128 Biochemical Society Transactions (2003) Volume 31, part 1

Arthropod semiochemicals: mosquitoes, midges

and sealice
A.J. Mordue Luntz1
Department of Zoology, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ, U.K.

Abstract
Signalling chemicals play an essential role in arthropod life cycles. They provide the means whereby
mates, host and oviposition sites are located and recognized. The identification of such semiochemicals (sex
pheromones, host location and habitat-related cues) in mosquitoes, midges and sealice of farmed salmon
is revealing ways in which such devastating pests may be monitored and controlled by behavioural means
using push–pull strategies to manipulate the pests away from hosts and into traps.

Introduction
Arthropods, such as insects and crustacea, that feed
on vertebrate hosts including man, transmit devastating
diseases, cause great distress to their hosts and cause
highly significant economic damage in animal husbandry.
Dealing with such pests traditionally has been based on
treatment of the host after biting/infection and control of the
vector with pesticides; however, knowledge of the chemical
ecology of these invertebrates is revealing the complexities
of chemical signalling mechanisms in mate location, host
location and oviposition, the components of which may
be manipulated for semiochemical-based control. Although
lagging behind detailed knowledge for insect–plant interac-
tions, arthropod–host interactions are revealing fascinating
parallels in behavioural processes, chemosensory mechanisms
and the chemicals involved. The identification of the essential
cues for locating a mate or a host, how such cues are
detected by antennal olfactory receptors and how they
are processed centrally to generate behaviour patterns
leading to successful mate and host recognition and location
involves studies of ecophysiology, behaviour and cellular
processing mechanisms. The production of odour cues in
the environment, the physical generation of plumes and
odour trails moving downwind in differing complex forms
depending upon wind speed, and the presence or absence of
vegetation etc., is beginning to be understood for haemato-
phagous insects, as it has been for phytophagous insect pests
of crops.
Such knowledge is required in order to successfully
manipulate the behaviour of pests for monitoring or control.
Species-specific traps have the potential for use in monitoring
the population density of vectors of disease and also the
proportion of that population carrying disease. Stimulo-
deterrent diversionary or push–pull strategies will also prove
useful in future for control of vectors and parasites by pushing
them away from their hosts with the use of repellents, non-
Key words: crustacean, host attractant, insect, oviposition cue, pheromone.
1
e-mail a.j.mordue@abdn.ac.uk
host cues or masking cues, while pulling them towards traps
containing pheromones together with host or oviposition
locating cues (Table 1).
Mate location
Sex pheromones in agricultural insect pests were the first
semiochemicals to be exploited in monitoring and control.
Sticky traps, containing female sex pheromone, into which
males are drawn form an essential monitoring role in inte-
grated pest management strategies. Flooding the environment
with sex pheromone provides direct control by disrupting
mating, as males cannot locate females. Haematophagous
insects and crustacea may also use sex pheromones to locate
a mate and such components may also be possible candidates
in control [1–5]. The first midge pheromone has recently
been identified at Aberdeen and Rothamsted Research. Tsetse
fly (the vector of trypanosomiasis in Africa) sex pheromone
and sandfly (the vector of leishmaniasis in the New World)
pheromone are also known (Figure 1). In these cases, mating
processes are associated with feeding cues such as host blood
volatiles, as mating occurs at the host site prior to blood
feeding.
Not all midges and mosquitoes have sex pheromones, as
recognition of females by males in many instances occurs by
auditory recognition of the female by the male using her wing
beat frequency as the females fly into male leks. To date, sex
pheromones have not been discovered in mosquitoes or the
Scottish biting midge (Culicoides impunctatus).
Insect pheromones are basically hydrocarbons with or
without double bonds, functional groups (such as alcohols,
esters, aldehydes), branched-chain or cyclic structures.
They vary in size from C3 to C37 so providing volatility
with sufficient structural variation for species specificity.
Geometrical isomerism provides active and inactive isomers
as insects readily perceive differences in geometry. Optical
isomerism (including enantiomers or diasteroisomers) often
occurs with the more complex molecules, providing further
species specificity in the form of active and inactive

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 Signalling the Future 129

Table 1 Semiochemicals that could be used in push–pull ratios in different species. Indeed, the sex pheromones of
strategies for pest control insects are rarely, if ever, single compounds. The pheromone
Push (away from host) Pull (to traps or targets) consists of one major component and one or more minor
components.
Repellents Pheromones Pheromones produced by female lepidoptera (butterflies
Non-host odours Host odours (kairomones) and moths) mainly consist of compounds with straight carbon
Host-masking chemicals Habitant attractants chains; however, the considerable diversity among them is
created by the interplay of chain length, functional group, po-
sition and number of double bonds, and their stereochemistry
optical isomers. In addition to chemical specificity, species (Figure 1). Using the same biochemical pathways for fatty-
recognition, in those with similar sex pheromones for acid synthesis the straight chain compounds (compounds 1, 2
example, can be achieved by the presence of mixtures of and 3; Figure 1) provide examples of monounsaturated acetate
closely related pheromones that may also occur in different (turnip moth), aldehyde (spruce budworm) and diunsaturated

Figure 1 Examples of arthropod semiochemicals to show the variety of structures

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130 Biochemical Society Transactions (2003) Volume 31, part 1
Figure 1 Continued
alcohol (silkworm moth) female sex pheromones. The first
two species are economically important pests of agriculture
and forestry. The tiger moth pheromone (compound 4 in
Figure 1) is an example of a simply branched hydrocarbon
where a single carbon lies off the main branch.
In Diptera (flies), the newly identified midge sex
pheromone n-heptadecane, (compound 5 in Figure 1) is
a simple hydrocarbon, whereas the tsetse sex pheromone
(compound 6 in Figure 1), is more complex with three
methylated branches (15,19,23-trimethylheptatriacontane)
and exists in four isomeric forms. The sandfly pheromone,
which varies in two different populations (compound 7 in
Figure 1) and which is produced by the male rather than the
female, consists of 9-methyl germacrene-B in Lapinha, Brazil
and 3-methyl-α-himachalene in Jacobina, Brazil [1,2].


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The Culex quinquefasciatus oviposition pheromone (com-
pound 8 in Figure 1) is produced by the female prior to laying
her eggs and volatilizes from the tips of the eggs in the egg
rafts over a 48-h period after oviposition. The pheromone,
which is a heterocyclic diasteroisomeric lactone mixture,
(5R,6S)-6-acetetoxy-5-hexadecanolide, attracts other females
to lay gregariously in the same larval habitat, which normally
consists of enriched organically polluted water such as
is found in cess pits or free standing marshy areas rich
in decaying vegetation. 3-Methyl indole (compound 18 in
Figure 1), a malodorous compound that signals organic
enrichment, is a highly effective attractant to gravid female
C. quinquefasciatus.
Terpenoid pheromones are built up via mevalonic acid
through the linkage of five-carbon isoprene units in a
 Signalling the Future 131

Figure 2 Four-way choice oviposition bioassays for C. quinquefasciatus comparing the attractiveness of oviposition pheromone
derived synthetically or via plant biosynthesis (using K. scoparia) [27], and the dirty water volatile skatole
(a) Synthetic pheromone and skatole at 10−10 g; (b) synthetic pheromone and skatole at 10−9 g; (c) plant-derived pheromone
and skatole at 10−10 g; (d) plant-derived pheromone and skatole at 10−9 g. Twenty gravid female mosquitoes were used in
each assay [n = 16 for (a) and (c); n = 12 for (b) and (d)]. Significant differences from controls: *P < 0.05, **P < 0.01 and
***P < 0.001. In (d) synergism is evident for skatole and pheromone supplied together, compared with the sum of skatole
or pheromone supplied separately (P < 0.01).

a c

b d

head-to-tail fashion. The aphid alarm pheromone (compound
9 in Figure 1) is an example of an acyclic sesquiterpenoid
pheromone, which is produced by many species of aphid
and is found also in the glandular hairs of wild potato as a
plant defence mechanism. The release of this compound on
attack by predators causes aphids to scatter away from the
pheromone source. Cyclic pheromones are also common in
insects; the polycyclic sex pheromone of the common wasp
(compound 10 in Figure 1) is an example of a spirocyclic
pheromone with two cyclic skeletons that share one
atom.
Host location
Blood-sucking insects and ectoparasites have an absolute
requirement for finding a scarce host in a vast environment.
Olfaction in these haematophagous arthropods may well be
the overriding sensory modality for host recognition and
host location. Host-finding mechanisms and the sensory cues
involved follow the same general rules from species to species
in insects and consist of activation and upwind orientation
stimulated and switched on by carbon dioxide.
The upwind-orientated flight behaviour is modulated and
synergized by a number of ‘generalist’ host odour cues from
breath, urine and skin emanations, such as 1-octen-3-ol, lactic
acid, acetone, alkyl phenols and butanone (compounds 11–
14 respectively in Figure 1) [6–10]. Large, day-flying insects
such as the tsetse fly use vision also. In an aquatic situation,
crustacean ectoparasites such as salmon lice also use host-
specific cues to switch on orientated behaviours (compound
17 in Figure 1). Such cues can be extracted from the water
in which salmon have been swimming [11,12]. The use of
more ubiquitous signals of respiration, such as the presence
of bicarbonate ions, in locating fish hosts has not yet been
investigated.
In addition to the generalist host cues, specific cues
relating to species of insect (anthropophilic versus zoophilic
mosquitoes), sex or physiological status within a species, are
beginning to be defined (A.J. Mordue Luntz, unpublished
work; [6,7,12–14]), all of which are giving rise to possibilities

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132 Biochemical Society Transactions (2003) Volume 31, part 1
of push–pull strategies using odour-baited traps and repel-
lents.
There are many volatile cues yet to be identified in
mammalian breath, in addition to the major cues of carbon
dioxide, 1-octen-3-ol and acetone that ‘switch on’ and
orientate biting flies to their hosts. Another very important
source of odour is skin emanations, and research indicates that
in humans, more than 300 compounds may be released and
may be used as kairomones by mosquitoes and midges [15].
Such host attractants are the subject of much research as likely
candidates for utilization in traps/targets. Compounds have
been shown to be similar among human subjects; however,
the relative amounts of different compounds vary between
subjects and there appears to also be subtle variation in
different compounds both between subjects and within the
same subject in different physiological states. Many of these
volatiles, such as the carboxylic acids (compounds 15 and 16 in
Figure 1), are the result of bacterial breakdown of sebum and
have the potential of varying with time of collection as well as
between individuals. The anopheline mosquitoes, vectors of
malaria, are especially tuned to human-specific odours rather
than carbon dioxide in locating their host [7].
Aggregation pheromones of insects or crustacea [5,16,17]
may also be used, in relation to finding a host, to aggregate
members of the same species together at a feeding site. The
evolutionary value of such pheromones lies in signalling
the presence of a scarce, but non-limiting, food resource
and in there being safety in numbers against host defence
reactions while feeding. Host-seeking parous females of C.
impunctatus, for example, have been shown to produce a
pheromone that acts as an attractant to other parous females
and that acts additively with 1-octen-3-ol, a host kairomone,
when presented in the relevant concentrations [16,17].
Oviposition
Haematophagous insects have aquatic or soil-dwelling larvae
and must be able to locate a suitable oviposition site for
their young, which may be some distance from their host.
Habitat-related semiochemicals, together with pheromones,
may play a large role here and may be successfully exploited
to produce ovitraps for monitoring and control. Apneumones
from the larval-habitat-signalling plant indicator species [18]
or organically enriched water sources [19–21] have been iden-
tified; for the mosquito C. quinquefasciatus have been used
together with the C. quinquefasciatus oviposition pheromone
[22,23] (compound 8 in Figure 1) to test ovitraps both in
the laboratory and in the field [21,24–26]. The oviposition
pheromone has been produced biosynthetically, by a cheap
and sustainable method, as a precursor using the summer
cypress plant Kochia scoparia (Chenopodiaceae), followed
by a final synthetic chemical alteration [27]. Behavioural
bioassays using the volatile skatole from dirty water,
together with the oviposition pheromone synthesized from
K. scoparia precursors reveals identical results to the synthetic
pheromone (Figure 2). Optimal concentrations of both
components provide additive or synergistic attractiveness to


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gravid C. quinquefasciatus when presented in a four-way
choice bioassay (Figure 2). Trials are being conducted at
present in the U.S.A., as West Nile virus is transmitted from
birds to man by C. quinquefasciatus.
Conclusions
Flies, midges and mosquitoes, that bite man, animals and
birds, and sealice (copepod crustacea) that feed on salmon
have been investigated by entrainment of air or water,
extraction and distillation, followed by GC–MS analysis to
identify components that switch on different behaviours. The
sex pheromone of the midge Culicoides nubeculosus coupled
with host blood volatiles in mating behaviour, host location
cues of the Scottish biting midge C. impunctatus, Aedes
aegypti (the mosquito vector of dengue fever) and the sealouse
Lepeophtheirus salmonis, and oviposition cues of the filiariasis
vector C. quinquefasciatus, have been described as examples
of the processes involved in chemical signalling and how such
cues may be used in developing push–pull control strategies.
I thank the Biotechnology and Biological Sciences Research Council,
the Natural Environment Research Council, Ormsary Estate, Argyll and
the Salmon Farming Industry for their support. I thank Mike Birkett
and Tim Olagbemiro, Rothamsted Research for providing pheromone
material, and Craig Rogers, University of Aberdeen, for help with the
bioassays.
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