Geographical Variation of Cuticular Hydrocarbon Profiles of Adult Flies and Empty Puparia Amongst Three Populations of Calliphora vicina (Diptera: Calliphoridae).

Blowflies (Diptera: Calliphoridae) are of great importance in forensic entomology and in determining the minimum post-mortem interval, as they may be the first group of insects to colonize decomposing remains. Reliable species identification is an essential prerequisite. Classically, morphological characters or DNA sequences are used for this purpose. However, depending on the species and the condition of the specimen, this can be difficult, e.g., in the case of empty fly puparia. Recent studies have shown that cuticular hydrocarbon (CHC) profiles are species-specific in necrophagous taxa and represent another promising tool for identification. However, the population-specific variability of these substances as a function of e.g., local climatic parameters has not yet been sufficiently investigated. The aim of this study was to determine the geographical variation of CHC profiles of the blowfly Calliphora vicina (Robineau-Desvoidy, 1830) depending on different countries of origin. Flies were reared in the United Kingdom, Germany, and Turkey in common garden experiments under ambient conditions. CHC profiles of the resulting adult flies and their empty puparia were analyzed using gas chromatography-mass spectrometry. Data were visualized by principal component analysis and clustered by population. The populations of the United Kingdom and Germany, both having similar climates and being geographically close to each other, showed greater similarities in CHC profiles. However, the CHC profile of the Turkish population, whose climate is significantly different from the other two populations, was very different. Our study confirms the high potential of CHC analysis in forensic entomology but highlights the need to investigate geographical variability in chemical profiles.

In vivo antihyperglycaemic and antihyperlipidemic activities and chemical constituents of Solanum anomalum.

Solanum anomalum is a plant used ethnomedically for the treatment of diabetes. The study was aimed to validate ethnomedical claims in rat model and identify the likely antidiabetic compounds. Leaf extract (70-210 mg/kg/day) and fractions (140 mg/kg/day) of S. anomalum were evaluated in hyperglycaemic rats induced using alloxan for effects on blood glucose, lipids and pancreas histology. Phytochemical characterisation of isolated compounds and their identification were performed using mass spectrometry and NMR spectroscopy. Bioinformatics tool was used to predict the possible protein targets of the identified bioactive compounds. The leaf extract/fractions on administration to diabetic rats caused significant lowering of fasting blood glucose of the diabetic rats during single dose study and on repeated administration of the extract. The hydroethanolic leaf extracts also enhanced glucose utilization capacity of the diabetic rats and caused significant lowering of glycosylated hemoglobin levels and elevation of insulin levels in the serum. Furthermore, triglycerides, LDL-cholesterol, and VLDL-cholesterol levels were lowered significantly, while HDL-cholesterol levels were also elevated in the treated diabetic rats. There was absence or few pathological signs in the treated hyperglycaemic rat pancreas compared to that present in the pancreas of control group. Diosgenin, 25(R)-diosgenin-3-O-α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranoside, uracil, thymine, 1-octacosanol, and octacosane were isolated and identified. Protein phosphatases along with secreted proteins are predicted to be the major targets of diosgenin and the diosgenin glycoside. These results suggest that the leaf extract/fractions of S. anomalum possess antidiabetic and antihyperlipidemic properties, offer protection to the pancreas and stimulate insulin secretion, which can be attributable to the activities of its phytochemical constituents.

Cuticular hydrocarbons for the identification and geographic assignment of empty puparia of forensically important flies.

Research in social insects has shown that hydrocarbons on their cuticle are species-specific. This has also been proven for Diptera and is a promising tool for identifying important fly taxa in Forensic Entomology. Sometimes the empty puparia, in which the metamorphosis to the adult fly has taken place, can be the most useful entomological evidence at the crime scene. However, so far, they are used with little profit in criminal investigations due to the difficulties of reliably discriminate among different species. We analysed the CHC chemical profiles of empty puparia from seven forensically important blow flies Calliphora vicina, Chrysomya albiceps, Lucilia caesar, Lucilia sericata, Lucilia silvarum, Protophormia terraenovae, Phormia regina and the flesh fly Sarcophaga caerulescens. The aim was to use their profiles for identification but also investigate geographical differences by comparing profiles of the same species (here: C. vicina and L. sericata) from different regions. The cuticular hydrocarbons were extracted with hexane and analysed using gas chromatography-mass spectrometry. Our results reveal distinguishing differences within the cuticular hydrocarbon profiles allowing for identification of all analysed species. There were also differences shown in the profiles of C. vicina from Germany, Spain, Norway and England, indicating that geographical locations can be determined from this chemical analysis. Differences in L. sericata, sampled from England and two locations in Germany, were less pronounced, but there was even some indication that it may be possible to distinguish populations within Germany that are about 70 km apart from one another.

Cuticular hydrocarbons for identifying Sarcophagidae (Diptera).

The composition and quantity of insect cuticular hydrocarbons (CHCs) can be species-specific as well as sexually dimorphic within species. CHC analysis has been previously used for identification and ageing purposes for several insect orders including true flies (Diptera). Here, we analysed the CHC chemical profiles of adult males and females of eleven species of flesh flies belonging to the genus Sarcophaga Meigen (Sarcophagidae), namely Sarcophaga africa (Wiedemann), S. agnata Rondani, S. argyrostoma Robineau-Desvoidy, S. carnaria (Linnaeus), S. crassipalpis Macquart, S. melanura Meigen, S. pumila Meigen, S. teretirostris Pandellé, S. subvicina Rohdendorf, S. vagans Meigen and S. variegata (Scopoli). Cuticular hydrocarbons extracted from pinned specimens from the collections of the Natural History Museum, London using a customised extraction technique were analysed using Gas Chromatography-Mass Spectrometry. Time of preservation prior to extraction ranged between a few weeks to over one hundred years. CHC profiles (1) allowed reliable identification of a large majority of specimens, (2) differed between males and females of the same species, (3) reliably associated males and females of the same species, provided sufficient replicates (up to 10) of each sex were analysed, and (4) identified specimens preserved for up to over one hundred years prior to extraction.

Chronic Atmospheric Reactive Nitrogen Deposition Suppresses Biological Nitrogen Fixation in Peatlands.

Biological nitrogen fixation (BNF) represents the natural pathway by which mosses meet their demands for bioavailable/reactive nitrogen (Nr) in peatlands. However, following intensification of nitrogen fertilizer and fossil fuel use, atmospheric Nr deposition has increased exposing peatlands to Nr loading often above the ecological threshold. As BNF is energy intensive, therefore, it is unclear whether BNF shuts down when Nr availability is no longer a rarity. We studied the response of BNF under a gradient of Nr deposition extending over decades in three peatlands in the U.K., and at a background deposition peatland in Sweden. Experimental nitrogen fertilization plots in the Swedish site were also evaluated for BNF activity. In situ BNF activity of peatlands receiving Nr deposition of 6, 17, and 27 kg N ha-1 yr-1 was not shut down but rather suppressed by 54, 69, and 74%, respectively, compared to the rates under background Nr deposition of ∼2 kg N ha-1 yr-1. These findings were corroborated by similar BNF suppression at the fertilization plots in Sweden. Therefore, contribution of BNF in peatlands exposed to chronic Nr deposition needs accounting when modeling peatland's nitrogen pools, given that nitrogen availability exerts a key control on the carbon capture of peatlands, globally.

Cuticular hydrocarbons as a tool for determining the age of Chrysomya rufifacies (Diptera: Calliphoridae) larvae.

Calliphoridae are one of the most important insect groups encountered as evidence collected from a crime scene. Age determination of the immature stages of these necrophagous flies is an important step toward estimating the time of colonization and inferring a minimum postmortem interval (PMImin ) in most instances. To determine if the cuticular hydrocarbons could be used to establish whether the development stages yield characteristics profiles, allowing for age estimation, hydrocarbons were extracted from 1st and 2nd, as well as feeding and post-feeding 3rd instar Chrysomya rufifacies, the hairy maggot blow fly. Extracted hydrocarbons were analyzed using gas chromatography coupled to mass spectrometry with the aim to investigate the changes in chemical profiles of each larval stage. A total of 23 compounds were identified with most of them being alkanes (65%) with carbon chain lengths of 9-33 carbons, alkenes (18%), and methyl-branched alkanes (17%). All the hydrocarbons except pentadecane (C15), hexadecane (C16), and nonacosane (C29) showed significant differences in their expression throughout larval development. For 1st instars, nonane was the most abundant (17% of the total hydrocarbons content) compound. Accounting for 11% and 10% of the cuticular hydrocarbons, tricosane and pentacosane, respectively, were the notable hydrocarbons in 2nd instars. For post-feeding 3rd instars, hentriacontane and tritriacontane were present with relative abundances 18% and 15%, respectively. On average, there was a shift from low to high molecular weight hydrocarbons as the larvae aged. These results indicate the change in hydrocarbons makeup as larvae age and could potentially be used to determine the age of immature C. rufifacies and hence aid in PMImin estimations. However, future research is needed to validate these results under natural conditions in the field.

Major Transitions in Cuticular Hydrocarbon Expression Coincide with Sexual Maturity in a Blowfly (Diptera: Calliphoridae).

In many animals, there is a prolonged pre-reproductive period prior to sexual maturity. To avoid premature mating attempts, it is common for phenotypic changes to occur during this period that signal the onset of reproductive viability. Among the insects, pre-reproductive phases can last for up to 50% of the adult lifespan, but little is known about the accompanying phenotypic changes that signal sexual maturity. Contact pheromones such as cuticular hydrocarbons (CHCs) may fulfil this role, as they are known to change rapidly with age in many insects. Despite this, few studies have investigated CHC development in the context of sexual maturity or considered differences in CHC development between sexes. The blowflies (Diptera: Calliphoridae) provide an ideal system for such studies because CHCs are known to change rapidly with age and likely play an important role in sexual behaviour. As such, using the small hairy maggot blowfly Chrysomya varipes, we investigate whether there are age- and sex-specific changes in CHCs over the course of adult blowfly maturation. We show that: (1) major qualitative transitions in CHC expression coincide with the onset of sexual maturity and (2) these changes occur more slowly in females - in line with their extended pre-reproductive phase. We suggest that CHCs may play an important role in signalling sexual maturity in the small hairy maggot blowfly and that this species will likely serve as a useful model for understanding the complex ontogeny of cuticular hydrocarbons in insects.

The evolution of sexually dimorphic cuticular hydrocarbons in blowflies (Diptera: Calliphoridae).

Cuticular hydrocarbons (CHCs) are organic compounds found on the cuticles of all insects which can act as close-contact pheromones, while also providing a hydrophobic barrier to water loss. Given their widespread importance in sexual behaviour and survival, CHCs have likely contributed heavily to the adaptation and speciation of insects. Despite this, the patterns and mechanisms of their diversification have been studied in very few taxa. Here, we perform the first study of CHC diversification in blowflies, focussing on wild populations of the ecologically diverse genus Chrysomya. We convert CHC profiles into qualitative and quantitative traits and assess their inter- and intra-specific variation across 10 species. We also construct a global phylogeny of Chrysomya, onto which CHCs were mapped to explore the patterns of their diversification. For the first time, we demonstrate that blowflies express an exceptional diversity of CHCs, which have diversified in a nonphylogenetic and punctuated manner, are species-specific and sexually dimorphic. It is likely that both ecological and sexual selection have shaped these patterns of CHC diversification, and our study now provides a comprehensive framework for testing such hypotheses.

Phenotypic Plasticity of Nest-Mate Recognition Cues in Formica exsecta Ants.

It is well established that many ant species have evolved qualitatively distinct species-specific chemical profile that are stable over large geographical distances. Within these species profiles quantitative variations in the chemical profile allows distinct colony-specific odours to arise (chemotypes) that are shared by all colony members. This help maintains social cohesion, including defence of their colonies against all intruders, including con-specifics. How these colony -level chemotypes are maintained among nest-mates has long been debated. The two main theories are; each ant is able to biochemically adjust its chemical profile to 'match' that of its nest-mates and or the queen, or all nest-mates share their individually generated chemical profile via trophollaxis resulting in an average nest-mate profile. This 'mixing' idea is better known as the Gestalt model. Unfortunately, it has been very difficult to experimentally test these two ideas in a single experimental design. However, it is now possible using the ant Formica exsecta because the compounds used in nest-mate recognition compounds are known. We demonstrate that workers adjust their profile to 'match' the dominant chemical profile within that colony, hence maintaining the colony-specific chemotype and indicates that a 'gestalt' mechanism, i.e. profile mixing, plays no or only a minor role.

Revealing chlorinated ethene transformation hotspots in a nitrate-impacted hyporheic zone.

Hyporheic zones are increasingly thought of as natural bioreactors, capable of transforming and attenuating groundwater pollutants present in diffuse baseflow. An underappreciated scenario in the understanding of contaminant fate in hyporheic zones is the interaction between point-source trichloroethene (TCE) plumes and ubiquitous, non-point source pollutants such as nitrate. This study aims to conceptualise critical biogeochemical gradients in the hyporheic zone which govern the export potential of these redox-sensitive pollutants from carbon-poor, oxic aquifers. Within the TCE plume discharge zone, discrete vertical profiling of the upper 100 cm of sediment pore water chemistry revealed an 80% increase in dissolved organic carbon (DOC) concentrations and 20-60 cm thick hypoxic zones (<2 mg O2 L-1) within which most reactive transport was observed. A 33% reduction of nitrate concentrations coincided with elevated pore water nitrous oxide concentrations as well as the appearance of manganese and the TCE metabolite cis-1,2-dichloroethene (cDCE). Elevated groundwater nitrate concentrations (>50 mg L-1) create a large stoichiometric demand for bioavailable DOC in discharging groundwater. With the benefit of a high-resolution grid of pore water samplers investigating the shallowest 30 cm of hypoxic groundwater flow paths, we identified DOC-rich hotspots associated with submerged vegetation (Ranunculus spp.), where low-energy metabolic processes such as mineral dissolution/reduction, methanogenesis and ammonification dominate. Using a chlorine index metric, we show that enhanced TCE to cDCE transformation takes place within these biogeochemical hotspots, highlighting their relevance for natural plume attenuation.

Correction to: The introduction history of invasive garden ants in Europe: integrating genetic, chemical and behavioural approaches.

Reinvestigation of the raw data revealed an unfortunate error in Ugelvig et al. 2008 [1].

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.

Adult fly age estimations using cuticular hydrocarbons and Artificial Neural Networks in forensically important Calliphoridae species.

Blowflies (Diptera: Calliphoridae) are forensically important as they are known to be one of the first to colonise human remains. The larval stage is typically used to assist a forensic entomologists with adult flies rarely used as they are difficult to age because they remain morphologically similar once they have gone through the initial transformation upon hatching. However, being able to age them is of interest and importance within the field. This study examined the cuticular hydrocarbons (CHC) of Diptera: Calliphoridae species Lucilia sericata, Calliphora vicina and Calliphora vomitoria. The CHCs were extracted from the cuticles of adult flies and analysed using Gas Chromatography-Mass Spectrometry (GC-MS). The chemical profiles were examined for the two Calliphora species at intervals of day 1, 5, 10, 20 and 30 and up to day 10 for L. sericata. The results show significant chemical changes occurring between the immature and mature adult flies over the extraction period examined in this study. With the aid of a Principal Component Analysis (PCA) and Artificial Neural Networks (ANN), samples were seen to cluster, allowing for the age to be established within the aforementioned time frames. The use of ANNs allowed for the automatic classification of novel samples with very good performance. This was a proof of concept study, which developed a method allowing to age post-emergence adults by using their chemical profiles.

Dietary geranylgeraniol can limit the activity of pitavastatin as a potential treatment for drug-resistant ovarian cancer.

Pre-clinical and retrospective studies of patients using statins to reduce plasma cholesterol have suggested that statins may be useful to treat cancer. However, prospective clinical trials have yet to demonstrate significant efficacy. We have previously shown that this is in part because a hydrophobic statin with a long half-life is necessary. Pitavastatin, the only statin with this profile, has not undergone clinical evaluation in oncology. The target of pitavastatin, hydroxymethylglutarate coenzyme-A reductase (HMGCR), was found to be over-expressed in all ovarian cancer cell lines examined and upregulated by mutated TP53, a gene commonly altered in ovarian cancer. Pitavastatin-induced apoptosis was blocked by geranylgeraniol and mevalonate, products of the HMGCR pathway, confirming that pitavastatin causes cell death through inhibition of HMGCR. Solvent extracts of human and mouse food were also able to block pitavastatin-induced apoptosis, suggesting diet might influence the outcome of clinical trials. When nude mice were maintained on a diet lacking geranylgeraniol, oral pitavastatin caused regression of Ovcar-4 tumour xenografts. However, when the animal diet was supplemented with geranylgeraniol, pitavastatin failed to prevent tumour growth. This suggests that a diet containing geranylgeraniol can limit the anti-tumour activity of pitavastatin and diet should be controlled in clinical trials of statins.

The potential use of cuticular hydrocarbons and multivariate analysis to age empty puparial cases of Calliphora vicina and Lucilia sericata.

Cuticular hydrocarbons (CHC) have been successfully used in the field of forensic entomology for identifying and ageing forensically important blowfly species, primarily in the larval stages. However in older scenes where all other entomological evidence is no longer present, Calliphoridae puparial cases can often be all that remains and therefore being able to establish the age could give an indication of the PMI. This paper examined the CHCs present in the lipid wax layer of insects, to determine the age of the cases over a period of nine months. The two forensically important species examined were Calliphora vicina and Lucilia sericata. The hydrocarbons were chemically extracted and analysed using Gas Chromatography - Mass Spectrometry. Statistical analysis was then applied in the form of non-metric multidimensional scaling analysis (NMDS), permutational multivariate analysis of variance (PERMANOVA) and random forest models. This study was successful in determining age differences within the empty cases, which to date, has not been establish by any other technique.

Species-Specific Cuticular Hydrocarbon Stability within European Myrmica Ants.

Recognition is a fundamental process on which all subsequent behaviors are based at every organizational level, from the gene up to the super-organism. At the whole organism level, visual recognition is the best understood. However, chemical communication is far more widespread than visual communication, but despite its importance is much less understood. Ants provide an excellent model system for chemical ecology studies as it is well established that compounds known as cuticular hydrocarbons (CHCs) are used as recognition cues in ants. Therefore, stable species-specific odors should exist, irrespective of geographic locality. We tested this hypothesis by comparing the CHC profiles of workers of twelve species of Myrmica ants from four countries across Europe, from Iberia to the Balkans and from the Mediterranean to Fennoscandia. CHCs remained qualitatively stable within each species, right down to the isomer level. Despite the morphological similarity that occurs within the genus Myrmica, their CHCs were highly diverse but remarkably species-specific and stable across wide geographical areas. This indicates a genetic mechanism under strong selection that produces these species-specific chemical profiles, despite each species encountering different environmental conditions across its range.

Age estimation of Calliphora (Diptera: Calliphoridae) larvae using cuticular hydrocarbon analysis and Artificial Neural Networks.

Cuticular hydrocarbons were extracted daily from the larvae of two closely related blowflies Calliphora vicina and Calliphora vomitoria (Diptera: Calliphoridae). The hydrocarbons were then analysed using Gas Chromatography-Mass Spectrometry (GC-MS), with the aim of observing changes within their chemical profiles in order to determine the larval age. The hydrocarbons were examined daily for each species from 1day old larvae until pupariation. The results show significant chemical changes occurring from the younger larvae to the post-feeding larvae. With the aid of a multivariate statistical method (Principal Component Analysis and Artificial Neural Networks), samples were clustered and classified, allowing for the larval age to be established. Results from this study allowed larvae to be aged to the day with at worst, 87% accuracy, which suggests there is great potential for the use of cuticular hydrocarbons present on larvae to give an indication of their age and hence potentially a valuable tool for minimum PMI estimations.

Cytotoxicity Effects and Apoptosis Induction by Bisbenzylisoquinoline Alkaloids from Triclisia subcordata.

Triclisia subcordata Oliv (Menispermeaceae) is a medicinal plant traditionally used for the treatment of various diseases in West Africa. The ethanol extract of T. subcordata and its fractions were screened for in vitro anti-ovarian cancer activities using the Sulforhodamine B assay. The crude alkaloids showed the strongest activity in cell growth assays on Ovcar-8 and A2780 cell lines (IC50 < 2.4 µg/mL). A bisbenzylisoquinoline alkaloid-cycleanine was isolated using HPLC and identified by mass spectrometry and nuclear magnetic resonance analyses. The IC50 values of cycleanine and tetrandrine (an alkaloid previously reported from this plant) ranged from 7 to 14 µM on Ovcar-8, A2780, Ovcar-4, and Igrov-1 ovarian cancer cell lines. The IC50 of cycleanine on human normal ovarian surface epithelial cells was 35 ± 1 µM, hinting at modest selectivity toward cancer cells. Both cycleanine and tetrandrine caused apoptosis as shown by activation of caspases 3/7 and cleavage of poly(ADP-ribose) polymerase to form poly(ADP-ribose) polymerase-1 by using western blot analysis. Flow cytometry analyses showed that the percentages of apoptotic cells and cells in subG1 phase increased after exposure of cycleanine and tetrandrine to Ovcar-8 cells for 48 h compared with control. Cycleanine, like its isomer tetrandrine isolated from T. subcordata, could be a potential new anti-ovarian cancer agent acting through the apoptosis pathway. Copyright © 2016 John Wiley & Sons, Ltd.

The Long and the Short of Mate Attraction in a Psylloid: do Semiochemicals Mediate Mating in Aacanthocnema dobsoni Froggatt?

Mating is preceded by a series of interdependent events that can be broadly categorized into searching and courtship. Long-range signals convey species- and sex-specific information during searching, while short-range signals provide information specific to individuals during courtship. Studies have shown that cuticular hydrocarbons (CHCs) can be used for mate recognition in addition to protecting insects from desiccation. In Psylloidea, four species rely on semiochemicals for long-range mate attraction. Psyllid mating research has focused on long-range mate attraction and has largely ignored the potential use of cuticular hydrocarbons (CHCs) as mate recognition cues. This study investigated whether CHCs of Aacanthocnema dobsoni have semiochemical activity for long- and short-range communication prior to mating. Using a solid sampler for solvent-less injection of whole psyllids into coupled gas chromatography/mass spectrometry, we found quantitative, sex- and age-related differences in CHC profiles. Males had higher proportions of 2-MeC28, 11,15-diMeC29, and n-C33 alkanes, while females had higher proportions of 5-MeC27, 3-MeC27, 5,15-diMeC27, n-C29 and n-C30 alkanes. In males and females, 84 and 68 % of CHCs varied with age, respectively. Y-tube olfactometer bioassays provided no evidence that males or females responded to odors emanating from groups of conspecifics of the opposite sex. Tests of male and female psyllids for attraction to branchlets previously occupied by conspecifics showed no evidence of attraction to possible semiochemical residues. Our short-range chemoreception bioassay showed that males were as indifferent to freshly killed individuals of either sex with intact CHC profiles as to those treated with hexane (to remove CHCs). Aacanthocnema dobsoni utilizes substrate-borne vibrations (SBVs) for communication. Therefore, our results indicate that SBVs are probably more important than semiochemicals for long-range mate attraction. Furthermore, CHCs are unlikely to mediate short-range mate recognition or provide mate assessment cues.

Changes in the chemical profile of cephalic salivary glands of Scaptotrigona postica (Hymenoptera, Meliponini) workers are phase related.

Most advanced eusocial bees recruit their nest mates to food resources. Recent studies in Meliponini species have revealed that the cephalic salivary (labial) glands (CSGs) are responsible for the production of scent trail pheromones. Studies on CSGs have shown that changes occur in worker glandular cell morphology from emergence from brood combs until forager phase, which may be correlated to changes in the composition of the CSG secretion. However, the composition of the CSG secretion and the chemical changes that occur in it according to the worker's life phase or tasks performed are unknown for many species, including Scaptotrigona postica. In this study, the chemical profile of CSG secretion in S. postica workers was studied. Glands were taken from specimens that were newly emerged (NE), working in the brood comb area (CA) and foraging (FO), and were analyzed by gas chromatography-mass spectrometry. The results showed that the glandular secretion consists of oxygenated compounds of middle volatility (acids, alcohols, aldehydes, ketones, esters and ether), and their quantity varies among the different life phases, increasing as the individual moves from intra- to extra-colonial activities. The NE phase contained the smallest variety and quantity of compounds. Because of the variability of compounds, the CA workers were separated into three subgroups according to the chemical constitution of their secretion. Forager workers showed the largest quantity and variety of chemical compounds. The major compounds in forager gland secretion were 7-hexadecen-1-yl acetate and 5-tetradecen-1-yl acetate. Statistical analysis indicates that the chemical composition of glandular secretion is phase related.