Is the Salivary Gland Associated with Honey Bee Recognition Compounds in Worker Honey Bees (Apis mellifera)?

Cuticular hydrocarbons (CHCs) function as recognition compounds with the best evidence coming from social insects such as ants and honey bees. The major exocrine gland involved in hydrocarbon storage in ants is the post-pharyngeal gland (PPG) in the head. It is still not clearly understood where CHCs are stored in the honey bee. The aim of this study was to investigate the hydrocarbons and esters found in five major worker honey bee (Apis mellifera) exocrine glands, at three different developmental stages (newly emerged, nurse, and forager) using a high temperature GC analysis. We found the hypopharyngeal gland contained no hydrocarbons nor esters, and the thoracic salivary and mandibular glands only contained trace amounts of n-alkanes. However, the cephalic salivary gland (CSG) contained the greatest number and highest quantity of hydrocarbons relative to the five other glands with many of the hydrocarbons also found in the Dufour's gland, but at much lower levels. We discovered a series of oleic acid wax esters that lay beyond the detection of standard GC columns. As a bee's activities changed, as it ages, the types of compounds detected in the CSG also changed. For example, newly emerged bees have predominately C19-C23n-alkanes, alkenes and methyl-branched compounds, whereas the nurses' CSG had predominately C31:1 and C33:1 alkene isomers, which are replaced by a series of oleic acid wax esters in foragers. These changes in the CSG were mirrored by corresponding changes in the adults' CHCs profile. This indicates that the CSG may have a parallel function to the PPG found in ants acting as a major storage gland of CHCs. As the CSG duct opens into the buccal cavity the hydrocarbons can be worked into the comb wax and could help explain the role of comb wax in nestmate recognition experiments.

Life history and chemical ecology of the Warrior wasp Synoeca septentrionalis (Hymenoptera: Vespidae, Epiponini).

Swarm-founding 'Warrior wasps' (Synoeca spp.) are found throughout the tropical regions of South America, are much feared due to their aggressive nest defence and painful sting. There are only five species of Synoeca, all construct distinctive nests that consist of a single sessile comb built onto the surface of a tree or rock face, which is covered by a ribbed envelope. Although locally common, research into this group is just starting. We studied eight colonies of Synoeca septentrionalis, a species recently been described from Brazil. A new colony is established by a swarm of 52 to 140 adults that constructs a colony containing around 200 brood cells. The largest colony collected containing 865 adults and over 1400 cells. The number of queen's present among the eight colonies varied between 3 and 58 and no clear association between colony development and queen number was detected. Workers and queens were morphologically indistinguishably, but differences in their cuticular hydrocarbons were detected, particularly in their (Z)-9-alkenes. The simple cuticular profile, multiple queens, large size and small number of species makes the 'Warrior wasps' an excellent model group for further chemical ecology studies of swarm-founding wasps.

Are Isomeric Alkenes Used in Species Recognition among Neo-Tropical Stingless Bees (Melipona Spp).

Our understanding of the role of cuticular hydrocarbons (CHC) in recognition is based largely on temperate ant species and honey bees. The stingless bees remain relatively poorly studied, despite being the largest group of eusocial bees, comprising more than 400 species in some 60 genera. The Meliponini and Apini diverged between 80-130 Myr B.P. so the evolutionary trajectories that shaped the chemical communication systems in ants, honeybees and stingless bees may be very different. The aim of this study was to study if a unique species CHC signal existed in Neotropical stingless bees, as has been shown for many temperate species, and what compounds are involved. This was achieved by collecting CHC data from 24 colonies belonging to six species of Melipona from North-Eastern Brazil and comparing the results with previously published CHC studies on Melipona. We found that each of the eleven Melipona species studied so far each produced a unique species CHC signal based around their alkene isomer production. A remarkable number of alkene isomers, up to 25 in M. asilvai, indicated the diversification of alkene positional isomers among the stingless bees. The only other group to have really diversified in alkene isomer production are the primitively eusocial Bumblebees (Bombus spp), which are the sister group of the stingless bees. Furthermore, among the eleven Neotropical Melipona species we could detect no effect of the environment on the proportion of alkane production as has been suggested for some other species.

Evidence for passive chemical camouflage in the parasitic mite Varroa destructor.

Social insect colonies provide a stable and safe environment for their members. Despite colonies being heavily guarded, parasites have evolved numerous strategies to invade and inhabit these hostile places. Two such strategies are (true) chemical mimicry via biosynthesis of host odor, and chemical camouflage, in which compounds are acquired from the host. The ectoparasitic mite Varroa destructor feeds on hemolymph of its honey bee host, Apis mellifera. The mite's odor closely resembles that of its host, which allows V. destructor to remain undetected as it lives on the adult host during its phoretic phase and while reproducing on the honeybee brood. During the mite life cycle, it switches between host adults and brood, which requires it to adjust its profile to mimic the very different odors of honey bee brood and adults. In a series of transfer experiments, using bee adults and pupae, we tested whether V. destructor changes its profile by synthesizing compounds or by using chemical camouflage. We show that V. destructor required direct access to host cuticle to mimic its odor, and that it was unable to synthesize host-specific compounds itself. The mite was able to mimic host odor, even when dead, indicating a passive physico-chemical mechanism of the parasite cuticle. The chemical profile of V. destructor was adjusted within 3 to 9 h after switching hosts, demonstrating that passive camouflage is a highly efficient, fast and flexible way for the mite to adapt to a new host profile when moving between different host life stages or colonies.

Sources of variation in cuticular hydrocarbons in the ant Formica exsecta.

Phenotypic variation arises from interactions between genotype and environment, although how variation is produced and then maintained remains unclear. The discovery of the nest-mate recognition system in Formica exsecta ants has allowed phenotypic variation in chemical profiles to be quantified across a natural population of 83 colonies. We investigated if this variation was correlated or not with intrinsic (genetic relatedness), extrinsic (location, light, temperature), or social (queen number) factors. (Z)-9-Alkenes and n-alkanes showed different patterns of variance: island (location) explained only 0.2 % of the variation in (Z)-9-alkenes, but 21-29 % in n-alkanes, whereas colony of origin explained 96 % and 45-49 % of the variation in (Z)-9-alkenes and n-alkanes, respectively. By contrast, within-colony variance of (Z)-9-alkenes was 4 %, and 23-34 % in n-alkanes, supporting the function of the former as recognition cues. (Z)-9-Alkene and n-alkane profiles were correlated with the genetic distance between colonies. Only n-alkane profiles diverged with increasing spatial distance. Sampling year explained a small (5 %), but significant, amount of the variation in the (Z)-9-alkenes, but there was no consistent directional trend. Polygynous colonies and populous monogynous colonies were dominated by a rich C23:1 profile. We found no associations between worker size, mound exposure, or humidity, although effect sizes for the latter two factors were considerable. The results support the conjecture that genetic factors are the most likely source of between-colony variation in cuticular hydrocarbons.

Weak patriline effects are present in the cuticular hydrocarbon profiles of isolated Formica exsecta ants but they disappear in the colony environment.

Chemical recognition cues are used to discriminate among species, con-specifics, and potentially between patrilines in social insect colonies. There is an ongoing debate about the possible persistence of patriline cues despite evidence for the mixing of colony odors via a "gestalt" mechanism in social insects, because patriline recognition could lead to nepotism. We analyzed the variation in recognition cues (cuticular hydrocarbons) with different mating frequencies or queen numbers in 688 Formica exsecta ants from 76 colonies. We found no increase in the profile variance as genetic diversity increased, indicating that patriline effects were absent or possibly obscured by a gestalt mechanism. We then demonstrated that an isolated individual's profile changed considerably relative to their colony profile, before stabilizing after 5 days. We used these isolated individuals to eliminate the masking effects of the gestalt mechanism, and we detected a weak but statistically significant patriline effect in isolated adult workers and also in newly emerged callow workers. Thus, our evidence suggests that genetic variation in the cuticular hydrocarbon profile of F. exsecta ants (n-alkanes and alkenes) resulted in differences among patrilines, but they were obscured in the colony environment, thereby avoiding costly nepotistic behaviors.