Insect Circulatory

System
 Transport
(Hormones,
Nutrients,
Storage Waste) Homeostasis
(Water and
(PH, Ionic,
Molecule
Fluid Balance)
Storage)

Hydraulics
Gas Exchange
(Moulting,
(Tracheal
Internal
Insect Ventilation)
Pressure)
Circulatory
System

Wound Thermoregula
Healing tion (Heat
(Phenolkinase Transfer and
based Freezing
Coagulation) Protection)
Immune Predator
Defence Defence
(Humoral (Reflex
Factors, Bleeding, Toxic
Haemocytes) Haemolymph)
• The insect circulatory system is at the center of most
physiological processes. It delivers nutrients and
hormones to cells and removes waste.
• The circulatory system also coordinates defense
mechanisms, modulates heat transfer, assists in gas
exchange, facilitates ecdysis, maintains homeostasis, and
more.
• In its most basic sense, the circulatory system is
composed of a fluid medium called hemolymph, a body
cavity called the hemocoel, and a series of muscular
pumps.
• The main driver of hemolymph circulation in the central
body cavity is the dorsal vessel, which is usually divided
into an aorta in the thorax and a heart in the abdomen.
Circulatory system & Circulation in insects
! Insects have an open circulatory system, the blood
(hemolymph) occupy all the body cavity (haemocoel), the
internal organs and tissues are exposed freely to it.
! The haemocoel is divided into three major sinuses by fibro–
muscular septa or diaphragms (Dorsal and Ventral); a dorsal
pericardia sinus, a perivisceral sinus and a ventral perineural
sinus.

Perivisceral sinus
" Circulation is produced by the activity of a dorsal longitudinal blood vessel.

" The dorsal blood vessel run a long the dorsal midline and is in the form of a
posterior heart and an anterior aorta.
" The dorsal vessel is open anteriorly and closed posteriorly except in
larval mayfly.
" The dorsal vessel is divided into two regions: a posterior heart in which
the wall of the vessel is perforated by incurrent and sometimes also by
excurrent openings (ostia), and an anterior aorta which is a simple
unperoforated tube.
• Peripheral circulation in the
appendages, however, is
driven by autonomous
pumps known as accessory
pulsatile organs, or
auxiliary hearts.
• Although insects have an
open circulatory system,
hemolymph does not
diffuse freely throughout
the hemocoel, and instead
flows along distinct
channel-like routes that are
created by the structural
organization of the internal
organs and by
fibromuscular septa or
diaphragms.
 The dorsal diaphragm

• It is a septum :extends across the abdominal cavity enclosing the

pericardial sinus , it contain the dorsal vessel.

• The dorsal diaphragm divides the haemocoel into the pericardial

sinus and the visceral sinus.
 Ventral diaphragm

• The ventral diaphragm forms a continuous ventral sheath .

• It extends from the prothorax to the end of the body .
• It encloses the perineural sinus.
• The perineural sinus encloses the nerve cord.
 Dorsal Vessel
! The dorsal vessel is a cylindrical structure that extends the entire length
of the insect and is organized by serial repetition of building blocks.
! It leaves the alimentary canal , just above the esophagus.

! It is divided into a posterior heart in which the wall is perforated and an

anterior aorta which is a simple unperforated tube.

! It collects blood from the abdominal cavity and discharges it in the

head .

! It is open anteriorly but closed posteriorly.

! The wall of the dorsal vessel is contractile. It consists of a single layer of a

pair of alary muscles, and connective tissue. These are composed of
cardiomyocytes.
The Heart

! It is often restricted to the abdomen or it may

extend as far as the prothorax .

! It is often a continuous tube that is not divided into

chambers, except in Orthoptera it is slightly enlarged
into ampullae, these are the points where the ostia
pierce the wall.

! The heart may be directly bound to the dorsal wall or

suspended from it by elastic filaments.

! A pair of alary musclesare attached laterally to the

walls of each chamber
• During early embryogenesis, the
cardioblasts move to the dorsal
midline and form two regular,
opposing rows (Figure 2a).
• The cardioblasts then extend
processes that meet their
contralateral counterparts in the
midline, thus forming a
cylindrical, spiral arrangement that
encloses a luminal space.
• In each segment, a pair of cardioblasts on each side of the body differentiate into
specialized cells that form inflow openings called ostia.
• Usually, each of these ostial cardioblasts develops a flap-like extension that
protrudes into the lumen of the dorsal vessel (Figure 2b).
• The ostial cells are indistinguishable from ordinary cardiomyocytes at the
ultrastructural level but express a suite of specific marker genes.
• In some insects, other cardioblasts form additional flap-like intracardiac valves
or muscular pads that regulate flow within the lumen of the dorsal vessel.
Excurrent Ostia
! These are ventro-lateral
openings in the wall of the
heart.
! No internal valves.
! Externally each opening is
surrounded by a papilla of
a spongy multinucleate
cells which expand during
systole, so the hemolymph
is forced out, and
contracts during diastole,
so that entry of blood is
prevented.
Incurrent Ostia
! They are vertical, slit-like
openings in the lateral wall
of the heart.
! The maximum number may
exist are 12 pairs, 9
abdominal and 3 thoracic.
! The anterior and posterior
lips of each ostium are
reflexed into the heart to
form a valve which permits
the flow of blood into the
heart at diastole (expansion
of the heart), but prevents
its outward passage at
systole.
Various kinds of ostia present in the dorsal vessel.
(a) Incurrent ostia with paired lips and an intracardiac valve.
(b) Incurrent ostia with elongate lips that function as a pouch valve.
(c) Two-way ostia with a single lip.
Left: Incurrent flow during the anterograde phase.
Right: Excurrent flow during the retrograde phase.
(a) Incurrent ostia (bottom), excurrent ostia with sphincter-like valves at the base of lateral
vessels (middle), and an unpaired excurrent ostium (top). The sphincter-like valve on the left
is shown in the open position, whereas the valve on the right is shown in the closed position.
Segmental vessels
! Most Blattodea and
Mantodea have no
excurrent ostia, but
the blood leaves the
heart via segmental
vessels that extend
laterally.
 Accessory pulsatile organs
! In addition to the dorsal vessel, insects have other pulsating
structures that maintain circulation through the appendages.
! A pulsatile organ drowning blood from the wings is present in
both wing-bearing segments of most adult insects.

! A blood space, or reservoir, beneath the posterior part of

the tergum which is largely or completely isolated from
the remaining hemocoel of the thorax connects with the
posterior veins of the wing via the axillary cord of the
wing.
! The ventral wall of the reservoir forms a muscular pump.
 by University of Sydney on 07/09/13. For persona the individualization of wing-hearts from the dorsal vessel (46). Thus the appear-
Annu. Rev. Entomol. 2000.45:495-518. Downloaded from w

ance of accessory pulsatile organs during insect evolution may be interpreted as

a result of alteration in other organ systems.
Conceivably, building blocks for new organs are recruited from various sys-
tems and assembled in new ways. Construction of a pump requires muscles as
well as elastic antagonists such as connective tissue structures or flexible cuticle.
Comparative investigation of the attachment sites, innervation, and ultrastructure
of accessory pulsatile organ muscles has revealed their heterogeneous provenance
(Table 1). Circulatory muscles may have been recruited by splitting some fibers
off a muscle and shifting their attachment sites, or by displacement of a muscle
portion. The former mode has probably been realized in some antenna-hearts (67),

Table 1 Survey of the accessory pulsatile organ components and their supposed provenance

Pulsatile organ
Appendage Contractile component Elastic antagonist Hemolymph flow conduit
Antenna Pharynx dilator Connective tissue Vessel
Mouthpart Skeletal muscle Flexible cutile Diaphragm
Leg Skeletal muscle Connective tissue Diaphragm
Wing 1. Open circulatory system
Myocardium Connective tissue Cuticular tube
Cercus a. The major
Rectum dilator portion of the "blood" or hemolymph is
Flexible cuticle not found within vessels.
Vessel
b. The hemolymph bathes the organs within the body cavity, the hemocoel.
Ovipositor Genital chamber muscle Flexible cuticle Diaphragm
c. Insects do not rely on the circulatory system for the transport of oxygen. This instead
is done by the tracheal system (see below).
d. Hemolymph enters the dorsal vessel or heart via small openings called ostia.
Accessory pulsatile organ components and their supposed provenance
e. The hemolymph is then pumped towards the head where it then returns to the
hemocoel.
!A pulsatile organ is also found at the base of each antenna.
!It consists of an ampulla from which a fine tube extends almost to
the tip of the antenna.

! In most insects , the ampullae have dilator muscles.

Compression drives hemolymph into the antenna.
!Most insects have a longitudinal septum in the legs which divides the
lumen into two sinuses and permits a bidirectional flow of blood within
the leg.
 Alary muscles

! These are muscles that are closely associated

with the heart.

! In Orthoptera ten abdominal and two

thoracic pairs are present.

! In other insect species the number is

reduced.

! They form integral part of the dorsal diaphragm

which spreads between them as connective tissue
membrane .
Heartbeat
! Systole

• It is the contraction phase of the heartbeat.

• It results from the contraction of the muscles in the heart wall

which starts posteriorly and spreads forward as a Wave.

! Diastole

• It is the relaxation phase of the heartbeat.

• It results from relaxation of the muscles assisted by the elastic

filaments supporting the heart.

• After diastole there is a third phase in the heart cycle known as

diastasis in which the heart remains in the expanded stat.

• Frequency with which the heart contracts varies from 14beats/

minute to 150 beats/minute
Circulation
! Normal circulation
At systole

• The blood is pumped forwards through the heart, entering the

perivisceral sinus through the anterior opening of the aorta in the head
and through the excurrent ostia where these exist.
• The valves on the incurrent ostia prevent the escape of blood through
these openings.
• The force of blood leaving the aorta anteriorly tends to push blood
backwards in the perivisceral sinus.
• The backwards flow is aided by the movements of the dorsal diaphragm
and by the inflow of blood into the heart, through the incurrent ostia, at
diastole.
• Movements of the ventral diaphragm presumably help to maintain
the supply of blood to the ventral nerve cord.
At Diastole

• Blood is drawn into the heart through the ostia.

 Modes of flow within the dorsal vessel and heart

Different types of circulatory organs and flow

modes in insects. Green arrows show inflow into
the dorsal vessel, and the dashed blue and red
arrows beside each insect indicate intracardiac
anterograde and retrograde flow, respectively.
• (a) Campodea augens (Diplura) represents an
ancestral condition, with antennal and cercal
arteries, a circumesophageal vessel ring in the
head, and bidirectional flow in the dorsal
vessel.

• (b) Schistocerca shoshone (Orthoptera) represents the condition in many basal pterygotes,
with anterograde flow in the dorsal vessel and numerous paired incurrent and excurrent ostia.
In the mesothorax and metathorax, the dorsal vessel has ampullary enlargements that suck
hemolymph from the wing veins through scutellar arms formed of tergal cuticle.
• (c) Drosophila melanogaster (Diptera) represents the derived condition with heartbeat
reversals, and thus, the dorsal vessel periodically alternates between contracting anterograde
(top) and retrograde (bottom). Hemolymph is supplied to the antennae and wings by separate
pulsatile organs.
Abbreviations: aa, antennal arteries; ah, antennal heart; cv, cercal vessel; cvr, circumesophageal vessel ring with ventral opening; eo, excurrent ostium; io, incurrent
ostium; iv, intracardiac valve; sa, scutellar arm; te, thoracic enlargement of dorsal vessel; wh, wing heart.
Circulation through the wings

• In most cases, blood is drawn in and out of the wings by the thoracic
pulsatile organs. Specific veins are contributed to this function.

• In Lepidotera, blood enters the wings along all the veins and go out
again during heartbeat reversal.
• The movement of blood out of the veins causes the tracheae inside
them to expand so that air is drawn into the wing.
• When heartbeat starts beating forwards again, the negative pressure
on the wing tracheae is removed and their elasticity causes them to
contract.

• This produces a negative pressure in the hemolymph space outside the

trachea and blood flows in.

• In insects lacking leg pulsatile organs, the flow of blood through the legs
is thought to be maintained by pressure differences at the base.
 Factors affecting the rate of the heartbeat

! Age and stage. Heartbeat is faster in adult than immature stages.

! Environmental factors such as temperature. Activity usually

stops above 45-50 ˚C and below 1-5 ˚C. Within this range, the rate is higher at

higher temperatures.

! Strong movement of the gut ( slow it or stop it). It is common for the heart to stop

beating sometimes for a few seconds.

! It is also common for the heartbeat to undergo periodic reversal, with waves of
contraction starting at the front. When this occurs, blood is forced out of the
incurrent ostia.
 Control of heartbeat

! No nerve cells are associated with the heart , and the heart muscles are the seat
of automation ( Myogenic beat ). Although the myogenic pattern maybe
modulated naturally or hormonally.

! The alary muscles are activated by the contraction of the heart and they exert
tension that modifies the heartbeat.

! The direction of a beat, from back to front or vice versa, maybe related to the
distribution of blood pressure. Also it may be related to the availability of
oxygen. In the absence of a good oxygen supply, the rate of heartbeat is
strongly reduced.
 Hemolymph
! Insect blood is known as hemolymph.

! It circulate round the body, bathing the tissue directly.

! It consists of a fluid plasma in which blood cells, hemocytes, are suspended.

! The hemolymph volume, expressed as a percentage of the total body weight of

the insect, varies with the type of insect.

! In the heavily sclerotized tenebrionid beetle, Onymachus, blood constitutes

about 11% of the beetle’s total mass; in mid-stadium larvae of Locusta, the figure
is about 18%, while in mature adults it is about 12%; in cockroaches it is about
17% and in caterpillars 35–40%.

! Hemolymph water comprises 20-25% of the total body water in adult insects,
but in caterpillars, the figure is close to 50%. This reflects the important
hydrostatic functions of the hemolymph in these larval forms
! It transports hormones, nutrients and wastes and has a role in, osmoregulation, temperature
control, Immunity, storage (water, carbohydrates and fats) and skeletal function.
! It also plays an essential part in the moulting process. An additional role of the hemolymph
in some orders, can be that of predatory defence.
! It can contain unpalatable and 106odorous chemicals that will act as a deterrent to predators.
CIRCULATORY SYSTEM, BLOOD AND IMMUNE SYSTEMS

a) blood volume ecdysis

250

blood volume (µl)

225
106 CIRCULATORY SYSTEM, BLOOD AND IMMUNE SYSTEMS
200

a) blood volume 175 ecdysis

250
-72 -48 -24 0 6

blood volume (µl)

Fig. 5.14. Changes in the blood volume (expressed as volume per
unit weight) during the development225 of Schistocerca. L3, L4 and b) hormone activity ecdysis
L5 refer to successive larval stages. Arrows indicate the time of
ecdysis (after Lee, 1961).
200

hormone activity
(arbitrary units)
antidiuretic diuretic
activity activity
5.2 HEMOLYMPH

The blood, or hemolymph, circulates175 round the body,

bathing the tissues directly. It consists of a fluid plasma in
which blood cells, hemocytes, are suspended. The plasma,
because of its function of maintaining the tissues -72
through- -48 -24 0 6
out the body, contains many chemicals. -72 -48 -24 0 6

time (hours)
Fig. 5.14. Changes in the blood volume (expressed as volume per
Changes in the blood volume (expressed 5.2.1 Hemolymph volume
L4 and volume, expressed as aRegulation of blood volume
ecdysis 1970).in relation to ecdysis in
unit weight) during the development of Schistocerca.The L3,hemolymph b) hormone
percentage of the activity
Fig. 5.15. Regulation of blood volume in relation to ecdysis in
as volume per unit
L5 refer to successive weight)
larval stages. during
Arrows indicate the theofweight of the insect, varies with the type of Periplaneta
totaltime
body
(after Mills & Whitehead, (a) Changes in blood

development of Schistocerca. L3, L4

ecdysis (after Lee, 1961). Periplaneta
insect. In the heavily sclerotized tenebrionid beetle,
andblood constitutes about 11% of the beetle’s suggesting
volume. (b) Antidiuretic and diuretic activity of the hemolymph,
that changes in hormonal activity regulate the changes
Onymachus,

L5 refer to successive larval stages. (a) theChanges

total mass; in mid-stadium larvae of Locusta, figure is in blood volume.
in volume.
hormone activity
(arbitrary units)

about 18%, while in mature adults it is about 12%;antidiuretic

in diuretic
Arrows indicate the time of ecdysis.cockroaches (b) Antidiuretic
it is about 17% and in caterpillars 35–40%. activity and
activity
cabbage butterfly, Pieris, diuretic
by about activity of the
70% in the hours follow-
5.2 HEMOLYMPH Hemolymph water comprises 20–25% of the total
hemolymph,
body water in adult insects, but in caterpillars, the figure is
suggesting that changes in
ing eclosion (Nicolson, 1980a).
Hemolymph volume is also affected by other factors.
The blood, or hemolymph, circulates roundclose the body,
hormonal
to 50%. This reflects the important hydrostatic func-
activity regulate the changes in
In Locusta, the hemolymph volume falls following
bathing the tissues directly. It consists of a fluid tionplasma in
of the hemolymph in these larval forms. This role is feeding, apparently because water moves into the gut.
further evidenced in other insects at the time ofvolume.
the molt Volume changes also result from desiccation. In the
which blood cells, hemocytes, are suspended. The whereplasma,
an increase in hemolymph volume occurs before desert tenebrionid, Onymacris, hemolymph volume is
 Constituents of
Plasma

Inorganic Metals Organic

Copper ( A
constituent of
Cations Anions Free Amino Acids Proteins Carbohydrate Lipid Organic Acid
tyrosinase), Iron,
Zinc, Manganese
Constituents of the plasma
In Plasma Inorganic & Organic Constituents
" Anion
1. Chloride is the most abundant
inorganic anion in insect blood.
• Conc. high ( hemimetabolous )
• low ( holometabolous ) lower
than 10% of total osmolar
concentration.
2. Phosohates & carbonates
" Cations
1 Sodium Na + is the most abundant
cation.
• Conc. Varies with taxonomic
position & diet.
• phytophagous : low
• predators : high
2 Potassium K & Magnisium Mg
• high in phytophagous
• low in carnivorous
• Blood –sucking e.g., Cimex &
Stomoxys high Na + low K ++
Na + and K ++: in the ionized form.
Ca & Mg : are bound to macromolecules
 Variation in cation concentration are wide

! e.g., in locust : K conc, increases before moulting.

! These changes affect the behaviour of insect , since ; neuromuscular

junctions are directly exposed to Hl. & low concentration in K ++ raises
the muscle resting potential.

! Mg : concentration is high in phytophagous insects ( as a result of its

high conc. In the diet since it is a constituent of chlorophyll .

! Mg conc. Is low in Lepidoptera when the larvae stop feeding.

! It’s conc. Is low in adult than the larvae

! In aquatic insects, the ionic composition of hemolymph affected by the

composition of water.
 Metallic traces

- Copper ( A constituent of tyrosinase)

- Iron

- Zinc

- Manganese

- These are found bound to organic complex & not found

as free ions .
Organic constituent
1-Amino Acids
• Very high level of free AA which constituting 33 –65 % about (
0.29 –2.43 gm), most of the known amino acids are present.
• They vary both quantitatively & qualitatively from one species
to another & in different stages of the same sp.
• They depend on those available in food.
• Insect blood plasma is characterized by very high levels of free
amino acids.
• The total concentration in plasma is usually more than 6
mg/ml in endopterygotes, but less than this in exopterygotes.
• Changes in A.A. concentration at different stages in the life
cycle.
 110 CIRCULATORY SYSTEM, BLOOD AND IMMUNE SYSTEMS

a) Locusta b) Periplaneta c) Calliphora larva Fig. 5.18. Amino acid conce

in the blood plasma of three
arginine Note the different scale for
histidine Periplaneta, where concentr

essential amino acids

isoleucine much lower (data from Irvin
leucine 1979). (a) Locusta, a phytop
lysine
hemimetabolous insect. (b)
methionine
Periplaneta, a detritivorous,
phenylalanine
hemimetabolous insect. (c)
threonine
Calliphora, a saprophagous,
tryptophan
holometabolous insect.
valine

alanine
aspartic acid
non-essential amino acids

cysteine/cystine
glutamic acid
glutamine
glycine
proline
serine
tyrosine

0 500 1000 1500 0 200 0 1000 2000

concentration (µM)

Amino acid concentrations

10!5 M. in
Thisthe blood because
is important plasmaglutamate
of three insects. Note
is a neuro- the different ecdysis
ecdysis scale for eclo
Periplaneta, where transmitter
concentrations are much
and high concentrations in thelower.
hemolymph (a) Locusta, feeding
a phytophagous,
spin pupa
hemimetabolous insect. (b) Periplaneta, a detritivorous, hemimetabolous
would impair this function (Irving, Wilson & Osborne, 3 insect. (c) Larval
larva

(mg.ml -1)
1979).
Calliphora, a saprophagous, holometabolous insect. glu
The concentrations of amino acids may change at
different stages of the life cycle. Tyrosine, for instance, 2
 0 500 1000 1500 0 200 0 1000 2000

concentration (µM)
The concentrations of amino acids may change at different stages of the life cycle.
10!5 M. This is important because glutamate is a neuro- ecdysis ecdysis eclosion
CUTICLE FORMATION 433
transmitter and high concentrations in the hemolymph feeding spin pupa
would impair a) tyrosine function (Irving, Wilson takes
thisstorage place primarily in the epidermis, althoughlarva
& Osborne, the prod-
ucts may be returned briefly to the3hemolymph. The first

concentration (mg.ml -1)

ecdysis ecdysis
1979). steps are its conversion to dihydroxyphenylalanine (dopa) glutamic acid
and then decarboxylation to dopamine (Fig. 16.20). From
The 10 concentrations of amino acids may change at
this, N-acetyldopamine and N-!-alanyldopamine, the two
different stages of the life cycle. Tyrosine, for instance, 2
concentration (mg.g -1 )

tyrosine glucoside compounds currently known to be important in sclerotiza-

commonly accumulates before each molttion,and are produced.
then
decreases 5
sharply as it is used in tanning and melanization glycine
1
of the new cuticle (see Fig. 16.19). Because free tyrosine is
tyrosine
not very 0
soluble, much of it is present as the more soluble aspartic acid

glucoside. Marked changes also occur in the silkworm, 0

Bombyx, b) when it produces its silken cocoon. Glycine is one 0 10 20 30
catecholamines
days
of the major amino acids in silk. Its concentration builds
hemolymph concentration (mM)

4
up towards the end of the feeding stage and then declines Fig. 5.19. Variations in the concentrations of some amino acids
used inVariations inthethe
cocoonconcentrations
of Bombyx. The rise in of some
3 N-β-alanyldopamine
during spinning (Fig. 5.19). Glutamine and asparagine are silk production for
not major 2
constituents of silk protein, but their concentra- aminoin the
concentrations acids used
early pupa in silk
probably resultproduction
from histolysis for the
of the tissues. In these experiments, glutamine and glutamate
tions in1 hemolymph
N -acetyldopamine fall sharply as they are taken up by the
were
cocoon
not separated, nor
of
were
Bombyx.
asparagine and
The
aspartate. They are
rise in
silk glands and converted to alanine, which is a major com-
shown concentrations in and
the earlyacid, pupa probably
0
in the figure as glutamic acid aspartic
ponent. Other2 amino
0 1 3 4 5acids 6 7 do 8 not9 1change in this way. After
days
the molt to pupa, the concentrations of glycine and gluta- result
respectively fromet al.,
(Jeuniaux histolysis
1961). of the tissues.
Fig. 16.19. Changes in the quantities of precursors of sclerotizing
Tyrosine,
mineagents(included
in the hemolymph for
with
of the lastglutamic instance,
larval stage of a acid commonly
in Fig.N-acetyldopamine
caterpillar 5.19) rise is probably the only tanning agent in
the cuticles of grasshoppers. In Manduca, N-acetyl-
sharply, possibly
(Manduca). (a) Tyrosinedue tolargely
is stored histolysis of the
as a glucoside, but tissues. The amino
accumulates
before ecdysis the glucoside before
is hydrolysed andeach
free tyrosine molt and molt, then
dopamine predominates at larva-to-larva molts and at the
acidincreases.
concentration in the
It is rapidly converted hemolymph
to a catecholamine so the probably rises after
pupa-to-adult but at the larva-to-pupa molt, N-!-
decreases sharply
amount of free tyrosine
feeding in many asvery
present is never
insects. Inithighis(afterused
Locusta, Ahmed, in alanyldopamine
tanning predominates
the concentration and (Fig. 16.19b). The latter
Hopkins & Kramer, 1983). (b) Catecholamines are only present has also been shown to be important in some flies.
melanization
in the hemolymph at the timeofof thethe new
molt. At the molt to the cuticle.
final Because
Immediately after ecdysis, these compounds are trans-
larval stage (left) N-acetyldopamine is most abundant, but at the ferred to the cuticle where they are oxidized to quinones by
freelarva/pupa
tyrosine molt (right)is
onlynot very soluble,
N-!-alanyldopamine is produced much ofIn it
phenoloxidases. is of Calpodes, a phenoloxidase is
the larva
(after Hopkins, Morgan & Kramer, 1984). produced as an integral part of the procuticle as it is laid
present as the more soluble glucoside. down, and this may be a common phenomenon. This phe-
insects (phenylalanine is also essential for protein synthesis). noloxidase is incorporated into the cuticle only in areas des-
Most insects store tyrosine over the intermolt period. tined to become sclerotized. The timing of sclerotization is
Sometimes it is stored in the fat body, but often the bulk is regulated, in Calpodes, by the availability of catecholamines as
2-proteins
- Numerous proteins are present in the hemolymph with a variety of
functions.

- They are not all present at the same time but, there are changes
through the life cycle.

- Proteins present in insect haemolymph are grouped as: storage

proteins, lipid transport proteins, vitelogenins, enzymes,
proteinase inhibitors, chromoproteins and a range of proteins
involved in the immune response of insects.
3- Other organic constituents
- Uric acid , urea and ammonia.

- Trehalose, a disaccharide, is the most characteristics sugar found

in insect hemolymph. Activity and starvation affect its conc. As it
is the main source of energy

- Glucose is also often present and other carbohydrates.

- Lipids. Marked increase may occur during development or flight ,

where lipid in fat body are mobilized as fuel for the flight.

- Organic acids
i.) e.g., Citrate : It is present in high concentration ( vary with
species and with development ).
ii.) Organic phosphates.
4- Pigments
Respiratory pigments do not occur in insect blood.
Except for : Chironomus larvae : Haemoglobin is present in solution in plasma.
Similarly the aquatic bug, Anisops, and the endoparasitic larvae of the bot fly,
Gasterophilus (Diptera) also contain Haemoglobin.
 HEMOLYMPH 115

Properties of the plasma

Organic acids are present in some quantity in the a) Orthoptera b) Neuroptera c) Lepidoptera
plasma. The major components are acids associated with
cations anions cations anions cations anions
the citric acid cycle, including citrate, !-ketoglutarate,
1- Osmotic pressure
succinate and malate. Citrate is usually present in high
concentration, although this varies considerably from one
The inorganic and organic solutes present in the
species to another. Organic phosphates are often present
hemolymph contribute to its and
in high concentrations osmotic pressure.
in some insects tyrosine is
Variation in presentplasma osmotic
as a phosphate. component
This is an in
alternative to glycosyla-
tion as a means of achieving a high concentration of tyro-
different orders. sine.
Three categories are present:

1. Na & Cl form
5.2.3most of the
Properties of theosmolar
plasma conc. cations anions
5.2.3.1 Osmotic pressure amino acids amino acids
The inorganic and organic solutes present in the hemo-
Basic type of insect blood & most of the
lymph contribute to its osmotic pressure. Sutcliffe (1963)
other components other components
magnesium
arthropods. grouped the insects into three broad categories on the basis
inorganic phosphate

of the osmotic components of the calcium chloride

e.g., Odonata. Ephemeroptera , plasma:
Homoptera,
potassium
Orthoptera. 1. Sodium and chloride account for a considerable pro-
portion of the osmolar concentration (Fig. 5.23a). This sodium
is probably the basic (in the evolutionary sense) type of Fig. 5.23. Osmotic components of the hemolymph in different
2. Cl is low relative toinsects
blood in Na because
whichit isconstitute
similar to that 21- Osmotic
in most components
groups of of theofhemolymph
insects expressed as percentages the total osmolar in
48 % of total other
osmolar arthropods.
conc,It and occursA. in A. are in different
Ephemeroptera, groups
concentration. of column
Each vertical insects expressed
represents 50% of the total as
Odonata, Plecoptera, Orthoptera and Homoptera. concentration (after Sutcliffe, 1963).
high conc. 2. Chloride is low relative to sodium, which constitutes percentages of the total osmolar
e.g., Diptera , Neuroptera&
21–48% of the mosttotalColeoptera.
osmolar concentration (Fig. concentration.
the osmotic effectors Each vertical
varied with hemolymph column
volume
(Fig. 5.24). It appears that they are stored in other tissues
represents
5.23b). Amino acids are also present in high concentra- 50% of the total concentration
when the hemolymph volume declines and returned to
tion. This type is found in Trichoptera, Diptera,
3. A.A. accountsMegaloptera,
for about 40% of the total
Neuroptera, Mecoptera and most the hemolymph when its volume increases. There is evi-
osmolar conc.Coleoptera.
The other substance never dence in Periplaneta that sodium, and perhaps also potas-
sium, are stored in urate granules in the fat body, but they
exceed 10 %3.. Amino acids account for 40% of the total osmolar may also remain in the hemolymph in an inactive form. In
e.g., Lepidoptera &concentration (Fig. 5.23c). There is a large category of
Hymenoptera.
unknown factors, but none of the other substances
the larva of the mosquito, Aedes, the osmotic activity of
sodium is lower than expected from its concentration,
 2- Hemolymph pH
• Definition: pH is defined as the negative logarithm of hydrogen ion concentration
of a solution. (pH= - log H+)
• In most insects the blood is slightly acidic, pH 6.4 – 6.8, although slightly
alkaline values have been recorded in a dragonfly larva and in the larva of midge
Chironomus.
• Most enzymes work within a limited pH range so its control is important .
• During normal activity the blood become acidic due to liberation of acid
metabolites (CO2), this tendency is buffered by substances in the blood.
• The buffering capacity of insect blood (its ability to prevent change in pH) is low
in the normal physiological range, but it increases sharply on either side of this
range.
• Bicarbonates and phosphates are the most important buffers.
• On the acid side of the range, carboxyl groups of organic acids are important.
• While on the alkaline side the amino groups of amino acids are most significant.
Proteins buffer over a wide range of pH.
Functions of Plasma
1. Nutritive materials are carried out from alimentary canal and storage tissues to
the sites at which they are metabolized.
2. Carrying excretory products from the place of origin to the malpighian tubules.
3. Transporting hormones from endocrine organs to the site of action. 4- The
plasma contains much more CO2 than O2, CO2 in solution accounts for about
20% of the total blood but the remainder is bound as bicarbonate. the greater
affinity of blood to CO2 than to O2 is important for the cyclic release of CO2
which occurs in some insects.
4. Store for some substances e.g., Trehalose as a source of energy and its supply
is achieved rapidly from fat body.
5. Water storage:
• An insect can maintain the level of its cell fluids if the food is dry.
• The water itself functions as a hydrostatic skeleton in forms such as larvae ,
where the cuticle is soft.
6. At molting the hydrostatic properties are used in expansion of appendages. ( The
amount of fluid present at this time influences the ultimate size of the insect. e.g.,
larvae of Lucilia with more moisture produce bigger adult).
7. Reflex bleeding results from the increase in hydrostatic pressure ( plasma is
forced through weak spots or pores in the cuticle . In species with this
phenomena the blood contain some repellant substances.
8. As a result of muscular activity; there is an increase in hydrostatic properties (
this is responsible for the eversion of the penis in male insects and the
osmeterium ( Fleshy bifurcate defensive device on the prothoracic segment in the
larvae of Papilinoidae) .

Osmeterium of Papilio xuthus Larva.

44
Insect Blood Cells (Hemocytes)

! The blood or hemolymph of insects consists of a

fluid plasma in which nucleated cells are
suspended , These cells are sessile. Several
different types of blood cells occur . Their functions
include: encapsulation , nodule formation and
phagocytosis which are considered as cellular
immunity , wound healing and intermediate
metabolism .
Rowley and Ratcliffe (1981) and Gupta (1997, 1985, 1991)
grouped
45 insect hemocytes into six cell types:

1. Prohemocytes.
2. Plasmatocytes.
3. Granulocytes (which are probably the same as
cystocytes or coagulocytes)
4. Spherulocytes.
5. Oenocytoids.
6. Adipohemocytes.
Another group, Brehelin and Zachary (1986) make the distinction
among nine types of hemocytes:

Prohemocytes,
Plasmatocytes,
Oenocytoids,
Spherule Cells,
Thrombocytoids, And
Four Granular Hemocytes Labeled
GH1, GH2, GH3, and GH4.
 age. Flow metrics such as velocity, acceleration, and heart pumping frequency h
a i ed f ad i e (Anopheles gambiae) and North American gra
(Schistocerca americana).

Fig. 5.4a: Types of haemocytes in insect.

Uttarakhand Open University

An illustration of the most common types of hemocytes from insect hemolymph. PR, prohemocy
te; PL, plasmatocyte; GR, granulocyte; SP, spherulocyte; CO, coagulocyte. Several different shape
s of plasmatocytes are shown in A, B, and C. The arrows indicate transformations of cells that ar
e believedto occur
Different types of hemocyte (a) after
Chiang, Gupta & Han, 1988; others after
Rowley and Ratcliffe, 1981): (a)
prohemocyte of Blattella;
(b) plasmatocyte of larval Galleria;
(c) granulocyte of larval Galleria;
(d) granulocyte (cystocyte) of Clitumnus.
Arrowheads indicate swollen perinuclear
cisterna;
(e) spherule cell of larval Galleria. The large
open areas, looking like vacuoles (and
labelled V), are probably caused by
extraction of spherules during preparation;
(f) oenocytoid from larval Galleria. Inset
shows size of nucleus relative to whole cell.
Abbreviations: G, granules; GO, Golgi
complex; IG, developing granules; M,
mitochondria; MT, microtubules; MVB,
multivesicular body; N, nucleus; PE,
protoplasmic extensions; PO, ribosomes;
PV, pinocytotic vesicles; R, ribosomes; RER,
distended cisternae of rough endoplasmic
reticulum; SP, spherules; V, vacuole.
Types of Hemocytes ( Ultrastructure identification)
1- Prohemocytes
• The prohemocytes are very variable in shape but usually small round cells
(6 to 13 µm in diameter) .
• Nucleus fills almost the entire cytoplasm. They are characterized by
undeveloped intracellular organelles .
• They rarely comprise more than 5% of the total hemocyte population.
• They are the stem cells from which most other hemocyte types are formed.
• Prohemocytes are known to divide and they may differentiate into
plasmocytes, which, in turn, may give rise to granulocytes, and these may
differentiate into sperulocytes.
• They contain ribosomes and mitochondria, but little endoplasmic
reticulum and Golgi membranes.
• They are not mobile and do not participate in phagocytosis.
• The main function of hemocytes may be to divide and give rise to new
hemocytes.
52

An illustration of the most common types of hemocytes from insect hemolymph. PR,
prohemocyte; PL, plasmatocyte; GR, granulocyte; SP, spherulocyte; CO, coagulocyte.
Several different shapes of plasmatocytes are shown in A, B, and C. The arrows
indicate transformations of cells that are believed to occur (according to Woodring,
1985).
 2. Plasmatocytes (phagocytes)
1. Plasmatocytes are polymorphic cells up to 40 to 50 μm in size, granular or
agranular usually round and oval .
2. They are the most abundant cell types, with a centrally placed, spherical
nucleus surrounded by well vacuolated cytoplasm.
3. Plasmatocytes are characterized by large nucleus containing a large nucleolus.
4. They possess well developed Golgi apparatus and endoplasmic reticulum as
well as many lysosomes.
5. The cells are capable of amoeboid movement and are phagocytic.
6. They are involved in phagocytosis and encapsulation of foreign organisms
invading insect so they are responsible for most of the cellular immunity of
the insect.
7. They may be binucleate. “Young” plasmatocytes can be confused with
prohemocytes.
8. They contain lysosomal enzymes and are usually the most numerous of
circulating cells.
3 ) Granulocytes
• Granulocytes are variable in size, spherical or oval, and up to 45 μm in size.
• Round or disc-shaped, with a relatively small nucleus surrounded by granular
cytoplasm, well developed granular rough endoplasmic reticulum and Golgi
apparatus.
• On the basis of histochemical tests, the granules are thought to be glycoproteins and
mucopolysaccharides.
• Granulocytes may arise from plasmatocytes.
• They discharge their contents on the surfaces of intruding organisms as early part of
the defense response.
• The precise function of granulocytes is unproven, but some researchers have suggested
that they serve storage and possibly secretory functions.
• They may be involved in cellular defensive functions in various insects, and may be
phagocytic in some insects, but in others neither of these functions is established.
4) Spherulocytes
55
• Spherulocytes are ovoid to round cells up to about
25 µm in length.

• They are characterized by the large, refractile

spherules which may occupy 90% of the
cytoplasm, stain for acid mucopolysaccharides .

• They are not common although they are found in

most of the studied species.

• Their function is unknown.

 5- Oenocytoids
56
• Mainly occur in Lepidoptera.
• Oenocytoids are variable in size, often large, may be binucleate,
and lyse easily, but do not cause hemolymph coagulation when
they lyse.
• They exhibit little development of rough endoplasmic reticulum or
Golgi complexes, but they have a complex array of microtubules.
• Their function is unknown.
• They are nonphagocytic.
• Some evidence indicates that they contain prophenoloxidase, an
inactive form of phenoloxidase.
• Oenocytoids should not be confused with oenocytes, cells found
among fat body cells and scattered among epidermal cells in many
insects.
• Oenocytes are not blood cells.
 6- Adipohemocytes
57

• Adipohemocytes may be small or large, spherical to

oval, and contain lipid droplets.
• They are characterized by containing lipid droplets.
• They contain well developed endoplasmic
reticulum and Golgi complexes.
• They might be plasmatocytes that are filled with
lipids under certain physiological conditions.
58

Many other hemocyte types are present and they vary

according to the different insect species and also to
the type of investigation
59
 Functions of Hemocytes
Phagocytosis
• The most common function of hemocytes is phagocytosis of
foreign particles, microorganisms and tissue debris that are
comparatively smaller in size than the cell.

• Many cells are capable of phagocytosis but plasmatocytes and

granular hemocyets are the most important.

• Endocytosis, especially phagocytosis, is important in both

metamorphosis and defense against disease, as well as in routine
cleaning up of dead or damaged cells.
 Wound healing and Coagulation
61

• The hemocytes collect at the site of injuries, forming a plug which helps to seal
the wound, proliferating and removing the dead tissues and discard it.
• The blood of various insects when collected in glass capillaries, usually clots
within 10 – 15 minutes.
• On the other hand the blood of other insect species never clots or coagulates.
• In Cockroaches , the apparent coagulum is merely a clump of hemocytes; the
blood cells become round, develop thread – like pseudopodia, and agglutinate to
form a plug which is the essential factor of the clot. When blood cells coagulates
due to the presence of hyaline hemocytes, these cells extrude thread – like
pseudopodial expansions which fuse to form a meshwork in which the other cells
are entangled.
 Nodule formation
63
• This occurs in response to invasion of the body by large quantities of
particulate matter (especially bacterial aggregates) that cannot be
removed effectively by phagocytosis.
• This process includes two phases:
• First: Within minutes of the foreign matter arriving in the body cavity, it
is surrounded either by granulocytes or by specific plasma proteins and by
material discharged from coagulocytes.
• In the second phase, which takes several hours to complete, plasmatocytes
surround the melanized core, become flattened, and form a multicellular
sheath comparable to that seen after encapsulation (Strand, 2002).
 Encapsulation
65
• Encapsulation is essentially nodule formation on a larger
scale.
• After the invader is initially coated with a thin layer of
granulocytes or plasma proteins, layers of plasmatocytes
surround it (Nappi, 1975; Lavine and Strand, 2001, 2002)
• This reaction on the part of hemocytes forms an insect’s most
important defense mechanism against metazoan
endoparasites such as nematodes and insects.
• An encapsulated organism almost always dies, as a result of
starvation, asphyxiation, and perhaps poisoning by quinones,
antibacterial peptides, hydrogen peroxide and nitric oxide.
 67 5- Connective tissue formation

• All tissues and organs of insects are separated from the body
cavity by connective tissue membranes and basement
membranes.
• Blood cells usually collect around foreign bodies and form
capsules. Usually the cellular character of these capsules
persists, but sometimes the nuclei disappear and a capsule of
connective tissue is produced. The hemocytes are concerned
in connective tissue formation.
 6- Metabolic and Homeostatic Functions
Hemocytes have been implicated in a variety of metabolic
and homeostatic functions.

Cells have been shown to contain, for example, glycogen,

mucopolysaccharide, lipid, and protein, it has been
suggested that hemocytes are important in storage of
nutrients and their distribution to growing tissues,
formation of connective tissue, synthesis of chitin, and
maintenance of hemolymph sugar level.
Cells might be important in hemolymph amino acid
homeostasis.
Certain hemocytes (spherule cells in Diptera, oenocytoids
in other insects) contain enzymes for metabolism of
tyrosine, derivatives of which are important in tanning
and/or darkening of cuticle.
 Numbers of Hemocytes
70

• Small insects have many fewer hemocytes than large

insects.
• Number of hemocytes are variable between species and
in different life stage in the same species.
• Count of the number of cells per unit volume of
hemolymph (usually called total hemocyte count) may
not reflect the total number of hemocytes in circulation
because the blood volume varies.
 Hemocyte profile
The relative abundance of different types of hemocytes
(called the hemocyte profile or differential hemocyte
count) is not constant.

Plasmatocytes and granulocytes are usually the most

abundant, often comprising about 80% of the total
hemocyte population.

There number varied again with different life stage and

physiological state.