DEPARTMENT OF AGRICULTURAL ENTOMOLOGY

UNIVERSITY OF AGRICULTURAL SCIENCES,

BENGALURU-560065

SEMINAR REPORT ON

“LIVING DOOR – Phragmosis”

Submitted to
Dr. M. Thippaiah
Professor and Head,
Dept. of Agril. Entomology,
UAS, GKVK, Bengaluru.
Dr. K. V. Prakash
Asst. Professor,
Dept. of Agril. Entomology,
UAS, GKVK, Bengaluru.
Submitted by
Mugundhan N
PALB8139,
Sr.M.sc.,
Dept. of Agril. Entomology,
UAS, GKVK, Bengaluru.

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 CONTENT

SI NO TOPICS Page No

1. Introduction 3

2. Phragmosis and its importance 3

3. Phragmotic Spider of the genus – Cyclocosmia 4-5

4. Phragmotic and trapping organ in Pachyteles larvae 5-7

5. Head-plug defense in a gall aphid 8-12

6. Phragmosis involved in a stabilization and as ecological 12-18

specialization in Cephalotes ant

7. Colony-level stabilization of soldier head width for head-plug 19-24

defense in Reticulitermes speratus

8. Conclusion 25

9. References 25

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Introduction:
Individual specialization within a larger group context is of central importance to the
organization of animal societies. The morphological specialization seen within the workforce
of social insects is the most striking example of this phenomenon. An ancestral and
functionally flexible worker phenotype has been elaborated upon such that additional and
usually more specialized forms are added to the workforce. Evolutionary shifts in ecology are
therefore likely to change the strength of selection for different worker phenotypes, and
ultimately the pattern and diversity of worker polymorphism within a lineage.
Percentage of insect decline in the last 50 years is nearly 41. Among this, nearly half
of the species are in rapid decline. This insect decline is two times more when compared to
vertebrates.

Phragmosis:
All insects are potential prey or host to many kinds of predators or parasitoids. To
make or keep safe from danger, attack or harm. (Sigmund Freud, 1894). So, insect has to
defend itself from predators or parasitoids for its survival. Specialized defence traits and
strategies are crucial in surviving enemy attacks and in resource acquisition.
Phragmosis is the method by which an animal defends itself in its burrow, by using its
own body as a barrier. This term was originally coined by W.M. Wheeler (1927), while
describing the defensive technique exhibited by insects.

Importance of Phragmosis:

 To protect itself from potential predators.

 To tolerate adverse climatic factors.

 To maintain constant conditions inside the chamber or nest.

 Also, involves in a cryptic concealment, thereby making it hard to find by predators

and others insects.

 Also, important during moulting and metamorphosis in Pachyteles larva.

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Examples of Phragmotic defence:

Phragmotic defense behaviour has been reported in many insects and also in other
animals like frogs (Corythomantis greening). It has been well studied and documented in
social insects like ants and Termites. Phragmosis also found in some aphid species and
Pachyteles larva.

1) Phragmotic Spider of the genus – Cyclocosmia

Spiders of the genus Cyclocosmia Ausserer, 1871, are fascinating because their
abdomen is abruptly truncated and ends in a hard, heavily sclerotized disc strengthened by
series of raised ribs separated by narrow grooves (Gertsch & Platnick, 1975). The spiders
usually live at steeply sloping banks of sandy clay, covered with moist leaf litter, where they
dig 7-15 cm deep vertical burrows with a hinged door attached to the entrance. Like all other
trapdoor spiders, Cyclocosmia leads an extremely sedentary existence.

Fig: Vertical burrow in the steeply sloping banks of sandy clay, covered with moist leaf litter.

It is distinct in having developed a morphological defense to cope with intruders.

When the spider retreats headfirst to the bottom of its burrow, the abdominal disc fits tightly
against the round walls of the burrow and forms an impenetrable false bottom. Due to the
successful camouflage of the trapdoor, Cyclocosmia is very difficult to find and collect, and
therefore it is often considered as one of the rarest spiders.

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Fig: Abdominal disc of Cyclocosmia fits tightly against the round walls
of the burrow forming an impenetrable false bottom.

2) Phragmotic and trapping organ in

Pachyteles larvae (Paussinae, Carabidae)

Paussinae, a carabid subfamily mainly distributed in tropical regions, includes about

600 species. Most of them are myrmecophilous and live in ant nests whereas others of the
genus Pachyteles Perty 1830, are non-parasitic and live in burrows in sandy banks or rotten
wood. Larvae of Paussinae are characterised by a hyperprognathous head, a U-shaped body
and the end of the abdomen transformed into a 6-lobed terminal disk.
The “discotelic” paussine larvae (Pachyteles digiulioi and Pachyteles vignai) show a
number of adaptive characters, including a large terminal disk heavily sclerotized and rich in
sensilla. The terminal disk is composed of the last three abdominal segments and the
branched urogomphi. The larva shows defensive and predatory behaviours which use their
terminal disk as both a phragmotic and trapping organ. Also, the larvae attract small
invertebrates by secreting certain substances and catch their prey by an ambush strategy.

Fig: Pachyteles digiulioi, second instar larva, right lateral view, habitus. td: terminal disk

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 Fig: Pachyteles vignai, second instar larva, terminal disk, posterior view; dp: dorsal plate,
consisting in the fusion of epipleurite IX and tergite VIII; lp: lateral plate, consisting in
the modified epipleurite VIII; ug: urogomphus.

Method:
About 20 larvae of Pachyteles vignai, mostly of first and second instars, and about 15
larvae of P. digiulioi of second and third instars were reared in the laboratory at 2 different
times. The larvae of this species were reared in transparent cylindrical containers, 10 cm in
diameter and 5 cm high. Each container was partially filled with soil collected together with
the larvae. The material was placed laterally throughout the cylinder, leaving the central area
clear.
To avoid cannibalism (observed both in larvae and adults during the rearing), no more
than two larvae were introduced into each cylinder. Temperature was set at 18-20 °C, while
humidity was always maintained at saturation level, by regularly moistening the soil without
flooding it. The food of the larvae consisted of small live insects (e.g. fruit flies and their
larvae) provided every 3 days.

Hunting and feeding behaviours:

Initially the larva perceives the presence of a prey in the surroundings of its burrow,
and immediately places the terminal disk to close the entrance. When the prey is situated
towards the middle of the disk, the larva turns and takes it with the mandibles, dragging the
prey inside the chamber.

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The phases of this sequence are summarized as follows,
(1) Mechanical stimulation of the terminal disk.
(2) Immediate closing of the disk (dorsal and lateral plates on branched urogomphi) on the
prey, already stuck on the secretions.
(3) Quick dorsal leaping up of the anterior half of the body (folding in the shape of a “V”)
(4) Dragging of the prey inside the chamber and again the entrance with the terminal disk
(avoiding possible escape of the prey).
(5) Feeding on fluids of the prey by piercing the body with the acute mandibles.
(6) Removal of undigested material.

Fig: Schematic reconstruction of the predatory sequence of a Pachyteles larva: the larva is in the
typical resting position, closing the entrance of the burrow using the terminal disk (a); the larva feels
the arrival of a prey (b) and gets ready to catch it (c); when the prey is in the middle of the disk, the
larva closes it and makes the dorsal predatory release (d) catching the prey with the mandibles (e);
then the larva drags the prey inside the chamber, closing the entrance with the disk (f).

3) Head-plug defense in a gall aphid

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 The aphid Astegopteryx sp. forms a banana-bunch shaped gall consisting of several
subgalls on Styrax benzoides in northern Thailand, and completes its life cycle on the tree,
without migrating to secondary hostplants. Phragmotic head-plug defense was showed by the
gall-forming aphid of the genus Astegopteryx. This species belongs to the tribe Cerataphidini
(Hormaphidinae), all of which form galls on Styrax trees and produce morphologically
distinct, sterile, second instar soldiers in their galls.

Fig: A mature gall of Astegopteryx sp. S, consisting of 11 subgalls, on Styrax benzoides

The soldiers had sclerotic, protruded heads with many spine-like setae, and that
several soldiers cooperate to plug the ostiole of the subgall with these heads. Many eggs and
sexuals were found within subgalls guarded by soldiers, and a number of males were found
trying to intrude into these subgalls.

Fig: Soldiers plugging the ostiole of

their subgall from inside, with the Fig: Eggs and Sexuals were
central soldier mounting on another. found within subgalls

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Functions of Sterile Soldiers:
These cerataphidine soldiers pierce enemies like moth larvae with their stylets and
inject a venomous substance into them (Kutsukake et al., 2004). They also push garbage out
of the gall. Of the tribe, species of Astegopteryx form a banana bunch shaped gall, which
consists of several, spindle-shaped subgalls. Each subgall is almost completely closed, but
with one or a few small ostioles, through which aphids can go out of the subgall. Soldiers
reside within the subgall and guard the ostiole(s) from inside against invaders.
Entrance- guarding and rushing-out” tactics:
When their subgall was disturbed, soldiers rush out onto the surface and excitedly
walk around, often raising the tip of the abdomen upward. Some soldiers soon go back into
the subgall, but, probably because of their inability to discriminate between clonemates and
non-clonemates inside soldiers often prevent outside soldiers from coming in, particularly
those that come back late, at the ostiole. The Astegopteryx species reported below has
developed the tactics to an extreme. Its soldiers defend their subgall in a similar way, but they
cooperate to plug the ostiole nearly completely with their sclerotic, spiny heads, which are
very likely to have evolved for that purpose.
Experiments:
Experiment 1 was carried out to determine whether soldiers that had rushed out of the subgall
could soon go back into the subgall. Experiment 2 was to observe how guarding soldiers
would behave toward soldiers that came back. Experiment 3 was to determine whether
guarding soldiers would prevent sexuals from invading the subgall.
Head-plug defense:
The 15 galls of Astegopteryx sp. observed in the field consisted of six to 20 (mean
12.1) subgalls. All subgalls but one had a single, small, round ostiole (ca. 0.4 mm in
diameter) on a longitudinal suture running from the apex toward the base. (One out of 182
subgalls had two ostioles.) Both sides of the suture subsided to form a V-shaped valley near
the apex. The ostioles were often completely plugged with the sclerotic heads of guarding
soldiers. Of the 182 subgalls, 10 were dead or heavily damaged. Of 173 ostioles of the
undamaged 172 subgalls, 157 (90.8 %) were completely plugged by soldiers. The soldier of
Astegopteryx sp. had an unusually protruded head with many spine-like setae, which are very
likely to function for defense against invaders.

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Interaction between outside and inside soldiers:
The soldiers of Astegopteryx sp. rushed out of the subgall soon began returning to
their subgall. However, some of them were blocked by guarding soldiers.
In experiment 1, a total of 140 soldiers initially came out of 10 subgalls. After five, 15
and 30 minutes, 118, 83 and 77 soldiers, respectively, were walking on the surfaces. Even
after 60 minutes, 54 (38.6 %) still remained outside.
In experiment 2, of 89 soldiers that rushed onto the surfaces of 10 subgalls, 42 (47.2
%) succeeded in entering their natal subgall within 30 minutes. In the latter experiment, we
recorded a total of 75 encounters between a soldier that tried to enter and guarding soldiers at
the ostiole. Of them, 22 entered the subgall without being blocked by guarding soldiers, but
53 were blocked by them. When blocked, an outside soldier pushed the heads of guarding
soldiers with its head and tried to thrust itself into the subgall. Of the 53 blocked soldiers, 20
managed to enter the subgall, and 32 failed and left the entrance. The remaining one soldier
was pierced and killed by guarding soldiers.

Table: Contents of 10 subgalls of Astegopteryx sp.

Interaction between
guarding soldiers and
sexuals:
The first-instar sexuals of nearly 12 were introduced onto a subgall, two males came
to the ostiole four minutes after the introduction, and began to thrust themselves into the

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subgall. Later, a third male joined them. Although they were repeatedly pushing the heads of
guarding soldiers, none of them had intruded into the subgall by the time we fi nished our
observations. One male once pushed its body half into the subgall, and another its entire body
except for the tip of the abdomen, but both were pushed back by guarding soldiers.

Interaction between guarding soldiers and other insects:

An encounter was recorded between plugging soldiers and a possibly predaceous
insect, a thrips, which was trying to invade a subgall in the field. The thrips was blocked by
guarding soldiers and went away. A twig harboring galls was cut off with scissors onto the
ground for collecting materials. Owing to this disturbance, a number of soldiers rushed out of
subgalls of a gall. There was a geometrid moth larva (6.6 mm long) attached to this gall. Four
soldiers attacked the larva. After depositing them in alcohol we confirmed, under a dissecting
microscope, that three of the soldiers were piercing the larva with their stylets.

Why do soldiers prevent colonymates and sexuals from coming in?

The Soldiers cannot discriminate between colonymates and non-colonymates. There
has been reported no case where aphid soldiers (or non-sterile defensive nymphs)
discriminated between kin and non-kin. By blocking soldiers from coming in, guarding
soldiers might consequently select still active, reusable soldiers. For the same reason, they
may prevent sexuals from invading their subgall. By blocking males from invading their
subgall, guarding soldiers might incidentally select strong males for sexual females that are
their colonymates.

4) Phragmosis involved in a stabilization and as ecological specialization in

Cephalotes ant:
Ecology can have a profound influence on social evolution. Ecology playing a vital
role in the evolution of specialized altruistic phenotypes or ‘castes’, a recurrent theme in
insect societies. Cephalotes ants nest in pre-existing arboreal cavities. Cephalotes use their
flat heads to block the colony’s nest entrances, thus preventing enemies from entering, and
the enlarged heads of soldiers are morphologically specialized for this task.
Ecology of Cephalotes ant:

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 Cephalotes normally inhabit pre-existing cavities (e.g. abandoned tunnels of
woodboring insects) in the canopy or understorey that they cannot greatly modify. Use of
such cavities provides highly defensible, hard wood nests with entrances that arthropod
enemies cannot easily open to circumvent the blocking defence. The limitation, however, is
that cavity capacity is relatively fixed, and each must be found in the environment and
occupied before it is used by other arthropods. Inter-specific differences in cavity use by
Cephalotes may be associated with differences in soldier morphology.

Fig: Cephalotes ants nest in pre-existing arboreal

cavities
Evolution of a
specialized soldier:
A morphologically distinct and functionally specialized soldier phenotype is a
common convergent pattern in the evolution of polymorphism, with independent origins in
the termites, thrips, polyembryonic wasps, multiple independent origins within the aphids and
ants. Cephalotes thus offers a unique opportunity to understand the role of ecology in a
recurrent and important theme in social evolution.
The four character states in Cephalotes soldier evolution, in order of increasing
morphological specialization, are

(i) No soldier (ancestral)

(ii) Soldiers with a simple domed head
(iii) Soldiers with an incomplete head-disc that retains
characteristics of the domed head
(iv) Soldiers with an elaborate and complete head-disc

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 C. atratus C. pusillus

C. depressus C. persimilis
NEST DEFENCE AND BEHAVIOURAL SPECIALIZATION
OF SOLDIERS:
Nest defence behaviour was experimentally tested to assess the types and range of
defensive strategies used by workers and soldiers. Preliminary observations revealed that all
focal Cephalotes species displayed stereotyped blocking behaviour whenever Azteca ants
(abundant and aggressive arboreal ants) were foraging nearby. Thus, in each trial, a large
Azteca worker was held with forceps at the focal Cephalotes nest entrance for 45 s, eliciting
an immediate defensive response. Attacks on the Azteca worker were noted, along with the
number and caste of individuals that blocked the nest entrance.
Ten trials were completed for each focal nest, with nine nests tested for each species.
Nests were selected to span the observed natural range of nest entrance area in each species
(previous section), with three centred around the population mean, three around the upper
whisker of an outlier boxplot (i.e. large entrances for the species) and three around the lower
whisker (small entrances).
The involvement of soldiers in foraging was also tested in C. pusillus, C. depressus,
and C. persimilis, to address broad differences in the behavioural repertoire of this caste
among species. Baits were placed approximately 1 m from a known nest entrance. The caste
of the ant that discovered the bait was recorded, along with the caste of the first 20

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individuals recruited to the bait. This procedure was repeated with 20 nests for each species.
Baits consisted of vertebrate urine on small cotton balls.

Nesting ecology:
All Cephalotes nests were established in what appeared to be the abandoned cavities
of wood-boring insects. The cavities occupied by Cephalotes were typically in trees with
substantial damage from wood-boring insects, and other ants often occupied cavities with
similar or identical entrances. Some cavities (≤ 5) occupied by each Cephalotes species
appeared to be newly inhabited because workers were actively removing frass from the
original occupant (dropped particles inspected with a hand lens). The entrance to each cavity
consisted of a tunnel (i.e. near-cylindrical exit tunnel of the original occupant) of variable
length passing through hard wood to the centre of the structure (e.g. branch, main limb, etc.).
This tunnel defined the true nest entrance for the Cephalotes because it was the area that they
blocked.

Ecological specialization:
There was a systematic increase in ecological specialization with increasing
morphological specialization of soldiers among species. The mean and variance of nest
entrance area was significantly different among species in both absolute terms and when the
data were corrected to the standardized scale of head-areas. The mean for absolute and
standardized entrance area was highest in C. atratus (no soldier, ancestral state) and
significantly higher than that in C. pusillus. The variance for absolute and standardized
entrance area was also highest in C. atratus, but the difference to that of C. pusillus was not
significant. The substantial absolute increase in ecological specialization between C. pusillus
and C. depressus (i.e. lower mean and variance for absolute entrance area) is particularly
noteworthy because these two species have soldiers of almost identical size, and thus only
differ in the degree of morphological specialization.
We also see that C. persimilis, with the most specialized soldier, had the most
specialized nesting ecology of the four focal species. In fact, the mean entrance area for C.
persimilis was exactly equal to the head area of a large soldier (Standardized mean of 1·0
head area), and both the variance and mean for standardized entrance area were significantly

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lower than for C. depressus, the species with the next most specialized ecology and soldier
morphology.

Fig b: Outlier boxplots of the standardized nest

Fig a: Outlier boxplots of entrance area (entrance area/max. head area)
nest entrance area

Behavioural specialization:
There was a clear dichotomy in the defensive strategies seen among species. In both
C. atratus (no soldier) and C. pusillus (domed-head soldier), some individuals attacked the
foreign ant while nestmates blocked the nest entrance. However, only the blocking defence
was seen in C. depressus and C. persimilis, the representatives of the two more specialized
and derived states of soldier morphology. Standard workers were involved in blocking in at
least some of the trials in all species except C. persimilis, where only the soldiers (complete
head-disc) blocked.
The agreement between the mean number of individuals that engaged in blocking and
the number predicted by the entrance area (i.e. standardized entrance area = entrance
area/maximum head area) was significantly different among species. The agreement between
predicted and actual number of blockers increased with increasing morphological
specialization and the associated increase in ecological specialization. This pattern can be
explained by the differences among species in the orientation of the head during blocking.

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 In C. depressus and C. persimilis, where the nest is often close to one head-area, the
blocker used the full area of their head by holding the dorsal surface parallel to the plane of
the entrance. However, when entrances were larger, as was common in C. atratus and C.
pusillus, more individuals were required to block them and they pushed the backs of their
heads together with the mandibles facing forward. This strategy reduces the area of the head
used to block, but frees the mandibles for biting and, in the case of C. pusillus soldiers, forms
a well-tessellated composite blocking structure.

C. pusillus was the only species where the soldiers acted as ‘scouts’, discovering rich

Fig: Typical nest entrance blocking in four species of Cephalotes ants, with one representative
for each of the four character states in the evolution of a specialized soldier caste.

food resources and then recruiting nestmates. Soldiers were recruited to the food in all three
species, but the relative representation decreased significantly with increasing morphological
specialization increasing of the soldier.
C. atratus C. pusillus C. depressus C. persimilis
Soldier morphology No soldier Domed head Incomplete Complete head-disc
head-disc
Defensive strategy Attack and block Attack and Block only (all Block only (all trials)
(all trials) block (all trials)
trials)
Caste of blockers Workers only Soldiers and Soldiers and Soldiers only
workers workers
Mean number of ants 2·0 (SD ± 0·0; 1·7 (SD ± 0·5; 1·0 (SD ± 0·0; 1·0 (SD ± 0·0; Pred. = 0·9)
blocking small Pred. = 1·0) Pred. = 0·7) Pred. = 0·6)
entrances 2·4 (SD ± 0·5; 2·3 (SD ± 0·5; 1·0 (SD ± 0·0; 1·0 (SD ± 0·0; Pred. = 1·0)
(3 nests per species, Pred. = 1·3) Pred. = 1·0) Pred. = 0·8)
10 trials per nest) 2·5 (SD ± 0·5; 2·4 (SD ± 0·5; 1·0 (SD ± 0·0; 1·0 (SD ± 0·0; Pred. = 1·0)

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 Pred. = 1·4) Pred. = 1·1) Pred. = 0·6)
Mean number of ants 4·6 (SD ± 0·8; 3·8 (SD ± 0·8; 1·0 (SD ± 0·0; 1·1 (SD ± 0·3; Pred. = 1·3)
blocking mean Pred. = 3·4) Pred. = 2·4) Pred. = 1·2)
entrances 5·5 (SD ± 0·5; 3·9 (SD ± 1·0; 2·2 (SD ± 0·4; 1·3 (SD ± 0·5; Pred. = 1·3)
(3 nests per species, Pred. = 3·8) Pred. = 2·6) Pred. = 1·3)
10 trials per nest) 8·8 (SD ± 0·6;
Pred. = 5·5)
4·2 (SD ± 0·8; 2·4 (SD ± 0·5; 1·3 (SD ± 0·5; Pred. = 1·3)
Pred. = 2·6) Pred. = 1·2)
Mean number of ants 8·8 (SD ± 0·6; 6·6 (SD ± 0·9; 2·0 (SD ± 0·0; 3·2 (SD ± 0·4; Pred. = 2·1)
blocking large Pred. = 5·5) Pred. = 5·6) Pred. = 1·8)
entrances 13·6 (SD ± 1·2; 8·2 (SD ± 1·3; 3·2 (SD ± 0·4; 3·4 (SD ± 0·5; Pred. = 2·2)
(3 nests per species, Pred. = 8·3) Pred. = 6·8) Pred. = 2·2)
10 trials per nest) 13·7 (SD ± 1·6; 7·5 (SD ± 0·9; 4·5 (SD ± 0·7; 3·3 (SD ± 0·5; Pred. = 2·0)
Pred. = 10·8) Pred. = 6·3) Pred. = 2·7)
Soldiers discovering N/A 3/20 trials 0/20 trials 0/20 trials
baits
Mean proportion of N/A 16·8% (SD ± 3·0% (SD ± 1·0% (SD ± 2·2)
soldiers recruited to 11·6) 4·4)
baits

Phylogenetic Analyses of Specialization:

Reconstruction of the ancestral states for standardized entrance area suggested a
systematic increase in ecological specialization (i.e. lower mean and SD) at each shallower
ancestral node, with each representing a more specialized state in soldier evolution. The
reconstructions thus support associated increases in ecological specialization and the
morphological specialization of the soldier caste. The smallest mean and standard deviation
for standardized entrance area (approx. 1·0 head-areas and 0·4 head-areas respectively), as
seen in C. persimilis, may have evolved only in association with the complete head-disc,
because the reconstructed values in the common ancestor of C. depressus and C. persimilis
are higher.
The identity of the root node is uncertain for the reconstruction of the mean and
standard deviation of standardized
entrance area, but in both cases it is
reconstructed as greater than (i.e.
less specialized) or equal to the next
shallower node. Reconstructions
for the use of an attack strategy
with the blocking strategy, and
involvement of standard workers

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in blocking, suggested the loss of these behaviours with particular transitions in the evolution
of the soldier caste. The attack strategy may have been lost in the transition from the
domedhead soldier morphology to the more specialized incomplete head-disc morphology
(Dark grey bar). While the loss of worker involvement in the blocking strategy may have
occurred with the evolution of the complete head-disc, the most specialized soldier
morphology (Light grey bar).

5) Fig: Ancestral character state reconstructions of nesting ecology and nest defence behaviours
in the ant genus Cephalotes using four terminal taxa

Colony-level stabilization of soldier head width for head-plug defense in

Reticulitermes speratus
Social insects have evolved a wide variety of defense mechanisms, ranging from nest
structure to morphological, physiological, and behavioral adaptations of individuals. Termite
soldier castes are probably the most specialized of all social insects. In most termite species,
soldiers specialize in defense and cannot reproduce, work, or feed themselves. Thus, the
soldier caste plays an essential role in colony defense at the cost of reproduction and self-
maintenance.
In Reticulitermes termites, the role of soldiers as colony defenders has been
questioned because soldiers are so few and do not use their cephalic sesquiterpenes against
ants (Zalkow et al. 1981). Waller and LaFage (1986) examined the relative effectiveness of
soldier defenses, and concluded that Reticulitermes soldiers were incompetentcompared with
Coptotermes soldiers.

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Chamber-by-chamber nest defense:
The nests of Reticulitermes speratus are divided into chambers connected by small
openings that allow only one termite at a time to pass through. Reticulitermes speratus Kolbe
soldiers use phragmotic defenses (blocking openings with their heads) that prevent enemies
from invading an undamaged part of the nest.

Fig: Typical nest structure of Reticulitermes speratus. The nests

are divided into chambers connected by small openings (arrowed).

Mandibular and Head-plug (Phragmotic) defense:

When logs infested with Reticulitermes speratus were broken open, individuals of
several ant species carried off struggling workers, whereas soldiers typically retreated to the
small openings between chambers. Soldier defenses included sticking their heads out of the
openings and snapping at approaching ants, and plugging up the openings with their heads to
prevent the ants from invading intact parts of the nest. R. speratus soldiers do not secrete
defensive fluid from the vestigial frontal pores. Thus, normal soldier defense consisted of a
combination of mandibular and head-plug (phragmotic) defenses that were complementary to
the structure of the nest.
Stabilizing selection:
Phragmotic defense requires soldiers to have heads wide enough to block the small
openings between each nest chamber. Soldiers whose heads are too narrow would allow
enemies to pass through, and those whose heads are too large would clog the openings. It

19
confirms that Reticulitermes soldier head width subjected to greater stabilizing selection than
would other morphological traits.
Soldier defense against ant predation:
Portions of four colonies of R. speratus were collected from rotten pine logs at
Takaragaike in the northern suburbs of Kyoto, western Japan, in March 2001. These colonies
were at least 50 m from each other. Termites were used for the experiment within 3 days after
collection. The ant B. chinensis was selected to be the predator in this experiment, because it
is sympatric to, and reported to be an important predator of R. speratus (Teranishi 1929). B.
chinensis nests were frequently found near termite nests in the field.
Experiments were conducted in modified Lüscher cells (Howard 1980), each
consisting of an 80×90 mm piece of window glass to which 2×70 mm strips of plastic plate
had been glued to form a 66×76 mm rectangular cell. Mixed sawdust bait was placed in the
upper third of the cell, and a cylindrical room (10 mm diameter) with an entrance (1.5 mm
width) was cut in the bait.
Two termite groups from the same colony were placed in the ants’ foraging arena.
One unit comprised ten workers and a soldier, the other ten workers only. When all the
termites in a group had entered the hole, it was covered with three 26×76 mm microscopic
slides. Ants were then allowed to enter the foraging area. The sequence of soldier defense
behavior was analyzed, and worker survival rates after 24 h were compared between cells in
which soldiers were present or absent.

Fig A: The experimental apparatus using modified Lüscher cells to compare relative defensive
efficiency between a soldier-present and soldier- absent group.
Fig B: Comparison of survival rate of workers between a soldier-present and soldier-absent group.20
 Significantly more workers were killed and carried off by ants in the absence than in
the presence of soldiers. When present, soldiers guarded the nest openings with their heads
and used their mandibles against ants, as observed in the field. As soon as soldiers detected
the ants via antennal contact, they stuck their mandibles out of the openings and frightened
the ants away. They succeeded in completely shutting out the ants, such that no soldiers were
killed, and only a few workers were killed when they were out of the hole. In the absence of
soldiers, workers frightened the invading ants by shaking their bodies and using them to
block off the openings. This defense was somewhat effective, but the ants expanded the
openings and had killed most of the workers by the end of the experiment.

Sizeblocking
Fig A: The soldier measurements and biometric
the ant by phragmosis. Fig B: The soldier threatening
analysis: the ants with the mandibley

Soldiers extracted from the wood were

immobilized with carbon dioxide and separated by sex, determined using external abdominal
characters. The following measurements were taken for 40 males and 40 females of each
colony: maximum head width (HW); head length (HL) measured from the base of the
mandibles to the posterior margin of the head; head volume (HV) estimated using the formula

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(HW/2)2π×HL; maximum pronotum width (PW); and fresh weight (FW) to the nearest 0.01
mg.
Frequency distribution:
In R. speratus, the HW CV was significantly smaller than CVs of other body parts,
whereas in Coptotermes and Nasutitermes soldiers this was not the case. The mean diameter
of small openings in colony C was 1.23±0.03 (SD) mm (n=20). The CV of the diameter of
small openings was 2.61, which was as small as the CV of soldier head width in R. speratus.
The frequency distributions of fresh weight, pronotum width, and head length showed
bimodal patterns, while that of head width showed an almost unimodal pattern.

Bimodal patterns: Frequency distribution of Pronotum width & Head length

How to Unimodal patterns: Frequency distribution of Head Width (HW)

determine Stabilization ?
Phragmotic defense requires that soldiers have heads wide enough to block the small
nest openings. This suggests that stabilizing selection acted on soldier head width.

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Comparison of the coefficient of variation of head width with that of other body parts showed
that head width was a stable character.
Analysis of the relative growth of head width and head length suggested that
stabilizing selection on soldier head width operates at the colony level. In contrast, soldier
head width was as variable as other body parts in non-phragmotic Coptotermes formosanus
Shiraki and Nasutitermes takasagoensis Shiraki soldiers. Within-colony uniformity of soldier
head width is likely an adaptation.

Fig: Comparison of CVs between the head width and other body parts of R. speratus soldiers
Bars indicate the CVs of pooled data of males and females.
Closed bars indicate significant difference from HW

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 Fig: Comparison of CVs between
Fig: Comparison of CVs between
the head width and other the head width and other body parts
body
parts of C. formosanus of N. takasagoensis

Evidences for Phragmosis:

The small openings in a Reticulitermes nest were of relatively uniform size in the
colony. Because soldiers move from place to place in the nest, within-colony uniformity of
soldier head width is imperative to phragmotic defense, by which soldiers can prevent enemy
invasions while being able to themselves pass through the small openings. The mean
diameter of small openings in colony C was 1.23±0.03 (SD) mm (n=20). The CV of the
diameter of small openings was 2.61, which was as small as the CV of soldier head width in
R. speratus. Thus, HW stabilization of R. speratus soldiers is most likely an adaptation for
phragmotic defense.
The use of phragmosis partially explains the small number of soldiers in
Reticulitermes colonies. Soldiers comprise 3.59±2.50% (mean±SD, n=108 colonies) of
termites in mature R. speratus colonies. Soldiers comprised approximately 2% of termites in
R. flavipes colonies. These soldier proportions are much smaller than in other Rhinotermitids
such as Coptotermes termites, whose colonies consist of up to 40% soldiers. However,
Coptotermes soldiers rush out and assault intruders, whereas Reticulitermes soldiers act
phragmotically, as ‘living plugs,’ complementing the built-in defense of nest structure.

Conclusion:
Specialized defence traits can be the key axes of adaptive evolution, because they
play a crucial role in surviving enemy attacks and in securing resources. For any organism,
specialized defence traits yield fitness gains through the combination of functional
characteristics and associated defence strategies. Selection is thought to favour the evolution
of novel phenotypes, because it improves functional specialization (i.e., performance of a
particular task) and ultimately yielding the fitness gains to the colony.

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References:
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(Coleoptera: Carabidae: Paussinae). Trop. Zool., 14: 157-173.

KUROSU, U., NARUKAWA, J., BURANAPANICHPAN, S. AND AOKI, A., 2006, Head-
plug defense in a gall aphid. Insect. Soc., 53: 86–91.

MATSUURA, K., 2002, Colony-level stabilization of soldier head width for head-plug
defense in the termite Reticulitermes speratus (Isoptera:Rhinotermitidae). Behav Ecol
Sociobiol., 51:172–179.

POWELL, S., 2008, Ecological specialization and the evolution of a specialized caste in
Cephalotes ants. Funct. Ecol., 22: 902–911.

XIN, X., CHEN, X., FAN, L., PHAM, D. AND DAIQIN, L., 2017, Trapdoor spiders of the
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