Labial Gland Secretions Research Articles

Atypical Secretions of the Male Cephalic Labial Glands in Bumblebees: The Case of Bombus (Rhodobombus) mesomelasGerstaecker (Hymenoptera, Apidae)

In bumblebees, the male secretion of the cephalic labial gland is species-specific. It is highly involved in the nuptial behavior, acting as a sexual attracting pheromone. Therefore, it is also used to accurately identify the species. In contrast to this common scheme, the secretions of Bombus mesomelas are found to be strongly reduced and do not include the most volatile compounds that are present in the secretions of all the other studied bumblebee species. These secretions correspond to cuticular hydrocarbons that can be found in all bumblebee species. This was also the case for another bumblebee species from the same Rhodobombus subgenus: Bombus pomorum. This atypical composition of the male cephalic labial gland secretions seems to indicate that, at least for these two species of Rhodobombus, these secretions are not used to attract virgin females from a long distance, as it is the case for all the other bumblebee species studied.

The feasibility of using infrared thermography to evaluate minor salivary gland function in euhydrated, dehydrated and rehydrated subjects

Infrared thermography technique (IRT) is utilized by a growing number of disciplines within medicine and dentistry. However, IRT has not been employed in the evaluation of salivary gland dysfunction. The purpose of this study was to examine the feasibility of using thermographic imaging in the evaluation of minor labial salivary gland function in subjects during euhydration, dehydration, and rehydration states. Ten subjects were studied. Upper labial minor salivary gland secretion was quantified whilst simultaneously visualizing lower minor salivary gland output thermographically during each state. A significant difference was observed in the minor labial salivary flow among euhydrated, dehydrated and rehydrated, states. Despite the lack of statistical difference in the thermographic findings, IRT images reflected noticeable differences among the three hydration states. The overall results of this study suggest that IRT could potentially provide a valuable non-invasive tool for evaluating the relationship between minor labial salivary gland function and hydration status.

Reduced cephalic labial glands in the male bumblebees of the subgenus Rhodobombus Dalla Torre (Hymenoptera: Apidae: Bombus Latreille)

Until now, all males of bumblebees are known to attract conspecific females by marking places with a chemical signal secreted by their cephalic labial gland. The specific combination of patrolling flight and scentmarking is of outmost importance to bumblebees as it is their main species specific recognition system. We report here the lack of that recognition system in species of the subgenus Rhodobombus by comparing the morphology and histology of the cephalic labial glands of Bombus (Rhodobombus) mesomelas with those of a well known species, B. (Bombus) terrestris (L.) The cephalic labial glands are much smaller in B. mesomelas than in other bumblebees species and most likely non-functional. This morphology is also observed in B. pomorum and B. brodmanni which also belong to the subgenus Rhodobombus. Our morphological observations are consistent with the chemical analyses of the secretions in B. mesomelas and B. pomorum which are very limited and of most unusual composition for a bumblebee. In addition, whereas other species are thought to use their barbae mandibularis to spread their secretion onto the substrate, these structures are absent in all Rhodobombus. All these observations would mean that the males of Rhodobombus do not attract females from far away by the use of their cephalic labial gland secretions.

Male Cephalic Labial Gland Secretions of Two Bumblebee Species of the SubgenusCullumanobombus (Hymenoptera: Apidae:Bombus Latreille) and Their Distribution in Central Europe

The cephalic labial gland secretion of Bombus semenoviellus males was analyzed, and its chemical composition is reported for the first time. The secretion functions as sex or marking pheromone. Eighty compounds were identified in the secretion, the main one being all-trans-geranylgeranyl acetate (48%). The same compound was shown to form 87% of the labial gland secretion of B. cullumanus males. Both species are closely related and belong to the subgenus Cullumanobombus.

Hexyl Decanoate, the First Trail Pheromone Compound Identified in a Stingless Bee, Trigona recursa

Foragers of many species of stingless bees guide their nestmates to food sources by means of scent trails deposited on solid substrates between the food and the nest. The corresponding trail pheromones are generally believed to be produced in the mandibular glands, although definitive experimental proof has never been provided. We tested the trail following behavior of recruits of Trigona recursa in field experiments with artificial scent trails branching off from natural scent trails of this stingless bee. First-time recruits (newcomers) did not follow these trails when they were laid with pure solvent or mandibular gland extract. However, they did follow trails made with labial gland extract. Chemical analyses of labial gland secretions revealed that hexyl decanoate was the dominant component (72.4 +/- 1.9% of all volatiles). Newcomers were significantly attracted to artificial trails made with synthetic hexyl decanoate, demonstrating its key function in eliciting scent-following behavior. According to our experiments with T. recursa, the trail pheromone is produced in the labial glands and not in the mandibular glands. Hexyl decanoate is the first component of a trail pheromone identified and proved to be behaviorally active in stingless bees.

Male labial gland secretions and mitochondrial DNA markers support species status of Bombus cryptarum and B. magnus (Hymenoptera, Apidae)

Spring queens of Bombus cryptarum and B. magnus from 2 localities in Brandenburg/Germany and Scotland/United Kingdom respectively were determined by morphological characteristics. The lateral border of the collare at the border of the pronotallobus or at the episternum proved to be an especially useful character. Artificial colonies were reared from safely determined spring queens and the cephalic part of the labial glands of males from these colonies were investigated by GC/MS. The investigation identified approximately 50 compounds, as a mixture of straight chain fatty acid derivatives (alcohols, esters and hydrocarbons). The labial secretions of B. cryptarum and B. magnus are significantly different. Mitochondrial cytochrome oxidase 1 (CO1) of two queens from each locality and species were sequenced. Each species from the different localities formed a cluster. Sequence divergence between B. cryptarum and B. magnus was about 30 base substitutions and approximately 0.04 in Tamura-Nei genetic distance. Bombus cryptarum and B. magnus were closer to each other than to B. lucorum and made the sister group in the topology of the tree. Both the CO1 sequences and the labial gland secretions of males of B. cryptarum from Brandenburg and of males from artificial colonies reared from safely determined spring queens from Scotland are identical. B. cryptarum has thus, for the first time, been identified as part of the British bumble bee fauna. The differences of both the labial gland secretions, used as species recognition signals, and the genetic differences established by sequencing CO1 confirm the morphological findings that B. cryptarum and B. magnus are distinct taxa which should be treated as distinct species.

Intra and interspecific variability of the cephalic labial glands' secretions in male bumblebees: the case of Bombus (Thoracobombus) ruderarius and B.�(Thoracobombus) sylvarum [Hymenoptera, Apidae

According to the species recognition concept of Paterson, the analyses of the secretions of the cephalic parts of the male labial glands confirm the conspecificity of Bombus (Thoracobombus) ruderarius ruderarius and B. (T.) r. montanus populations from the Pyrenees. These secretions were compared in B. ruderarius and B. sylvarum. We identified the same 7 major compounds as previously known for these species. We also identified 69 minor compounds. These minor compounds emphasise the close relationship between both species. Principal Component Analyses (PCA) were carried out on standardised peak areas of GC-MS chromatograms. The first PCA component is discriminant and shows no overlap between both species. Their secretions differ mostly by the relative concentration of their compounds rather than by their qualitative composition. On the contrary, PCA is unable to separate montanus from ruderarius. The larger variance in the secretions of B. ruderarius results from the very low concentration of the main compound (9-hexadecenol) in some specimens. Bombus sylvarum sylvarum / Bombus ruderarius ruderarius / Bombus ruderarius montanus / sexual pheromone / cephalic labial gland secretion / chemosystematics

Analysis of the Labial Gland Secretions of the Male Bumblebee Bombus griseocollis (Hymenoptera: Apidae)

The labial gland secretions from males of the bumblebee Bombus (Separatobombus) griseocollis De Geer, a bumblebee exhibiting perching behaviour, were analysed by gas chromatography/mass spectrometry (GC/MS) in the electron impact and positive ion chemical ionization mode. The major compound of the complex mixture of alkenols, acetates, hydrocarbons, wax type esters and steroids is tetradecyl acetate, considerable amounts of hexadecyl, geranyllinaloyl, geranylgeranyl, docosyl, tetracosenyl and hexacosenyl acetate were also found. 1,3-Tetradecanediol diacetate, detected as a minor component, has not yet been identified in male bumblebee labial gland secretions. Besides small amounts of primary alcohols (tetradecanol and hexadecanol) the tertiary alcohol geranyllinalool (3,7,11,15-tetramethyl-hexadeca-1,6,10,14-tetraene-3-ol) was also present. The primary alcohols were also present as esters of butanoic, dodecanoic, tetradecanoic, and hexadecanoic acid. Besides the usual mixture of un- and mono-unsaturated straight chain hydrocarbons, the labial gland contains the isoprenoid hydrocarbons beta-springene [(6E, 10E)-7,11,15-trimethyl-3-methylene-hexadeca-1,6,10,14-tetraene] and two isomers of a-springene [(3Z,6E,10E)- and (3E,6E,10E)-3,7,11,15-tetramethyl-hexadeca-1,3,6,10,14-pentaene]. The close relationship in chemical composition in male bumblebees with perching and flight pass behaviour is discussed.

Marking pheromones of the cuckoo bumblebee males (Hymenoptera, Apoidea, Bombus Latreille): compositions of labial gland secretions of six species found in the Czech Republic

Labial gland secretions of males of six cuckoo bumblebee species ( Bombus vestalis, Bombus rupestris, Bombus campestris, Bombus sylvestris, Bombus bohemicus, and Bombus maxillosus) were analysed and components of their extracts were identified. The specimens of two species collected in the Czech Republic had a significantly different composition of the labial gland secretions than those described earlier in Scandinavia ( B. rupestris, B. campestris). Czech populations of three other species studied ( B. vestalis, B. sylvestris, and B. bohemicus) produced similar components as those known from the literature. Marking pheromone of B. maxillosus is here reported for the first time. This paper describes detailed analyses of the labial gland secretions including minor and trace components and a determination of the double bond positions in unsaturated compounds.

Analysis of the labial gland secretions of the male bumble bee Bombus perplexus Cresson (Hymenoptera: Apidae) from North America.

The labial gland secretions from males of the bumble bee Bombus (Pyrobombus) perplexus Cresson were analysed by gas chromatography/mass spectrometry (GC/MS) in the electron impact and positive ion chemical ionization mode. The major compound of the complex mixture of alkenols, alkenals, fatty acids, hydrocarbons, wax type esters and steroids is 3,7,11,15-tetramethyl-2,6,10-hexadecatrien-1-ol (geranylcitronellol), considerable amounts of hexadecan-1-ol and Z-9-hexadecen-1-ol were also found. All alcohols were present as esters of the detected acids. In older samples both the acids and the alcohols sometimes could not be detected in the GC; therefore, the possibility to check the detected acid-alcohol pattern by interpreting the wax type ester peaks is very instructive. Moreover, the labial gland contains a rich mixture of mono- and di-unsaturated straight chain hydrocarbons. The similarity in composition of the labial glands of the North American B. (Pyrobombus) perplexus with the Eurasian species B. (Pyrobombus) hypnorum corroborates the assumption that the two species are conspecific. The likely supposition that the hydrocarbons could play an essential role in the chemical communication in bumble bees is discussed.

Gelelectrophoretic studies on labial gland secretions of immature blackflies (Simuliidae, Diptera)

Any contact of blackfly larvae and pupae to the substrate they colonise relies on the adhesion of the secretion which is produced in their large labial glands. This secretion anchors their filter-feeding position, any locomotive activity, and the pupal cocoon as well. The secretion works as a biological adhesive which adheres to nearly any substrate surface — even to wood or the integument of other aquatic animals. We began analysing the proteinaceaous compounds of the labial gland secretions of Simulium ornatum (complex) Meigen. According to the SDS-PAGE and amino acid analyses, blackfly gland secretion consists of a variety of proteins ranging from high to low molecular weights. The most prominent proteins form broad bands at about 70, 40 and 20 kDa. These main proteins were present in secretions from young to old larvae as well as in pupal cocoon secretion, but their relative amounts change with larval development. Results of N-terminal sequencing of prominent proteins revealed no obvious homologies to protein sequences deposited in the databases up to now.

Composition of the labial gland secretion of the bumblebee males Bombus pomorum.

Labial gland secretions of 22 males of the bumblebee Bombus pomorum, collected in the Czech Republic, were analysed separately for each individual. The secretions contained 70 compounds among which saturated and unsaturated hydrocarbons strongly dominated. The proportion of hydrocarbons in the secretion was unusually high (85-100%) compared to other bumblebee species studied so far (3-15%). Methyl and ethyl esters of fatty acids, known from many other bumblebee species, formed only minor components (less than 1% in sum) of the secretions of several B. pomorum individuals. No terpenic compounds, typical for males' marking secretion of many bumblebee species, were detected in B. pomorum. The absolute quantities of hydrocarbons present in the labial gland extracts were comparable with those usually present in other species. The composition of hydrocarbons found in the labial glands was different from the profile of the cuticular hydrocarbons. Despite our expectations in species exhibiting a regular patrolling and scent-marking behaviour, the labial gland extracts obtained from B. pomorum males were unusually low concentrated and their chemical composition was atypical with respect of the proportions of hydrocarbons when compared with other patrolling species. This is the first report on the analysis of the labial gland secretion of the B. pomorum males.

Chemotaxonomical characterisation of males of Bombus lucorum (Hymenoptera: Apidae) collected in the Czech Republic

Labial gland secretions of 26 males of the bumblebee Bombus lucorum (L.), collected in the Czech Republic, were analysed. The secretions consisted of 60 compounds; ethyl (Z)-9-tetradecenoate was the main component (average 53%). Although the males varied in colour, their labial gland secretions were similar in composition, which indicated they belonged to one species. Chemically the B. lucorum occurring in the Czech Republic correspond to the earlier described blonde form of this species.

Thin-layer chromatography assessing feeding stimulation by labial gland secretion compared to synthetic chemicals in the subterranean termite Reticulitermes santonensis.

The labial gland of the French subterranean termite Reticulitermes santonensis De Feytaud contains a polar, heat-resistent, and persistent chemical signal that is released onto the food during food exploitation and stimulates feeding in nestmates. Separation of the labial gland secretion by thin-layer chromatography on cellulose plates revealed that the secretion contains components with reducing and amino groups. In feeding bioassays conducted on the cellulose plate after TLC, termites preferred the area between Rf 0.46 and 0.88 (biologically active zone) for feeding, indicating the location of the feeding-stimulating signal. Thirty-five synthetic chemicals with similar chemical properties as the feeding-stimulating signal were analyzed with TLC. None of them covered the biologically active zone. Therefore, all chemical classes tested, such as sugars, amino acids, and salts, are unlikely as possible sources for the signal structure. In feeding choice tests with synthetic chemicals, termites showed clear feeding preference only for sugarlike components with physiologically excessive concentrations of 10 mmol and 100 mmol. Amino acids induced only light feeding preference. The intensity of feeding stimulation by the natural signal from the labial gland as compared to synthetic phagostimulants is discussed.

Food exploitation in termites: indication for a general feeding-stimulating signal in labial gland secretion of isoptera.

The paired labial glands are located in all termite species in the thorax. During food exploitation workers of the French termite Reticulitermes santonensis and the African termite Schedorhinotermes lamanianus release the secretion of their labial glands directly onto the food. The secretion carries a water-soluble, heat-resistant, nonvolatile signal that stimulates gnawing and feeding and leads to aggregations of feeding workers. In a feeding bioassay, extracts of the labial glands of 11 termite species from five families all proved to have this feeding-stimulating effect both on R. santonensis and S. lamanianus. The heat resistance of the feeding-stimulating signal also could be shown for selected species from all termite families tested. A combined thin layer chromatography-feeding bioassay on cellulose TLC plates showed that after chromatography of labial gland secretion, the feeding-stimulating signal is located in all 11 species in the same area from Rf 0.46 to 0.88. An extract of labial glands of cockroaches stimulated feeding in R. santonensis and S. lamanianus as well, but was not active after heat treatment and after TLC. This points towards a general feeding-stimulating signal having evolved only in the labial gland secretion of termites.

SCENT MARKING IN MALE PREMATING BEHAVIOR OF Bombus confusus

Premating behavior in the species Bombus confusus Schenck, 1859 was studied. The marking habits of bumblebee males were revealed and the use of their labial gland secretion in scent marks was proved chemically. Identical compounds were present both in the labial gland secretion and on male-marked objects. This chemical proof is reported for the first time from natural conditions of a B. confusus locality. Males' flight activities, site tenacity, and male–male interactions are described in detail. Contrary to literature reports, male B. confusus behavior does not basically differ from that of already known perching bumblebee species.

PREMATING BEHAVIOR OF Bombus confusus MALES AND ANALYSIS OF THEIR LABIAL GLAND SECRETION

Premating behavior in the bumblebee Bombus confusus was studied. Visual searching for females is not the only premating strategy of this species, as was believed earlier. Males of B. confusus have a normally developed labial gland and its secretion is used to mark a perch from which they visually search for females. The labial gland secretion contains geranylcitronellol and (Z)-9-octadecenyl acetate as the main components.

Immunoelectron microscopic identification of lysozyme-expressing cells in human labial salivary glands.

Although human labial gland secretions contain serous components such as the bactericidal enzyme, lysozyme, the presence of serous cells in this gland has yet to be clearly visualized under the electron microscope. The present study identifies lysozyme-expressing cells of the labial glands using microwave-fixed, Epon-Araldite-embedded specimens, which showed excellent preservation of both ultrastructural detail and antigenicity for post-embedding immunogold labeling of lysozyme. Ultrastructurally, all of the secretory cells of the glands appeared to be a mucous-type and have a serial maturation relationship, consistent with a previous report by TANDLER et al. (1969a): their secretory granules were electron-lucent and exhibited reactivity for mucus staining by the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method. We classified them into two immature types (I, II) and two mature types (I, II). Their distinctive features were the following: 1) relatively small (0.5-1 microm) secretory granules and well-developed basal rough endoplasmic reticulum for the immature types; 2) larger (1-2 microm) secretory granules and well-developed Golgi apparatus, which showed intense PA-TCH-SP reactivity in the 2-3 trans-cisterns, for the mature types; 3) few secretory granules in the immature type I; and 4) the darkest appearance for the mature type II. Immunogold labeling with anti-lysozyme showed specific labeling of the two immature-type cells, in which gold particles were found mainly over the secretory granules and Golgi apparatus, and moderately over the rough endoplasmic reticulum. In the secretory granules, the labeling was distributed throughout the contents and was present even if they showed strong PA-TCH-SP reactivity; in the Golgi area, it was seen over the stacked cisternae, trans-Golgi networks, and condensing vacuoles. No specific labeling was seen in the mature-type cells or in the duct cells. These immature- and mature-type cells were almost equivalent to the "serous demilune or acinus" and "mucous tubule" cells, respectively, at the light-microscopic level. These results indicate that the traditional "immature mucous-type cells" of the human labial glands produce lysozyme and should be classified as seromucous cells.

Abgrenzung der Hummel-Arten Bombus cryptarum und B lucorum mittels männlicher Labialdrüsen-Sekrete und morphologischer Merkmale (Hymenoptera, Apidae)

Three taxa of the subgenus Terrestribombus Vogt 1911 [genus Bombus Linnaeus 1758] have been reared in the laboratory and assigned by the main component (MC) of the male labial gland secretions to the species terrestris Linnaeus 1758 (MC = 2,3-Dihydrofarnesol), lucorum Von Linne 1761 (MC = Myristoleic acid ethyl ester) and cryptarum Fabricius 1775 (MC = Lauric acid ethyl ester). Individuals of terrestris can be identified by proboscis-length, individuals of lucorum and cryptarum by the Rf-values of Phosphoglucomutase on PAGE-gel. Coloration and boundary of the collare at the edge of the pronotolobus of queens are reliable characters to discriminate cryptarum and lucorum. 39 compounds including mainly hydrocarbons, alcohols and esters are identified from the male labial glands of cryptarum.

Feeding Stimulating Signal in Labial Gland Secretion of the Subterranean Termite Reticulitermes santonensis

The paired labial glands of the French subterranean termite Reticulitermes santonensis Feytaud are located in the thorax. In the head, the glandular ducts join with those of the water sacs. In feeding choice tests with two semicircles of moist filter paper as food, workers of R. santonensis preferred the semicircle treated with labial gland extract compared to the semicircle treated with water (control). The labial gland secretion carries a signal that stimulates gnawing and feeding by termite workers during food exploitation. The extract of the labial gland even elicited feeding behavior when applied without food (on glass plates). The content of the water sacs was not effective as a feeding stimulus and neither were different body parts except for the fat-body. Water sacs are thus not a reservoir for the labial gland secretion, but their contents may serve as a solvent or carrier for the phagostimulant signal. The signal is highly polar, heat-resistant, nonvolatile, and thus very persistent. There is evidence that the signal from the labial gland is not species specific. Instead, it might be part of a general strategy by termites to exploit food sources.