings, one posteriorly through the occipital foramen to the prothorax, the other to the mouthparts. It
bears one pair of compound eyes located dorsa-laterally and sometimes, with light-sensitive simple eyes
Figure 7. Morphology of a generalized insect 6
Chapter 1 – The Concept of Insect Physiology and Toxicology FF
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called ocellus (plural: ocelli) arranged in triangle at the anterior vertex. Insect eyes may either be: a.)
dichoptic – eyes are separated b.) holoptic – eyes are fused into each other. It also bears a pair of antenna
placed medially.
Figure 8. Front and back view of a typical winged insect.
Regions on the head:

 The posterior horseshoe-shaped posterior cranium (dorsally the occiput) links the vertex dorsally
and the genae (singular: gena) laterally; the vertex adjoins the frons anteriorly and more anteriorly
lies the clypeus, both of which may be fused into a frontoclypeus.
 The regions of an insect head are weakly delimited showing external grooves or lines on the head
called sutures (sulcus = shallow grooves or furrows):
o (1) remnants of original segmentation, generally restricted to the postoccipital suture
o (2) ecdysial lines - where the head capsule of the immature insect splits at molting, often
prominent inverted “Y”
 epicranial suture
 frons (also called frontal sutures)
o (3) Grooves showing the underlying internal skeletal ridges (frontoclypeal or epistomal
suture) that delimits the frons from the more anterior clypeus.
 Several invaginated ridges and arms (apophyses, or elongate apodemes) on the head
endoskeleton.
o two pairs of tentorial arms (anterior and posterior)
o Some of these arms may be absent or fused to form the tentorium, an endoskeletal support.
 Pits on the surface of the cranium where tentorial arms invaginate
There are two different types of head structures based on the inclination of the long axis of the head
and orientation of mouthparts (Fig. 9):
1. Prognathous type
- “pro” means in front; “gnathous” means jaw
- The head lies at similar axis as the body and mouth parts (e.g. beetle)
2. Hypognathous type
- “hypo” means below; “gnathous” means jaw
- The head remains vertical and is at right angle to the long axis of the body
- Mouthparts are ventrally located and oriented downwards
- E.g. grasshopper, cockroaches
3. Opisthognathous type
- “opistho” means behind; “gnathous” means jaw
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- Almost similar as the prognathous but the mouthparts are directed backwards and held in
between the forelegs
- E.g. Hemiptera
A B C
Figure 9. Types of insect heads according relative to body axis and orientation of the mouthparts. (A) Prognathous (B)
Hypognathous and (C) Opistognathous. (Photo source: jscienceclass.blogspot.com)
The Mouthparts
From the appendages of the head segments such as the labral, mandibular, maxillary and labial, the
insect mouth is formed. Modifications in the morphology of the mouthparts are due to the feeding
mechanisms of modern insect species, incomparable with the omnivorous insects such as crickets,
cockroaches and earwigs possessing biting or chewing mouthparts (mandibulate), a representative design
of the ancient pterygote insect mouth.
There are five basic components of the mouthparts (Fig. 10):

1. Labrum – the upper lip; with ventral surface called the epipharynx;
2. Hypopharynx – tongue-like structure;
3. Mandible – or jaw;
4. Maxilla – (pl. maxillae); pincers but less powerful than the
mandibles; and
5. Labium – lower lip
The labrum forms the roof of the pre-oral cavity and mouth
covering the base of the mandibles. Projecting from the back is
the hypopharynx, a lobe assumed to be associated with the
mandibular segment. In earwig and nymphal mayflies, the
hypopharynx bears lateral lobes or the superlingua (pl.
superlinguae) dividing the cavity into a dorsal food pouch
(cibarium) and ventral salivarium from which the salivary duct
opens. The mandibles, maxillae, and labium are paired
appendages and varies from one insect order to another.
Mandibles are responsible in cutting and crushing solid
food and may be used in defense at times. The mandibles consist
of the apical cutting edge and the basal molar area for food
grinding. Approximately, mandibles read a rating of 3 on the
Moh’s scale of mineral hardness, characterized as extremely
hard structures. This is evidently described on how termites and
beetles can easily bore through woods and foils made from
metals such as copper, lead, tin and zinc. The maxillae lies
behind the mandibles consisting of a basal part composed of the Figure 10. Insect mouthparts.
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proximal cardo and more distal stipes. Attached to the stipes are the two lobes, the mesal lacinia (pl.
lacinae), the lateral galea (pl. galeae) and the lateral segmented maxillary palp or palpus (pl. palps or palpi).
The main function of the maxillae is to assist the mandibles in the processing of food. The pointed and
sclerotized lacinae hold and macerate the food while the galeae and palpi serve as mecho- or
chemoreceptors (taste-test of food before ingestion). In earwigs, the labium is attached to the ventral
surface of the head through the postero-ventral surface of the head via a sclerotized plate, the gula. The
main parts of the labium are: postmentum, prementum, labial palpi, glossa (pl. glossae) and paraglossa
(pl. paraglossae). The lobes of the prementum forms the floor of the pre-oral cavity (lower lip) while the
labial palpi are sensory structures with functions similar to the maxillary palpi.
Evolution resulted to an array of morphological

changes on the feeding structures of the basic
mandibulate mouthparts. These feeding structures is a
distinguishable character useful in both taxonomic
classification and identification as well as ecological
diversity and mechanisms of insects. Fig. 11 shows the
different specialized mouthparts according to feeding
habit.
Figure 11. Specialized mouth parts of insects. (A) mandibulate/chewing

type of grasshopper, (B) lapping type of bee, (C) siphoning
type of a butterfly and (D) piercing-sucking type of a female
mosquito. Legend: a, antennae; c, compound eye; lb,
labium; lr, labrum; md, mandibles; mx,
maxillae hp hypopharynx.
The Antenna
Aside from the eyes, another sensory structure of an insect is their antenna (pl. antennae). Antennae
are mobile, segmented, paired appendages of the insect’s head. Typically, an entire insect antenna
consists of (Fig.12):
a. Scape – first and largest among the segments;

basal stalk
b. Pedicel – second segment; usually containing the
sensory organ called the Johnson’s organ which
responds to the distal movement of the antenna
relative to the pedicel
c. Flagellum (pl. flagella) – last segment; often
filamentous and multi-segmented (flagellomeres);
may be reduced or modified.
Figure 12. Structure of Antenna
Several sensilla (sing. sensillum) such as hairs, pegs, pits, or cones are in the antenna functioning
as chemo-, mechano-, thermo- and hygroreceptors.
Main Types of Antenna (Fig. 13)
a. Aristate – with enlarged third segment bearing a bristle (e.g. most Diptera, true flies)
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b. Capitate – (Latin “caput” = head); distinctly clubbed or terminates abruptly forming a club (e.g.
butterflies)
c. Clavate – (Latin “clava” = club); antenna terminate into a gradual club (e.g. some beetles)
d. Filiform – (Latin “filum” = thread); thread-like with untampered segments and uniform widths; linear
and slender (e.g. grasshoppers)
e. Flabellate – (Latin “flabellum” = fan); the terminal segments extends laterally, with long parallel lobes
that lie flat against one another. (e.g. orders Hymenoptera, Coleoptera and Lepidoptera)
f. Geniculate – (Latin “genu” = knee); antenna is bent or elbowed (e.g. ants and bees)
g. Lamellate – (Latin “lamella” = thin plate or scale); segments at the tips are flattened and nested
looking like a folded fan (e.g. scarab beetle)
h. Moniliform – (Latin “monile” = necklace), like a string of beads; segments are usually spherical and
uniform in sizes (e.g. termites)
i. Pectinate – (Latin “pectin” = comb-like); the segments are longer on one side, giving each antennae
a comb-like shape; bipectinate looks like two-sided combs (e.g. beetles and sawflies)
j. Plumose – (Latin “pluma” = feather); segments have fine branches; bearing whorls of setae (e.g.
mosquitoes and moths)
k. Serrate – (Latin “serra” = saw); segments are notched and angled on one side like a saw blade (e.g.
some beetles)
l. Setaceous – bristle-like with segments becoming thinner towards the top (e.g. cockroaches)
m. Stylate – (Latin “stylus” = pointed instrument); the final segment terminates in a long, slender point
called style. The style may be hair-like but will extend from the end and never on the sides (e.g.
Brachycera, horse flies)
(j)
a b c d e f g
h i j k l m
Figure 13. Main types of insect antenna.
The Thorax
Insects have three thoracic segments: prothorax, mesothorax, and metathorax (Fig. 14a) and
each bear one pair of legs. Each section has a dorsal sclerotized region (tergum or notum), a ventral
region (sternum) and laterally on each side, a pleuron all joined together by non-sclerotized membranous
cuticle (Fig.14b). These may be fused or subdivided into sclerites (e.g. tergites, sternites and pleurites). A
pair of spiracles or openings into the respiratory system are found between the pro- and mesothorax and
between the meso- and metathorax.
In addition to movement by appendages, most insect adults are able to fly enabling them to drive to
almost any habitat. In winged insects, the mesothorax and the metathorax forms the pterothorax (wing-
bearing), which are bigger relative to the prothorax. Almost all nymph and adult insects have three pairs
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PHYSIO&TOXI
of legs, each pair located on one thoracic segment. A pair of spiracles or openings important in the gas
exchange of the tracheal system are present on the lateral sides of the meso- and metathorax.
Figure 14. The insect thorax (a). Dorsal sclerotized region (tergum or notum), a ventral region (sternum) and lateral pleuron (b).
The Wings
Insect wings are only present (fully developed) in adult insects (exception is the presence of wings
in the subimago of Ephemeroptera, e.g. mayfly). Typically, an insect has two pairs of well-developed and
functional wings which are flap-like cuticular projections supported by sclerotized veins. A pair of forewings
are attached on the mesothorax while the pair of hindwings are attached to the metathorax, both dorsa-
laterally. In some insects the meso- and metathorax fuse for increasing strength. The major regions of the
wings are shown in Fig 15a.
Wing venation is based on a combination of speculation and fossil data called the archedictyon (Fig.
15b). According to current dogma, the archedictyon contained 6–8 longitudinal veins named according to
the system developed by John Comstock and George Needham called the Comstock–Needham system.
- Costa (C) – the leading edge of the wing
- Subcosta (Sc) – second longitudinal vein (behind the costa), typically unbranched
- Radius (R) – third longitudinal vein, one to five branches reach the wing margin
- Media (M) – fourth longitudinal vein, one to four branches reach the wing margin
- Cubitus (Cu) – fifth longitudinal vein, one to three branches reach the wing margin
- Anal veins (A1, A2, A3) – unbranched veins behind the cubitus
a. b.
Figure 15. Major regions of an insect wing and venation.
The hindwings are greatly reduced in Diptera and are called halteres (Fig. 16a) – used for balance,
so there is only one pair of wings attached to the highly developed mesothorax. Insect wings are basically
membranous and transparent however, variations occur in the size, shape, and degree of sclerotization of
the fore- and hindwings. The following are the types of insect wings (Fig. 16b):
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PHYSIO&TOXI
a. b.
Figure 16. Houseflies have reduced hindwing or haltere shown by the yellow pointer (a). Types of insect wings: (from upper
left to lower right) Tegmen (pl. tegmina), Elytron (pl. elytra), Membranous, Scaly and Hemelytron (pl. hemelytra) (b).
The Legs
Insects have three pairs of segmented legs: the

fore-, mid- and hind legs each attached on the pro-,
meso-, and metathorax, respectively; and each leg has
six segments, from proximal to distal: coxa, trochanter,
femur, tibia, tarsus and pretarsus (or post-tarsus)
(Fig. 17). Additionally, found in some fossils of insects
and arachnids, and in Ephemeroptera and Odonata are
prefemur, patella and basitarsus.
The tarsus appears to be subdivided into smaller

Figure 17. The hind leg of a cockroach, Periplaneta
segments however, there is only one tarsal muscle
americana.
(tarsomere) hence more appropriately called as a
pseudosegment. The first segment is sometimes called the basitarsus but should not be confused with the
“true” basitarsus segment. On the underside of the tarsomere are ventral pads, either hairy or smooth pads
called pulvilli necessary in adhesion to surfaces. The pretarsus located terminally bears a pair of lateral
claws (called ungues) and a median lobe (called arolium). In Diptera such as the house fly, their pretarsus
consists of a spine-like or pad-like empodium and a pair of lateral pulvulli which allow flies to walk on wall
and ceilings. Variations on these structures such as the presence of adhesive setae facilitates climbing of
beetles.
The femur and tibia are the longest segments but vary in length and robustness relative to their
functions. The following are types of insect legs based on specialization (Fig. 18):
1. Walking (gressorial) and running (cursorial)

- have well-developed femora and tibiae on all legs (E.g. cockroaches)
2. Jumping (saltatorial)
- disproportionately developed hind femora and tibiae. (E.g. grasshoppers)
3. Swimming (natatorial)
- the tibiae and/or one of the tarsi of one or more legs are modified. (E.g. aquatic beetles, and bugs)
4. Digging (fossorial)
- the tibiae of the fore legs are enlarged and modified for digging. (E.g. ground-dwelling insects such
as mole crickets, nymphal cicadas and scarab beetles)
5. Seizing of prey (raptorial)
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PHYSIO&TOXI
- found in fore legs of predatory insects. (E.g. preying mantis)

6. Collecting and carrying
- the tibia and basal tarsomere modified for collection and carriage of pollen (E.g. honeybees)
Preying mantis -
raptorial
Honeybee - collecting
Figure 18. Insect leg modifications.
The Abdomen
A primitive insect abdomen has 11

segments although segment 1 may be
reduced or fused into the thorax and the
terminal segments are usually modified and/or
diminished. More generally, the first seven
abdominal segments (pre-genital) of an adult
insect contain no appendages except for
apterygotes such as bristletails and silverfish
as well as immature aquatic insects which Figure 19. Section of Abdomen
contain rudimentary appendages called styles
showing ventral surface of the pretarsus and the last tarsomere.
and one or two pairs of protractible vesicles on some abdominal segments. Each segment is made up of a
dorsal sclerite (tergum) and a ventral sclerite (sternum). These are joined to side pieces (pleurons) by
flexible non-sclerotized cuticle.
The External Terminalia
Terminalia, a collective term for the anal-genital parts of the insect abdomen, starts from segment 8
or 9 up to the abdominal apex. Articulating laterally on segment 11 is a pair of cerci, commonly annulated
or filamentous (Fig. 20). The structures comprising the external terminalia of an adult female insect is
necessary for oviposition or egg-laying. The ovipositor is the egg-laying tube mostly present in adult female
insects, except in female termites, parasitic lice, many Plecoptera and most Ephemeroptera. In male
insects, the external genitalia include a
spermatophore, the organ used to transfer the
spermatozoa, involving structures that grasp and
hold the female during mating.
Figure 20. The external genitalia starts from the 8th segment of the insect abdomen (A). Structures of the external genitalia is
important in the identification of insects such as the presence and absence of style in cockroaches (B).
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PHYSIO&TOXI
The Integument
The epidermis-cuticle complex is called the integument, the outer layer of the insect body (Fig.
21).
Roles and Functions of the integument (epidermis and cuticle formed together):
1. Determines the habit of the insect body (form, surface markings)
2. Protects the internal structures against harmful external effect (mechanical, physical, chemical and
biological)
3. Keeps water, ionic and thermal balance; and gas exchange
4. External skeleton (exoskeleton) providing places for muscle attachments within the body
(endoskeleton)
5. Other functions of the integument are: formation of fore- and hindgut walls, external genitalia,
trachea system and sensory organs.
Importance to insecticide toxicology: (1) site of penetration of contact insecticides and (2) prone to
desiccation by detergents.
Parts of the integument

1. Basement membrane (membrana basalis) - the innermost component, porous acellular layer,
0.5 μm thickness, and main components: proteins, glycoproteins, collagen produced by the
epidermal cells.
2. Epidermis - more or less continuous single layer (one-cell-thick), with polygonal or hexagonal
arranged epidermal cells
Procuticle
Figure 21. Structure of the Integument

3. Cuticle
The cuticle is a vital contributor to the success of insect life. The cuticle is an inert layer which
provides not only strength to the exoskeleton of the body and limbs, apodemes (internal support and muscle
attachment), and wings but also protection of the internal structures against harmful effects of the
environment including mechanical, physical, chemical and biological. Likewise, restriction of water loss is
a critical function of the cuticle in insects. The cuticle may range from rigid and armor-like such as in beetles,
to thin and flexible such as in larvae.
General features of the cuticle

1. Two-fold character
a. lipophile, hydrophobic epicuticle
b. hydrophile, lipophobic procuticle
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PHYSIO&TOXI
2. Functional asymmetry- water can easier get into the cuticle than get out
Composition of cuticle
A. Chitin
Chitin is an unbranched polymer of high molecular
weight, an amino polysaccharide mainly consisting of N-
acetyl-D-glucosamine (C8H13NO5) monomers linked in β-1,4
configuration (Fig. 22). Chitin molecules (18 to 20 chitin
chains) are bundled and assembled covalently into flexible
microfibrils embedded in and intimately linked to a protein
matrix giving rise to great tensile strength. The most common Figure 22. Structure of part of a chitin chain
arrangement of chitin microfibrils is a sheet wherein showing two-linked units of N-acetyl-
microfibrils are arranged in parallel. D-glucosamine.
The tensile strength of the cuticle is derived from the
hydrogen bonding of the chitin molecules and additionally, from sclerotization – an irreversible process
resulting to a darkened exocuticle and insolubility of proteins in water.
B. Proteins
- 40-80% of the dry weight of the procuticle
- lots of different proteins, more than 100; in a soft type of cuticle they are more hygrophilous
- many of them are soluble in water (arthropodin)
- they can make linkages with chitin chains (chitinoproteins); linkage can be loose (H-bonding) or
close (quinone)
- special protein: resilin, which is rich in glycine amino acid and elastic like rubber, colorless and
transparent
4. Epidermal layers
Despite being a thin layer, the structure of cuticle is complex and still of research importance. A layer
of cell beneath the cuticle where it is secreted is the epidermis, which consists of a procuticle overlaid
with thin epicuticle. The epidermis is subject to molting or shedding of the old cuticle.
A. Epicuticle
Ranging from 1 µm to 4 µm, the epicuticle is a thick layer usually consisting of three levels: the inner
epicuticle, an outer epicuticle (or cuticulin layer), a lipid or wax layer, and a cement layer (Fig. 21).
The inner epicuticle is a loose, thick (0.5 μm–2 μm) comprising of tanned lipoproteins. The outer
epicuticle is a thin (5 nm–20nm), compact and darker layer comprising of lipoproteins and lipids. The most
external layer of the cuticle is the wax layer mainly composed of lipids and waxes with a variable discrete
cement layer atop. The wax layer consists of saturated aliphatic carbohydrates, aliphatic alcohols (12-50
C atoms), esters and free fatty acids (12-34 C atoms) and is a product of oenocytes. Following this free-
wax layer is the superficial layer composed of glycoproteins. The cement layer is a hard, generally thin
and discontinuous layer consisting of mucopolysaccharides and lipids. Present only in certain species, the
cement layer serves as a protective layer and excretion of special epidermal gland cells.
The chemical composition of these outermost layers is essential in preventing dehydration, due to
the water-repelling property of lipids, waxes, and protein-bound lipids. Besides water retention, waxes on
the surface of the insects may deter predation, provide patterns for mimicry (camouflage), repel excess
rainwater, reflect solar and ultraviolet (UV) radiation or provide species-specific olfactory signals.
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PHYSIO&TOXI
B. Procuticle
The inextensible and unsupportive properties of the epicuticle is supported by the underlying
chitinous layer or the procuticle. Later, the procuticle differentiates into thicker endocuticle covered by a
thinner exocuticle (thinner due to sclerotization of the endocuticle) (Fig. 21). The procuticle range from 0.5
µm to 10 µm thick mostly made up of chitin and protein (in contrast with the non-chitinous epicuticle.)
Procuticle layers:
a. Exocuticle: mostly rigid, hardened, dark colored, pigmented upper layer
Properties: produced before molting; sclerotized and tanned after molting; former exocuticle is
removed during molting.
b. Endocuticle: generally soft, flexible, bright, pigment free lower layer
Properties: produced after molting; will be dissolved during next molting and its ingredients will be
utilized
c. Mesocuticle: produced rarely, similar to exocuticle
Properties: produced after molting; tanned and sclerotized
Pore Canals (wax canals)

- helically tend upwards through the procuticle; branch out within the epicuticle
- their cross-section can be round or flat, with1 μm in diameter
- there are 30-200 canals above one epidermal cell, which means 15000 canals/mm 2
- they contain plasma fibers, wax and protein filaments
- transport lipid (waxes) from the epidermis to the epicuticular surface
External appendages of the integument (Fig. 23)

a. Microtrichia - tiny, false hairs - produced by special epidermal
cells
b. Macrotrichia (setae) - bigger true hairs; produced by seta
forming (trichogen) cells; surrounding the macrotrichia are
socket constructing (tormogen) cells
c. Scales – produced by special scale constructing cells
Special cuticle formations

- outer extensive appendages or dents grooves, ribs, furrows. Figure 23. Appendages of the
- inner appendages, skeletal elements serve for the inner integument.
structure, attachment of muscles, organs (endophragma, apodema, apophysis)
Glands of the insect integument, dermal glands (glandulae)

Classification of glands according to their structure
a. unicellular glands
b. complex glands: a group of adjacent unicellular glands both can occur within the epidermis (intra-
epithelial) or under the epidermis (sub-epithelial)
c. polycellular glands always occur sub-epithelial; their structure is nodular or tubular
Classification of glands according to their function

- wax glands, enamel glands, cement glands, lubricant producing glands, adhesion helping glands,
pheromone producing glands, odor glands, stink glands, venom producing glands
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PHYSIO&TOXI
Penetration of compounds with different characteristics through the cuticle:

1. water-soluble compounds - the epicuticle hinders and restricts penetration owing to the wax layer
the procuticle is selectively permeable undissociated forms of compounds can get in easier
2. fat-soluble compounds - procuticle hinders and restricts penetration epicuticle is permeable
3. The integument acts as a physical barrier to decrease the rate of entry of different compounds.
Color of the integument

a. Chemical or objective colors
Pigments are within the epidermis (after the death they break up rapidly) or within the procuticle
(exocuticle) (they remain longer after death).
Most common pigments:
- melanin pigments (brown or black)
- pteridin, pterin pigments (white, yellow, orange, red)
- ommochrome pigments (yellow, red)
- bile pigments (red, green, blue)
- carotenoids (yellow, orange, red, violet) of plant origin
b. Physical colors (subjective colors)

- results of special optical phenomena occurring on the uneven surfaces (e.g. reflection, interference,
diffraction) possible colors: white, brilliant green or blue, flashing metallic color , iridescent (Schiller
color)
Lesson 3. Principles of Toxicology

Insect toxicology plays an important role in managing insect pests both in agriculture and public
health. The word toxicology is derived from two Greek words, toxicos meaning “poisonous” and logos
meaning, “to study”. Toxicology is a broad field involving multidisciplinary sciences related to adverse
chemical effects on living organisms - including humans. Toxicology is defined as:
- “Toxicology is the branch of the medical science that deals with the nature, properties, effects and
the detection of poison. It is, therefore, the science of poisons” (Du Bois and Geiling, 1959).
- “Toxicology is the study of limits of the biological effects of a chemical or mixtures of chemicals”.
(Fogleman, 1963).
- “Science of doses” (Matsumura, 1985).
The specialized field of insect toxicology deals with such poisons which are used in killing insect
without appreciable effect on mammals. Insecticide toxicology differs from its parent discipline, medical
toxicology, in that it does not include clinical diagnoses or treatment of human patients who are affected by
insecticides. However, insecticide toxicology does include efforts to determine tolerance levels of pesticides
in man and is concerned with establishing a logical basis for selective toxicity, in order to kill insects without
affecting mammals.
Main Principles of Insect Toxicology
1. Insecticides be able to strike the weakest link of the pest.

 Life cycle
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PHYSIO&TOXI
 Behavior of the pest be known

2. Assessment of loss, nature and extent of damage and the economics involved in it be properly
calculated.
3. Proper selection of selection of insecticide or combination of insecticides be made prior evaluation.
4. Insecticides be of such nature and quality that they bring about least disruptions in the eco-system
and remain restricted in the area where they are used.
Pesticides
Pesticides are biocidal substances utilized to attract and later destroy any pest. Pesticides are most
commonly used as control against pests of agricultural crops but can also be used to manage urban and
domestic animal pests. The term pesticide is collective referring to any chemical substances used to control
weeds (herbicide), rodents (rodenticide), insects (insecticide), nematodes (nematicide), mollusks (e.g.
golden apple snail) (mulloscicide), bacteria (bactericide), fungi (fungicide), insect growth regulators,
repellents, disinfectant and sanitizer. The Food and Agriculture Organization (FAO) defined pesticide as:
“any substance or mixture of substances intended for preventing, destroying, or controlling any
pest, including vectors of human or animal disease, unwanted species of plants or animals, causing
harm during or otherwise interfering with the production, processing, storage, transport, or
marketing of food, agricultural commodities, wood and wood products or animal feedstuffs, or
substances that may be administered to animals for the control of insects, arachnids, or other pests
in or on their bodies. The term includes substances intended for use as a plant growth regulator,
defoliant, desiccant, or agent for thinning fruit or preventing the premature fall of fruit. Also used as
substances applied to crops either before or after harvest to protect the commodity from
deterioration during storage and transport.”
Insecticide – is a pesticide used to kill or eliminate insect pests in agriculture, household and
industries. Insecticides have high toxicity to non-target organisms and capability to develop resistance
through widespread use, and have high potential to significantly affect and alter ecosystems. Many are
toxic to humans and animals, and can accumulate in the food chain and water resources, giving rise to
serious environmental contamination and pollution.
Brief history of pesticide application
Since the beginning of agriculture 10,000 years ago in the Fertile Crescent of Mesopotamia, crops
and farm animals suffer from pests and diseases causing large losses in yield and alarm to food security.
This events caused the surge in the use and continued formulation of pesticides. Table 1 shows the
important chronological events of pesticides application in agriculture and public health.
Table 1. Important historical events in using pesticides as pest control

Period Insecticide usage and insect control methods
Approx. 4,500
Sulfur dusting was used by Sumerians to control insects and mites
years ago
Approx. 3200
Chinese use mercurial and arsenic compounds to control body lice
years ago
Approx. 2000 Pyrethrum, derived from the dried flowers of Chrysanthemum cinerariaefolium
years ago “Pyrethrum daisies”, has been used by Persians to control pests of stored grain
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PHYSIO&TOXI
Toxic inorganic chemicals, for example, arsenic, lead, and mercury, were applied
15th century
to crops.
17th century Nicotine sulfate extracted from tobacco was used as an insecticide.
Pierre- Marie-Alexis Millardet formulated and discovered the use of Bordeaux
1860s
mixture to control powdery mildew of grapes
1940-41 Methyl bromide was used as a fumigant against stored product pests.
DDT and other organochlorines with a wide spectrum of toxicity, that were
inexpensive and had a persistent residual effect, eventually gave rise to serious
Mid-1940s
environmental problems. Organophosphates (OPs) with high toxicity acting on the
nervous system were introduced in 1944.
Carbamates were first introduced in 1956. They had high insecticidal toxicity, were
1950s
less toxic to humans, and had relatively faster breakdown.
Pyrethrins (botanical insecticides) and male annihilation (combining a
1960s
lure/attractant and an insecticide-usually OPs).
Juvenile hormone analogs/mimics (insect growth regulator, IGR) and insect
1970s
pheromones/semichemicals (“attract and kill”).
Introduction of the world’s greatest selling herbicide, glyphosate, the low use rate
1970s – 1980s sulfonylurea and imidazolinone (imi) herbicides, as well as dinitroanilines and the
aryloxyphenoxypropionate (fop) and cyclohexanediones (dim) families
Synthetic pyrethroids, insect growth regulators (inhibitor of chitin synthesis),
1980s
biological control, and integrated pest management (IPM) was introduced
New families of agrochemicals have been introduced to the market such as the
1990s triazolopyrimidine, triketone and isoxazole herbicides, the strobilurin and azolone
fungicides and chloronicotinyl, spinosyn, fiprole and diacylhydrazine insectides
1990s Neonicotinoids, area-wide male annihilation, sterile insect technique.
2000s Phenyl pyrazoles, IPM.
Classification of Insecticides
Insecticides can be classified according to:

1. Target insect stage of development for example: ovicides, larvicides, and adulticides kill insect
eggs, larvae, and adults, respectively.
2. Application technique, for example, dusting, fumigant, spray, residual, and topical.
3. Modes of action
- Physical poisons—dusts, fumigants, and oils. These poisons kill insects by asphyxiation by
blocking the flow of oxygen through the insect tracheal (respiratory) system.
- Protoplasmic poisons are inorganic chemicals that physically destroy cells.
- Metabolic inhibitors either interfere with metabolic pathways or inhibit certain enzymes.
- Neuroactive agents affect the transmission of nerve impulses or the neurotransmitter.
- Insect growth inhibitors disrupt growth and the development or malformation of cuticle.
4. Active group in the insecticide, for example, carbamate, organochlorine, organophosphate.
5. Chemical nature (e.g. arsenical, botanical, organochlorine or carbamate insecticides)
Evaluation of Toxicity of Insecticides
The LD50 (median lethal dose) test was introduced in 1927 by J. W. Trevan to estimate the dose of
a test substance that produces 50% death in a given species of animals. It is usually the first test conducted
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PHYSIO&TOXI
for every chemical before further toxicity tests are carried out. It is used for estimating the potential hazards
of chemicals on humans. Although its major endpoint is death, non-lethal acute effect may occur as signs
of toxicity depending on the chemical being tested.
Figure 24. Graphical representation of LD50.
The LC50/LD50 represent the concentration (LC50) or dose (LD50) at which 50% of the population
responds. For example, consider comparing the toxicity of two different pesticides to aphids, pesticide A
and pesticide B. If the LC50 of pesticide A is 50ug/L and the LC50 of pesticide B is 10ug/L, pesticide B is
more toxic than A because it only takes 10ug/L to kill 50% of the aphids, versus 50ug/L of pesticide B.
Table 2. Classification of LD50 based on dose range.

LD50 Classification
<5 mg/kg Extremely toxic
5–50 mg/kg Highly toxic
50–500 mg/kg Moderately toxic
500–5,000 mg/kg Slightly toxic
5000-15,000 mg/kg Practically non-toxic
>15,000 mg/kg Relatively harmless
LC50 – The lethal concentration of toxic compound mixed in external medium ie. water that kills half of the
population of test animals.
KD30 – It represents the median knockdown dose sufficient to kill 50% population of test animals.
LT50 - This term represents the lethal time required to kill 50% population of test animals at a certain dose
or concentration.
KT50 – It represents the median knockdown time required to kill 50% of test animals for a given dose.
ED50 - The dose of such a chemical which brings sterility in 50% population in test animals will be the value
of ED50.
EC50 – It is the concentration of chemical resulting sterility in 50% of test animals.
Other important terms in toxicological studies:
Toxicity- ability of a chemical to bring about changes in the biological system of the target animal.
Acute Toxicity – It is the acute stage of poisoning due to the application of a single dose.
Chronic Toxicity – It is the condition of toxicity which lasts for the entire life of the target animal and has
the accumulating effect of small repeated doses.
Hazard – It is the probability of being harmed due to the use/exposure/handling of the toxic substance.
Risk- It is the degree of physical, biochemical and histochemical changes acceptable in terms of usefulness
of a chemical and its possible effects on Public Health.
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PHYSIO&TOXI
SUMMARY
 Insects are the most successful organism on earth evident of the biological and ecological nature.
They have developed structures that enable them to feed on almost any food and fly from one
place to another.
 Insects are important because:
 they are the primary pests affecting both animal and crops
 some insects are natural enemies to other pests
 entomophagy
 can be an alternative material in animal feed formulation
 soil enhancers
 some species produce unique products such as silk and honey
 All species belonging to Class Insecta have:
 Three pairs of jointed legs
 Segmented body with appendages
 Exoskeleton with chitin
 Ventral nervous system
 Dorsal heart
 Familiarity of the external and internal structures of insects are essential in proper identification
and in the study of insect physiology.
 Insecticide toxicology is vital in managing agricultural pests. Toxicology refers to the study of the
nature, properties, effects and mode of action of chemicals used in living organisms. Insecticide
toxicology deals with the use of chemicals without detrimental effects to mammals.
 The main principles of insecticide toxicology include:
 Efficacy of pesticides to the weakest link of the target organism
 Assessment of loss and nature of damage
 Proper selection of pesticides to be used
 Keeping in mind the effects of pesticides to the environment
 Pesticides are biocidal substances or mixture of substances intended for preventing, destroying,
or controlling any pest. Among the kinds of pesticides, insecticides are the most broadcasted to
manage insect pests in agriculture.
 Insecticides can be classified according to:
a. Target insect stage of development for example: ovicides, larvicides, and adulticides
kill insect eggs, larvae, and adults, respectively.
b. Application technique, for example, dusting, fumigant, spray, residual, and topical.
c. Modes of action
 Physical poisons—dusts, fumigants, and oils. These poisons kill insects by
asphyxiation by blocking the flow of oxygen through the insect tracheal
(respiratory) system.
 Protoplasmic poisons are inorganic chemicals that physically destroy cells.
 Metabolic inhibitors either interfere with metabolic pathways or inhibit certain
enzymes.
 Neuroactive agents affect the transmission of nerve impulses or the
neurotransmitter.
 Insect growth inhibitors disrupt growth and the development or malformation of
cuticle.
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PHYSIO&TOXI
d. Active group in the insecticide, for example, carbamate, organochlorine,

organophosphate.
e. Chemical nature (e.g. arsenical, botanical, organochlorine or carbamate insecticides)
 The toxicity of insecticides is usually expressed in terms of lethal dose or LD50 which is the amount
of toxin per unit weight which kills 50% of the test population.
REFERENCES
Chapman, R.F. (2013). The Insects – Structure and Function (5th ed.). UK: Cambridge University Press.
Gullan, P. J. & Cranston, P. S. (2014). The Insects: An outline of Entomology. 5th Ed. West Sussex, UK:
John Wiley and Sons, Ltd.
Imms, A.D. (1925). A General Textbook of Entomology. London: Methuen & Co. Ltd.
Freeman. Nation, J.L. Sr. (2016). Insect Physiology and Biochemistry (3rd ed.). USA: Taylor and Francis
Group, LLC.
Triplehorn, C.A. & Johnson, N.F. (2005). Borror and DeLong’s Introduction to the study of insects (7 th ed).
USA: Brooks/Cole.
Yu, S.J. (2015). The Toxicology and Biochemistry of Insecticides (2nd ed.). USA: Taylor and Francis Group,
LLC.
QUIZ 1
A. Multiple choice. Choose the BEST answer among the choices. Answer honestly.
1. The insect body is segmented forming three major units such as _________________.
a. Head, cephalothorax and abdomen
b. Head, pterothorax and abdomen
c. Head, thorax and abdomen
d. Head, thorax and exoskeleton
2. An insect head is a sclerotized capsule bearing the feeding, and sensory organs. A
grasshopper has two sets of eyes, a pair of compound eyes located dorsa-laterally and the
three simple eyes or __________.
a. Ocelli
b. Antennae
c. Tympanum
d. None of the above
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PHYSIO&TOXI
3. It is the mouthpart which enables termites and beetles to bore through woods and foils
easily.
a. Labrum
b. Hypopharynx
c. Mandible
d. Maxilla
4. Morphological changes on the feeding structures of the basic mandibulate mouthparts has
become an adaptive feature of insects making them diverse feeders. A black aphid
(Pentalonia nigronervosa) and fleas (--), transmitting bunchy top virus and infesting
ruminants respectiviey have this kind of specialized mouthparts.
a. Piercing-sucking
b. Lapping
c. Chewing
d. Siphoning
5. __________ refers to the last segment of an insect antenna.
a. Scape
b. Sensillum
c. Pedicel
d. Flagellum
B. Identification
1. Identify the type of antenna of the following insects:
a. Termite -
b. Cockroach -
c. Silkworm -
d. Dragon fly -
e. Fire ant –
2. A pair of forewings are attached on the mesothorax while the pair of hindwings are attached
to the metathorax, both dorsa-laterally. Collectively, the wing-bearing meso- and
metathorax is called ___________.
3. An insect has three pairs of legs each divided into six segments. What is the segment
proximal to the insect body?
4. The insect abdomen is composed of segments made up of a dorsal tergum and a ventral
_______ and pleural membrane joined together by flexible non-sclerotized cuticle.
5. _______________ is a collective term for the anal-genital parts of the insect abdomen,
starting from segment 8 or 9 up to the abdominal apex.
6. The _______ is the egg-laying structure of female insects.
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PHYSIO&TOXI
Name: _____________________________________ Schedule: _____________________

Student Number: _____________________________ Instructor: _____________________
Program: ___________________________________ Score: ________________________
LABORATORY ACTIVITY 1
External Anatomy and Morphology of Insects
Objectives:
1. To be familiarized with the basic external structures of a generalized insect

2. To describe the morphology of insect structures used in feeding and flight
I. Identify the parts and segments of a grasshopper specimen.
3
1
2
11 12
0
10
0 6
8
9
7
13 14 15
0 0 0
II. Based on the model above, identify the following:

1. Type of mouthpart
2. Type of antennae
3. Type of wings
4. Is it a male or female?
5. What insect order does grasshoppers belong?
Image source: https://www.pinterest.ph/pin/101471797830954933/
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PHYSIO&TOXI
Chapter 2
INSECTICIDE USE PATTERN
OVERVIEW
Pesticides are widely applied everywhere in the world. Pesticides are chemical substances used for
controlling pests. In the United States, approximately 1,200 active ingredients were approved for use and
over 20,000 pesticide products were formulated and available in the markets.
The world population reached 7 billion in 2011 and is estimated to reach 8.04 billion by the year 2025.
To feed and clothe this ever increasing human population, increase in the production of food and fiber crops
is a must. Agricultural experts believe that global food demands can be met however, requires an increase
of using pesticides. There are two things to consider in using pesticides: (1) most pesticides are non-
selective especially insecticides which may harm non-target organisms including humans and (2) pesticide
application is not always efficient, wherein 45% reaches the crop, 4% reaches the target insect pest while
the remaining 50% are wasted and will pollute the environment. Therefore, to reach the global demand for
food, balance between benefits of pest control and risks to human health and the environment must be
taken into consideration.
This chapter tackles the importance of pesticide use in agriculture, health and home, as well as the
patterns of insecticide use, trends and risk factors focusing on the Philippine setting. Moreover, a brief
discussion on mass rearing of insects and preparation of standardized test materials are included as part
of laboratory exercise. Insect rearing is a knowledge and skill necessary for entomologists or
insect/insecticide researchers.
LEARNING OUTCOMES
1. Comprehend the reasons why pesticides are indispensable in agriculture and community.
2. Describe the patterns of pesticide use.
3. Enumerate the strategies to reduce pesticide use
4. Discuss the importance, types and procedures used to rear and prepare standardized test insects.
5. Identify the limiting factors in rearing insects.
6. Conduct a hands-on insect rearing.
MOTIVATION
“If all mankind were to disappear, the world would regenerate back to the rich state of equilibrium that
existed ten thousand years ago. If insects were to vanish, the environment would collapse into chaos.”
- E. O. Wilson
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PHYSIO&TOXI
Lesson 1. The Need for Pesticides
Food production
The major benefits of pesticides is in agriculture – the protection of crop yields and livestock health.
Pesticides can save farmers’ money by preventing losses against insects and other pests. One study in US
states that removing pesticides would cause decline in crop production as much as 50%. It can be
concluded that pesticides are inevitable tools in diverse pest management tactics.
Approximately one-third of agricultural crops are destroyed by pests during growth, harvesting and
storage worldwide. As cited by Yu (2008) Losses are even higher in developing countries:
 In Latin America, 40% of crops are lost because of pests.
 In Ghana, cocoa production was tripled by the use of insecticides to control just one insect pest
species.
 Sugar production in Pakistan was increased one-third because of the use of insecticides.
 FAO estimated that 50% of cotton production in developing countries would be destroyed without
the use of insecticides.
 In the United States, crop losses due to pests are estimated to be 30%, or $33 billion.
According to Kan-Rice (2019), pathogens and pests are causing wheat losses of 10% to 28%, rice
losses of 25%to 4%, maize losses of 20% to 4%, potato losses of 8% to 2%, and soybean losses of 11%
to 3%. In the Philippines, PhP 473 million worth of production loss due to pest infestation were incurred in
Negros Occidental in 2019 (Nicavera, 2019). These losses are due to damages cause by rats, insects and
leaf blights.
Public health
There are many diseases that insect, mites and ticks are able to transmit to humans. Insects are
vectors of human pathogens that makes them of great importance in securing health of the community.
Table 3 shows the list of most common insect-vectored diseases of humans (adapted from Ware and
Whitcare (2004) by Yu (2008).
Table 3. Common diseases transmitted by insects, mites and ticks.

Disease Vector
African sleeping sickness Tsetse flies
Anthrax Horse flies
Bubonic plague Arat flea
Dengue fever Two mosquitoes
Dysenteries Several flies
Encephalitides Several mosquitoes
Endemic typhus Oriental rat flea
Filariasis Several mosquitoes
Hemorrhagic fevers Several mites and ticks
Lyme disease Ixodes spp. ticks
Malaria Anopheles mosquitoes
Onchocerciasis Several black flies
Leishmaniasis Sandflies
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PHYSIO&TOXI
Q fever Ticks
Relapsing fevers Several ticks
Rocky Mountain spotted fever Two ticks
St. Louis encephalitis Culex pipiens
Trypanosomiasis Several flies
West Nile virus Several mosquitoes
Yellow fever Several mosquitoes
The World Health Organization (WHO) recognized that the use of synthetic insecticides can markedly
reduce the risk of insect-borne diseases. For example, annual death rate for malaria was decreased from
6 million in 1939 to 2.5 million in 1965 and 1 million in 1991. Similarly, the use of insecticides controlled
yellow fever, sleeping sickness, and Chagas’ disease. Cockroaches are also associated with the incidence
of bronchial asthma, particularly, in children. Cockroach’s body parts, cast skin, egg shells, and fecal
material contain several major and minor allergens involved in triggering an asthma attack.
Home and business
In the US, wood destroying insects such as termites and wood boring beetles causes $1 billion of
property damage each year. Despite their ecological role, termites locally known as “anay” are formidable
threat to home structures. Based on Rentokil (2021), an average house repair costs an estimate of Php
205, 000 due to termite infestation. The Philippines is a tropical country with high humidity which is favorable
to growth of termites.
Apart from economic reasons, some insects cause distress at home or in business establishments,
such in the case of cockroaches. Not only that they are carriers of infectious diseases such as
salmonellosis, dysentery, gastroenteritis and typhoid, their body parts, casts, egg shells and fecal matter
can trigger eczema and asthma. Cockroaches produces obnoxious odor that stain food and objects. Most
commonly found bed bug species in the Philippines that readily bite humans are Cimex
lectularius and Cimex hemipterus. Public accommodations such as hotels, dormitories and cinema are
more at risk with bedbug infestations. But bed bugs can be brought home through your luggage, clothes,
and bags and by carrying just 1 single surviving female bed bug home can cause infestation. Rats are
persistent and are able to thrive in dark, narrow and unclean habitats. Rats can cause diseases, damage
structures and belongings. In food establishments, rats can cause immediate loss of trust from customers
and employees (bad company reputation)
Lesson 2. Patterns and Trends of Pesticide Use

Pesticides are used worldwide. According to FAO, a total of 4.12 million tons of pesticides were
broadcasted in 2018 globally (Fig. 25). Likewise, for agricultural purposes alone, the countries such as
China (1.40 million tons), USA (406 thousand tons), Brazil (221 thousand tons), Argentina (126 thousand
tons), USSR (89 thousand tons), France (82 thousand tons), Italy (76 thousand tons), Japan (67 thousand
tons), Colombia (52 thousand tons) and Canada (50 thousand tons) belong to the top 10 biggest pesticide
users from 1990 to 2018, on average (Fig. 26). In the Philippines, an increase in pesticides usage was
observed from 1990 to 2014. However, in 2015, there is a sudden decline in pesticides use (Fig. 27).
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PHYSIO&TOXI
Figure 25. Total pesticides use worldwide.
Figure 26. Top 10 countries according to pesticide use Figure 27. Trend of pesticide use in the Philippines from
from 1990 to 2018 (FAO, 2019) 1990 to 2018 (FAO, 2019)
From 2005 to 2010, a study was conducted by Lu (2010) providing an analysis of type of pesticide
used, exposure factors, and the pesticide-related concerns among the vegetable farmers in Benguet
province. Their study revealed the top three most commonly used pesticides by Benguet farmers: Tamaron
(methamidophos), an organophosphate, followed by Dithane (mancozeb), a dithiocarbamate and
Sumicidine (fenvalerate), a pyrethroid (Fig. 28).
Figure 28. Percentage distribution of pesticides used by Benguet farmers from 2005-2010.
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PHYSIO&TOXI
Lesson 3. Strategies to Reduce Pesticide Use
1. Integrated Pest Management (IPM)
- “the careful consideration of all available pest control techniques and subsequent integration of
appropriate measures that discourage the development of pest populations and keep pesticides and other
interventions to levels that are economically justified and reduce or minimize risks to human health and the
environment. IPM emphasizes the growth of a healthy crop with the least possible disruption to agro-
ecosystems and encourages natural pest control mechanisms.” (UN’s Food and Agriculture Organization)
2. Biopesticides or ‘biological pesticides’
 “A form of pesticide based on micro-organisms or natural products”. (European Union)

 “Include naturally occurring substances that control pests (bio-chemical pesticides), microorganisms
that control pests (microbial pesticides), and pesticidal substances produced by plants containing
added genetic material (plant-incorporated protectants) or PIPs”. (US Environmental Protection
Agency)
 They are obtained from organisms including plants, bacteria and other microbes, fungi, nematodes,
etc.
 An important component of integrated pest management (IPM) programs, and have received much
practical attention as substitutes to synthetic chemical plant protection products (PPPs).
Biopesticides can be classified into these classes:

• Microbial pesticides which consist of bacteria, entomopathogenic fungi or viruses (and sometimes
includes the metabolites that bacteria or fungi produce). Entomopathogenic nematodes are also
often classed as microbial pesticides, even though they are multi-cellular.
• Bio-derived chemicals. Four groups are in commercial use: pyrethrum, rotenone, neem oil, and
various essential oils are naturally occurring substances that control (or monitor in the case of
pheromones) pests and microbial diseases.
• Plant-incorporated protectants (PIPs) have genetic material from other species incorporated into
their genetic material (i.e. GM crops). Their use is controversial, especially in many European
countries.
• RNAi pesticides, some of which are topical and some of which are absorbed by the crop.
Examples:
a. Bacillus thuringiensis, a bacterial disease of Lepidoptera, Coleoptera and Diptera. The toxin from
B. thuringiensis (Bt toxin) has been incorporated directly into plants through the use of genetic
engineering.
b. Entomopathogenic fungi (e.g. Beauveria bassiana, Paecilomyces fumosoroseus, Lecanicillium
spp., Metarhizium spp.)
c. Plant disease control agents: include Trichoderma spp. and Ampelomyces quisqualis (a hyper-
parasite of grape powdery mildew); Bacillus subtilis is also used to control plant pathogens.
d. Beneficial nematodes attacking insect (e.g. Steinernema feltiae) or slug (e.g. Phasmarhabditis
hermaphrodita) pests
e. Entomopathogenic viruses
f. Weeds and rodents have also been controlled with microbial agents.
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PHYSIO&TOXI
3. An insect growth regulator (IGR)

 A substance (chemical) that inhibits the life cycle of an insect. IGRs are typically used as
insecticides to control populations of harmful pests, such as cockroaches or fleas.
 Targets juvenile harmful insect populations while causing less detrimental effects to beneficial
insects.
 Prevents an insect from reaching maturity by interfering with the molting process.
4. Insecticidal soap
 based on potassium fatty acids
 insecticidal soap only works on direct contact with the pests, it is sprayed on
 have a low mammalian toxicity
 based on long-chain fatty acids (10–18 carbon atoms), because shorter-chain fatty acids tend to
be damaging for the plant (phytotoxicity)
Possible mechanisms of insecticidal soaps:

• Soap, which enters via the insect’s trachea, may disrupt cell membranes, resulting in the cell
contents leaking from the damaged cells (cytolysis).
• Soap may dissolve the wax layer on the cuticle which leads to water loss by evaporation.
• Soap may block breathing openings or trachea, which leads to suffocation.
• Soap may interfere with growth hormones.
• Soap may affect insect metabolism.
Lesson 4. Mass Rearing and Preparation of Test

Insects
Today, literary thousands of insect species can be reared through multiple life cycles and/or parts of
their life cycle. The goal of organized insect rearing is to provide reliable, affordable sources of high quality
insects for intended purposes. In this work, the provision of fresh food or host material or the development
of suitable nutritious diet that induces feeding is the most difficult task. Precautions must be taken especially
during the starting phase where insect colonies must be well-maintained to maintain lower mortality rates.
This will prevent genetic bottlenecks caused by inbreeding of insects, contamination with other species and
diseases. As the number of insects increase, rearing must organized efficiently to incorporate the necessary
procedures for each stage of the insect’s life being reared. Facilities must be designed and constructed to
maintain the desired environmental conditions, which is the most common cause of mortality in rearing
experiments.
Purposes of insect rearing

1. To screen newly formulated chemical insecticides
2. Resistance screening of plants against insects or diseases transmitted by insects
3. Screening of non-target insects and plants as possible hosts of non-native natural enemies before
releasing commercially
4. To provide specimens for the understanding of insect’s biology, physiology and epidemiology (e.g.
orientation, dispersal and migration)
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PHYSIO&TOXI
5. To mass produce and release potential natural enemies or biological control as part of integrated
pest management (e.g. predators and parasitoids) or for autocidal control (sterile techniques)
6. To improve wild and domestic populations of insects producing commercial products such as
silkworms and honeybees
7. Conservation of depleting populations of rare and endangered insect species
8. Because of their aesthetic values (personal collections, static displays, insect zoos or even
household pets)
Kinds of insects that are reared

Almost all free-living insects can be reared partially (egg to larva or nymph and adult) or completely
(egg to egg) but considerable knowledge about the insect’s life cycle, habitat and nutrient requirement, as
these are the limitations of rearing activities. These may involve mimic of soil or aquatic habitat, symbiotic
relationships and specialized foods or host plants; some insects are temperature and photoperiod sensitive.
Because of these, recommended insects to be reared should be relatively small, produce one generation
per year, plant feeding, terrestrial species with wide host range and no special environmental requirements.
Examples include those that infest common crops, landscape plants, or stored products.
Nutritional provisions
Natural diet
Immature plant-feeding insects can be fed by clean, fresh cut plant materials on which they fed
naturally. For example, larval silkworms feed on mulberry leaves; boll weevils feed on cotton bools; tropical
fruit flies on papaya fruits and monarch butterflies on milkweed leaves. Veterinary or medical insects such
as adult mosquitoes and biting flies can be fed on their hosts or suitable alternatives such as rodents,
sheep, goats or pigs. Considerable difficulty is the nutritional requirement of natural enemies since the
scheme must composed of three trophic levels (plant, insect host and natural enemy).
Artificial diets
The following are some of the formulated diets for rearing insects:
1. Henry Richardson (1932)
- Bran, alfalfa meal, yeast and diamalt formula for rearing houseflies
2. (CSMA medium; Chemical Specialties Manufacturer’s Association)
- A commercial diet made of wheat bran (33.3%), alfalfa meal (26.7%), and brewer’s grain (40%)
for rearing house fly larvae
3. M. H. Haydak (1936)
- Grain flower, milk powder, honey, and glycerin diet for stored-grain insects, such as mealworms
and flower moths.
4. Gelled diets were developed for rearing insect larvae that require large quantities of contained
water in their diets
- Pearl (1926) diet for Drosophila spp.
- Botger’s (1942) larval medium for the European corn borer (Ostrinia nubilalis)
- gelled with agar, a polysaccharide derived from seaweed, that was previously developed for use
in bacteriology
- Ground plant fibers, such as sugarcane bagasse (pulp remaining after the sugar is extracted),
corncobs, and carrot powder are used to rear tropical fruit fly larvae
5. David Taylor of the USDA, Agricultural Research Service (ARS) (1988)
- a starch polymer-gelled diet for rearing screwworm larvae, Cochliomyia hominivorax
-
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Categories of insect rearing

1. Single species rearing – immature stages are usually fed on host plants or animals or artificial
diets; natural environments and oviposition substrates are duplicated from nature and all rearing
operations can be performed by a single individual.
2. Multiple-species rearing - usually accomplished in centralized facilities to support research. There
is economy of scale in the rearing operations, i.e. diet preparation, egg treatment, larval rearing,
harvesting of pupae, and adult colony maintenance can be combined for similar species; common
in research laboratories (e.g. those used for insecticide or transgenic crop development; and is
typically performed by a small staff)
3. Mass rearing - involves a single species produced for biologically based pest management that is
reared in factory-like facilities with controlled environments, artificial diets and oviposition
substrates, mechanized equipment, and operations performed by work units.
Below is an example procedure on rearing rice brown planthoppers, but can be adopted on other
herbivorous insects.
Collection of Insect
1. Identify a suitable rice field; take note its location and geographic positions
2. Collect at least 50 healthy unparasitized adult female brown planthoppers or 100 nymphs.
Planthopper adults, preferably short-winged, should be collected from the base of the rice plants
using an aspirator (by mouth or suction bulb) and placed into test tubes covered with nylon mesh.
Alternatively, a sweep net can be used (Fig. 29).
Figure 29. Collection equipment: (A) a mouth aspirator and test tube covered with nylon mesh; (B) a sweep net .
3. Transfer the collected insects immediately onto clean potted plants enclosed with circular or
rectangular mylar cages (Fig. 30). These plants and cages should be prepared in the research
center before going on the collection trip. Alternatively, collected insects can be transferred to
rearing cages with clean potted plants. Collection cages must be labeled with the respective
collection dates, location names, and geographic positions.
Figure 30. Collection cages: (A)

potted plant with circular mylar cage,
61 cm high and 10.5 cm in diameter;
(B) rectangular mylar cage,
measuring 29 cm × 21.5 cm × 56.5
cm, with a potted plant; (C) aluminum
rearing cage, measuring 56.5 cm ×
56.5 cm × 91.5 cm, with potted plants;
(D) test tubes with rice seedlings; (E)
box with rice seedlings.
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PHYSIO&TOXI
4. The insects can be reared in a greenhouse or insectary maintained with a temperature of 27±2°C
and 12 hours of light.
Rearing Methods
a. Aluminum cages
- one month before the collection date, 10-day-old seedlings should be planted in clay pots 10 cm in
diameter. One week prior to collection, the outer leaf sheaths or infested tillers of the rice plants
should be removed.
- In the greenhouse or insectary, the adult males and females (at 1:1 ratio) should be transferred into
the oviposition (egg-laying) aluminum cages and labeled with the respective collection dates and
locations. The oviposition cages should be provided with 35-day-old clean potted plants (replaced
daily to have uniform populations). For insecticide testing, the preparation of standardized insects is
discussed later.
- Remove the adult planthoppers from the oviposition cage and allow the nymphs to emerge. Each
rearing cage can accommodate six to eight potted plants that can sustain 600–800 hoppers. The
standard test insects (1- to 2-day-old female adults) are collected from these daily rearing cages. The
plants are replaced every 2 days (or as needed) and transferred to new rearing cages (labeled with
egg collection dates and locations).
b. Flexi-glass cages
The three sides of the cage should be covered with fine-mesh nylon cloth for ventilation. Planthoppers
are reared on rice seedling mats (measuring about 22 cm x 28 cm) grown in nutrient solution, adapted from
Yoshida et al (1976). Approximately 12 g of seeds can be sown per seedling mat. The procedures for the
preparation of a seedling mat are as follows:
1. Select clean and healthy seeds of any susceptible variety.
2. To minimize fungal growth, soak seeds in hot water (70 °C) for 10 minutes.
3. After the hot water has been drained, soak seeds for another 2–3 days in a glass container and
cover them with a paper towel.
4. After soaking, thoroughly wash the germinated seeds (3-4 times) with sterile distilled drinking water
to avoid any further contamination.
5. Then, line the flexi-glass trays with two layers of moistened gauze to keep the seeds in place.
6. Place a flexi-glass guide on top of the gauze and sow the seeds in rows.
7. Remove the guide and add enough rice nutrient solution to cover the seeds.
8. Cover the prepared seedling mat with flexi-glass to protect the seeds from infestation by other
pests.
9. Water the seedling mats daily or as needed with filtered drinking water to maintain enough moisture
and keep the seedling mats wet.
10. Add nutrient solution again after 3–4 days or when the seedlings are yellowish in color.
Figure 31. Aluminum rearing/oviposition cage (56.5 cm × 56.5 cm × 91.5 cm) with aluminum wire mesh on three sides, top, and
doors for ventilation. Flexi-glass cage (30 cm × 25 cm × 30 cm) with seedling mat.
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PHYSIO&TOXI
When the seedlings have grown (about 5–7 days after sowing) and the roots are entangled in the
gauze, the mats can be transferred into a rearing cage. Fifty adult insects are introduced for oviposition and
removed after 1 day. The cage can be inverted and another seedling mat put into the cage as nymphs
emerge. A seedling mat can accommodate about 1,000 late-instar nymphs. Instead of gauze, peat moss
can be used: about 20 g of seed for one seedling mat are pre-germinated for 3–4 days and sown on
moistened peat moss on flexi-glass trays. The mats are covered with rectangular flexi-glass and are also
watered as needed to keep them wet. One week after seeding, the mats can be transferred into a rearing
cage. Adult insects (100–200 pairs) are introduced for oviposition and removed after 1–2 days. The nymphs
are allowed to emerge and the seedling mat is replaced weekly until the plant hoppers become adults.
Likewise, the rearing cages are placed in an insectary with a controlled temperature of 27 + 2 °C and 12
hours of light.
Preparation of Standardized Test Insects
Insecticide bioassays in the laboratory need to have consistent and accurate results. This requires
standardization of the test insects to be used for each treatment. Age, sex, and physiological condition of
the insects affect their susceptibility to insecticides. After field collection, bioassays can be done using plant
hoppers from the second generation up to the fifth generation
Newly emerged adult insects are generally more susceptible. Thus, 1-day-old to 2-day-old adults
should be used in insecticide bioassays. To have approximately the same insect age, adults of the same
sizes are collected from daily oviposition cages. Either brachypterous or macropterous adult female plant
hoppers can be used for insecticide treatments but they should not be mixed in one set of treatments. In
addition to standardization, the preparation and rearing of plant hoppers and plant materials to be used for
the bioassays can be planned based on the life cycle to synchronize their availability.
SUMMARY
 Pesticides are necessary in agriculture, public health and maintenance of home and business
structures. Judicious use is a MUST.
 According to FAO, there is an increase in pesticide use based on the data from 1990-2018. Among
all the types of pesticides, insecticides are widely broadcasted worldwide.
 In the Philippines, the use of pesticide constantly increases but a considerable decline was
observed from 2015-2016.
 Alternatives to pesticides have been developed to lessen the impact of synthetic chemicals on the
environment, non-target species and man.
 Some of the strategies to reduce pesticides’ use are:
 Integrated Pest Management (IPM)
 An ecological strategy to manage agricultural pests through a combination of
one or more methods such as physical, biological, cultural methods and genetic
resistance, and chemical method as the last option to maintain damage to crops
below economic threshold level.
 Biopesticide or biological pesticide
 Pesticide based on microorganisms and natural products
 Includes the modification of genetic material if the plant
 Insect growth regulator
 A substance (chemical) that inhibits the life cycle of an insect.
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PHYSIO&TOXI
 Insecticidal soap
 based on potassium fatty acids; requires direct contact with pests
 Insect rearing is an important aspect and skill in entomological research, pest management, and
pesticide assessment. Insect rearing aims to provide reliable, affordable sources of high-quality
insects for their intended purposes.
 There are three types of insect rearing: single species, multiple species and mass rearing.
REFERENCES
Adams, D. (2017). Insecticides and pesticides: Techniques for crop protection. New York: Library Press.
Food and Agriculture Organization of the United Nations. (2019). FAOSTAT database. Rome, Italy: FAO.
Retrieved March 7, 2019 at http://www.fao.org/faostat/en/#data/RP/visualize
Kan-Rice, P. (2019 Febraury 11). Pests and Pathogens Place Global Burden on Major Food Crops. UC
Agriculture & Natural Resources. URL: https://californiaagtoday.com/pests-diseases-cause-
worldwide-damage-crops/
Leppla, N.C. (2009). Rearing of Insects. pp. 866-869.
Nicavera, E. (2019, April 23). NegOcc incurs P473-M losses due to dry spell, pests. Philippine News
Agency. Retrieved March 9, 2021 from https://www.pna.gov.ph/articles/1067924
Rentokil. (2021). Retrieved March 9, 2021 from https://www.rentokil.com/
Riddick, E.W. and Chen, H. (2014). Production of Coleopteran Predators (Chapter 2). Mass Production of
Beneficial Organisms. USA: Elsevier Inc.
Yu, S.J. (2015). The Toxicology and Biochemistry of Insecticides (2nd ed.). USA: Taylor and Francis Group,
LLC.
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QUIZ 2
Explain your answers concisely.
1. In your opinion, is it necessary to use pesticides? Why or why not? (2 points)
2. What are the strategies implemented to reduce the use or effects of pesticides in the
environment? Give three and describe each. (3 points)
Name: ____________________________________ Schedule: _____________________

Student Number: ____________________________ Instructor: _____________________
Program: __________________________________ Score: ________________________
Insect Rearing*
Objectives:
1. To know the proper steps in mass rearing of insects
2. To do the actual proper steps in mass rearing of insects
Answer the following questions:

1. What are the factors to consider when rearing insects?
2. What are the types of insect rearing? Describe each briefly.
3. Summarize the steps in rearing plant hoppers using a schematic diagram.
*A video tutorial/handout will be provided for your appreciation of the laboratory activity.
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PHYSIO&TOXI
Chapter 3
INSECT PHYSIOLOGY
OVERVIEW
Insect physiology is the study of the functional properties of insect tissues and organs. It requires
knowledge on anatomy, morphology and biochemistry. Although diverse, insects are quite similar in overall
design, internally and externally. Insects evolved adaptations that allow them to live in a variety of
ecosystems.
The basic body plan comprises a hard exoskeleton forming a protective covering over internal
tissues. The interior of the exoskeleton is referred to as the hemocoel. Insects have an open circulatory
system with hemolymph (blood) freely moving within the hemocoel and bathing all of the internal tissues.
Insect’s ontogeny involves the entire process of transformation into a mature adult from a fertilized egg.
Insect species have different developmental stages – the ametabolous, hemimetabolous and
holometabolous. In insects, the transport of oxygen to the cells occurs through a tracheal system where
external surfaces are invaginated into the body cavity to provide an oxygen pipeline from the outside. The
endocrine and the nervous systems serves as channels of information transfer between cells.
Communication, either inter- or intra- is essential in insect survival and reproduction. Chemical, tactile,
acoustical, and visual signals have been identified as being the primary means of communication in insects.
The efficient movement of insects on land, in the air, and in water is a major factor in their domination of
terrestrial ecosystems.
The study of insect physiology is usually divided into a systems approach. These systems are:
digestive, excretory, circulatory, immune, muscular, nervous, and reproductive. A general overview of the
internal structure and physiology of the insect is discussed in this chapter, including sensory organs,
temperature control, flight, and molting. Insect physiology is vital in toxicological studies of pesticides.
LEARNING OUTCOMES
At the end of the lesson, the students will be able to:
1. Determine the basic parts and functions of the insect’s circulatory, digestive, excretion,
nervous, endocrine, respiratory and reproductive systems
2. Describe the physiological processes of an insect, from reproduction, embryogenesis to
morphogenesis.
3. Explain the physiological events in the circulatory system, digestion and excretion of food,
integration, and hormonal secretions.
4. Identify the sensory systems, modes of communication and locomotion of insects.
MOTIVATION
“Our treasure lies in the beehive of our knowledge. We are perpetually on the way thither, being by nature
winged insects and honey gatherers of the mind.” – Friedrich Nietzsche
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Lesson 1. Insect reproduction, growth and

development
Insects multiply in a variety of ways. Most of the time, adult insects lay eggs but some reproduce
during their immature stages. Some insects give birth to live young while other reproduce by laying
unfertilized eggs. Likewise, insects lay not just one but many eggs, which differs from one species to
another! The reproductive organs of insects are similar in structure and function to those of vertebrates -
male's testes produce sperm and a female's ovaries produce eggs (ova). Both types of gametes are haploid
and unicellular, but eggs are usually much larger in volume than sperm.
Modes of reproduction
 Sexual
a. Oviparity – lays eggs
b. Viviparity – gives birth to young
c. Ovoviviparity – eggs retained in bursa copulatrix until hatching
 Asexual
a. Theletoky - Unfertilized female offspring
b. Arrhenotoky – unfertilized male offspring
c. Deuterotoky or amphitoky – unfertilized male/female offspring
 Immature reproduction
a. Paedogenesis – precocious reproductive maturity
b. Neoteny – adult retained larval character
 Polyembryony - where a single fertilized egg divides into many and in some cases thousands of
separate embryos
 Parthenogenesis - female can reproduce and give birth without having the eggs fertilized by a
male
Reproductive strategy
1. Semelparity – produce all their offspring in a single event
2. Iteroparity – reproduce in a successive breeding seasons
Mating systems
a. Monogamy
b. Polygamy
c. Lek
d. Gift-based
Male Reproductive System

The male reproductive system consists of a pair of testes, a duct system (vas efferens (short),
seminal vesicle, and vas deferens, the ejaculatory duct and accessory glands (Fig. 32). All these
structures produce secretions that facilitates the transfer of the sperm to the female. Each testis is
subdivided into follicles where sperm are actually produced. A typical testis may contain hundreds of
follicles, generally aligned parallel to one another. The male reproductive system has one or more pairs of
accessory glands (e.g. Conglobate and Mushroom gland in cockroach) which produces the seminal fluid
that sustains and nourishes the sperm. The accessory glands also produces the spermatophores or
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PHYSIO&TOXI
pouch-like structures made of protein which encloses and protect the sperm when delivered to the female
during copulation. Aedeagus (located externally) is the copulatory organ of male insects.
Figure 32. An illustration of the male reproductive system from (a) the American cockroach, Periplaneta americana and
(b) the milkweed bug, Oncopeltus fasciatus. (Nation, 2016)
Spermatogenesis
Zones of maturation stages of sperm exist
along the length of a typical follicle (Fig. 33). The distal
part of a testicular tubule is a germarium where
repeated mitotic divisions give rise to undifferentiated,
diploid, spermatogonia. In a growth zone (Zone I),
the spermatogonia divide by mitosis into many diploid
spermatocytes enclosed within a cyst or capsule of
somatic cells. The spermatocytes may undergo more
mitotic divisions, but eventually in the “zone of
maturation” (Zone II) meiosis and haploid
spermatids are produced. A spermatid has
completed its meiotic divisions, but is an immature
sperm. Normally four sperm are produced from each Figure 33. (a) (Source: Nation, 2016)
spermatocyte. In Zone III (the region of
transformation), the mature sperm develop.
Transfer of sperm
 Some insects transfer packets or bundles of sperm to the female reproductive tract by insertion of
the aedeagus into the reproductive tract of the female.
 Many insects produce a spermatophore that contains the sperm and is transferred to the female.
Accessory glands secrete spermatophorins, proteins that form the spermatophore.
 Spermatophorin production is stimulated in the mealworm by 20-hydroxyecdysone, but
juvenile hormone (JH) stimulates production in the hemipteran, R. prolixus
 In R. prolixus, the spermatophore consists of a pear-shaped mass of transparent mucoprotein
in a sol or gel state depending on the pH
 Spermatozoa are released from the spermatophore after it is deposited in the bursa copulatrix of
the female
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 Mechanical abrasion of the spermatophore or the action of proteolytic enzymes, or both, may play
a role in releasing sperm.
Female reproductive system

The female's reproductive system (Fig. 34) consists of a pair of ovaries. When the insect is actively
reproducing, these organs swell with developing eggs and may nearly fill the abdomen. Each ovary is
subdivided into ovarioles each containing a string of “egg-shaped” egg chambers called follicles. The
eggs are actually produced in the ovarioles. The base of the ovariole is called the calyx. The female
accessory glands (one or more pairs) supply lubricants for the reproductive system and secrete a protein-
rich egg shell (chorion) that surrounds the entire egg. These glands are usually connected by small ducts
to the common oviduct or the bursa copulatrix, where the spermatophore is deposited. The female's
spermatheca, a pouch-like chamber acts as a temporary storage of sperm. Sperm may live in the
spermatheca for weeks, months, or even years.
Figure 34. (a) The internal reproductive structures of a female insect. (b) Two types of insect ovarioles.
Insect ovarioles depending on whether there are nurse cells associated with the developing egg
(meroistic ovaries) or no nurse cells (panoistic ovaries) (Fig. 35): Meriostic ovaries are further divided
into polytrophic and telotrophic, depending on the location of the nurse cells.
a. Polytrophic ovarioles - each oocyte is closely associated with nurse cells in its follicle or an adja-
cent follicle. E.g. most of the Holometabola, some Coleoptera (Adephaga), Dermaptera,
Psocoptera, Anoplura, and Mallophaga.
b. Acrotrophic or telotrophic ovarioles - nurse cells are located at the distal apex of the ovariole, in
the germarial region, and long, connecting nutritive chords extend from the nurse cells to each
developing oocyte. E.g. Hemiptera and some Coleoptera.
Oogenesis
Each ovariole consists of a distal germarium and a more proximal vitellarium. The germarium
contains pre-follicular tissue and a stem line of oogonia. When these oogonia divide, one of the daughter
cells remains a functional stem line cell, while the other becomes an oogonium that will develop into an
oocyte. Oocytes continues to enlarge as they pass back down the ovariole. As the oogonia leaves the
germarium, it is clothed by the pre-follicular tissue forming the follicular epithelium. As the oocyte continues
to grow, the follicular epithelial cells continues to divide until cuboid or columnar. During yolk accumulation,
growth of the oocyte is very rapid, but the follicle cells ceases to divide instead, it is stretched over the
oocyte as a flattened squamous epithelium. The functions of the follicle cells change during oocyte
development: At first they produce minor yolk proteins and some of the enzymes that will later be involved
in processing the yolk. The follicle cells also produce ecdysone (or a precursor of ecdysone), which will
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accumulates in the oocyte. In the later stages of oogenesis they produce the vitelline envelope and the
ligands responsible for determination of the terminals of the embryo and its dorsal–ventral axis. Finally,
they produce the egg shell, or chorion.
Fertilization
 A female receives sperm from her male partner during the act of mating.
 She can store that sperm for long periods of time in a special part of her reproductive system, the
spermatheca.
 As a developing egg moves past the opening to the spermatheca, a few sperm are released onto
its surface.
 The sperm swim toward the micropyle, the first one to reach its destination enters and injects its
nucleus into the egg. The sperm nucleus quickly fuses with the egg nucleus (syngamy) to form a
diploid zygote - a one-celled embryo. This event is known as fertilization.
Oviposition
Insects lay eggs through one of the following:
- The last segments of the abdomen are elongated into an ovipositor. The egg then passes out from
the ovipositor and is deposited on the surface of a leaf or other substrate
- If there are no structures for oviposition (e.g. ovipositor), an egg ready to be laid passes through
the lateral oviducts and into the common oviduct, and is deposited on some substrate.
- The ovipositor is hardened and pointed and can be inserted inside another insect
- The ovipositor of many insects have been shown to have mechanoreceptors and chemoreceptors
that provide sensory information about the substrate they probe with the ovipositor.
The Egg
Figure 35. Insect eggs (a). Diagrammatic representation of insect egg structures
Most insects produce large eggs relative to their own size due to a majority of insects packaging their
eggs with large amounts of yolk, which serves as the source of nutrients for growth and development of the
embryo. At the time of oviposition, the egg cytoplasm forms the periplasm and an irregular reticulum within
the yolk. The oocyte nucleus is usually located at the center within the egg, but in some species may be
more anterior or posterior.
The eggshell of insects consists of three distinct layers (vitelline membrane, wax layer and
chorion), which are formed during the late stage of oogenesis. The innermost layer of the eggshell is the
vitelline membrane which is also the first layer of the eggshell to be made by the follicular epithelium and
its deposition usually begins late in oogenesis when vitelline membrane proteins accumulate on the surface
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of the oocyte in small vesicles called vitelline bodies. The vitelline membrane surrounds the periplasm. A
wax layer is sometimes but not always present on the outside of the vitelline membrane. The wax layer is
formed due to the accumulation of lipid-filled vesicles on the surface of the vitelline envelope, creating a
water-impermeable layer. The chorion forms the outermost layer of the eggshell, and is produced by the
follicle cells while the egg is in the ovary. The chorion is primarily composed of sclerotized proteins. The
hardening process is due to peroxidase-catalyzed protein crosslinking.
The sperm, which are released from the spermatheca as the egg passes down the common oviduct,
have to enter the egg through a small, usually twisted channel, the micropyle passing through the various
layers of the chorion.
Gender determination
Insects have at least three chromosomal systems for gender determination, with variations existing
within the types:
1. Type 1- Probably the most primitive mechanism, the male is heterogametic, or male XY and female
XX. A variation within this type is the loss of the Y chromosome, so that the male is XO
2. Type 2 - The female is heterogametic (female ZW, male ZZ)
3. Type 3 - Females are diploid while males are haploid
The genetic mechanisms by which the several chromosomal patterns lead to gender determination
are variable and poorly known. Two broad mechanisms are known: the ratio of sex chromosomes to
autosomes and the presence of sex-determining genes.
Embryogenesis
After the egg is fertilized, it undergoes a period of rapid growth and development known as
embryogenesis. Usually, the egg nucleus is diploid until the entry of the sperm stimulates meiotic division
leading to the haploid egg nucleus. The union of a sperm nucleus with the egg nucleus produces the
zygote. Complete cleavage of the zygotic yolk and cytoplasm occurs in eggs of some species during the
first few divisions, but yolk cleavage stops after a few divisions. In most species, cleavage of yolk and
cytoplasm is incomplete from the beginning. Ultimately, zygotic divisions in all insect eggs produce large
numbers of nuclei lacking cell membranes but each surrounded with a small field of cytoplasm called
energids. Energids gradually migrate into a single layer near the periphery of the egg, forming the
blastoderm. Cell membranes become complete after blastoderm formation. A few cells, the pole cells,
aggregate at the posterior end of the egg and will become the gametes (a futuristic character). Cells on the
ventral side of the blastoderm enlarge and become committed as the germ band - the cells that will become
the embryo. Organogenesis leads to formation of the organ systems of the embryo (Fig. 36). Insects with
complete development retain within the larval body small embryonic clusters of cells called imaginal discs
that divide, differentiate, and grow into adult structures during pupation.
Table 4. Example of an insect embryogenesis (Drosophila)

Morphological events (25°C) Hours*
Stage 1 25 min Cleavage divisions 1 and 2 0:25
Stage 2 40 min Divisions 3–8 occur 1:05
Stage 3 15 min Pole bud formation, division 9 occurs 1:20
Final four divisions, syncytial blastoderm formed, stage 4 ends at
Stage 4 50 min 2:10
beginning of cellularization
Stage 5 40 min Cellularization occurs 2:50
Stage 6 10 min Early stages of gastrulation 3:00
Stage 7 10 min Gastrulation complete 3:10
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Formation of amnioproctodeal invagination and rapid germ band

Stage 8 30 min 3:40
elongation
Transient segmentation of mesodermal layer, stomodeal
Stage 9 40 min 4:20
invagination
Stage 10 60 min Stomodeum invaginates, germ band growth continues 5:20
Growth stage, no major morphogenetic changes, parasegmental
Stage 11 120 min 7:20
furrows develop
Stage 12 60 min Germ band shortens 9:20
Stage 13 60 min Germ band shortening complete, head involution begins 10:20
Stage 14 60 min Closure of midgut, dorsal closure 11:20
Stage 15 30 min Gut forms complete tube and encloses yolk sac 13:00
Stage 16 3h Intersegmental grooves evident, shortening of ventral nerve cord 16:00
Stage 17 Stage 17 extends to hatching
Figure 36. Schematic summary of

insect embryogenesis to completion
of organogenesis. The origins of
different cell and tissue types, along
with the major organs or cells they
give rise to are shown.
Post-embryonic development of insects

 Once an insect hatches from the egg it is usually able to survive on its own, but it is small, wingless,
and sexually immature.
 Its primary role in life is to eat and grow.
 If it survives, it will periodically outgrow and replace its exoskeleton (a process known as molting).
 In many species, there are other physical changes that also occur as the insect gets older (e.g.
growth of wings and development of external genitalia).
 Collectively, all changes that involve growth, molting, and maturation are known as
morphogenesis.
Eclosion (egg hatch) First molt Second molt Third molt

Embryogenesis 1st instar 2nd instar 3rd instar ……..etc
Metamorphosis
Each time an insect molts, it gets a little larger. It may also change physically in other ways -
depending on its type of metamorphosis: ametabola, hemimetabola or holometabola.
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Types of Metamorphosis (Fig. 37)
1. Ametabola
Ametabolous insects undergo little or no structural change as they grow older. Immatures are called
young; they are physically similar to adults in every way except size and sexual maturity.
2. Hemimetabola
Hemimetabolous insects exhibit gradual changes in
body form during morphogenesis. Immatures are called
nymphs or, if aquatic, naiads. Maturation of wings, external
genitalia, and other adult structures occurs from molt to molt.
Wings may be completely absent during the first instar,
appear in the second or third instar as short wing buds, and
grow with each molt until they are fully developed and
functional in the adult stage. Developmental changes that
occur during gradual metamorphosis are usually visible
externally as the insect grows, but adults retain the same
organs and appendages as nymphs (eyes, legs, mouthparts,
etc.).
3. Holometabola
Holometabolous insects have immature forms (larvae)
that are very different from adults. Larvae are "feeding
machines", become larger at each molt but do not acquire
any adult-like characteristics. When fully grown, larvae molt
Figure 37. Types of insect metamorphosis.
to an immobile pupal stage and undergo a complete
transformation. The adult stage usually bears wings adapted for dispersal and reproduction.
Table 5. Larval Forms

Common
Appearance Larval type Description Example
name
Flattened body with
Neuroptera
long legs usually with
Campodeiform Crawler Trichoptera
filaments of the end of
Dytiscidae
the abdomen
similar to above, but
Chrysomelidae
Carabiform Crawler legs are shorter and
Carabidae
filaments lacking
cylindrical, well-formed
Lepidoptera
Eruciform Caterpillar head, thoracic legs,
sawflies
and abdominal prolegs
C-shaped, well-formed
Scarabaeiform White grub head and thoracic legs Scarabidae
(no prolegs) weevils
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cylindrical,smooth, and
Elateridae
Elateriform Wireworm tough skinned with
Tenebrionidae
short legs
broad and flat w/ legs Syrphid fly

Platyform None
short or absent blister beetle
Diptera
cylindrical and
Vermiform Maggot Hymenoptera
elongate lack legs
Siphonaptera
Table 6. Pupae Form

Pupal Common
Appearance Description Example
type name
Developing appendages held
tightly against the body by a shell-
Obtect Chrysalis Lepidoptera
like casing. Often found enclosed
within a silken cocoon
All developing appendages free Coleoptera

Exarate None
and visible externally Neuroptera
Body encased within the hard

Coarctate Puparium exoskeleton of the next-to-last Diptera
larval instar
Lesson 2. The Digestive System, Nutrition and

Excretion
The structure and function of the alimentary canal, usually called the gut, coevolved with the food
habits of insects. Both insects and their alimentary canals are diverse, and the alimentary canal is modified
in special ways for solid vs. liquid food and animal vs. plant food. Insects often have to adapt to a dwindling
food supply or change in the quality of their food.
Structures and their functions
The insect alimentary system is divided into three basic divisions—the foregut, midgut, and hindgut
(Fig. 38). Sphincters (valves) control food–fluid movement between regions. The foregut (stomodeum) is
concerned with ingestion, storage, grinding and transport of food to the next region, the midgut
(mesenteron). In the midgut, digestive enzymes are produced and secreted, and absorption of the products
of digestion occurs. The material remaining in the gut lumen, together with urine from the Malpighian
tubules, then enters the hindgut (proctodeum), where absorption of water, salts and other valuable
molecules occurs, prior to elimination of the faeces through the anus.
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PHYSIO&TOXI
Figure 38. Alimentary System
Foregut = stomodeum
- lined with intima (endocuticle + epicuticle)
- little digestion occurs
- often has spines or teeth (denticles)
- grinding (gizzard-like function as in case of birds)
- sorting (e.g.: honey bee)
The foregut is subdivided into a pharynx, an oesophagus (esophagus) and a crop (food storage
area), and in insects that ingest solid food there is often a grinding organ, the proventriculus (or gizzard).
At the anterior end of the foregut, the mouth opens into a pre-oral cavity, which is bounded by the bases of
the mouthparts and often divided into an upper area, or cibarium, and a lower part, or salivarium. The
paired labial or salivary glands vary in size and arrangement from simple elongated tubes to complex
branched or lobed structures.
Midgut = Mesenteron
- site of digestion and absorption
- not lined with intima
Most digestion occurs in the midgut, where the epithelial cells produce and secrete digestive enzymes
and also absorb the resultant food breakdown products. The generalized midgut has two main areas – the
tubular ventriculus and blind-ending lateral gastric caeca (ceca). Most cells of the midgut are structurally
similar, being columnar and with microvilli (finger-like protrusions) covering the inner surface.
Hindgut = Proctodeum
Typically, the beginning of the hindgut is defined by the entry point of the Malpighian tubules, often
into a distinct pylorus forming a muscular pyloric sphincter, followed by the ileum, colon and rectum. The
main functions of the hindgut are the absorption of water, salts and other useful substances from the feces
and urine.
Fat body
The fat body tissue is a conspicuous internal component forming a pale tissue comprising loose
sheets, ribbons or lobes of cells lying in the haemocoel. The structure of this organ is ill-defined and
taxonomically variable. The fat body has multiple metabolic functions such as:
- the metabolism of carbohydrates, lipids and nitrogenous compounds
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- the storage of glycogen, fat and protein

- the synthesis and regulation of blood sugar
- the synthesis of major haemolymph proteins (such as haemoglobins, vitellogenins for yolk
formation, and storage proteins)
- an endocrine organ producing growth factors and hydroxylating the ecdysone synthesized in the
prothoracic glands to20-hydroxyecdysone (the active molting hormone)
Digestion of food
Carbohydrates and protein polymers are the primary food source of insects. These polymers are
digested by enzymes into monomers. The midgut has a pH of 6-7.5, but more basic pH occur in plant-
feeding insects which feed on hemicelluloses of plant cell walls, contrary to acidic pH of Diptera midgut.
High pH may prevent or reduce the binding of dietary tannins to food proteins, thereby increasing the
digestibility of ingested plants. Commonly, the midgut epithelium is separated from the food bolus by the
peritrophic membrane which constitute a very efficient high flux sieve. The passage of small molecules
while restricting the larger ones is due to the pores or small perforations. The peritrophic membrane also
acts as protection of insects from ingesting allelochemicals such as tannins. For some insects, digestion
occurs inside the peritrophic membrane (in the endoperitrophic space). In some, only the initial digestion
occurs in the endoperitrophic space then the smaller food molecules diffuse into the ectoperitrophic
space, where it is further digested. The final phase of digestion occurs on the surface of the midgut
microvilli.
Excretion
– The removal from the body of the waste products of metabolism, especially nitrogenous
compounds. Figure 39 summarizes the flow waste elimination in insects.
The main organs of excretion and osmoregulation in insects are the Malpighian tubules and the
rectum and/or ileum, acting together. Malpighian tubules consist of long, thin tubules formed of a single
layer of cells surrounding the lumen. They range in number from as few as two in most scale insects
(coccoids) to over 200 in large locusts. Generally, they are free, waving around in the haemolymph, where
they filter out solutes. Aphids are the only insects that lack Malpighian tubules, although they are reduced
to papillae in Strepsiptera. The anterior hindgut is called the ileum, the generally narrower middle portion
is the colon, and the expanded posterior section is the rectum. In many terrestrial insects, the rectum is
the only site of water and solute resorption from the excreta, but in other insects (e.g. desert locust
Schistocerca gregaria (Orthoptera: Acrididae)), the ileum makes some contribution to osmoregulation.
Figure 39. Generalized excretory system showing the path of elimination of wastes.
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PHYSIO&TOXI
Lesson 3. The Circulatory System

In most but not all insects, the abdomen is divided by
the dorsal and the ventral diaphragm (or septum) into three
partitioned regions: the pericardial sinus, the perivisceral
cavity, and the perineural sinus (Fig. 40).The dorsal
vessel lies on the upper surface of the dorsal diaphragm.
Loose clusters and strings of cells, called pericardial cells,
often occur on the external surface of the heart, and are
attached to the dorsal diaphragm at various places in the
pericardial sinus. The alimentary canal, reproductive
organs, and some of the fat body lie in the central or peri-
visceral sinus below the dorsal diaphragm. There often is a
ventral diaphragm separating the perineural sinus Figure 40. Cross-section of the internal abdomen of
containing the ventral nerve cord and ganglia from organs insects.
in the perivisceral sinus, but the ventral diaphragm usually
does not extend into the thorax.
The insect circulatory system is an open system called the hemocoel. The principal organ of whole-
body circulation in insects is a tubular vessel lying just beneath the dorsal body wall that generally runs the
length of the insect. The “blood,” usually called hemolymph, typically enters the abdominal portion, the
heart, through paired ostial openings, and is pumped anteriorly through the thoracic portion, the aorta. The
aorta sends out branches in a few insects, but generally it is a simple tube.
Structures of the circulatory system

The only closed portion of the circulatory system is the dorsal blood vessel referred to as the heart
and aorta (Fig. 41b). Sometimes the dorsal blood vessel is referred as the heart alone. The heart is the
abdominal portion of the vessel, extending into the thorax in some insects. The presence of alary muscles
and incurrent ostia determines the areas of the heart and aorta. It is open in the heart area by paired lateral
valved openings called ostia. In the relaxed stage, the ostia open and hemolymph enters. When the heart
contracts, the valves of the ostia are closed by pressure and the hemolymph is moved forward and pumped
into the aorta. The heart is also assisted in contractions by a special set of muscles known as the alary
muscles (Fig. 41a). Blood circulation is aided by the presence of a dorsal and ventral diaphragms.
a) b)
Figure 41. A diagram of the dorsal vessel, alary muscles, and branches of the dorsal vessel (a). Dorsal blood vessel
showing the aorta and heart.
Ostia
- small, slit-like, paired openings in the dorsal vessel that allow hemolymph to enter or leave the vessel.
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PHYSIO&TOXI
- Incurrent ostia allow hemolymph to enter during diastole and excurrent ones permit hemolymph to
exit.
- Pairs of ostia are usually located laterally, with one on each side of the heart, but some are ventrally and
dorsally located.
- openings tend to occur at the base of shallow pockets or at deeper, funnel-shaped invaginations in the
wall of the dorsal vessel, which often give the heart a chambered appearance
Alary muscles
- wing or delta shape in many insects, form part of the dorsal diaphragm.
- The muscles probably provide support for the heart. The muscle fibers typically fan out from a small point
of origin on the lateral wall of the dorsum to a broad insertion on the heart in many insects, presenting
the typical delta appearance.
Hemocytes
Hemocytes are blood cells. The seven most common types of hemocytes found in insects are:
1. Prohemocytes - the smallest hemocytes; known to divide and they may differentiate into plasmocytes,
which, in turn, may give rise to granulocytes, and these may differentiate into spherulocytes.
2. Plasmatocytes - small-to-large, polymorphic cells up to 40–50 μm in size, granular or agranular, and
round-to-spindle shaped in wet suspensions; may be binucleate.
3. Granulocytes - variable in size, spherical or oval, and up to 45 μm in size; nucleus is usually small and
the cytoplasm is granular; the granules are thought to be glycoproteins and mucopolysaccharides.
4. Spherulocytes - ovoid to round cells up to about 25 μm in length; may contain few to many small
spherical inclusions that stain for acid mucopolysaccharides.
5. Adipohemocytes - be small or large, spherical to oval, and contain lipid droplets
6. Oenocytoids - variable in size, often large, may be binucleate, and lyse easily, but do not cause
hemolymph coagulation when they lyse; non-phagocytic; contain prophenoloxidase, an inactive form
of phenoloxidase (PO)
7. Coagulocytes - also called hyaline hemocytes and cystocytes; rupture within seconds after injury
Functions of Hemocytes
- Hemocytes play major protective roles against invading microorganisms and the eggs or larvae of
parasitoids by production of antibiotics, encapsulation, PO production and release, and phagocy-
tosis.
- Hemocytes participate in wound healing
- Certain hemocytes, particularly coagulocytes and possibly granulocytes in some insects,
participate in the coagulation of plasma to help plug a wound.
- Some hemocytes contain enzymes that aid in detoxification, including detoxification of some
insecticides.
Hemolymph
The circulating fluid in insects is called hemolymph. It comprises 5-40% of the total body weight but
due to dehydration, physiological stage of development, and daily with food and water availability,
fluctuations in the volume is common. The hemolymph does not transport oxygen in insects. It contains
many dissolved inorganic and organic substances, colloidally suspended proteins, and lipoproteins. It is
about 90% water and 10% solids.
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PHYSIO&TOXI
Functions of hemolymph
- Lubricant for different tissues
- Hydraulic medium (newly emerged butterflies expand their wings by forcing hemolymph through
them)
- Transport of nutrients and wastes
- Protection (blood cells)
 Phagocytosis = ingestion of foreign particles such as bacteria, debris
 Encapsulation = clumping around invading organisms, e.g. internal parasites
 Detoxification - e.g. decomposing insecticides non-toxic hemolymph clottings
- Cryoprotectant effect: A number of freeze-avoiding insects (e.g.: moths, beetles), produce
antifreeze proteins (AFP-s) that can lower the freezing point of water, yet have no effect on its
melting point. AFPs in the hemolymph and gut inhibit the ice nucleators that seed ice crystal
formation and also help stabilize cell membranes. These proteins are high in threonine and cysteine
and are able to bind to ice crystals and prevent them from growing.
- Thermoregulation
 Heat for flying
 Heat for brooding eggs and larvae
 Heat to kill a predator
 Heat for maintaining hive temperature in winter
Hemoglobin
Some species of chironomid larvae, Chironomus tentans and others (Diptera: Chironomidae); horse
bot larvae, G. intestinalis (Diptera: Tendipendidae); and three bugs, Buocnoa margaritacea, Anisops
producta, and Macrocorixa geoffrey (Hemiptera) have hemoglobin as a suspended colloid in the plasma of
the hemolymph. The hemoglobins of the chironomids consists of as many as 12 monomers with molecular
weights of about 15,900 each; each monomer may be coded by its own gene. In C. tentans, the
hemoglobins account for up to 40% of the total proteins in hemolymph.
Heartbeat
The heartbeat is a wave of contractions (systole) generally originating at the posterior end of the heart
and traveling anteriorly. The rate of contractions or beats is highly variable in different insects and varies
with physiological conditions, temperature, species, and stage of development, nervous activity, and
neurosecretions. The rate may be as slow as 15 beats/min in the larva of Lucanus cervus (Coleoptera), to
rates near or higher than 100 beats/min in several insects. Proctolin, a neuropeptide produced in
motoneurons, interneurons, and neurosecretory cells stimulates heart rate. 5-Hydroxytryptamine
(serotonin) is a neurotransmitter in the nervous system, causing vigorous increase in heartbeat rate even
at very low concentrations.
Lesson 4. The Respiratory System

Respiration or breathing and gas exchange, is a function of the tracheal system, a tubular network
that originates at spiracular openings on the body surface and radiates to all parts of the insect body. The
respiratory system functions to provide the cells and tissues with oxygen, eliminate carbon dioxide a product
of cellular respiration and work in conjunction with the circulatory system in providing oxygen to the flight
muscle system
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Structure of the respiratory system

Tracheae and tracheoles have an epicuticular lining,
composed primarily of a cuticulin layer that is continuous with
the external cuticle. Larger tracheal trunks have an
endocuticle layer that gives more strength to the tubular
structure. A hydrophobic substance is secreted on the lumen
surface of tracheae that helps prevent water from entering
the tracheae and reduces evaporative water loss from the
humid, extensive internal tracheal surfaces.
The major distinction between tracheae and tracheoles
is size. Tubes down to about 1 μm in size are called tracheae
(sing. trachea), while those smaller than 1 μm are trache-
oles. Tracheoles typically have fluid in their terminal endings,
their amount believed to be related to metabolic demand for
oxygen delivery to cells near the endings.
Figure 42. Cross-sectional structure of the insect’s
respiratory system.
Spiracles
The openings of the tracheal system at the body surface are called spiracles. Spiracles usually occur
on the pleural surfaces of the body, typically one on each side of each segment, but numerous variations
have evolved. Most terrestrial insects can close the spiracles as an adaptation for water conservation.
Types of respiration or gaseous exchange in insects

1. Cutaneous respiration-gaseous exchange directly through the cuticle. Occurs to a limited extend
in all insects. Occurs in Protura and those Collembola that lack a tracheal system (ancient primitive
insects)
2. Gaseous exchange in terrestrial insects
a. air-tube diffusion
b. tissue diffusion
- oxygen diffuses in air 100,000 times faster than it does in water
- carbon dioxide travels much faster through tissues than oxygen
In many species, air movement, plus spiracular opening and closing, is coordinated with the
ventilatory movements of the abdomen so that air is pushed out when the abdominal muscles contract and
sucked in when they relax.
Ventilation
• Ventilation is the use of muscles to assist in air movement
• It varies and depends on the insect
• In well flying insects:
a. Thoracic pumping in locusts, large moths and beetles, and dragonflies
b. Abdominal pumping in hymenopterans and dipterans
Discontinuous ventilation
The spiracles remain closed for a period of time. Movement of gases occurs in discrete bursts. This
type of ventilation usually occurs when the insect is at rest and it also occurs in pupae. In diapausing pupae
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of Hyalophora the interburst period may be 8 hours in which time the spiracles remain closed. During
closure, relatively little gaseous exchange takes place.
Lesson 5. The Muscular System, Locomotion and Flight

Insects possess only striated muscles characterized as overlapping thicker myosin and thinner
actin filaments that give the appearance of cross-banding under a microscope. Each striated muscle fiber
comprises many cells, with a common plasma membrane and sarcolemma, or outer sheath.
Basic structure of a muscle

A muscle is made up of a number of long multinucleate fibers. These muscle fibers are composed
of myofibrils embedded in the cytoplasm extending from one end of the muscle fiber to another. Myofibrils
are composed of sarcomeres, each of which contains actin and myosin, which through their cyclical ATP-
dependenet interactions generate the contractile forces and movements that shortens each sarcomere and
overall myofibril thereby producing muscle contraction.
Functions of the muscular system

- Support of the body
- Helps maintain posture
- Movement of the limbs, including ovipositor
- Movement of the wings (insects are the only invertebrates that fly)
- Movement of the viscera
- Locomotion
- Closure and opening of spiracles
- Operation of various pumps such as cibarial pump and the pumping of the poison glands
- Generation of heat by “shivering”
Types of insect muscles

1. Visceral muscle
- move the viscera and have only one or, commonly, no attachment to the body wall (e.g. muscles
around the gut and ducts of the reproductive system).
- resembles skeletal muscle in contrast to the “smooth” visceral muscle of vertebrates
2. Cardiac muscle
- insect heart usually consists of a simple tube that contains a layer of contractile myocardial cells.
These are usually mononucleate cells with striated longitudinal and circular myofibrils.
3. Skeletal muscles
- can be differentiated functionally into synchronous and asynchronous muscles.
Synchronous and asynchronous muscles

Skeletal muscles of insects can be divided into two groups based on the contraction rate per nerve
input. Contraction frequency in synchronous muscles is directly controlled in a 1:1 manner by the output
of nerve impulses from the Central Nervous System. In asynchronous muscles, the contraction frequency
is a property of the muscle itself and its anatomical arrangement in the musculo-skeletal system.
Asynchronous wing muscles, the dorsal longitudinal and dorsoventral, do not attach directly to the wings,
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but are attached to the thoracic cuticle. Asynchronous muscles oscillate and produce several to many
contractions in response to a single neuronal stimulus.
Mechanics of muscle contraction

Muscle contraction results from the thin and thick filaments sliding relative to each other so that each
sarcomere is shortened. This is driven by the ATP-dependent cyclical biochemical/biomechanical activities
of individual myosin motor domains.
Muscle Attachments to the Exoskeleton

Skeletal muscles are usually anchored to the epicuticle layer of the exoskeleton. A large 2.5 MDa
extracellular matrix protein (Dumpy protein) encoded by the dumpy (dp) gene in D. melanogaster connects
the outer membrane of muscle fibers to the basal membrane of epidermal cells through a cross-linking zona
pellucida domain and a transmembrane anchoring sequence. Dumpy protein provides a strong attachment
for muscles to cells and cuticle, allowing high mechanical tension and preventing the tearing of muscle
away from the cuticle. Bundles of intracellular microtubules originate at the junction with the epidermal cells,
pass through the epidermal cells, and, at the cuticle, are cemented to fibers of chitin (tonofibrillae) that are
formed extracellularly and embedded in the cuticle.
Muscles and development

• Muscle development and maintenance in hemimetabolous insects differs greatly from that in
holometabolous insects.
• In the former, the transition is gradual and the same muscle sets are present in the adult that were
in the larva.
• In the holometabolous insects, most immature muscles degenerate (i.e., histolyzed) and new ones
are developed.
• Basically there are four ways in which muscles undergo modifications in holometabolous insects:
1. Larval muscles may pass unchanged into the adult
2. Existing larval muscles are reconstructed
3. Larval muscles may be destroyed and not replaced
4. New muscles, not represented in the larva such as flight muscles and
leg muscles, are formed from imaginal discs.
*Muscles are resistant to molting fluids, thus they remain functionally attached to the
cuticle at apolysis and ecdysis.
Locomotion
a. Walking
Movement with six legs at slow to moderate speed requires continuous contact
of a tripod fore and hind legs one side and midleg on the opposite sides thrusting
rearwards (retraction) while each opposite legs move forward (protraction). This tripod
acts as the center of gravity, providing great stability. Movement is initiated through
action of the thoracic muscles on the leg bases with transmission to extend or flex via
the internal muscles. Anchorage is necessary as lever to propel the body, is through
pointed claws and adhesive pads (the arolium or, in flies and some beetles,
pulvilli). For faster movement, the following are alternatives: (1) increase in the Figure 43. A ground beetle
(Coleoptera: Carabidae:
frequency of the leg movement by shortening the retraction period; (2) increase
Carabus) walking in the
direction of the dashed line.
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PHYSIO&TOXI
in the stride length; (3) alteration of the triangulation basis of support to move two- or four-legged while
other legs are held above.
b. Crawling
Crawling is observed in most soft-bodied larvae with hydrostatic skeletons. Muscular contraction in
one part of the body gives equivalent extension in a relaxed part elsewhere on the body. In legless larvae,
waves of contractions and relaxation run from head to tail. Forward motion is facilitated by the adhesive
hooks that grips and detach from the substrate. For naiads, lateral waves of contraction against the
hydrostatic skeleton can give a sinuous, snake-like, swimming motion, with anterior-to-posterior waves
giving an undulating motion. For caterpillars (with thoracic legs) posterior to anterior waves of turgor muscle
contraction was developed. Their movement is described as cycles of successive detachment of the
thoracic legs, reaching forwards and grasping the substrate.
c. Swimming
When swimming, water contact is maintained during drawing out. In swimming, contact with water
should be maintained and the insect must thrust to a rowing motion than the recovery stroke to advance.
This is achieved by expanding the effective leg area during retraction by extending fringes of hairs and
spines. Specialist insects with water-repelling cuticles and fringes can move in surface film of water.
Mechanism of Insect Flight

The mechanism responsible for the beating of wings is simple. The wing-bearing segments can be
imagined as a box: the pleura and sternum rigidly fused and the wings connected to the pleura by flexible
membranes. The membranous attachment and wing hinge are composed of resilin giving elasticity to the
thoracic box. There are two kinds of muscle arrangement powering flight, the direct muscles connected
to the wings and indirect muscles where there is no muscle to wing connection, but rather muscle action
deforms the thoracic box in order to move the wings such as in the case of Odonata and Blattodea. Direct
muscles produce the upward stroke by contraction of muscles attached to the wing base inside the pivot
point (Fig. 44a) while the downward wing stroke is produced through contraction of muscles that extend
from the sternum to the wing bas outside the pivot point (Fig. 44b). The indirect muscles are attached to
the tergum and sternum. Contraction causes the tergum, and with it the very base of the wing, to be pulled
down which in turn levers the outer main part of the wing to go up (Fig. 44c). The downbeat is pulled through
contraction of the second set of muscles (running from back to thorax), forming a box and lifts the tergum
(Fig. 44d). At each stage in the cycle, energy is conserved when flight muscles relax as the elasticity of the
thorax restores its shape.
Figure 44. Direct flight mechanisms: thorax during (a) upstroke and (b)
downstroke of the wings. Indirect flight mechanisms: thorax during (c)
upstroke and (d) downstroke of the wings
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Solutions to problems of turbulence
 Flap hind wing first

 Reduced forewing stroke (Orthoptera, Coleoptera)
 Flap wings simultaneously
 Wing loss Diptera, Strepsiptera Muscles
 Wing coupling (Lepidoptera, Hymenoptera, Trichoptera, Hemiptera)
- the fore- and hindwings are linked anatomically so that they move together as a single unit
- involves lobes or spines at the wing base.
Types of wing coupling:

a. Jugal wing coupling – exhibited by more primitive forms of moths. They have enlared lobe-like area
near the basal posterioi margin, or at the base of the forewing called jugum that folds under the hindwing
during flight
b. Frenulo-retinacular wing coupling – some moths possess frenulum on the hindwing that hooks under
a retinaculum on the forewing.
- Retinaculum – hook on the underside of the forewing in moths
- Frenulum – spine at the base of the forward or costal margin of
hindwing forming coupling mechanism
c. Amplexiform wing coupling – (e.g. butterflies and Bombycoidea)
- An enlarged humeral area of hindwing is broadly overlapped by the forewing
- the forewing’s power stroke pushes down the hindwing simultaneously
d. Hamulate – a row of small hooks on the coastal margin of the hind wing called hamuli and folded lower
margin in the forewing (e.g. honeybees)
- during flight, the hamuli lock onto fold at the lower margin of the forewing and function as one unit
- both wings move together with utilizing body energy in efficient way to fly higher and faster
Figure 45. Wing coupling mechanisms involving the jugal and humeral regions of the wings
Lesson 6. Coordination and Integration

Similar to vertebrates, the basic component of the nervous system is the nerve cell (neuron),
composed of a cell body with two projections: (1) the dendrite, which receives stimuli; and the (2) axon,
which transmits information, either to another neuron or to an effect or organ such as a muscle. Insect
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PHYSIO&TOXI
neurons release a variety of chemicals at synapses to either stimulate or inhibit effector neurons or
muscles.
Types of neurons
- sensory neurons receive stimuli from the insect’s
environment and transmit them to the central nervous
system
- interneurons (or association neurons) receive information
from and transmit it to the neurons
- motor neurons receive information from interneurons and
transmit it to muscles
- neurosecretory cells (neuroendocrine cells)
Figure 46. Nerve cell and its parts.
Basic functions of neurons:
a. generation of electrical response
b. conduction of electrical response
c. transmission of electrical response
The cell bodies of interneurons and motor neurons

are aggregated, with the fibers interconnecting all types
of nerve cells to form nerve centers called ganglia.
Simple reflex behavior has been well studied in insects,
but insect behavior can be complex, involving integration
of neural information within the ganglia.
Figure 47. Diagram of a simple reflex mechanism of an insect.
Original number of ganglions per tagma

head - brain – 3 pairs - suboesophageal ggl. – 3 pairs
thorax - 3 pairs (pro-, meso- and metathoracic)
abdomen - 8 pairs (usually less)
The Central Nervous System (CNS)

The CNS is the principal division of the nervous system, and consists of series of ganglia joined by
paired longitudinal nerve cords called connectives. Two ganglia of each thoracic and abdominal segment
are fused into a single structure and the ganglia of all head segments are joined to form two ganglionic
centers–the brain and the suboesophageal (subesophageal) ganglion. The chain of thoracic and
abdominal ganglia found on the floor of the body cavity is called the ventral nerve cord. The brain, or the
dorsal ganglionic center of the head, is composed of three pairs of fused ganglia (from the first three
head segments):
- protocerebrum - the eyes and thus bearing the optic lobes
- deutocerebrum - the antennae
- tritocerebrum - handling of the signals that arrive from the body
Joined ganglia of the three mouthpart-bearing segments (mandibular, maxillary and labial) form the
suboesophageal ganglion, with nerves emerging that innervate the mouthparts. The visceral (or
sympathetic) nervous system consists of three subsystems: the stomodeal (or stomatogastric) nervous
system (which includes the frontal ganglion); the ventral visceral nervous system; and the caudal
visceral nervous system. Together, the nerves and ganglia of these subsystems innervate the anterior
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PHYSIO&TOXI
and posterior gut, several endocrine organs (corpora cardiaca and corpora allata), the reproductive organs,
and the tracheal system including the spiracles.
Figure 48. The central nervous system of various insects showing the diversity of arrangement of ganglia in the ventral nerve cord (a).
The CNS showing the different regions (b) CNS in close up.
Protocerebrum
The protocerebrum is associated with the
compound eyes, ocelli, and some integument sensory
receptors. The two large lateral lobes containing dense
groupings of neurons and neurosecretory cells within the
area known as pars intercerebralis.
Initial nerves: 3 (2 paired, 1 unpaired) nerves:

a.) nervus opticus to the ocelli of frons and vertex Figure 49. Structure of the Protocerebrum
b.) nervus connectivus to the a stomatogastric system
c.) nervi corporis cardiaci to the corpori cardiaci
Its most important regions:

1. Optic lobes
- process and analysis of optic information:
Properties:
• inversion of pictures
• retinotopic mapping
• pattern recognition with “small-field” neurons
• general motion recognition horizontally and vertically with “wide-field” neurons
Structure of optic lobes (lobus opticus)

1. lamina (lamina ganglionaris)
2. medulla (medulla externa)
3. lobula (medulla interna)- in some insects it has two parts: (a. lobula b. lobula plate)
4. nerve-fibre intercrossings – (outer chiasma and inner chiasma)
2. Mushroom bodies (corpus pedunculatum)

- cognitive and associative functions relating to olfaction and vision
- centers for learning and memory
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State of development:
a.) big – social insects Hymenopterans, termites
b.) intermediate – Coleopterans, Lepidopterans
c.) small – Heteropterans, Dipterans
Parts: a.) cup (calyx) b.) handle (pedunculus) c.) lobes (lobi): α-, β- and (sometimes) γ-lobus
3. Central body
- Composed of a lot of nerve-fibre intrecrossings, rich arborisation of axons and dendrites
responsible for the integration of information between the two hemispheres of brain
Deutocerebrum
- bilateral innervations of antennae – nervus
olfactorius
a. olfactory glomeruli
b. connection with sensillae of antennae
c. neuromuscular junctions with muscles of
antennal segments
Olfactory glomerulus:
- ball or cap like shape (50-100 μm diameter) Figure 50. Structure of the Deutocerebrum
- rich dendrite and axon terminals
- their number is typical for an insect species
- their neurons have special interconnections (synapses) with certain neurons:
1. local interneurons
2. projective neurons (uni- and multiglomerular)
3. so called “incoming neurons”
Most important physiological role of deutocerebrum:

- processing and transmission of olfactory and partly mechanical information coming from antennae
- motorial control of antennal segments
Tritocerebrum
- its two parts are connected with a commissure
- is connected to the stomatogastirc system (ganglion frontale) with two slight nerves
- innervated regions: mostly pharynx and labrum
Suboesophageum
- its three parts innervates the three parts, segments of the mouth (mandibular, maxillar and
labial)
- contains a lot of nerve-fibres
- its sensory and/or motorial nerves innervate: maxillar and labial palpi, mouth parts, salivary
glands, muscles of neck region
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PHYSIO&TOXI
Sympathetic nervous system

This element of the nervous system, innervates the gut, heart, and endocrine glands and forms a
network of peripheral ganglia that is associated with digestive processes. These ganglia innervate the
muscles of the oral cavity, foregut, and midgut and regulate food ingestion, transport, digestion, and the
excretion of wastes.
• provides innervations for visceral organs with its sensory and motorial nerve fibers
• its nerve fibers create rich arborisation in the walls of visceral organs
• contains paired and unpaired ganglions nearby the innervated regions
Parts:
1. Stomatogastric system (Fig. 51) - specifically innervates the foregut and midgut
- The frontal ganglion that arises from the frontal connectives issued by the tritocerebrum.
- The frontal ganglion innervates the foregut
and controls crop emptying. It gives rise to
the recurrent nerve that passes underneath
the brain and expands into the
hypocerebral ganglion that innervates the
corpora allata, corpora cardiaca, and the
fore- and midguts.
2. unpaired ventral nerve-trunk and its
connections
3. Caudal autonomic sympathetic system:
consists of those nerves that innervate the
hindgut and genitalia, usually originating in
the compound terminal abdominal
ganglion.
Figure 51. Lateral of the brain showing the major ganglia

of the stomodeal nervous system.
The Peripheral Nervous System
The PNS consists of all of the motor neuron axons that radiate to the muscles from the ganglia of the
CNS and stomodeal nervous system plus the sensory neurons of the cuticular sensory structures (the
sense organs) that receive mechanical, chemical, thermal or visual stimuli from an insect’s environment.
Lesson 7. Molting and Formation of New Cuticle

The insect grows but not its exoskeleton. The external skeleton gradually becomes too small for the
growing body tissues of an immature insect so it must molt its cuticle. Molting is a vulnerable time for
insects; they are easy prey for predators and subject to environmental hazards, particularly desiccation.
The muscles that move the body must be detached from the old cuticle, but they are detached only
immediately before the ecdysis and new muscle attachments are made quickly to the new epicuticle. The
new cuticle must harden sufficiently to resist the pull of the muscles or else, may cause skeletal deformation
resulting to permanent restriction of movement, especially flight. Figure 52 summarizes the steps involved
in molting of insects.
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PHYSIO&TOXI
Step 1: Apolysis or
Step 6. Epidermis Step 7: Ecdysis or
the separation of old
secretes new shedding of the old
exoskeleton from
procuticle exo- and epicuticle
the epidermis
Step 2: Secretion of Step 5: Digestion and

Step 8: Expansion of
inactive molting fluid absorption of old
new integument
by epidermis endocuticle
Step 3: Production of Step 9: Tanning or

Step 4: Activation of
cuticulin layer for sclerotization of new Figure 52. Events in insect
molting fluid
new exoskeleton exocuticle molting
Hormonal regulation of molting
Molting is regulated by the endocrine and nervous systems. Stretch receptors in the body probably
are stimulated by the increasing growth of body tissues and, when it attains the critical size, the brain
secretes prothoracicotropic hormone (PTTH). PTTH is released into the circulating hemolymph,
circulates around the body, and binds specific receptor proteins on the surface of prothoracic gland (PGL)
cells. Multiple biochemical reactions are initiated that result in the synthesis of ecdysone by the PGL.
Ecdysone is released into the circulating hemolymph and converted into 20-hydroxyecdysone by
epidermal cells as well as other tissues. Epidermal cells respond to 20-hydroxyecdysone by separating
from the old cuticle (apolysis) and by mitotic activity that produces new cells to spread over the larger body
surface that must be enclosed within the new epidermal cell layer and new cuticle. Apolysis, or separation
of the epidermal cells from the old cuticle, marks the beginning of a molt and of a new instar, and the animal
within the loosened, but not yet shed, cuticle is the pharate next instar (or stage, if the next form is the pupa
or adult).
Molting fluid
Molting fluid is first evident as droplets of polar lipids secreted by the epidermal cells into the
apolysial space. As a result of coalescence of these droplets, an ecdysial membrane soon appears and
persists during the pre-molt period but later will be shed along with the old exuvium. Molting fluid contains
both proteinases and chitinases that digest the proteins and chitin, respectively, in the old endocuticle.
Ecdysis
Ecdysis has been divided into phases: pre-ecdysis I, pre-ecdysis II, ecdysis (eclosion), and post–
ecdysis. The process of molting from one stage to another and eventual metamorphosis to the adult form
is coordinated by a suite of neuropeptides: corazonin, pre-ecdysis-triggering hormone (PETH), edysis-
triggering hormone (ETH) eclosion hormone (EH) and CCAP and bursicon and the hormones PTTH,
ecdysone, and juvenile hormone (JH).
Prior to ecdysis, the brain neurosecretory cells release PTTH that targets the prothoracic gland cells,
inducing the synthesis and release of ecdysone into the hemolymph. Ecdysone is then converted to 20-
hydroxyecdysone that triggers the epidermal cells to prepare for the new synthesis of cuticle; juvenile
hormone is also secreted from the corpora allata which will determine whether the new cuticle produced
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will be larval, pupal, or adult. The high level of 20-hydroxyecdysone in the hemolymph induces receptors in
the central nervous system for the ecdysis-triggering neuropeptides (PETH and ETH) and stimulates
production of ETH in the Inka cells. These neuropeptides cause pre-ecdysis I contractions which occur
synchronously in thoracic and abdominal segments. These pre-ecdysis contractions occur every 10–12 s
with a contraction lasting 5–7 s. Slowly PETH and ETH accumulate in the hemolymph, leading to pre-
ecdysis II characterized with more vigorous and frequent contractions. Eclosion hormone (EH) will be
released as a result of accumulated ETH.
The final ecdysis contractions are a series of peristaltic waves of contractions that originate in the
most posterior segment and pass anteriorly for about 10 min until ecdysis is complete.
Figure 53. (a) Ecdysis of the adult cicada Neocicada hieroglyphica from the last nymphal cuticle. (b) The adult with partially expanded
wings. (c) The adult Neocicada hieroglyphica with fully expanded wings, but cuticle not yet hardeneded and darkened. (d)
Adult Neocicada hieroglyphica with sclerotized and darkened cuticle.
Post-ecdysis
- Hardening (sclerotization) and darkening of the cuticle and wing expansion (Figure 4.9d) are
under control of the tanning hormone, bursicon
- Following actual shedding of the old cuticle, lipids and cement are secreted onto the new
epicuticle, muscle reattachments become firmly fixed to the epicuticle, and the cuticle begins
to sclerotize.
Sclerotization of Cuticle
Sclerotization is the process of cross-linking protein-to-protein chains, chitin-to-chitin chains, and
possibly protein-to-chitin chains also called tanning or simply, hardening of the cuticle. Tanning refers to
the cross-linking process itself and not to a color change, although sclerotization often is accompanied by
tan, brown, or black colors. The phenols associated with sclerotization easily undergo autoxidation (phenols
to quinones) and quinones readily polymerize, processes usually leading to melanin and tan or brown to
black colors. Hardening (sclerotization/tanning) and darkening are two different processes, and cuticle can
become sclerotized without darkening (e.g. compound eyes).
Only protein-to-protein sclerotization occurs in the epicuticle because there is no chitin, but in other
layers of the cuticle all the combinations may exist. Sclerotization gives strength and rigidity to the cuticle.
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N-β-alanyldopamine has been implicated as the principal tanning agent in the pupal cuticle. Bursicon is
a neuropeptide that promotes sclerotization and specifies how much cross-linking of molecules will occur.
Lesson 8. Sensory systems

Vision
Almost all insects are able to detect light energy. The basic mechanism of stimulation is very likely
the same: the solar energy striking the photo sensory cell is absorbed by pigments in the cell. The pigment
undergoes as light conformational change that causes a momentary increase in permeability of the receptor
cell membrane and, thereby, the initiation of nerve impulses that travel to the optic lobes and CNS.
Two main types of insects' eyes:

1. compound eyes
2. simple eyes
Figure 54. Cross-section of the (a) compound and (b) simple eye.
Compound eyes
 Occur in most adult and hemimetabolous larvae on either side of the head capsule
 Absent in ancient insect orders (e.g. Protura, Diplura), wingless parasite adults (lice, fleas),
subterranean termites, female scale-insects and cave-dwelling forms (sensitivity to light over the
general body surface)
 Consists of 1 (ants) to 28.000 (dragon flies) cone-shaped sensory cartridges, facets = ommatidia
(sing. ommatidium); in hexagonal arrangement on the surface
Basic functions of insect compound eyes

o image formation (visual acuity, mapping of forms)
o mechanism of vision
o color vision, differentiating of colors
o perception of light intensity changes, photo adaptation, perception of flickering and motion
o perception of light polarization
o spatial vision, perception of distances
Mechanism of vision:
- Starts with activation of rhodopsin and the resulting membrane depolarization.
- The unstable cis-retinal is converted to its trans form when it absorbs a photon of light.
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- Its transformation to the stable form of metarhodopsin activates G proteins that ultimately causes the
membrane depolarization.
- The metarhodopsin loses its ability to activate the G-proteins after it is phosphorylated and binds with
arrestin proteins to assume its unstable form.
- Light converts this metarhodopsin to an inactive rhodopsin that releases its arrestin and again becomes
capable of responding to light.
- Regeneration of the photopigments is a rapid process (0.7 ms) transretinal - 11-cis-retinal conversion.
Simple eyes
Two basic types:
1. Stemmata (in lateral position)
- the only visual organs of larval holometabolous insects (absent in Siphonaptera, Hymenoptera,
certain Diptera)
- located on both side of the head capsule of caterpillars 6-6; Tenthredinidae larvae 1-1
- functions for rough mosaic vision, poor color vision, movement perception and minor spatial vision
2. Ocelli
- typical of winged adult and hemimetabolous larvae - location: one medial ocellus on the frons and
two on the vertex in a triangular arrangement
- Functions for: perception of rapid changes in light intensity, poor color perception
Extraocular (non-visual) photoreception

• with the help of simple photoreceptors in the epidermis
• in a diffuse arrangement e.g. larvae of flies and larvae of Tenebrio molitor
• at the end of the abdomen on both side of the genitalia so called genital photoreceptors which have
wide importance in mating and egg laying
Mechanoreception
 well-developed accessory units helping perception of stimuli
 displacement, deformation or movement of the sensillum trigger the receptor potential
Basic mechanoreceptors:
1. tactile hair (trichoid sensilla)
2. bell shaped sense cone (campaniform sensilla)
3. stringed or chordotonal sensilla
4. free nerve endings forming nerve nets
Tactile hair
- reacts to deflection
- pore free hair
- narrows towards its tip
Parts:
- cuticular hair - bipolar receptor cell (with tubular dendrites)
- seta forming cell (trichogen cell)
- articulation membrane forming cell (tormogen cell)
- dendrite sheath forming cell (thecogen cell)
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PHYSIO&TOXI
- articulates on a flexible membrane ring - often forms hair pates
Campaniform sensilla
- reacts to expansive and compressive forces of the cuticle
Parts:
• bipolar receptor cell
• trichogen and thormogen cell
Main types of insects’ mechanoreception:

1. tactile sense
2. static sense
3. sense of vibration and hearing
4. sense of internal tensions
Forms of static sense:

• posture and position sense
• sense of force of gravity
• pressure sense
• sense of drift - sense of acceleration
Sense of vibrations
• vibrations can be perceived on the boundaries of mediums e.g. solid - air, air water
• relatively low frequency periodic stimuli
• can be perceived only from a short distance
• generally has alarming effect or plays role in communication
Hearing (phonoreception)
• perception of vibrations spreading in the medium (air or water)
• changes in pressure effect as stimuli
• frequency is high, it can be measured generally in kHz
Johnston’s organs
• large complex chordotonal organs
• occurs within the second segment of antennal flagellum (pedicellus) of all adult and many larval
insects
• consists of numerous scolopidium in radiant arrangement within two rings to perceive air-borne
vibrations
Tympanal organs
• the most elaborate sound reception system in insects; occur always in pairs
• typical parts: eardrum a cuticular membrane (tympanum), wall of drum cavity, drum cavity,
sometimes another drum (anti-tympanum)
• drum cavity has air sack origin (there are air storage sacks within the respiratory system)
• sometimes from numerous scolopidium (~60 pc) a hearing crest (crista accustica) is formed
• receptible frequency differs according to species: 0,1-100 kHz
Types of tympanal organs:

a. located in the thorax (thoracal tympanal organ); Occurs: Noctuidae (metathorax)
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b. located in the abdomen (abdominal tympanal organ); Occurs: Acrididae, Cicadidae,

Geometridae
c. located in the fore legs (tibial tympanal organ); Occurs: Gryllidae, Tettigoniidae
Thermoreception
• Insects actively seek out a “preferred” temperature
• insects evidently detect variation in temperature, yet the function and location of receptors is poorly
known
• most studied insects have antennal sensing of temperature (mosquitoes)
• transduction mechanism for thermoreceptors remains unclear
Hygroreception
Many observations on their behavior indicate that insects are able to monitor the amount of
water/vapor in the surrounding air. Insects actively seek out a “preferred” humidity or sources of liquid.
Humidity detectors are typically located on the antennae, though they occur on the spiracles and their
surroundings.
Properties:
- typical sense organ: sensillum coeloconicum with 1 thermosensitve and 2 hygro-sensitive receptor
cells
- There are two types of hygroreceptors: “moist” receptors respond to increasing humidity, while “dry”
receptors are stimulated by a decrease in humidity.
Chemoreception
1. smell, olfaction (olfactory chemoreception)
2. tasting, gustation (gustatory chemoreception)
3. general chemical sense
Olfaction
• sense of materials derived from near or further sources
• sense of gases or other volatile compounds
• generally lower stimulus (membrane) threshold
Gustation
• requires direct contact for sense (contact chemoreception)
• perception of liquid or water-soluble compounds
• generally higher membrane threshold
Common chemical sense

• aroused by exciting, irritant materials and pervasive essential oils
• they effect generally in higher amounts, concentrations
• several types of sensory neurons participate in a non-specific way
• they trigger always self -defending mechanisms
Types of sensilla in case of chemoreception

a. sensillum trichodeum
b. sensillum basiconicum
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c. sensillum placodeum
d. sensillum coeloconicum
Lesson 9. Insect Communication

Insects also have many ways to communicate but, unlike humans, their "language" is almost entirely
innate. Each individual is born with a distinctive "vocabulary" that is shared only with other members of its
own species. An insect may send a communication signals by doing something (e.g. make a noise, release
a chemical, or flash a light) or the signal may simply be an inherent part of the insect's physical makeup
(e.g. wing pattern, body color, or surface chemistry). Some form of intraspecific communication is a
prerequisite for any behavior that involves the participation or cooperation of two or more individuals.
Insects may also communicate with members of other species (interspecific communication). The adaptive
value of these communication signals may include:
– Recognition of kin or nest mates
– Locating or identifying a member of the opposite sex
– Facilitation of courtship and mating
– Giving directions for location of food or other resources
– Regulating spatial distribution of individuals - aggregation or dispersal; establishing and maintaining
a territory
– Warning of danger; setting off an alarm
– Advertising one's presence or location
– Expressing threat or submission (agonistic behaviors)
– Deception / mimicry
Tactile Communication
"Keep in touch!"
• Since many insects have poor vision and sound perception, physical contact provides an important
avenue of communication.
• Antennal tapping is also an essential component of communication in both ants and termites. It's
not clear exactly what information may be exchanged, but it certainly involves nest mate recognition
and leads to exchange of food through trophallaxis.
• Antennal tapping on the hind legs is used during tandem running in both ants (Fig. 55) and termites.
This is a "follow-the-leader" behavior in which the tapping informs the leader that she has not lost
her disciple. If tapping stops, the leader instinctively turns around and searches in ever-widening
circles until she re-establishes contact with the follower.
Figure 55. Antennal tapping in ants
• The "dance" language of honeybees is largely a tactile communication system, performed in total
darkness on the vertical surface of the honeycomb.
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• A "round dance" signals to nest mates the presence of a nectar source in close proximity to the
hive (usually less than 80 feet). It consists of a series of circular runs with more or less frequent
changes in direction. The greater the frequency of direction changes, the better the quality of the
nectar source.
• The "waggle dance" is used for longer distances. It involves a figure eight pattern with a series of
abdominal waggles on a straight run after each half-circle turn (see figure at left). Distance is
indicated by the duration of the straight run and the frequency of the waggles. Direction is indicated
by the angle of the straight run (relative to vertical) and corresponds to the horizontal angle between
the sun and the direction of the food source.
o Advantages:
 Instantaneous feedback
 localized area
 Individual recipient
 Effective in the dark (e.g. caves, wood galleries)
o Disadvantages:
 Not effective over distance
 Organisms must stay in direct contact
 Message must be repeated to each recipient
 Vibration signals can be intercepted by predators
Acoustic Communication
• Sound is produced when a physical object vibrates rapidly, disturbs nearby air molecules or other
surrounding medium, and generates compression waves that travel in all directions away from the
source.
• Some insects produce and detect sounds that are above the frequency range of humans.
• Some grasshoppers and moths, for example, produce
ultrasonic sounds as high as 80,000 Hertz.
• Most insects detect sound with a tympanic membrane
in the abdomen (e.g. grasshoppers and moths) or in
the tibiae of the front legs (e.g. crickets and katydids).
• Mosquitoes have antennal hairs that resonate to
certain frequencies of sound.
• Some species of ants, bees, termites, and treehoppers
can sense substrate vibrations with mechanoreceptors
(chordotonal organs) in their legs. Since these signals
are "felt" rather than "heard", they are usually regarded
as a form of tactile communication.
Figure 56. Mechanisms of sound production in insects
.
Mechanisms producing sound and vibrations
 Percussion - refers to vibrations produced by the impact of part of the body against the substrate
or by clapping two parts of the body against each other. A single tap on a solid surface produces a
complex
- E.g. Booklice (Psocoptera) and some stoneflies (Plecoptera) of both sexes generate
substrate vibrations through abdominal movements. Some species have evolved a small
cuticular knob on the ventral surface of the abdomen, which they rhythmically drum against
the ground.
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 Stridulation - production of vibrations by moving a cuticular ridge (called the scraper or plectrum)
on one part of the body over a toothed ridge (known as the file or strigil) on another. Repeated
contacts of the scraper against the teeth of the file cause part of the body to vibrate.
- E.g. In male Grylloidea, each tegmen has a file on the underside of the second cubital vein
near its base, while the ridge forming the scraper is on the edge of the opposite tegmen - the
tegmina are raised at an angle of 15–40_ to the body and rhythmically open and close so
that the scraper rasps along the file to produce sound. Each wing closure produces one
syllable of sound by vibrations of both tegmina.
 Timbal
- A timbal is an area of thin cuticle surrounded by a rigid frame. Vibrations are produced when
the timbal buckles, meaning that it gets alternately distorted and relaxed by a muscle
attached to its inner surface. Timbals are found in Hemiptera, especially in the Homoptera,
and in some adult Lepidoptera, primarily in Arctiidae.
 Signal produced by flight muscles
- oscillation of the flight muscles produces thoracic and wing vibrations which may be used in
communication by a number of insect species. Vibration of the wings in flight produces a
sound with a fundamental frequency the same as the frequency of the wingbeat.
- E. g. Lepidoptera with very low wingbeat frequencies (20 Hz), produce a flight tone that is
inaudible to humans, but insects with higher wingbeat frequencies produce clearly audible
- sounds and these may sometimes provide relevant signals to the insects.
 Air expulsion – relatively rare mechanism
- E.g. The death’s head hawk moth, Acherontia, repeatedly draws in and expels air through
the mouth by dilating and contracting its pharynx. Through vibrations of the epipharynx the
intake of air produces a series of sound pulses and the subsequent contraction of the pharynx
with the epipharynx held erect expels the air, producing a whistle.
Visual Communication
• The color patterns and other markings on the wings of butterflies and moths facilitate species
recognition
• Some insects use bright colors, eyespots, or other distinctive patterns to scare away predators, to
advertise their ability to sting, or to mimic the appearance of another unpalatable species.
• Other insects use dance-like body movements to attract a mate or to communicate with nest mates.
• Most of these signals are effective only as long as they are visible in daylight. But a few insects
(fireflies, for example) can generate their own light and use visual signals that can be seen at night.
• Many insects have the ability to see ultraviolet light. Female cabbage butterflies, for example, have
ultraviolet reflecting scales on the dorsal wing surface. When they fly, each down stroke of the
wing creates a brief "flash" of UV that males apparently recognize as the flight signature of a
potential mate. Missing scales reduce the wings' reflectivity - a sign of aging that impairs a male's
ability to attract a mate.
Chemical Communication
• Insects rely more heavily on chemical signals called semiochemicals or infochemicals, than on
any
• Insects may be highly sensitive to low concentrations of these chemicals - a few molecules may be
enough to elicit a response.
• Semiochemicals can be divided into two groups based on who "sends” a message and who
"receives" it:
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1. Pheromones - carry information from one individual to another member of the same species.
These include sex attractants, trail marking compounds, alarm substances, and many other
intraspecific messages.
2. Allelochemicals - travel from one animal to some member of a different species. These include
defensive signals such as repellents, compounds used to locate suitable host plants, and a vast
array of other substances that regulate interspecific behaviors.
Chemicals elicit certain responses

 Arrestants – chemicals that cause an organism to aggregate or stop when in contact with it.
 Attractants – chemicals that cause an organism to make oriented movements towards a source.
 Repellents – chemicals that cause an organism to make oriented movements away from a chemical
 Stimulants – chemicals that cause insect to disperse rapidly, begin feeding, mating or oviposition
 Deterrents – chemicals that inhibit feeding, mating or oviposition when in a place where an
organism would, in its absence feed, mate or oviposit.
Pheromones - intraspecific chemicals where both the emitter and receiver benefits (+,+)
 Sex pheromones to attract mates
 Alarm pheromones in aphids
 Egg marking in parasitoids and fruit flies
 Territory marking in ants
 Trail markers in ants
 Aggregation pheromones in bark beetles
Table 6. Classes of pheromones that mediate interactions between conspecific individuals.

Class Characteristics
Released by one or both sexes; benefit both emitter and receiver, but
Sex pheromones not always, because pheromones also may be used to assess the quality
of the individual as a mate.
Anti-aphrodisiac Used by a male to mark a mated female, to render her unattractive to
pheromones other males; benefits male by helping to ensure his paternity, benefits
female by reducing harassment by courting males.
Epideictic or Deposited on the surface or inside a resource, such as a fruit or
marking parasitized host egg, to indicate that it has already been exploited;
pheromones benefits the marker by reducing competition for its progeny.
Used by social and semi-social species to indicate the location of
Trail pheromones exploitable resources with respect to the position of the colony; benefits
the colony by increasing the efficiency of foraging.
Emitted in response to danger, such as an attack by a predator; benefits
Alarm pheromones receivers as they modify their behavior to defend the colony, or to reduce
the probability of being captured.
Results in aggregations of conspecifics; potentially beneficial to all
Aggregation individuals because it decreases the risk of predation or parasitism,
pheromones improves the exploitation of a resource and/or increases the probability
of locating a mate.
Anti-aggregation Emitted when a resource is in danger of being over-exploited; benefits
pheromones both emitters and receivers because it reduces intraspecific competition.
Pheromones of Numerous pheromones produced to coordinate activities within the
social colony. For example, the cuticular lipids contain signals that serve in
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insects recognition of nest mates, caste, reproductive status and even whether
an individual is alive or dead. Queen pheromones serve many roles,
including the suppression of worker reproduction and the organization of
workers to feed and groom the queen.
SUMMARY
 Insect physiology is the study of the functional properties of insect tissues and organs. It requires
knowledge on anatomy, morphology and biochemistry. Insect physiology is vital in toxicological
studies of pesticides.
Female reproductive organs Male reproductive organs
 Paired ovaries composed of ovarioles  Paired testes composed of follicles
(ovarian tubes) (testicular tubes)
 Paired oviducts (ducts leading from  Paired vasa deferens (ducts leading from
ovaries) testes)
 Egg calyces (if present, reception of  Seminal vesicles (sperm storage)
eggs)
 Common (median) oviduct and vagina  Median ejaculatory duct
 Accessory glands (ectodermal origin: Accessory glands (two types):
colleterial or cement glands)  ectodermal origin
 mesodermal origin
 Bursa copulatrix (copulatory pouch) and  No equivalent
spermathecal (sperm storage)
 Insect circulatory system is open. It is composed of: dorsal vessel, which mediates the circulation
of the hemolymph (blood) around the insect body, and multiple accessory pulsatile organs that
promote blood flow through appendages, including legs, wings and antennae. Hemolymph is
generally pumped forwards from the abdomen to head by peristalsis of the dorsal vessel. The
hemolymph contains hemocytes (blood cells).
 The alimentary tract comprises three regions: the foregut, midgut and hindgut. The foregut of many
insects includes a crop, where food is stored and proventriculus for mechanical grinding of food
particles. The midgut is the principal site of digestion and absorption marked by the gastric ceaca.
The hindgut includes the ileum and rectum, and mediates the uptake of inorganic ions, water and
other solutes derived from the Malpighian tubules.
 The main organs of excretion and osmoregulation in insects are the Malpighian tubules and the
rectum and/or ileum, acting together.
 Overall insect muscle structure, sarcomeric proteins, contraction mechanisms and regulation are
homologous to those of all other animals. Insect muscles allow insects to walk, crawl, swim, jump
(often prodigiously), pounce, “sing” and fly.
 The cuticle has only a limited capacity for expansion thus must be shed regularly during molting, a
process that involves the partial degradation of the old cuticle by chitin lytic and proteolytic enzymes
and the biosynthesis of the new cuticle.
 The central nervous system is composed of neurons, which are cells specialized for
communication. Neurons communicate with each other and with effectors at specialized structures
called synapses. The brain is composed of protocerebral, deutocerebral and tritocerebral
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segements. It receives and processes sensory inputs from the major sense organs of the head –
the eyes and antennae
 Numerous hormonal substances control the physiological, developmental and behavioral activities
of insects. These belong to three major chemical classes: steroids (molting hormones),
sequiterpenes (juvenile hormones) and polypeptides. Hormones are secreted by specialized
glandular tissues in various body regions and by modified neurons (neurosecretory cells).
 Many insects produce air-borne sounds and vibrations to transmit information to conspecifics, other
insects or even non-insect species.
 Insects use visual color signals to recognize mates and conspecific competitors, and to deter
predators
 Insects communicate with each other and obtain information about their environment with
infochemicals. Insects use pheromones for intraspecific communication classified according to their
function (e.g., sex pheromone, trail pheromone or alarm pheromone).
REFERENCES
Chapman, R.F. (2013). The Insects – Structure and Function (5th ed.). UK: Cambridge University Press.
Gullan, P. J. & Cranston, P. S. (2014). The Insects: An outline of Entomology. 5th Ed. West Sussex, UK:
John Wiley and Sons, Ltd.
Imms, A.D. (1925). A General Textbook of Entomology. London: Methuen & Co. Ltd.
Freeman. Nation, J.L. Sr. (2016). Insect Physiology and Biochemistry (3 rd ed.). USA: Taylor and Francis
Group, LLC.
Triplehorn, C.A. & Johnson, N.F. (2005). Borror and DeLong’s Introduction to the study of insects (7 th ed).
USA: Brooks/Cole.
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QUIZ 3
I. Modified True or False. If the statement is correct, write TRUE. Otherwise, write FALSE if the
statement is incorrect AND change the underlined word to make the statement true.
1. Each testis is subdivided into follicles where sperm are actually produced.
2. The accessory glands produces the spermatecha or pouch-like structures made of protein
which encloses and protect the sperm when delivered to the female during copulation.
3. A spermatid has completed its meiotic divisions, but is an immature sperm.
4. The female's bursa copularis, a pouch-like chamber acts as a temporary storage of sperm.
5. The union of a sperm nucleus with the egg nucleus produces the zygote.
6. In aquatic insects, immatures are called nymphs.
7. Oviparity is an asexual mode of insect reproduction.
8. During spermatogenesis, meiotic divisions producing haploid spermatids occur in Zone III.
9. When the sperm is delivered to the female, the male accessory glands secrete
spermatophorins which is stimulated in the mealworm by juvenile hormone.
10. Holometabolous insects undergo little or no structural change as they grow older.
II. Matching type. Match the structures in column A with its corresponding function or definition in
column B.
Column A Column B
1. Fat body a. Protection of insects from

2. Peritrophic membrane ingesting allelochemicals such as
tannins
3. Ileum b. Dorsal blood vessel
4. Insect blood c. Regulates the entry and exit of
5. Heart hemolymph in the dorsal blood
vessel
6. Ostia d. Anterior hindgut
7. Hemocytes e. The storage of glycogen, fat and
protein
8. Spiracles f. Hemolymph
9. Cutaneous respiration g. Respiration through the cuticle
10. Proctolin h. Stimulates heart rate
i. Openings of the tracheal system at
the body surface
j. Blood cells
III. Multiple Choice. Choose the BEST answer among the choices.
1. __________ is a basic component of the nerve cells which transmits information, either to
another neuron or to an effector or organ such as a muscle.
a. dendrite
b. axon
c. spiracles
d. ostia
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2. What is the functional and structural units of nervous system?

a. axon
b. dendrites
c. neuron
d. receptor
3. The only visual organs of larval holometabolous insects is the _________.

a. stemmata
b. ocelli
c. axon
d. chorion
4. It refers to any exchange of information between individuals.

a. communication
b. reduction
c. oxidation
d. audition
5. The following are reasons why insects communicate, except:

a. Recognition of kin or nest mates
b. Locating or identifying a member of the opposite sex
c. Facilitation of courtship and mating
d. Attraction often differ from those used in courtship
IV. Answer the following questions concisely.
1. Describe the mechanism of insect flight.

2. Give three functions of the muscular system of insects.
3. Why is ventilation important to insects? Give an example of how an insect ventilate. (2
points)
4. Summarize the events involved in molting. (3 points)
5. Why do insect need to molt?
6. Describe the antennal tapping of ants.
7. What is the difference between apolysis and ecdysis?
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Name: ____________________________________ Schedule: _____________________

Program: __________________________________ Score: ________________________
Insect Internal Anatomy and Morphology
Objectives:
1. To distinguish among different types of tissues within an insect’s body.
2. To be familiar with the structures and locations of the digestive, circulatory and reproductive
systems of a typical insect.
Materials:
 Dissecting Kit: insect pins, dissecting scissors, fine-point forceps
 Dissecting dish or tray
 Dissecting microscope/stereomicroscope
 Cockroach (freshly-killed)
 Lab gown, surgical mask and gloves
 Sterile distilled water (in atomizer)
Procedures:
1. Obtain a freshly killed cockroach (Periplaneta americana) (can be done using chloroform or
drowning in water) and determine its gender. Remove the legs and wings at their bases with
dissecting scissors.
2. Carefully slit the abdominal and thoracic terga longitudinally near the lateral margins of the
insect. Leave the last abdominal tergum and the head intact. Pin the insect ventral side down
in a dissecting dish filled with enough tap water to cover the body.
3. Remove the terga from posterior to anterior ends using forceps and insect pins.With forceps
and insect pins remove the terga from the posterior end forwards. Leave the last abdominal
tergum intact.
4. Along the midline of the body, a dorsal blood vessel flanked laterally by silvery-white tracheae
and air sacs of the respiratory system. In the abdomen, this dorsal blood vessel is called the
heart with segmental “chambers” or the ostia. In the thorax, the blood vessel is just a simple
tube called the aorta – there are no chambers, muscles, or valves.
5. Pull away the heart, dorsal tracheae, and associated membranes and muscles to expose the
translucent thoracic muscles and strips of opaque white organs, the fat body. Remove the fat
bodies.
6. Cut through the digestive system just behind the head. The foregut includes the esophagus
(short narrow tube), the crop (large and brown filling the abdomen), and the proventriculus
(conical bulge behind the crop). The midgut is a simple tube, marked by the gastric caeca.
Throughout the abdominal cavity, there are a multitude of very thin, spaghetti-like structures
that are often pale yellow-green in color called Malpighian tubules. The hindgut is subdivided
into an ileum, a colon, and a rectum.
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7. In females, the ovary contains ovarioles and each ovariole contains a linear array of eggs with
the most mature egg nearest the base. The mass of female accessory glands are more
spaghetti-like in shape and pure white in color.
8. In males, the accessory glands are a white ball of anemone-like tubules near the base of the
last abdominal tergite. These accessory glands surround and obscure the testes and seminal
vesicles. An unpaired accessory gland (similar in appearance to fat body) lies along the ventral
wall of the abdomen and joins the ejaculatory duct near the base of the aedeagus.
9. Remove all the digestive and reproductive strictures from the abdominal cavity to find the nerve
cord lying along the ventral wall of the body. It consists of segmental ganglia joined to one
another longitudinally by intersegmental connectives. Use the point of an insect pin to separate
the two parallel nerves within each connective.
10. Clean up your dissection. Pour your excess liquid into the sink and wrap the body parts in a
paper towel before throwing them in the trash.
Reference:
NC State University. (2015). ENT 425 - General Entomology Lab 6 handout. Retrieved March 9, 2021 at
https://genent.cals.ncsu.edu/wp-content/uploads/2015/05/intanat1.pdf
Assume you have performed the dissection, locate the structures of the digestive,
reproductive and circulatory (heart and ventral cord) system of a cockroach. Label only the
pointed parts with yellow boxes
A. Heart
1. A cockroach has 13 hearts. The heart of cockroach is
in fact the dorsal blood vessel divided into 13
chambers. Oxygen enters each chamber sequentially
through a pair of slit-like openings called ____.
B. Digestive system:
Determine which is the foregut, midgut and hindgut. Label which is the crop, proventriculus
and gastric caeca, esophagus, Malphigian tubules, ileum, colon, and rectum.
5 4
8 3
6 2
7
9
12
10
11
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C. Reproductive system:
a. Determine the sexes (male or female) of cockroaches in #13 and #14.
b. Label the ovary, female accessory gland, testes and male accessory gland and the
unpaired accessory gland
13 14
18
17
15
16
D. Nervous system:
Label which is the ventral nerve cord and ganglia
19
20
Study questions:
1. Describe the mechanism how walking and crawling insects move.

2. Give two (2) insect hormones and identify where they are synthesized and their functions.
3. Summarize the steps of insect molting. What is the importance of molting to pesticide
application?
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Chapter 4
INSECTICIDE RESEARCH, DEVELOPMENT,
AND BIOASSAY
OVERVIEW
This chapter contains the discussion of insecticide research that generally involves comparing the
level of toxicity of different compounds or comparing the susceptibility of different insect species or the
same species from different environments. A useful way to make comparisons is to determine doses that
have equal toxicity and there are three general ways to bioassay compounds to obtain the critical doses
(Finney 1964). First is through direct assaying to measure the exact doses necessary to kill individual
animals by gradually increasing the doses up to the critical point. For insects, these methods are not
practical. The other two ways involve indirect assaying and this is performed by exposing batches of
individuals to standard doses and recording the responses, which may be death, knockdown, deformity, or
discoloration, depending on the expected effects of the compound on the insect species. Bioassays may
be based on quantitative responses, such as time of survival, but there are technical difficulties in
determining survival times and thus this method is not useful for testing insecticides. The third method is to
use quantal response bioassays. The binary quantal response with one explanatory variable is the simplest
and most common bioassay test used in insecticide research. In such dose-response or concentration-
response bioassays, the explanatory variable is a range of dosages or concentrations and the response is
an all-or-nothing observation, such as dead or alive, knocked down or remaining standing, deformed or not
deformed, and discolored or not discolored. The other two quantal response bioassays are more complex,
time-consuming, and less frequently used. Details can be found in Robertson et al (2007).
LEARNING OUTCOMES
1. Identify the chronology of insecticide research and development process.

2. Identify the important parameters in the data sheets.
3. Practice of bioassay and probit analysis
MOTIVATION
“Our farmers feed the world. They are up before dawn and work till well after sunset. They face any
number of challenges they can't control, from too much rain to not enough, from disease to insect
infestations.” - Luther Strange
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Lesson 1. Research and Development Process

Agrochemicals such as pesticides are indispensable in crop protection, productivity and reduction
of farm operations. However, insecticides for example, are getting less effective over a short period of
time due to resistance of insects upon wide application. Public concerns on the health and
environmental impacts of chemicals became the primary conditions for the development of novel or
reformulation of pesticides. Likewise, aside from safety and efficacy, new product discovery and
development should make profit.
Discovery phase: Finding the few among the many

The process by which a molecule is discovered, developed, and formulated into a pesticide
product is tenuous and risky. It takes an average of one to two years to determine if a molecule has
useful pesticidal properties. A three-year predevelopment phase follows for compounds that show
sufficient promise, then a three-year development phase and a commercial launch phase. New
pesticidal chemistry is identified by altering existing compounds or discovering new ones.
Innovation of existing chemistry

- to develop new products from existing compounds is to enhance certain critical characteristics—
spectrum, residual qualities, selectivity
- Modify compounds to improve their efficacy, spectrum of control, and chemical profile.
- E.g. the first synthetic pyrethroid was based on the structure of pyrethrum derived from
chrysanthemums. Useful innovations of that original molecular structure have been
created to increase soil persistence and decrease toxicity to non-target organisms.
Synthesis of compounds
- Compounds can be purchased from other commercial laboratories, pharmaceutical companies, and
universities.
- Other molecules may be selected by computer on the basis of characteristics determined internally.
Modelling target sites

- Design molecules that have the desired efficacy, safety profiles, and physical and environmental
properties.
- The physiological properties (most vulnerable) of target weeds, insects, and diseases must be known
(where to target control).
- Creation of a three-dimensional visualization of the structure of an enzyme or receptor to determine
how bioactive molecules interact and inhibit the target enzyme of the pest
- predicting which molecular structures may interact with target sites known for becoming resistance
mechanisms
- fine-tuning of chemicals to improve environmental or human safety
- use of genomic techniques (knowledge on the genetic code of plants, insects, and diseases specific
sites sensitive to interruption by novel pesticides)
Screening
When potential chemicals are selected, their biological activity is determined or screened.
- screen against “indicator” insects, weeds, diseases, or other organisms known to be sensitive to low
levels of biologically active pesticides - must be cheap to produce uniformly in large numbers, and they
must be easy to rear in the laboratory (e.g. house flies, fruit flies, and mosquito larvae
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- detailed testing in a simulated real-world situation (greenhouse screening)
Non-target toxicity test

- Examined for the compound’s acute toxicity to laboratory animals, potential to leach or run off into
surface water, and its eco-toxicological profile for wildlife.
- early indicators that the molecules will or will not need to be altered to reduce potential adverse effects
to humans, wildlife, and water
Field testing
- how well they perform under field conditions
- The volume of active ingredient necessary to conduct field trials and additional toxicological and eco-
toxicological studies increases from milligrams to kilograms
Lesson 2. Data Generation and Value Safety

Pesticides are agricultural weapons in preventing, controlling and managing insect pests,
diseases, weeds and other plant pathogens to reduce yield losses and maintaining high quality of
produce. Although these chemicals are developed under crucial regulation processes to minimize
impacts on environment, and human health, exposure and residues are relevant concerns. Agricultural
workers are constantly exposed to chemicals in the greenhouse or open fields while the general public
are risked to contamination of drinking water and food and bodies of water, soil and air are in constant
contamination including the wildlife, fish, plants, non-target organisms and natural enemies.
Exposure depends on toxicity of pesticides, adsorption, application methods, dosage applied,
prevailing weather conditions and how long the residue persist on the environment. Therefore, the risk
assessment of the impact of pesticides either on human health or on the environment is not an easy
and particularly accurate process because of differences in the periods and levels of exposure, the
types of pesticides used (regarding toxicity and persistence), and the environmental characteristics
Types of Data or Studies required (adapted from the United States Environmental
Protection Agency)
Pesticide Chemistry
Requirements related to product chemistry (e.g. odor to flammability, information about impurities)
Pesticide Performance
A mechanism to ensure that pesticide will control the pests in accordance to the label and that
unnecessary pesticide exposure to the environment will not occur as a result of the use of ineffective
products.
Hazard to Humans and Domestic Animals

- to assess hazards to humans and domestic animals are derived from a variety of acute, subchronic,
and chronic toxicity tests, and tests to assess mutagenicity and pesticide metabolism.
 Acute Studies
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Determination of acute oral, dermal, and inhalation toxicity that will provide information on health
hazards likely to arise soon after, and as a result of, short-term exposure. Data from acute studies serve as
a basis for classification and precautionary labeling. For example, acute toxicity data are used to calculate
farm worker reentry intervals and to develop precautionary label statements pertaining to protective clothing
requirements for applicators.
- provide information used in establishing the appropriate dose levels in sub-chronic and other
studies;
- provide initial information on the mode of toxic action(s) of a substance
- determine the need for child-resistant packaging
- determine the need to restrict use of the pesticide to trained applicators or in other ways to minimize
human and environmental hazards.
 Subchronic Studies
Subchronic tests provide information on health hazards that may arise from repeated exposures over
a limited period of time. They provide information on target organs and accumulation potential.
 Chronic Studies
Chronic toxicity studies (usually conducted by feeding the test substance to the test species) are
intended to determine the effects of a substance in a mammalian species following prolonged and repeated
exposure. Under the conditions of this test, effects that have a long latency period or are cumulative should
be detected.
Hazard to Non-target Organisms

The information required to assess hazards to non-target organisms is derived from tests to
determine pesticidal effects on birds, mammals, fish, terrestrial and aquatic invertebrates, and plants. These
tests include short-term acute, subacute, reproduction, simulated field, and full field studies arranged in a
hierarchical or tier system that progresses from the basic laboratory tests to the applied field tests (Table
6).
Table 6. The three level tests to assess pesticide toxicity on non-target organisms
Tier 1 Tier 1 Tier 1
Species
Acute toxicity Reproduction test Field test
Birds (bobwhite quail LD50 (8–14 days) Fish life cycle study
or mallard ducks)
Freshwater fish LC50 (96 h) Effects on spawning
(rainbow trout or
minnows)
Aquatic invertebrate LC50 (48 h) Full life cycle
(Daphnia, shrimp)
Non-target
LD50 (48 h) Effects of residues on
invertebrate (honey Pollination field test
foliage
bee)
Non-target
LC50 (14 days) Effects of residues
invertebrate
on foliage
(earthworms)
Aquatic plants (algae) LC50 (96 h) Plant vigour
Other beneficial
LD50 (48 h)
species
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 Acute and Subacute Studies

The short-term acute and subacute laboratory studies provide basic toxicity information that serves as a
starting point for the hazard assessment. These data are used to:
- establish acute toxicity levels of the active ingredient to the test organisms;
- compare toxicity information with measured or estimated pesticide residues in the environment in
order to assess potential effects on fish, wildlife, plants, and other nontarget organisms; and
- indicate whether further laboratory and/or field studies are needed.
 Chronic and Field Studies

- estimate the potential for chronic effects, taking into account the measured or estimated residues
in the environment; and
- determine if additional field or laboratory data are necessary to further evaluate hazards.
- Simulated field and/or field data are used to examine acute and chronic adverse effects on captive
or monitored fish and wildlife populations under natural or near-natural environments. Such studies
are required only when predictions as to possible adverse effects in less extensive studies cannot
be made, or when the potential for harmful effects is high.
Post-Application Exposure Studies

Data required to assess hazard to farm employees resulting from reentry into areas treated with
pesticides are derived from studies on toxicity, residue dissipation, and human exposure. Monitoring data
generated during exposure studies are used to determine how much pesticide people may be exposed to
after application and to establish how long workers must wait before reentering a treated area.
Applicator/User Exposure Studies
to evaluate the potential risks to people applying the pesticide, i.e., those who may be exposed to higher
concentrations of the pesticide through handling, including mixing or applying.
Pesticide Spray Drift Evaluation

- to evaluate pesticide spray drift based on the range of droplet sizes and spray drift field evaluations
- to provide data to help determine the need (and appropriate wording) for precautionary labeling to
minimize the potential harm to nontarget organisms.
Environmental Fate
- assess the presence of widely distributed and persistent pesticides in the environment that may result
in loss of usable land, surface water, ground water, and wildlife resources;
- assess the potential environmental exposure of other nontarget organisms, such as fish, wildlife, and
plants, to pesticides; and
- help estimate expected environmental concentrations of pesticides in specific habitats where
threatened or endangered species or other wildlife populations at risk are found.
Residue Chemistry
- to estimate the exposure of the general population to pesticide residues in food and for setting and
enforcing tolerances for pesticide residues in food or feed.
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Valuing safety
Pesticide Registration and Safety
Pesticide registration is a scientifically-based, legal, and an administrative process, where a wide
variety of effects associated with the use of a pesticide product and its potential effect on human health and
the environment is assessed. It is an important step in the management of pesticides as it enables
authorities primarily to determine permissible products or chemicals, to what purposes are these chemicals
are intended and also to exercise control over quality, usage rates, claims, labelling, packaging and
advertising of pesticides, thus ensuring that the consumers and environment are well protected. Also, the
registration process is restricted to the assumption that pesticides are only used for their intended function
proving that such use does not promote unreasonable effects either on human health or on the
environment. Therefore, before any pesticide can be used commercially, several tests are conducted that
determine whether a pesticide has any potential to cause adverse effects on humans and wildlife, including
endangered species and other non-target organisms, or potential to contaminate surface waters and
groundwater from leaching, runoff, and spray drift. Effects in any non-target species may translate into
ecosystem unbalance and food-web disruption that ultimately may affect human health and edible species.
Human health and exposure

Pesticide exposure primarily occurs in agricultural field workers and urban pest exterminators where
they are directly exposed to these chemicals during handling, storing, and application. However, the
presence of these chemicals in the environment constitutes potential hazard. The workers who mix, load,
transport and apply formulated pesticides are at high risks on acute exposure. The following are factors to
consider:
- Form of formulation may affect extent of exposure
- Types of packaging
- Weather conditions
Table 7. Acute toxicity of pesticides according to WHO classification.

LD50 for the rat (mg/kg b.w.)
Class Classification Oral Dermal
Solids Liquids Solids Liquids

Ia Extremely hazardous <5 <20 <10 <40
Ib Highly hazardous 5–50 20–200 10–100 40-400
II Moderately hazardous 50–500 200–2,000 100–1,000 400-4,000
III Slightly hazardous >501 >2,001 >1,001 >4,000
Unlike to present acute

U >2,000 >3,000 – -
hazard
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Lesson 3. Economic and Legal Aspects of Insecticide Use

The Fertilizer and Pesticide Authority
In the Philippines, the Fertilizer and Pesticide Authority (FPA) under the
Department of Agriculture is responsible on the regulation of pesticides. FPA is
mandated to assure that there are adequate supplies of fertilizer and pesticide
at reasonable prices, to protect the public from risks of use of pesticides, and
to educate the agricultural sector on proper handling and application of these
chemicals. The creation of the FPA is in response to the need to regulate
fertilizer and pesticide imports and exports and ensure their adequate supply in
the country.
The following details are adopted from the FPA website, discussing the history of the agency, latest
implementing guidelines and list of registered, banned and restricted pesticides in the Philippines:
Brief history of FPA
1972-1973 - The Philippine government created the Fertilizer and Industry Authority (FIA) by virtue of
Presidential Decree No. 135 signed by Ferdinand Marcos on 22 February 1973. It serves as a response to
the decrease in rice production caused by fertilizer shortage, series of natural calamities and prevailing oil
crisis in the 1970s.
Establishment
1977 – Since application of pesticide products goes side by side with fertilizers in agricultural production,
the government thought of expanding its regulatory powers to pesticide industries. The FIA was then
replaced by the Fertilizer and Pesticide Authority (FPA) which was created on 30 May 1977 by virtue of
Presidential Decree 1144. This is also attributed to the fact that fertilizer and pesticide industries possesses
the same clientele, distribution channels, system of application in farmers’ fields and technical supervision
by the same farm management technicians under the government's food production program.
Under the new law, FPA acquired broader powers with absolute control over the fertilizer and
pesticide sector. FPA’s mandate can be categorized into regulatory, monitoring, evaluation, and promotion.
Growth
1986- The fertilizer industry was liberalized as part of the government’s policy to open up domestic
agriculture to foreign competition. This included the abandonment of quantitative restriction for fertilizer and
with it the price-setting function of FPA. Import duties were also reduced to one to three percent through a
series of tariff reduction executive issuances.
1997- The Agriculture and Fisheries Modernization Act (AFMA) allowed the duty-free importation of fertilizer
enterprises engaged in agriculture. The law exempts fertilizer subsidiaries from payment of the 12% Value-
Added Tax.
May 5, 2014- FPA was transferred to the Office of the Presidential Assistant for Food Security and
Agricultural Modernization (OPAFSAM) through Executive Order No. 165 signed by President Benigno
Aquino III.
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September 17, 2018 - FPA was brought back to Department of Agriculture’s fold by virtue of Executive
Order 62 signed by President Rodrigo Roa Duterte
Powers and Functions
Specific to Pesticides and other Agricultural chemicals

1. To determine specific uses or manners of use for each pesticide or pesticide formulation;
2. To establish and enforce tolerance levels and good agricultural practices for use of pesticides in
raw agricultural commodities;
3. To restrict or ban the use of any pesticide or the formulation of certain pesticides in specific areas
or during certain periods upon evidence that the pesticide is an imminent hazard, has caused, or
is causing widespread serious damage to crops, fish or livestock, or to public health and
environment;
4. To prevent the importation of agricultural commodities containing pesticide residues above the
accepted tolerance levels and to regulate the exportation of agricultural products containing
pesticide residue above accepted tolerance levels;
5. To inspect the establishment and premises of pesticide handlers to insure that industrial health and
safety rules and anti-pollution regulations are followed;
6. To enter and inspect farmers’ fields to ensure that only the recommended pesticides are used in
specific crops in accordance with good agricultural practice;
7. To require if and when necessary, of every handler of these products, the submission to the FPA
of a report stating the quantity, value of each kind of product exported, imported, manufactured,
produced, formulated, repacked, stored, delivered, distributed, or sold;
8. Should there be any extraordinary and unreasonable increases in prices, or a severe shortage in
supply of pesticides or imminent dangers or either occurrences, the FPA is empowered to impose
such controls as may be necessary in the public interest, including but not limited to such
restrictions and controls as the imposition of price ceilings, controls on inventories, distribution, and
transport, and tax-free importation of such pesticides or raw materials thereof as may be in short
supply.
Registration and Licensing

No pesticides, fertilizer, or other agricultural chemical shall be exported, imported, manufactured,
formulated, stored, distributed, sold or offered for sale, transported, delivered for transportation or used
unless it has been duly registered with the FPA or covered by a numbered provisional permit issued by
FPA for use in accordance with the conditions as stipulated in the permit. Separate registrations shall be
required for each active ingredient and its possible formulations in the case of pesticides or for each fertilizer
grade in the case of fertilizer.
No person shall engage in the business of exporting, importing, manufacturing, formulating,
distributing, supplying, repacking, storing, commercially applying, selling, and marketing, of any pesticides,
fertilizer and other agricultural chemicals except under a license issued by the FPA.
The FPA, in the pursuit of its duties and functions, may suspend, revoke, or modify the registration
of any pesticide, fertilizer and other agricultural chemicals after due notice and hearing.
Penalties
Pesticides – Any person who violates any of the provisions of this Decree or any of the provisions of
the rules and regulations issued or promulgated by FPA on pesticides, shall be liable to a penal servitude
of not in excess of one year or a fine of P5,000.00 but not more than P10,000.00 provided that if the
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violation is committed by a corporation, firm, partnership, cooperative, association or any other entity, the
penalty shall be imposed upon the guilty officials or officers of such entities.
Implementing Rules and Guidelines for Pesticides

A separate handout will be provided as additional learning material
List of Banned Pesticides in the Philippines
Name of chemical Details of banning

1-Naphthylthiourea (ANTU) Banned as per FPA Circular No. 04, Series of 1989.
2, 4, 5-T Banned as per FPA Circular No. 04, Series of 1989.
Banned as per FPA Circular No. 4, Series of 1989 (Banned since
Aldrin
1989. There are alternatives to aldrin as prescribed by UNEP).
Azinphos Ethyl Banned as per FPA Resolution No. 01, Series of 1993.
Chlordane Banned as per FPA Resolution No. 01, Series of 1999.
Chlordimeform Banned as per FPA Circular No. 04, Series of 1989.
Copper(II) Acetoaresenite (Paris Green) Banned as per FPA Circular No. 04, Series of 1989.
DBCP Banned as per FPA Circular No. 04, Series of 1989.
DDT Banned as per FPA Board Resolution No. 04, Series of 2005.
Dieldrin
1989. There are alternatives to dieldrin as prescribed by UNEP).
Elemental Phosphorus (White & Yellow) Banned as per FPA Circular No. 04, Series of 1989.
Endosulfan Banned as per FPA Board Resolution No. 01, Series of 2015.
Endrin
1989. There are alternatives to endrin as prescribed by UNEP).
EPN Banned as per FPA Circular No. 04, Series of 1989.
Ethylene Dibromide (EDB) Banned as per FPA Circular No. 04, Series of 1989.
Gophacide Banned as per FPA Circular No. 04, Series of 1989.
HCH/BHC Banned as per FPA Circular No. 04, Series of 1989.
Heptachlor
1989. There are alternatives to heptachlor as prescribed by UNEP).
Leptophos Banned as per FPA Circular No. 04, Series of 1989.
Mercuric Fungicides Banned as per FPA Circular No. 04, Series of 1989.
Nitrofen Banned as per FPA Circular No. 04, Series of 1989.
Organotin Compounds Banned as per FPA Resolution No. 01, Series of 1993.
Parathion-ethyl Banned as per FPA Circular No. 04, Series of 1989.
Parathion-methyl Banned as per FPA Resolution No. 01, Series of 1993.
2-Fluoroacetamide (1081) Banned as per FPA Circular No. 04, Series of 1989.
Sodium Fluoroacetate Banned as per FPA Circular No. 04, Series of 1989.
Strychnine Banned as per FPA Circular No. 04, Series of 1989.
Thallium Sulfate Banned as per FPA Circular No. 04, Series of 1989.
Toxaphene/Camphechlor
1989. There are alternatives to toxaphene as prescribed by UNEP).
Triphenyltin Banned as per FPA Circular No. 04, Series of 1989.
List of Restricted Pesticides
Name of Chemicals Details of restriction

(e.g. reason for control action, remaining allowed uses)
(Severely Restricted)
Pentachlorophenol Restricted as per FPA Circular No. For use in wood treatment only by FPA
(PCP) 04 Series of 1989 Accredited wood treatments plants and
institutions.
Restricted as per FPA Circular No. Importation not allowed except in cases of
Aldicarb
04 Series of 1989 emergency as determined by the authority.
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Adequate time for aeration is required after

Restricted as per FPA Circular No.
Carbon Disulfide treatment before commodities are processed into
04 Series of 1989
food or feed.
Carbon Tetrachloride treatment before commodities are processed into
04 Series of 1989
food or feed.
Restricted as per FPA Circular No. Importation not allowed except in cases of
Chlorobenzilate
04 Series of 1989 emergency as determined by the authority.
Chloroform treatment before commodities are processed into
04 Series of 1989
food or feed.
Banned as per FPA Memorandum
Deltamethrin Banning of the use of Deltamethrin in Banana
Circular No. 01, Series of 2019
Entropop For use in banana plantations only.
04 Series of 1989
Ethyl formate treatment before commodities are processed into
04 Series of 1989
food or feed.
Banned as per FPA Memorandum Banning of the use of Fipronil in Banana through
Fipronil
Circular No. 29, Series of 2018 bud injection
HCN Generating Restricted as per FPA Circular No.
treatment before commodities are processed into
Materials 04 Series of 1989
food or feed.
Inorganic Arsenicals Restricted as per FPA Circular No. For use by FPA Accredited wood treatment and
(Arsenic Trioxide) 04 Series of 1989 wood preserving plants only.
Restricted as per FPA Circular No. The only allowed use to date is on pineapple
Lindane (Gamma-BHC)
04 Series of 1989 plantations by soil pre-plant application.
Methidathion 04 Series of 1989 For use in banana plantations only.
Restricted as per FPA Circular No. Adequate time for aeration is required after
Methyl Bromide 04 Series of 1989 treatment before commodities are processed into
food or feed.
Restricted as per FPA Resolution Allowed use is for beanfly control on legumes
Monocrotophos
No. 01, Series of 1993. only.
Restricted for institutional Use Only. Approval of
Restricted as per FPA Circular No. use will be based on strict compliance by the
Paraquat
04 Series of 1989 imported/end-user of the requirements act for its
use.
Phenamiphos For use in banana and pineapple plantations.
04 Series of 1989
Phosphine Generating Restricted as per FPA Circular No.
treatment before commodities are processed into
Compounds 04 Series of 1989
food or feed.
Lesson 4. Types of Bioassay and the Factors to Consider

Principles of bioassay
- Compare potency of unknown substance with standard (including assessment of errors).
- Standard and test sample should have same pharmacological effect and mode of action.
- The test and standard should be compared using a specified pharmacological technique.
- Method selected should be sensitive, reproducible and should minimize errors, biological variations
and methodology.
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Types of bioassay
a. Graded
1. Matching
2. Bracketing
3. Interpolation
4. Multiple Point
b. Quantal
1. Direct end point assay (DEPA)
2. LD50 Determination
Methodology of graded bioassay

1. Checking of apparatus for proper functioning.
2. Prepare physiological salt solution.
3. Arrange the instrument and adjust the water bath.
4. Balance the lever.
5. Tissue selection
6. Surgical process and collection of required tissue.
7. Tissue attachment to the water bath.
8. Relaxation time given to the tissue.
9. Prepare the standard drug (serial dilution).
10. Select lowest possible measurable cone.
11. Prepare DRC for the standard drug.
12. Prepare DRC for the test drug (serial solution).
13. Select an assay method (3 point or 4 point assay).
14. Calculation
Graded Bioassay may: a. intermittent dose method b. cumulative dose method
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Multiple point assays:

- Adv: reduced error, reduced variability.
- Dis: Lenghty, large amount of test sample required
Quantal bioassay
a. Direct End-Point Assay
- Threshhold dose producing a required response is measured on each animal.
- E.g. Bioassay of Digitalis in Cats, Hypoglycemic convulsions in mice.
- Threshold dose=Period of infusion x rate
Concentration of test = TDS X CSD
TDT
Bioassay of antagonist (Determination of the type of drug antagonism)

a. parallel shift of the log DRC
b. double reciprocal (Linewaver & Burk) plot
c. Schild Plot and pA2 value.
Advantage:
- chemical assay to complex
- If difference between results; bioassay given more importance
- Toxicity of new drug
disdvantages:
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- Time consuming
- requires much skill
- biological variations exist
Errors in bioassays
a. Biological variation
- loss of tissue sensitivity
- different species/sex/age/weight/health status
- laboratory condition may be variable
- housing and handling of animals
b. Methodological error
- lack of standardization of procedure
- set up of apparatus
- tissue isolation/preparation for experiment
- drug preparation or dilution
Lesson 5. Probit Analysis

Probit analysis is a specialized type of regression used to analyze binomial response variables. It
transforms the sigmoid dose-response curve to a straight line that can then be analyzed by regression
either through least squares or maximum likelihood. Probit analysis can be conducted by one of three
techniques:
 Using tables to estimate the probits and fitting the relationship by eye,
 Hand calculating the probits, regression coefficient, and confidence intervals, or
 Having a statistical package such as SPSS do it all for you.
The idea of probit analysis was originally published in Science by Chester Ittner Bliss in 1934. He
worked as an entomologist for the Connecticut Agricultural Experiment Station and was primarily concerned
with finding an effective pesticide to control insects that fed on grape leaves. By plotting the response of
the insects to various concentrations of pesticides, he was able to visualize that each pesticide affected the
insects at different concentrations. However, no available statistical method was appropriate at that time,
and the best alternative is to fit a regression of the response versus the concentration or dose, and compare
between the different pesticides but regression was applied only on linear at that time, while the relationship
of response to dose is sigmoid. Therefore, Bliss developed the idea of transforming the sigmoid dose-
response curve to a straight line. In 1952, David Finney, a professor of statistics at the University of
Edinburgh took Bliss’ idea and wrote a book called Probit Analysis. Today, probit analysis is still the
preferred statistical method in understanding dose-response relationships.
Remember that regression is a method of fitting a line to your data to compare the relationship of the
response variable or dependent variable (Y) to the independent variable (X).
𝑌 = 𝑎 + 𝑏𝑋 + 𝑒 ; Where: a = y-intercept; b = the slope of the line and e = error term
Probit Analysis is commonly used in toxicology to determine the relative toxicity of chemicals to living
organisms done by testing the response of an organism under various concentrations of each of the
chemicals in question and then comparing the concentrations at which one encounters a response. As
discussed above, the response is always binomial (e.g. death/no death) and the relationship between the
response and the various concentrations is always sigmoid. Probit analysis acts as a transformation from
sigmoid to linear and then runs a regression on the relationship. Once a regression is run, the researcher
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can use the output of the probit analysis to compare the amount of chemical required to create the same
response in each of the various chemicals. There are many endpoints used to compare the differing
toxicities of chemicals, but the LC50 (liquids) or LD50 (solids) are the most widely used outcomes of the
modern dose-response experiments.
How does probit analysis work? How to get from dose-response curve to an LC50?
Below you will find a step by step guide to using probit analysis with various methods. The easiest
by far is to use a statistical package such as SPSS, SAS, R, or S, but it is good to see the history of the
methodology to get a thorough understanding of the material.
Step 1: Convert % mortality to

probits (probability unit)
Method A: Determine probits by

looking up those corresponding to the
% responded in Finney’s table
(Finney 1952): For example, for a
17% response, the corresponding
probit would be 4.05. Additionally, for
a 50% response (LC50), the
corresponding probit would be 5.00.
Method B: Hand calculations (Finney and Stevens 1948): The probit Y, of the proportion P is defined by:
The standard method of analysis makes use of the maximum and minimum working probits:
And the range 1/Z where
Method C: Computer software such as SPSS, SAS, R, or S convert the percent responded to probits
automatically.
Step 2: Take the log of the concentrations.
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This can either be done by hand if doing hand calculations, or specify this action in the computer program
of choice. For example, after clicking Analyze, Regression, Probit, choose the log of your choice to
transform:
Step 3: Graph the probits versus the log of the concentrations and fit a line of regression.
Note: Both least squares and maximum likelihood are acceptable techniques to fitting the regression, but
maximum likelihood is preferred because it gives more precise estimation of necessary parameters for
correct evaluation of the results (Finney 1952).
Method A: Hand fit the line by eye that minimizes the space between the line and the data (i.e. least
squares). Although this method can be surprisingly accurate, calculating a regression by hand or using
computer program is obviously more precise. In addition, hand calculations and computer programs can
provide confidence intervals.
Method B: Hand calculate the linear regression by using:
the following method (Finney and Stevens 1948):

First set the proportion responding to be equal to p = r/n and the complement equal to q = 1-p. The probits
of a set value of p should be approximately linearly related to x, the measure of the stimulus, and a line
fitted by eye may be used to give a corresponding set of expected probits, Y. The working probit
corresponding to each proportion is next calculated from either of the following equations:
Next a set of expected probits is then derived from the weighted linear regression equation of working
probits on x, each y being assigned a weight, nw, where the weighting coefficient, w, is defined as:
The process is repeated with the new set of Y values. The iteration converges to give you a linear
regression. Method C: Use a computer program. SPSS uses maximum likelihood to estimate the linear
regression. To run the probit anaylsis in SPSS, follow the following simple steps:
Simply input a minimum of three columns into the Data Editor
• Number of individuals per container that responded
• Total of individuals per container
• Concentrations
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For example in the following screen, a_mort is the number of individuals that responded per container,
a_total is the total number of individuals per container, and a_conc are the concentrations. Row 7 in the
following example is data from the control where 0 out of 10 responded at a concentration of 0.
After you columns are set, simply go to analyze, regression, probit:
Screen 2:
Then set your number responded column as the “Response Frequency”, the total number per container
as the “Total Observed”, and the concentrations as the “Covariates”. Don’t forget to select the log base
10 to transform your concentrations.
Screen 3:
If you run the above example, you will see that SPSS determines an optimal solution after 18 iterations.
Step 4: Find the LC50

Method A: Using your hand drawn graph, either created by eye or by calculating the regression by hand,
find the probit of 5 in the y-axis, then move down to the x-axis and find the log of the concentration
associated with it. Then take the inverse of the log and voila! You have the LC50.
Method B: The LC50 is determined by searching the probit list for a probit of 5.00 and then taking the
inverse log of the concentration it is associated with.
Step 5: Determine the 95% confidence intervals:
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PHYSIO&TOXI
Method A: Hand calculate using the following equation:

The standard error is approximately: ± 1/b √(Snw)
• b = estimate of the slope of the line • Snw = summation of nw • w = weighted coefficient from table III =
Z²/PQ Finney, 1952
Method B: SPSS and other computer programs calculate this automatically
Notes of Interest for Probit Analysis
• Probit analysis assumes that the relationship between number responding (not percent response) and
concentration is normally distributed. If data are not normally distributed, logit is preferred.
• Must correct data if there is more than 10% mortality in the control
Logit vs. Probit:

Logit is another form of transforming binomial data into linearity and is very similar to probit. Logit
functions by taking the log of the odds: logit(P) = log P/ (1-P). Yet, the relationship between logit and
probit is almost indistinguishable: Logit ≈ (π/√3) x probit. In general, if response vs. dose data are not
normally distributed, Finney suggests using the logit over the probit transformation.
SUMMARY
 The research and development process of pesticides include a systematic steps:

1. Discovery phase: Finding the few among the many
2. Synthesis of compounds
3. Modelling target sites
4. Screening
5. Non-target toxicity test
6. Field testing
 Although pesticides are developed through very strict regulation processes to function with
reasonable certainty and minimal impact on human health and the environment, serious concerns
have been raised about health risks resulting from occupational exposure and from residues in food
and drinking water
 In the Philippines, the Fertilizer and Pesticide Authority (FPA) under the Department of Agriculture
is responsible on the regulation of pesticides. FPA is mandated to assure that there are adequate
supplies of fertilizer and pesticide at reasonable prices, to protect the public from risks of use of
pesticides, and to educate the agricultural sector on proper handling and application of these
chemicals.
 There are many existing bioassay for pesticides, all of which share similar objectives:
o Compare potency of unknown substance with standard (including assessment of errors).
o Standard and test sample should have same pharmacological effect and mode of action.
o The test and standard should be compared using a specified pharmacological technique.
o Method selected should be sensitive, reproducible and should minimize errors, biological
variations and methodology
 Probit Analysis is a type of regression used with binomial response variables. It is very similar to
logit, but is preferred when data are normally distributed. Probit analysis can be done by eye,
through hand calculations, or by using a statistical program.
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PHYSIO&TOXI
REFERENCES
Abbott, W.S. (1925). A method of computing the effectiveness of an insecticide. J. Econ. Entomol. 18:265-
267.
Damalas, C.A. and Eleftherohorinos, I.G. (2011). Pesticide Exposure, Safety Issues, and Risk Assessment
Indicators. Int. J. Environ. Res. Public Health, 8, 1402-1419.
United States Environmental Protection Agency [EPA]. (2021). Data Requirements for Pesticide
Registration. Retrieved March 12, 2021 from https://www.epa.gov/pesticide-registration/data-
requirements-pesticide-registration
Fertilizer and Pesticide Authority [FPA]. Retrieved March 12, 2021 from fpa.da.gov.ph.
Fertilizer and Pesticide Authority [FPA]. (2020). Pesticide Regulatory Policies and Implementing Guidelines.
Retrieved March 12, 2021 from fpa.da.gov.ph.
QUIZ 4
True or False.
1. The Fertilizer and Pesticide Authority can enter and inspect farmers’ fields to ensure that only
the recommended pesticides are used in specific crops in accordance with good agricultural
practice
2. Endosulfan, an organochlorine insecticide and acaricide, is allowed in the Philippines
3. Entropop can be applied to control insect pests of tomatoes.
4. In bioassay of compounds, control treatments are necessary to compare the efficacy of the test
compounds.
5. Probit analysis transforms the sigmoid dose-response curve to a straight line that can then be
analyzed by regression.
6. The Fertilizer and Industry Authority was replaced by the Fertilizer and Pesticide Authority
(FPA) 30 May 1977 by virtue of Presidential Decree 1144.
7. In 2015, the FPA is an attached agency of the Department of Agriculture.
8. Methyl bromide can be used as long as adequate time for aeration is achieved after treatment
before commodities are processed into food or feed.
9. The types of packaging of pesticide products can also affect potential exposure.
10. Standardization of materials must be done to avoid errors in bioassays.
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PHYSIO&TOXI
Name: ____________________________________ Schedule: _____________________

Program: __________________________________ Score: ________________________
Probit analysis
Objectives:
1. To determine how toxicity of pesticides are measured quantitatively
2. To conduct a hands-on probit analysis using SPSS statistical software
Procedure:
A video tutorial on how probit analysis is done using the SPSS statistical software will be
uploaded as reference material.
Study questions:
1. What is probit analysis? How is it applied to compute for toxicity of pesticides?
2. What are the variables needed to conduct a probit analysis?
3. Draw the resulting curve of mortality vs. concentration showing the LD 50.
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PHYSIO&TOXI

GENERAL PHYSIOLOGY & TOXICOLOGY

CatSU College of Agriculture and Fisheries

Module Cover Image Source: https://unsplash.com/photos/J-hYMYa2jY8

Definition of Key Terms

Anterior – near or toward the head (or part most nearly

Apolysis - separation of old exoskeleton from epidermis.

I. The science of insects

Choose the BEST answer among the choices.

Coconut rhinoceros Oriental migratory Dragonfly (Anisoptera) Black bean aphid

II. External anatomy of insects

Identify the external structures of a grasshopper.

III. Internal anatomy of insect

IV. Insect toxicology

THE CONCEPT OF INSECT PHYSIOLOGY

At the end of this chapter, the students shall be able to:

1. Identify the importance of insect physiology to pesticide toxicology.

Lesson 1. Understanding insects

Entomology and entomologists

Importance of insects in agriculture and human life

2. Insects as human food (entomophagy)

3. Insects as feed for domesticated animals

7. Production of honey, beeswax, silk and shellac

Insect nomenclature and classification

evident in their great number of species. Arthropods are believed to have

Figure 6. Phylogeny of insect orders.

Lesson 2. External Insect Structures and Functions

Figure 8. Front and back view of a typical winged insect.

Regions on the head:

There are five basic components of the mouthparts (Fig. 10):

Evolution resulted to an array of morphological

Figure 11. Specialized mouth parts of insects. (A) mandibulate/chewing

a. Scape – first and largest among the segments;

Figure 13. Main types of insect antenna.

Figure 15. Major regions of an insect wing and venation.

Insects have three pairs of segmented legs: the

The tarsus appears to be subdivided into smaller

1. Walking (gressorial) and running (cursorial)

- found in fore legs of predatory insects. (E.g. preying mantis)

Figure 18. Insect leg modifications.

A primitive insect abdomen has 11

The External Terminalia

Parts of the integument

Figure 21. Structure of the Integument

General features of the cuticle

Pore Canals (wax canals)

External appendages of the integument (Fig. 23)

Special cuticle formations

structure, attachment of muscles, organs (endophragma, apodema, apophysis)

Glands of the insect integument, dermal glands (glandulae)

Classification of glands according to their function

Penetration of compounds with different characteristics through the cuticle:

Color of the integument

b. Physical colors (subjective colors)

Lesson 3. Principles of Toxicology

Main Principles of Insect Toxicology

1. Insecticides be able to strike the weakest link of the pest.

 Behavior of the pest be known

Brief history of pesticide application

Name: _________________ Schedule: _

Name: ________________ Schedule: _