SCHOOL OF MEDICAL ENTOMOLOGY

AND PARASITOLOGY

ANOPHELES MOSQUITOES
ENTO 2102

An Introduction to Malaria Entomology

Senabulya George William

PRINCIPAL TUTOR
 MALARIA ENTOMOLOGY:
 Classification, morphology, biology, behavior
and ecology of mosquitoes that transmit
malaria

 Identification of Malaria mosquitoes

 Relationship between vector biology, its

ecology and behavior, the parasite and the
host

 To develop and implement effective vector

control strategies 2
 Malaria entomology…..
 Malaria is an infection caused by Plasmodium
parasite
 Five species of Plasmodium cause malaria in
humans: P. falciparum, P. vivax, P. malariae, P.
ovale and P. knowlesi
 Malaria parasites are transmitted by female
Anopheles mosquitoes
 Females depend on blood meals for
oviposition
 Also feed on sugar as males to obtain energy
 Males are not vectors

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 Genus Anopheles:
 Greek an (“not”) + ophelos (“benefit”) = harmful
(with no benefit)
 528 species world wide
 70 can transmit malaria
 40 species are of major importance
 Most common in the tropics and subtropics, but also
distributed in temperate climates and may extend to
summer arctic distributions
 About 39 different known anopheline mosquito
species in Uganda
 Most are not vectors and do not enter houses
 Some are forest species and while some others are
highland species
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Anopheline Mosquitoes in Uganda
 3 species are incriminated as major vectors
Major Vectors:
1. Anopheles gambiae s.s.
2. Anopheles funestus
3. Anopheles arabiensis

Minor Vectors:
1. An. pharoensis
2. An. nili
3. An. moucheti moucheti
4. An. bwambae

Note: Minor vectors may become major vectors

once the major vectors are eliminated in an area
through IRS/LLINs
INDIVIDUAL COURSE WORK ENTO 2102
1. a) Describe the life cycle of malaria
transmission in the mosquito vector

b) Explain the role of entomological

studies in malaria control

 Due date: Exactly 2 weeks from the date

of issue.

 Typed & printed

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 Taxonomy
 Phylum Arthropoda
 Class: Insecta
 Order: Diptera
 Family: Culicidae (mosquitoes)
 Subfamily: Anophelinae (Anophelines),
Culicinae (Culicines)
 Species:
 A. albimanus
 A. arabiensis
 A. barberi
 A. bellator
 A. crucians
 A. cruzii 7
 Species
 . culicifacies  A. moucheti
 A. darlingi  A. nili
 A. dirus  A. punctipennis
 A. earlei  A.
 A. freeborni quadrimaculatus
 A. funestus  A. stephensi
 A. gambiae(Giles  A. subpictus
1902)  A. sundaicus
 A. introlatus  A. walkeri
 A. latens  Etc…
 A. maculipennis
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 BEHAVIOR AND BIOLOGY OF ANOPHELINES
 Anopheles mosquitoes can be found anywhere in
the world with the exception of Antarctica.
 The body of the adult is dark brown to black in
color and has 3 sections which are the head,
thorax and abdomen.
 When resting, the stomach area of the Malaria
Mosquito species points upward, rather than being
even with the surrounding surface like most
mosquitoes.
 The female Anopheles mosquito will mate several
times in her short lifespan, producing eggs after
she has found a blood meal.
 Although she only lives a few weeks to a month at
most, she will have been able to produce
thousands of eggs during that time. 10
 Behaviour and biology
• Breeding Habits: The female will deposit up to
200 eggs individually onto the water’s surface.
• Each of the single eggs remains on the water by
the use of floats.
• The eggs take anywhere from two days to three
weeks to hatch, depending on the temperature of
the region.
• Eggs are laid in a wide range of locations including
fresh water or salt-water, vegetative or non-
vegetative, shady or sunlit. Ground pools, small
streams, irrigated lands, freshwater marshes,
forest pools, and any other place with clean, slow-
moving water are all considered prime Malaria
mosquito breeding grounds for egg-laying.
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 Host preferences: Anthropophagic: prefer
human blood, e.g. An. gambiae s.s & An.
Funestus, zoophagic: feed on animals, e.g. An.
Amharicus (Ethiopia), anthropozoophagic: feed
on both humans and animals, e. g. An.
arabiensis, An. culifacies (in India)
 Fertilized females, may survive winter by
hibernating in caves, which means the malaria
breeding cycle can virtually last year-round in
some locations.
 Eggs are capable of surviving cold
temperatures; however, freezing usually kills
eggs.
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 Behaviour and biology..
• Nocturnal biting habit - Most anopheline
mosquitoes bite at night, Some bite at dusk
while others bite late, or early morning
• Feeding site preferences: endophagic feed
indoors, exophagic: bite outdoors
• Resting Preferences: endophilic: rest on
indoor surfaces, exophilic : rest outdoors
- The life span of a female is 3-4 weeks (in
tropical countries), longer in temperate
climates.
- Males usually have a shorter life-span than
females
- Water is a detrimental factor for the
development of the immature stages
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 LIFE CYCLE OF ANOPHELINE MOSQUITOES
 Four stages - Holometabolus

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 Life cycle of Anopheline mosquitoes
 EGG STAGE

 A female Anopheline requires blood meal to

oviposit eggs

 She takes her first blood meal after mating

 Mating takes place after completing

maturation for hours even up to 2 days

 Males form swarms during mating

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 Life cycle….
 A female swings together with a male
followed by mating
 Male deposits in the female a life long
sperm together with mating plug (mass
of protein: Matrone)

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 Life cycle….
• The mating plug contains proteins and seminal
fluids
-This is to ensure correct retention of sperm in
female spermatheca
•The female usually mates only once
• Males may mate more than once

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 Life cycle…egg
- Initial batch of eggs often develops after 2 blood
meals, while the successive batches usually
require only 1 blood meal

- After feeding the female rests either indoor or

outdoor for 2 - 3 days until eggs develop
(tropical countries).
- In colder temperature climate, hatching may
take 2-3 weeks

- Then she searches for suitable oviposition site

containing water (rain pools, swamps, marshes,
canals, streams, rivers etc.,)

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 Life cycle egg…
- The female oviposits 50- 200 eggs at a time
(small brown or blackish boat-shaped) on
water surface
- Most females will lay 1-3 batches, some may
lay up to 7 batches
- Eggs are oviposited singly, boat-shaped,
have floats
- Eggs are not resistant to drying

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 Larvae Stage
 Larvae are known as wigglers since they seem to
move in that manner.
 4 instars and the first instar hatch within 1-2 days
 larval development takes 8-10 days: food and
temperature dependent
 Anopheles larvae do not have a siphon and they
lay parallel to the water surface.
 They lie parallel to the water’s surface in order to
feed on fungi, bacteria and other tiny organisms.
 The larva feed on micro-organisms and organic
matter in the water.
 They are filter-feeders, feed on bacteria, yeasts,
protozoans and other microorganisms.
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Larvae of Anopheles

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 Anopheles larvae……
 Once disturbed, larvae swim quickly to the
bottom of the water, but resurface some
seconds later
 Less permanent habitats: ricefields, wells,
borrow pits, small and temporary breeding
places (puddles, hoofprints, discarded tin
cans, water storage pots, etc…
 In general, Anopheles is found in clean
and unpolluted waters, absent from
habitats containing rotting plants and
faeces
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 Anopheles larvae (cont’d)
 In tropical countries, larval period lasts for
about 7 days, but in cooler climates this
stage can last for 2-4 weeks.
 In temperate areas, some Anopheles
overwinter as larvae, may live for months
 These larvae will undergo growth
throughout the four instars of this stage,
after which they become pupae.

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 Pupal Stage
 Pupae are known as tumblers because of the way
they seem to “tumble” through the water.
 Their rounded, comma-like shape makes this mode of
movement easy.
 These pupae come to the surface of the water in
order to breathe using tiny “trumpets,” but they do not
eat during the 1-2 days in which they will become an
adult mosquito.
 Pupa transforms to adult after 2 days
 Adults emerge as male and female

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Life cycle of anopheline mosquitoes…

 Duration of development depends on

temperature and nutritional factors

 Development is more rapid at higher

temperatures

 Gonotrophic cycle: is the period from

blood feeding to oviposition

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Mosquitoes of Medical Importance in Uganda
Anopheles Culex

Aedes Mansonia
Differentiating Mosquitoes of Medical Importance
STAGE MOSQUITO SPECIES
ANOPHELES AEDES CULEX MANSONIA
Eggs Laid Singly Laid Singly Laid in Rafts Laid Singly
Has Floats Has No Floats Has No Floats Has No Floats but with
spine to attach to water
vegetation
Larvae Rest parallel to the Rest at an Angle to Water Rest at an angle to Rest at an angle to
water surface Surface water Surface water Surface
Air tube: Rudimentary Air tube: Short, stout Air tube: Long slender Air tube: In form of a
breathing tube breathing tube with one pair breathing tube with spine to attach to
of hair tufts several pairs of hair tufts water vegetation

Pupae Differ only slightly Differ only slightly Differ only slightly Differ only slightly,
except for a spine to
attach to water
vegetation
Adults Rest at an angle to Rest parallel to resting Rest parallel to resting Rest parallel to resting
resting surface. surface. surface. surface.
Proboscis and the Proboscis and body at an Proboscis and body at Proboscis and body
body in the same angle to one another. an angle to one at an angle to one
straight line. Maxillary palps shorter another. another.
Maxillary palps as than the proboscis. Maxillary palps shorter Maxillary palps shorter
long as proboscis. Wings generally uniform. than the proboscis. than the proboscis.
Wings sported Tip of female abdomen Tip of female abdomen Tip of female abdomen
pointed blunt blunt
Identification Features of Larval and Adult Anopheline
and Culicine Mosquitoes
1. Is it an anopheline or culicine mosquito?

Culicines have short palpi; palpi of female

anophelines are as long as the proboscis
Identification of Anopheline Mosquitoes
2. Is it a Male or Female Anopheline mosquito?

Males have long and bushy antenna; hairs on females are

small and short
Dev. Differentiating characters
stage
Anophelines Culicines
Eggs •Oviposited singly (Aedes) or together in
•Oviposited singly
a “raft” (Culex)
•Boat shaped •Oval or cylindrical
•Lack floats
•Have floats
Larvae •Rest parallel to the •Subtend or hang
surface of water •Possess siphon (longer in Culex than in
•Lack siphon Aedes)
Pupae •Comma-shaped •Comma-shaped
•Breathing trumpet is •Breathing trumpet is long and slender
short and has wide with a narrow opening
opening
Adults •At rest, they form an •Rest parallel to resting surface
angle with the surface •Palps are short (in females)
•Palps are as long as •Palps are longer than proboscis with
tapered tips (males)
proboscis (females)
•Palps are as long as
proboscis and club-
shaped at tip (males) 33
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Identifying Anopheline species

Rational: accurate identification

Method:

Use identification keys based on

morphological feature:

on the head, wing, legs and

abdomen

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Head bears:
 proboscis
 a pair of
compound eyes
 a pair of
antennae
 Pair of palps: 5
segments
covered with
scales of
different colors

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Main features
PALPS
a) Scales on
palps
(smooth or
shaggy)
b) number of
pale bands
(3, 4 or none)

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Thorax bears:
Thorax : pro, meso
and metathorax
 A Pair of wings
 A Pair of halters
 Three pairs of legs
Wings
 Have several veins
covered with brown,
black, white or cream
scales
 Each vein is given
a number and/or
name
 Veins 2,4 and 5 are
forked
 Back edge has fine scales
Figure 2.4 Anophele’s wing 41
◦ Presence/absence of pale and dark spots
is one character for species identification

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Legs:
 Coxa, femur,
tibia
and tarsus
 Tarsus has 5
segments

 Colored
scales

 Scale
arrangments
differs
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44
Abdomen

 Eight visible

segments

 Upper plates-

Tergites

 Lower plates-

sternites

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• Presence/absence
of scales
• If present, their
position

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External anatomy of
Anopheles larva
Body divided in to head, thorax
and
abdomen
Head:-
 A pair of antennae
 A pair of mouth brushes
 Several hairs: position and
shape
Thorax
 Prothorax, mesothorax and
metathorax
 Prothoracic, mesothoracic
and metathoracic hairs

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b) External anatomy of Anopheles (Cellia) Abdomen
Anopheles larva
Abdomen
 Has eight similar and Anterior Tergal Plate
Posterior Tergal Plate

Accessory Tergal Plate

two modified segments Palmate Hair

 The 9th segment has

a pair of spiracles
 The 10th is the anal
part
 Palmate hairs
(segment IV-VI)
 Tergal plates
 Saddle hairs
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Identification keys and how to use them

Dichotomous or couple key

1. Wing scales are dark........................................................................ 2

Wings with pale and dark scales...................................................... 3

2. Legs with dark scales only............................................ Species A

Legs with pale and dark scales.................................... Species B

3. Legs with dark scales only.......................................... Species C

Legs with pale and dark scales (speckled)............................. 4

4. Palps all dark........................................................ Species D

Palps with pale scales on apical half.................... Species E

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Sibling species or complexes
Sibling (cryptic) species (species complexes):
are morphologically identical
But vary in:
1. Biology and ecology
2. Geographic distribution
3. Larval habitat characteristics
• Breeding in saline, fresh or mineral spring water
4.Behavior
• Host preference
• Feeding behavior
5. Capacity to transmit malaria
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 Methods for identification sibling
species

1. Cytogenetics:
banding patterns
of polytene
chromosome and
inversions: 4th
instar larva or
ovarian nurse
cells

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2.Molecular methods
– Ribosomal DNA (Scott et al., 1993) for members
of the An. gambiae : part of the mosquito such as
a segment of a leg or wing

Fig. Lane 1, I kb ladder, Lane 2 negative control,

Lane 3 and 4 An. gambiae s. s, Lane 5 An. arabiensis
positive control, Lanes 6-25 An. arabiensis from
Ethiopia
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Malaria vectors in Africa
1. Members of Anopheles gambiae
8 named and one unnamed species
 An. gambiae and An. coluzzi (formerly M and S
forms)[An. gambiae s.s): more anthropohilic and
endophilic, breeds in fresh water, major vector in
west, southern and some parts of east Africa
 An. arabiensis: more zoophilic and exophilic,
breeds in fresh water,major vector all over Africa
 An.melas (vector in coastal west africa): breeds in
saline water
 An. merus (vector in coastal east Africa): breeds in
saline water
 An.bwambae (vector in Uganda): mineral spring
 An. quadriannulatus and An. amharicus (are
zoophilic and non-vectors)
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 Malaria vectors in Africa
2. Anopheles funestus group
Consists 11 members
 An. funestus: main vector in many African
countries
 An. rivolurum: vector in Tanzania

Other vectors
 An. nili
 An. pharoensis
 An. moucheti

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