R.E. Page, C.Y.-S.

Peng / Experimental Gerontology 36 (2001) 695±711 695

Experimental Gerontology 36 (2001) 695±711

www.elsevier.nl/locate/expgero

Aging and development in social insects with

emphasis on the honey bee, Apis mellifera L.
Robert E. Page Jr.*, Christine Y.-S. Peng
Department of Entomology, University of California, One Shields Avenue, Davis, CA 95616, USA

Abstract
Honey bee co`lonies typically consist of about 20±40 thousand workers, zero to few thousand
males (drones), depending on the time of year, and a single queen, the mother of the colony. Workers
typically live 3±6 weeks during the spring and summer and can live about 4 months during the
winter. Queens are longer lived. Anecdotes of queens living 2±3 years are not unusual, though they
normally live less than a year in commercial hives. Little is known about the life span of drones.
Queens develop from fertilized eggs that are not different from the eggs that develop into workers.
Queens are, however, twice as large, have specialized anatomy, live much longer, and develop faster
from egg to adult. All of these differences are derived from differences in larval rearing environment,
primarily nutrition. The developmental trajectory of a female larva from worker into a queen can be
determined as late as the third day of larval development, after this time the developmental pathway
is ®xed for a worker phenotype. The total time of larval development is only 5±6 days, therefore, just
2±3 days of differential feeding can lead to profound differences in development, and longevity.
Workers undergo age development after they become adults. Workers usually initiate foraging
behavior when they are 2±3 weeks old. The age at which a worker initiates foraging is a strong
determinant of her length of life. This is presumed to be a result of the hazards of foraging, but
natural senescence also occurs. Some bees remain in the nest and are never observed to forage,
thereby outliving their forager sisters. Corresponding to this behavioral development are changes in
the sizes of glands and the production of glandular products, increases in biogenic amine titers within
the brain, an increase in the volume of speci®c regions of the brain, and changes in the neural system
that affect perception of stimuli, and learning and memory. These age-related changes in behavior
are regulated by intrinsic and extrinsic factors. Genetic variation has been demonstrated for many of
these life history and behavioral traits. Selection and genome mapping studies have demonstrated
relationships between the neural system, behavior, and life history traits. q 2001 Elsevier Science
Inc. All rights reserved.

Keywords: Aging; Development; Social insects; Honey bee; Apis mellifera

* Corresponding author. Tel.: 11-530-752-0492; fax: 11-530-752-1537.

E-mail address: repage@ucdavis.edu (R.E. Page Jr.).

0531-5565/01/$ - see front matter q 2001 Elsevier Science Inc. All rights reserved.
PII: S 0531-556 5(00)00236-9
696 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711

1. Introduction

Advanced insect societies are characterized by overlapping generations, where

offspring, members of the `worker' caste, remain in the nest and contribute to the repro-
ductive success of their parents, the reproductive caste, at a cost to their own reproduction
(Wilson, 1971). Therefore, longevity of the reproductives was a key enabler of the
evolution of social insects. The maternal parent, the `queen', is usually anatomically
adapted to high output egg production, is larger in size, and has a longer length of life
than her non-reproductive worker offspring. Workers are often anatomically distinct from
the reproductives, and sometimes anatomically differentiated into subclasses of workers
(subcastes). These differences between queens and their worker offspring, and among
worker anatomical subcastes, are usually trophogenic in origin, a result of differential
feeding during larval development.

2. How long do social insects live?

To address this question, we must distinguish between how long they `do' live and how
long they `can' live. Wilson (1971) and HoÈlldobler and Wilson (1990) list numerous
published accounts of observations of length of life for different species of social insects:
ants (29 species), wasps (4), bees (8), and termites (9). For queen ants, maximum recorded
life spans vary from 9±10 weeks for Monomorium pharoanis to 30 years for Pogono-
myrmex owyheei. Queens of 15 ant species were reported to live more than 10 years. Ant
workers were reported to live a maximum of more than 3 years for Aphaenogaster rudis
while workers of eight other species were reported to live more than 2 years. A. rudis
queens have been reported to live up to 13 years yielding a 10 year difference in maximum
reported life spans of queens and workers (Table 1).

Table 1
Maximal life spans of some social insects

Caste Maximal life span Reference

Termites (Isoptera) Kings and Queens 12 years Wilson, 1971

Workers 5 years
Wasps (Hymenoptera) Queens 88 days Wilson, 1971
Workers 97 days
Ants (Hymenoptera) Queens 30 years HoÈlldobler and Wilson, 1990
Workers 3 years
Honey bees Queens 8 years Bozina, 1961
(Hymenoptera: Apis
mellifera)
3 years Seeley, 1978
61% .10 months Gordon et al., 1995
28% .1 year GuzmaÂn-Novoa et al., 1998
Workers 320 days Ribbands, 1953
Drones 59 days Howell and Usinger, 1933
Sperm 8 years Bozina, 1961
 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711
Fig. 1. A queen honeybee (Apis mellifera L.) is attended and cared by a `retinue' of her daughter worker bees throughout her life time (Photographed by K. Locenzlu).

697
698 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711

The reported ages of bees are less than for ants. Among bees, honey bee (Apis mellifera)
queens have the longest reported life span, 8 years as reported by Bozina (1961). However,
this reported life span is far greater than those reported in other similar studies. A
maximum of about 3 years is more likely (Seeley, 1978). If that is the case, then honey
bees queens are outlived by the halictid bee, Euylaeus marginatus, which have been
reported to live up to 5 years (see Table 21-1 in Wilson, 1971). Honey bee workers
have been reported to live up to 320 days (reported in Ribbands, 1953).
Wasp queens and workers of the species Mischocyttarus drewseni have been reported to
live 88 and 97 days, respectively, the longest lived reported in Wilson (1971). Only queens
survive and build colonies in the Hymenoptera (wasps, ants, and bees) while termite kings
and queens cofound nests and live together for up to 12 years. Termite workers have been
reported to live more than 5 years (Table 1).
These studies represent a combination of laboratory and ®eld observation, and anecdote.
It is very dif®cult to study how long social insects live because they often live deep in the
ground or inside trees or other solid structures. Queens of some species apparently can live
decades, making long-term studies of survival unlikely. In addition, comparative studies
between species and between reproductives and non-reproductives of the same species are
confounded by different mortality risks associated with different life histories and
behavior. For example, foraging by workers is a very risky behavior compared with
queen egg laying. Many social insects, such as honey bees, cannot be maintained in
laboratories, where risks can be adequately controlled to determine how long they `can'
live; we only know how long they `do' live under a given set of uncontrolled conditions.
In the following sections, we focus on the honey bee, the best studied of all social
insects. We review caste determination, aging, and length of life.

3. Honey bees

Honey bees typically live in colonies consisting of a single queen, approximately 10±30
thousand ªsterileº female workers, and from zero to a few thousand males, depending on
the time of year (Fig. 1). Workers perform all of the tasks associated with colonial living
while drones ¯y daily from the nest-seeking mates. Queens mate with many males while in
¯ight, soon after they develop into adults. They store the sperm from these many mates in a
specialized structure, the spermatheca, for the rest of their egg-laying life.

4. Honey bee queens

Queens require just 16 days to develop from egg into an adult, 3 days as an egg, 6 days
as a feeding larva (has 5 instars during the larval stage), 7 days as a pre-pupa and pupa. At
emergence, they weigh about 178±292 mg (see Winston (1987) for review).

4.1. How long do queens live?

It is generally believed that queens may survive 1±3 years. Bozina (1961) found that
35% of queens in normal colonies survived 4±6 years, and noted that three queens lived
 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711 699

8 years or more. These are upper estimates of how long they do live, and far higher
estimates than other reported studies. The average life expectancy is probably much
lower. Seeley (1978) reported that 79% of queens survived for one year in unmanaged
colonies, 26% for 2 years, and no queen survived 3 years. One controlled study of queen
survival in commercial colonies in California demonstrated that only 61% of established
queens lived longer than 10 months (Gordon et al., 1995). A similar study conducted in
Mexico under very different conditions showed that only 28% survived more than
12 months (GuzmaÂn-Novoa et al., 1998). The average life expectancy of a feral colony
(which could be longer, but not shorter, than the life span of a queen) in California
declined from an estimated mean of 3.5 years to less than 1 year following the introduction
of the parasitic mite Varroa jacobsoni in 1987 (Kraus and Page, 1995). From this we
conclude that queens can live a long time, and sometimes do, but most live one year or less
(Table 1).

4.2. Causes of queen mortality

It would be extremely dif®cult to determine how long queens can live. Pathogens kill
not only adult queens (Bailey and Ball, 1991), but also their offspring. Aging queens are
often replaced with a daughter queen, raised by workers, then killed. Often the aging queen
is still functioning and appears normal to the observer. But, apparently, the workers detect
changes in signals or cues produced by the queen that result in her supersedure (Butler,
1957). These changes may only signal changes in her reproductive status, such as egg-
laying potential or the quantity and condition of her stored sperm, not her capacity to live
and survive (Winston, 1987). Queens can only survive in a colony environment where her
worker offspring, not the researcher, control her fate.

4.3. Differentiation of queens and workers

Honey bee queens and workers are differentiated into distinct castes exhibited in their
differences in growth and developmental time, life span, morphology, anatomy, physiol-
ogy, and behavior. The mechanism(s) of female caste determination and differentiation
have been investigated by numerous researchers over several decades (see reviews by
Jung-Hoffmann, 1966; Weaver, 1966; de Wilde, 1976; Beetsma, 1979; de Wild and
Beetsma, 1982). Female larvae less than three and a half days old are bipotent, they can
develop either into queens or workers (Shuel and Dixon, 1959; Shuel, 1960; Jay, 1964).
Their destined fate in the caste differentiation process depends on the quantity and quality
of larval food fed to them by young worker bees, `nurse' bees, during the next two instars
(Haydak, 1943; Jay, 1964). At this critical time, those larvae that continue to be fed with a
surplus of glandular secretions, the royal jelly, gain weight at faster rate, grow much larger
(ca. 120 h in larval stage, the queen larvae are 60% heavier) (Weaver, 1955, 1957; Wang,
1965), and develop into the queens. The larvae that are fed a mixture of glandular
secretions (Liu and Jay, 1976; Peng and Jay, 1977, 1979), pollen, and honey (modi®ed
worker jelly) show a slower growth rate, weigh less and develop into the worker bees.
From the ®fth instar onward, a queen continues to develop faster by having a shorter pupal
stage, and emerges as an adult 5 days earlier than the workers. This is remarkable because
queens develop faster, are heavier, and live longer than the workers.
700 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711

Although worker and queen caste differentiation is clearly trophogenic, the food
components that result in the caste differentiation process have not yet been identi®ed.
Studies have shown that adding sugars, e.g. glucose and fructose, in larval food stimulates
food consumption and consequently more queens and queen-like intercastes are reared
(Asencot and Lensky, 1988). A recent study on the development of stomatogastric nervous
system revealed that this neuronal pathway is likely involved in transmitting information
on food quality, such as feeding stimulants and sugars, in larval food (Boleli et al., 1998).
Other qualitative and quantitative difference in larval food may also play an important role
in regulation of caste differentiation (Rembold, 1967).
The role of hormones in regulation of nutritional effects on caste differentiation has
been studied extensively (see review by de Wild and Beetsma, 1982; Rembold, 1987;
Hartfelder and Engels, 1998; Hartfelder, 2000). By the end of the third instar, the queen-
destined larvae have larger corpora allata (CA) and higher titres of juvenile hormone (JH).
The fourth instar generally marks the divergence of the two developmental pathways in
caste differentiation, a process that proceeds continuously under the regulation of JH.
Differentiation of reproductive organs is well established during the ®fth instar. Labora-
tory rearing studies have shown that larvae fed royal jelly beginning from younger ages
(less than third instar) develop into queens with more ovarioles, and larvae fed royal jelly
only during the ®fth instar developed into intercastes (Dedej et al., 1998).
Rachinsy and Hartfelder (1995) demonstrated that JH can turn on ecdysterioids during
caste development and hypothesized that an elevated ecdysteroid titer early in the pre-
pupal phase may act directly at the DNA level, regulating transcription of genes which
shift differentiation toward the queen pathway. Consequently, the differentiation of adult
queen characters is ultimately regulated by ecdysteriods. In addition to JH and ecdyster-
iods, prothoracicotropic hormone (PTTH) secreted by the protocerebral complex during
the larval stage, may also play a role in regulation of caste differentiation (Rachinsky,
1994, 1995±96; Simoes et al., 1997).
Recent studies have provided insights into the complexity of hormonal regulation of
neurogenesis during female caste differentiation in honey bees. Using BrdU (5-bromo-2-
deoxy-uridine) to label neuroblasts and ganglion mother cells in the mushroom bodies and
antennal lobes in the brains, Vitt and Hartfelder (1998) demonstrated signi®cant differ-
ences in neurogenesis in the protocerebrum compared to deutocerebrum during the ®fth
instar and pre-pupal stage. They also demonstrated that mitotic activity in the brain during
the last instar was coordinated with a low titre of JH and high titres of ecdysteriods.
Using FrdU and TUNEL (terminal deoxynucleotidyl transferase-meidated dUTP nick
end labeling) labeling techniques, it was demonstrated that JH affects DNA synthesis and
apoptosis of the larval honey bee ovaries during caste differentiation (Capella and
Hartefelder, 1998). While ovary apoptosis was evident in the germ cell regions of the
ovarioles in worker larvae during the late ®fth instar, ovary apoptosis in the queen larvae
was not observed. Apparently, an elevated JH titer in the early ®fth instar had prevented
the induction of apoptotic DNA degradation, a natural condition that occurs in the larvae
destined to become queens, as well as in JH treated worker larvae. The ovaries of a queen
are large and consist of 150±180 ovarioles in each ovary, whereas the worker ovaries
contain only 2±12 ovarioles in each ovary. In addition, worker bees have only non-
functional rudimentary spermathecae and mating genital chambers.
 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711 701

Recent studies have provided evidence that caste differentiation is also regulated by
transcription activity of m-RNA (Severson et al., 1989), and differential gene expression
(Evans and Wheeler, 1999; Corona et al., 1999). Evans and Wheeler concluded that one
locus is homologous to a class of proteins that bind lipids and other hydrophobic ligands,
and the other locus shows sequence similarity to a DNA-binding domain in the Ets family
of transcription factors. The remaining 5 loci appear to be involved with downstream
changes inherent to queen or worker-speci®c developmental pathways. Differential
expression of mitochondrial genes, including a gene homologous to the nuclear-encoded
mitochondrial translation initiation factor (AmIF-2mt), cytochrome oxidase subunit 1
(COX-1; mitochondrial-encoded), and cytochrome c (cyt c; nuclear-encoded) have been
demonstrated (Corona et al., 1999). The cyt c transcript is more abundant in queen larvae
and throughout the development of the queen bees.

4.4. Sexual maturation and oviposition of queens

Sexual maturity of the queen is reached at 5±6 days after emergence (Ruttner, 1956).
Oogenesis occurs 3±7 days after mating (Snodgrass, 1925; DuPraw, 1967; Kaftanoglu and
Peng, 1982). JH, nutritional condition, as well as age of the queen affect vitellogenesis
(Engels, 1974; Lensky and Skolnik, 1980). Corresponding to sexual maturity is the
maturation of the mandibular glands. The various functions of the mandibular glands of
drones, queens and workers are known to re¯ect the pronounced changes in their ultra-
structure as a consequence of age, physiological condition, and the tasks they perform
(Lensky et al., 1985; Cassier and Lensky, 1991, 1992; Vallet et al., 1991). In queens, these
glands produce pheromones that have profound effects on the behavior of workers and the
maintenance of the social organization of the colony. The mandibular glands of the queen
have been extensively studied (Winston and Slessor, 1998).

4.5. Mating and sperm storage

Virgin queen honey bees mate while in ¯ight within a few days after emerging as adults.
During this time they may take 1±3 mating ¯ights where they may mate with 0±13 males
on a single ¯ight (Tarpy and Page, 2001). They will never mate again. During mating, the
endophallus of the drone breaks off, and consequently, the drone falls to the ground and
dies. The mated queen returns to her nest where over the next 24 h, sperm which are
deposited in the common and lateral oviducts during mating, migrate into the storage
organ, the spermatheca. In the spermatheca, the sperm, probably maintaining in a
quiescent stage, retain their viability and fertility for the next 3±5 years, one of the longest
case in the animal kingdom. How sperm cells are maintained for such long periods of time
in the social Hymenoptera remains a mystery to biologists.

5. Honey bee drones

Drones are derived from unfertilized eggs laid by the queen. They are haploid, inherit-
ing just one set of chromosomes from their mother. Drones require 24 days to develop
from egg to adult. Eggs hatch after 3 days, larvae feed for about 6 days, and the pre-pupal
702 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711

and pupal stages last about 15 days. Drones weigh about 196±225 mg when they emerge
as adults, making them about the same size as queens and about twice as large as workers
(see Winston, 1987).

5.1. How long do drones live?

Estimates of average life span for drones vary from about 20 to 40 days. Fukuda and
Ohtani reported that drones survive for 21±32 days (Fukuda and Ohtani, 1977) during the
summer. The average life span of drones was reported as 21.1 days by Witherell (1972) and
23.5 and 36.2 days by Jaycox (1956) for two different sets of experimental conditions.
Others reported that the average drone life span in July was 23 days, whereas 3% lived
up to 40 days (Drescher, 1969), or 50% lived 21 days, 25% lived 30 days, and 5% lived
51 days (Kepena, 1963). Jaycox (1956) reported a maximum life span of less than 57 days
while Howell and Usinger (1933) reported a maximum life span of 59 days (Table 1).
The main dif®culty in determining how long drones `can' live is that they must be
con®ned in order to extend their life span. Their daily mating ¯ights can be lethal (see
Section 5.2); however, con®nement itself places major stress factors on them. Drones that
have not ¯own have rectums full of feces which might directly affect their survival. Drones
and workers must ¯y in order to empty their recta. Queens defecate inside the nest and the
workers remove it. Life span studies are also seasonally limited because they do not
survive after the end of summer, regardless of age.

5.2. Causes of drone mortality

Like queens, adult drones can die from pathogenic diseases (Bailey and Ball, 1991).
However, they have additional mortality factors. Drones ¯y from the nest each afternoon
in the spring and summer in search of ¯ying queens. Besides the risks associated with ¯ight
(predation, misorientation, exhaustion, etc.), the act of copulating results in death. Drones
are usually expelled from the nest and killed by their sibling workers late in the summer
when foraging resources become scarce. Like queens, drones require workers to feed
them, protect them from predators in the nest, and keep them warm. As a consequence,
the fate of drones is controlled by their nestmate workers, leaving little experimental
control to the researcher. About all we can say is that some drones under some conditions
can live at least 59 days.

5.3. Sexual maturation of drone honey bees

Changes occur in glandular functions as drones age. The secretory activity of the
mandibular glands of drones begins in 0±3 day old drones, and the glands reach full
development by the seventh day. After 9 days the glands begin to degenerate; but the
product is stored in the lumen and emitted secretions attract drones to the congregation
area during drone ¯ights (Lensky et al., 1985).
The development of testes and spermatogenesis occur during the pupal stage of a drone.
Spermatogenesis begins at the eighth day during the pupal stage (Hachinohe and Onishi,
1952), and spermiogenesis occurs before emergence (Bishop, 1920; Hoage and Kassel,
1968). The migration of ¯agellated spermatozoa from the testes to the seminal vesicles
 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711 703

Table 2
The effect of drone age on sperm viability and motility (Different letters indicate signi®cantly different at
P ˆ 0.05 by LSD test, n ˆ 5.)

Drone age (weeks) Viability (%) Motility (%)

1 85.2 ^ 1.41 ab 2.81 ^ 0.16 a

2 86.2 ^ 1.12 a 2.85 ^ 0.13 a
4 81.4 ^ 1.62 bc 2.55 ^ 0.15 a
6 80.1 ^ 2.01 c 2.65 ^ 0.17 a

takes place 2±3 days after adult emergence. By the eighth day after emergence, the
seminal vesicles are ®lled with spermatozoa and the testes have degenerated to about 1/
4 their original size. During the next 2±3 days, the spermatozoa attach themselves by the
head to the epithelial cells of the seminal vesicles where the spermatozoa undergo a
`ripening' process and later become more motile. The epithelial cells of seminal vesicles,
on the other hand, greatly reduce their size (Ruttner, 1976). Parallel to testes development,
shortly after adult emergence, the paired male accessory glands (the mucus glands) begin
to secrete mucus into the gland lumen; initially from the glandular cells located at the
distal ends of the glands and progressing later to the cells at the basal ends. Five to six days
after emergence, the mucus glands are ®lled with mucus ¯uid (Ruttner, 1976).
A newly emerged young drone can be induced to evert its endophallus but without the
ejaculation of mucus and semen. Drones 5±6 days old may ejaculate ¯uid without semen;
the ¯uid contains lumps of white mucus and a watery substance. From the eighth to the
tenth day, the ejaculate contains creamy colored semen, which is mixed in a similar ¯uid
with mucus. By the 12th day, the drones can maximally evert their endophalli and their
ejaculate contains creamy colored semen on top of the homogenous white mucus (Ruttner,
1976)-indicating that they are sexually mature.
Little is known about the duration of sexual potency in drones. However, drones older
than 3 weeks have a noticeable decrease in semen volume as well as an increase in semen
viscosity (Peng and Marsten, unpublished data). Woyke and Jasinski (1973) reported that
as drones increase in age, fewer sperm are transferred into the spermathecae of queens
during arti®cial insemination. Also, with older drones, queens are more likely to retain
semen residues in their lateral oviducts, a condition which can cause infertility and death
in the queens (Vesely, 1970). Signi®cant differences in sperm motility and viability were
observed between drones of different age groups (Locke and Peng, 1993) when sperm was
collected from ejaculates and assessed for their motility and viability with supravital
staining, using Hoecht 33342 ¯uorochrome and propidium iodide (Locke and Peng,
1990; Peng et al., 1990). Sperm viability reached a maximum in ejaculates of 2-week
old drones, and declined with the advancement drone age to 4±6 weeks old (Table 2).
However, the effect of a drone's age on the fertility of sperm is not known.

6. Honey bee workers

We know far more about the life history of worker honey bees than any other group of
704 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711

social insects. Worker honey bees require 21 days to develop from egg to adult: 3 days as
an egg, 6 days as a larva, and 12 days as a pre-pupa and pupa. They weigh about 81±
151 mg when they emerge as adults (Winston, 1987). After they emerge as adults they go
through an age-correlated progression of behavioral changes that has been called
behavioral development. The youngest bees typically clean the nest and feed glandular
secretions to larvae. Middle-aged bees perform nest construction, process food, and guard
the entrance, while older bees forage outside the nest for food, water, and nest construction
materials.

6.1. How long do workers live?

Reports of the average life expectancy of workers vary from 12 (Winston, 1979) to
320 days (reported in Ribbands, 1953) depending on geographic location, season, and
genotype (see Section 6.2). Summer bees live for an average of 15±38 days while workers
can survive for 140 days and longer during the winter (Ribbands, 1953; Fukuda and
Sekiguchi, 1966; Winston et al., 1981, 1983). Winter bees do not have the risks associated
with foraging. In addition, winter bees have more fat-body stored and do not feed larvae,
another mortality factor for workers (Ribbands, 1953). The longest reported survival of
320 days (reported in Ribbands, 1953) demonstrates that workers can live much longer
than the reported averages and can live about as long as the `normal' life expectancy of
queens (Table 1).

6.2. Causes of worker mortality

Worker honey bees suffer from several adult pathogenic diseases (Bailey and Ball,
1991), but, the survival curve is very similar to that of humans (see Fig. 21-2 in Wilson,
1971). Most workers survive until they initiate foraging, when they are about 2±3 weeks
old. Relatively few bees die inside the nest, most just fail to return from foraging trips
(Gary, 1960), which certainly results in death. Mortality then comes primarily as a result of
wear and tear and foraging risks. Wing damage results from foraging and may eventually
result in a bee failing to return to the nest. Neukirch (1982) proposed that worker honey
bees have a ®nite, limited energy reserve in their ¯ight muscles. When this reserve is used,
they can no longer ¯y and cannot return to the nest, resulting in death.
It appears that natural senescence, independent of foraging risks and wear and tear, also
occurs. GuzmaÂn-Novoa et al. (1994) showed that bees that initiated foraging later in life
lived longer than more precocious foragers, but had signi®cantly shorter foraging lives. In
addition, the length of foraging life did not correlate signi®cantly with the number of
foraging trips taken by foragers. Therefore, the length of life is not just a consequence of
events after foraging is initiated, and wear and tear does not explain all of the mortality
associated with foraging. Instead, it appears that bees that develop faster senesce faster.
Using the Gompertz model, Finch (1990) estimated that the mortality rate doubling time
(MRDT) was 0.02 year for the summer bees (worker bees) and 0.03 year for the winter
bees, calculated from the mortality tables of Sakagami and Fukuda (1968). The mortality
rate doubling time of A. mellifera approximate that of the fruit¯y, Drosophila melano-
gaster (Finch, 1990).
 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711 705

6.3. Genetic variation for length of life

The life expectancy of bees varies with genotype. GuzmaÂn-Novoa et al. (1994) tested
bees derived from 6 different genetic sources that were raised together in a common
colony. They found signi®cant variation among sources for the mean ages at which
bees initiated foraging (16.0±18.5 days), length of foraging life (7.0±11.6 days), and
total life span (24.0±28.7 days). Milne (1980) demonstrated variation in length of life
of bees maintained in cages in the incubator. The bees came from different genetic sources.
However, this study only demonstrated variation for how low bees live under the very
stressful conditions of being caged and probably has little relevance for understanding how
long bees can live.

6.4. Age-related changes

Division of labor is the hallmark of insect societies. In honey bees, workers change the
tasks they perform as they age (see Winston, 1987, for review). Associated with changes in
age are changes in the anatomy and physiology of the bee.
In the summer, newly emerged bees have small glands with little secretory activity. At
3±5 days old, the hypopharyngeal glands located in the head begin to produce the enzyme
invertase, used to process nectar into honey. By 6±11 days old the glands are fully
developed. Coinciding with the glandular development, worker bees of this age group
often feed young larvae secretions from these glands. By 17±20 days, the glands show
primarily invertase activity (Maurizio, 1965; Simpson et al., 1968) corresponding to a
principal task performed by bees of this age, processing honey. Ohashi et al. (1999)
demonstrated that genetic homologues of a-amylase and glucose oxidase are expressed
in the hypopharyngeal glands of foraging worker bees. Transcripts of m-RNA for both
enzymes were detected in the hypopharyngeal glands of the foragers but not in younger
bees that feed larvae. Their data suggest that expression of the genes for these two
enzymes (used for processing nectar into honey) is associated with the age-dependent
behavioral changes of the worker bees.
During the aging of worker bees, performing tasks inside the nest (house bees) and
foraging (®eld bees), the general organization of the mandibular glands does not change,
nor do they degenerate, however, the mitochondrial system shows progressive changes.
These changes are most apparent in 10-day old workers, and in older forager bees, and are
correlated with the secretion of the alarm pheromone 2-heptanone (Shearer and Boch,
1965; Boch and Shearer, 1966; Cassier and Lensky, 1991).
The wax glands are located on the sterna of abdominal segments IV, V, VI and VII of
worker bees. These glands secrete wax used in nest construction. In a normal colony, wax
glands reach to their full development in bees that are 10±15 days old, corresponding to
the age of bees that perform the nest construction tasks. Generally, wax secretion by the
wax glands is maximal in bees about 12±18 days old (Hepburn, 1986).
Glands associated with the sting apparatus increase their production of alarm
pheromone with age, a change that corresponds to changes in the defensive behavior of
bees. When bees begin guarding and foraging, about 2±3 weeks old, the sting glands are
producing their maximal quantities of alarm pheromone and the mandibular glands have
706 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711

changed from producing brood food secretions to producing 2-heptanone, another alarm
pheromone (Shearer and Boch, 1965; Boch and Shearer, 1966).
Changes also occur with age in the volumes of some central neuropiles of the brain
(Brandon and Coss, 1982; Durst et al., 1994; Fahrbach et al., 1995; 1997; Withers et al.,
1993). In addition to brain volume changes, titres of brain neuromodulating biogenic
amines (Fuchs et al., 1989; Harris and Woodring, 1992, Harris and Woodring, 1995;
Bozic and Woddring, 1998; Schulz and Robinson, 1999; Wagener-Hulme et al., 1999),
m-RNA transcripts of the period (per) gene (Toma et al., 2000), and blood titres of the
insect growth hormone, JH (Huang et al., 1991), all increase with age. The functional
signi®cance of these changes, if any, is unclear or unknown.

6.5. Does juvenile hormone pace behavioral development?

Many studies have demonstrated increasing blood titres of JH with age (Fluri et al.,
1982; Huang et al., 1991, 1994; Giray et al., 1999). In general these studies show that JH
correlates with age and whether a bee performs tasks within the nest, or forages. In
addition, titres appear to be affected by genotype (Giray et al., 1999), season (Huang
and Robinson, 1995), and demographic manipulation of colonies (Huang and Robinson,
1996; Huang et al., 1998). It was proposed that JH sets the pace, or activates, the observed
age-related changes in behavior. JH titres increase with age and reach a threshold level that
results in the activation of foraging behavior. However, these studies are based on correla-
tions between JH titres and behavior and cannot distinguish between cause and effect. For
example, JH titres may change with stress which is induced by changes in tasks performed,
or the demography of the colony; or, JH titres and behavior may both change as a result of
changes in a common causal factor.
The crucial test of the JH hypothesis was performed by Sullivan et al. (2000). JH is
produced in the CA, paired endocrine glands associated with the brain. They removed the
CA from very young adult workers, resulting in no detectable levels of JH. Allotectomized
workers underwent age-related changes in behavior and initiated foraging behavior at the
same time as the controls, demonstrating that JH is not activating foraging behavior. Sham
treated bees had the surgical procedure performed, but did not have the CA removed. In
most trials they foraged signi®cantly earlier than the controls (probably a consequence of
the effects of stress) but they did not have signi®cantly higher levels of JH than the
controls, again demonstrating that JH titres were not setting the pace of behavioral
development.

6.6. Genetic variation for behavioral development

Genetic variation for the age of onset of foraging behavior has been demonstrated
repeatedly. Page and Fondrk (1995) selected strains of bees for differences in foraging
behavior. Workers from one strain were more likely to forage for pollen while the other
was more likely to collect nectar. Associated with these changes in behavior were changes
in the ages they initiated foraging. The high pollen foraging strain initiated foraging about
10 days earlier than the low strain (Pankiw and Page, unpublished data). These results are
similar to those of Calderone and Page (1988, 1991, 1996) who showed the same relation-
ship for high and low pollen foraging strains derived from an independent selection
 R.E. Page, C.Y.-S. Peng / Experimental Gerontology 36 (2001) 695±711 707

program. Subsequent studies have demonstrated phenotypic and genetic correlations

between foraging behavior, the perception of sugar, and the age of initiation of foraging
behavior (Page et al., 1998; Pankiw and Page, 2000; Page et al., unpublished data). These
strains should be useful for determining the underlying physiological basis of age related
changes in behavior.

7. Conclusion

What do we know about aging in social insects? (1) Queens of some species of ants and
termites live a very long time. (2) It appears that workers usually do not live as long as
their queens. (3) Individual sperm cells live much longer than the males that produce them.
(4) Differences between honey bee queens and workers result from differential feeding
that results in differential gene expression that results in differential humeral control of
development. (5) Processes of maturation continue after queens, drones, and worker honey
bees become adults. (6) Worker honey bees change the tasks they perform as they age, but
the physiological regulators are not understood. (7) Worker honey bees senesce. (8) Rates
of behavioral change are heritable and correlate with the perception of sugar and with
foraging behavior.

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