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.     

Chemical reproductive traits of diploid Bombus terrestris males: Consequences on bumblebee conservation

The current bumblebee decline leads to inbreeding in populations that fosters a loss of allelic diversity and diploid male production. As diploid males are viable and their offspring are sterile, bumblebee populations can quickly fall in a vortex of extinction. In this article, we investigate for the first time a potential premating mechanism through a major chemical reproductive trait (male cephalic labial gland secretions) that could prevent monandrous virgin queens from mating with diploid males. We focus our study on the cephalic labial gland secretions of diploid and haploid males of Bombus terrestris (L.). Contrary to initial expectations, our results do not show any significant differentiation of cephalic labial gland secretions between diploid and haploid specimens. Queens seem therefore to be unable to avoid mating with diploid males based on their compositions of cephalic labial gland secretions. This suggests that the vortex of extinction of diploid males could not be stopped through premating avoidance based on the cephalic labial gland secretions but other mechanisms could avoid mating between diploid males and queens.     

The cephalic labial gland secretions of two socially parasitic bumblebees Bombus hyperboreus (Alpinobombus) and Bombus inexspectatus (Thoracobombus) question their inquiline strategy

Social parasitic Hymenopterans have evolved morphological, chemical, and behavioral adaptations to overcome the sophisticated recognition and defense systems of their social host to invade host nests and exploit their worker force. In bumblebees, social parasitism appeared in at least 3 subgenera independently: in the subgenus Psithyrus consisting entirely of parasitic species, in the subgenus Alpinobombus with Bombus hyperboreus, and in the subgenus Thoracobombus with B. inexspectatus. Cuckoo bumblebee males utilize species‐specific cephalic labial gland secretions for mating purposes that can impact their inquiline strategy. We performed cephalic labial gland secretions in B. hyperboreus, B. inexspectatus and their hosts. Males of both parasitic species exhibited high species specific levels of cephalic gland secretions, including different main compounds. Our results showed no chemical mimicry in the cephalic gland secretions between inquilines and their host and we did not identify the repellent compounds already known in other cuckoo bumblebees.     

Exocrine gland secretions of virgin queens of five bumblebee species (Hymenoptera: Apidae, Bombini)

Secretions of three different glands (mandibular gland, labial gland, and Dufour's gland) of virgin queens of five bumblebee species (Bombus lucorum, B. lapidarius, B. hypnorum, B. pascuorum, and B. terrestris) were analysed. Around 200 compounds were identified in the secretions. The compositions of the secretions of labial and mandibular glands were species-specific. Dufour's gland of all species produced mainly hydrocarbons, both saturated and unsaturated, the proportions of which differed quantitatively between the species studied.     

Labial gland marking secretions of male Bombus lucorum bumblebees from Europe and China reveal two separate species: B. lucorum (Linnaeus 1761) and Bombus minshanicola (Bischoff 1936)

Differences in male bumblebee labial gland secretions can be used to separate species pairs. Cephalic labial gland secretions from male Bombus lucorum Linnaeus bumblebees from Europe, and male ‘B. lucorum’ bumblebees from China, were analyzed by gas chromatography/mass spectrometry (GC/MS). In the European B. lucorum L., ethyl tetradec-9-enoate was identified as the major compound (58% peak area), along with a complex mixture of straight-chain alkenols, acetates, hydrocarbons, and wax type esters. In contrast, the main component of the labial gland secretions of the Chinese ‘B. lucorum’, ethyl dodecanoate (31%), did not dominate the secretion, which additionally contained large amounts of 3,7,11-trimethyldodeca-6,10-dien-1-ol (=2,3-dihydrofarnesol, 24% peak area), ethyl octadec-9-enoate (15% peak area) and a mixture of acyclic diterpenes (alcohols, aldehydes, and acetates). Furthermore, in B. lucorum, 3,7,11-trimethyldodeca-6,10-dien-1-ol was only detected in trace amounts (0.05%) and ethyl octadec-9-enoate corresponded to only 0.8% of the mixture. These results indicate that ‘B. lucorum’ from China is a separate taxon from B. lucorum L. Further analysis revealed that ‘B. lucorum’ from China has been previously described as Bombus minshanicola Bischoff 1936 from Gansu/China. Differences in the chemical composition of male bumblebee labial gland secretions are discussed in comparison with other known bumblebee species pairs.     

Methods for species delimitation in bumblebees (Hymenoptera,Apidae, Bombus): towards an integrative approach

Delimitation of closely related species is often hindered by the lack of discrete diagnostic morphological characters. This is exemplified in bumblebees (genus Bombus). There have been many attempts to clarify bumblebee taxonomy by using alternative features to discrete morphological characters such as wing shape, DNA, or eco‐chemical traits. Nevertheless each approach has its own limitations. Recent studies have used a multisource approach to gather different lines of speciation evidence in order to draw a strongly supported taxonomic hypothesis in bumblebees. Yet, the resulting taxonomic status is not independent of selected evidence and of consensus methodology (i.e. unanimous procedure, majority, different weighting of evidence). In this article, we compare taxonomic conclusions for a group of taxonomically doubtful species (the Bombus lapidarius‐group) obtained from the four commonly used lines of evidence for species delimitation in bumblebees (geometric morphometric of wing shape, genetic differentiation assessment, sequence‐based species delimitation methods and differentiation of cephalic labial gland secretions). We ultimately aim to assess the usefulness of these lines of evidence as components of an integrative decision framework to delimit bumblebee species. Our results show that analyses based on wing shape do not delineate any obvious cluster. In contrast, nuclear/mitochondrial, sequence‐based species delimitation methods, and analyses based on cephalic labial gland secretions are congruent with each other. This allows setting up an integrative decision framework to establish strongly supported species and subspecies status within bumblebees.     

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.     

Distribution and flower visitation records of bumblebees in Lebanon (Hymenoptera: Apidae)

Summary

West Palearctic bumblebees are common wildflowers and crop pollinators that are well studied in their central and northern distribution ranges, but fewer information is available on their southern distribution areas. Lebanon falls on the southern limit of their distribution and no published information is available on the local bumblebees. Our study aims to produce a data baseline of the local bumblebee species. In order to do so we grouped available old records of bumblebees in Lebanon with recent author collections and produced preliminary distribution maps. We listed four species: Bombus terrestris, B. argillaceus, B. niveatus vorticosus and B. melanurus. Preliminary distribution shows that Bombus terrestris and B. argillaceus are widespread and have a large foraging range, whereas B. niveatus vorticosus and B. melanurus have a restricted distribution to altitudes above 1800 m with a smaller foraging range. The male cephalic labial gland secretions analysis of local Bombus terrestris specimens provides preliminary evidence that the local subspecies could be Bombus terrestris calabricus. Therefore, we highlight the importance of regulating foreign Bombus terrestris subspecies importation for agriculture purposes, as well as monitoring B. niveatus vorticosus and B. melanurus that are rendered vulnerable by their isolated populations.     

First Chemical Analysis and Characterization of the Male Species‐Specific Cephalic Labial‐Gland Secretions of South American Bumblebees

The evolution of signals and reproductive traits involved in the pre‐mating recognition has been in focus of abundant research in several model species, such as bumblebees (genus Bombus). However, the most‐studied bumblebee reproductive trait, the male cephalic labial gland secretions (CLGS), remains unknown among bumblebee species from South America. In this study, the CLGS of five South American bumblebees of the subgenera Thoracobombus (Bombus excellens and B. atratus) and Cullumanobombus (B. rubicundus, B. hortulanus, and B. melaleucus) were investigated, by comparing the chemical compositions of their secretions to those of closely related European species. The results showed an obvious interspecific differentiation in both subgenera. The interspecific differentiation among the species of the Thoracobombus subgenus involved different compounds present at high contents (main compounds), while those of the Cullumanobombus subgenus shared the same main components. This suggests that among the species of the Cullumanobombus subgenus, the differentiation in minor components could lead to species discrimination.     

Male cephalic labial gland secretions of two bumblebee species of the subgenus Cullumanobombus (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.     

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

Summary. 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.Received 25 March 2004; revised 13 June 2004; accepted 15 June 2004.     

Age-dependent changes in structure and function of the male labial gland in Bombus terrestris

The cephalic region of the labial gland in the buff-tailed bumblebee, Bombus terrestris, consists of numerous acini (formed by associated secretory cells and a central lumen) and connecting ducts. Age-dependent changes in secretion production (both qualitative and quantitative) are associated with changes in the amount of rough endoplasmic reticulum (RER), Golgi apparatus, and smooth endoplasmic reticulum (SER). The main secretory organelle is RER in the youngest individuals (pharate, and less-than-a-day old males), Golgi apparatus in 1-day-old males, and SER in males older than 2 days. Secretory cell death starts at 5 days of age, with maximal longevity to 10 days. Pheromone production starts immediately after eclosion, with pheromone quantities increasing until day 7. 2,3-dihydrofarnesol, the main component of the male-marking pheromone, appears in 1-day-old male glands, and reaches a maximum at 7 days of age, when its presence in the gland starts to decrease gradually. Older glands contain compounds not present in young ones. Variation in pheromone quantity and composition are reflected sensitively in the response of the queen antennae. Though queen antennae responded to gland extracts of all ages examined, maximum sensitivity was observed in response to extracts of glands 2-10 days old, while extracts of older glands gradually lose their effectiveness. Both major and minor components of the labial gland secretion extract elicited queen antennal responses suggesting that the pheromone is a multicomponent blend. Age-dependent changes in pheromone production, accumulation and tuning of pheromone activity are all synchronized approximately with male flight from the hive.     

Absolute configuration of chiral terpenes in marking pheromones of bumblebees and cuckoo bumblebees

The absolute configurations of citronellol, 2,3-dihydrofarnesol, and 2,3-dihydrofarnesal in male marking pheromones of seven species of bumblebees and cuckoo bumblebees were determined by enantioselective gas chromatography on a capillary column coated with 60% heptakis(2,3-di-O-acetyl-6-O-TBDMS)-beta-cyclodextrin in polysiloxane PS 268. Pure (-)-S-enantiomers of all three terpenes were found in the labial glands of all investigated specimens of the following species: Bombus (Bombus) terrestris, B. (Bombus) lucorum, B. (Pyrobombus) pratorum, B. (Pyrobombus) pyrenaeus, B. (Pyrobombus) jonellus, B. (Pyrobombus) impatiens, and the cuckoo bumblebee B. (Ashtonipsithyrus) bohemicus. Within species, specimens were collected at different localities and in different years. Except for 2,3-dihydrofarnesol in B. terrestris, this is the first report on the absolute configuration of terpenes in marking pheromones of bumblebees.     

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.     

Infestation of commercial bumblebee (Bombus impatiens) field colonies by small hive beetles (Aethina tumida)

Abstract.  1. The small hive beetle, Aethina tumida , is a parasite of honeybee ( Apis mellifera ) colonies native to sub-Saharan Africa and has become an invasive species. In North America the beetle is now sympatric with bumblebees, Bombus , not occurring in its native range. Laboratory studies have shown that small hive beetles can reproduce in bumblebee colonies but it was not known whether infestations occur in the field.
2. For the first time, infestation of bumblebee colonies by small hive beetles was investigated in the field. Commercial Bombus impatiens colonies ( n = 10) were installed in proximity to infested apiaries. Within 8 weeks, all colonies that were alive in the 5-week observation period ( n = 9) became naturally infested with adult small hive beetles and successful small hive beetle reproduction occurred in five colonies.
3. In four-square choice tests, the beetles were attracted to both adult bumblebee workers and pollen from bumblebee nests, suggesting that these odours may serve as cues for host finding.
4. The data indicate that bumblebee colonies may serve as alternative hosts for small hive beetles in the field. To foster the conservation of these essential native pollinators, investigations on the actual impact of small hive beetles on wild bumblebee populations are suggested.     

Lipophilic compounds in femoral secretions of male collared lizards,Crotaphytus bicinctores (Iguania,Crotaphytidae)

In many lizards, chemical compounds from the femoral gland secretions are used in intraspecific communication, but most studies describing these chemicals are for lizard species included in the Scleroglossa clade, whereas lizards within the Iguanian clade have been much less studied, probably because these lizards were considered to rely more on visual cues. However, many iguanian lizards have abundant femoral secretions and are able of chemosensory conspecific recognition, which might be based on compounds secreted by femoral glands. By using GC–MS analyses, we found 58 lipophilic compounds in femoral gland secretions of male Great Basin collared lizard, Crotaphytus bicinctores (Iguania, Crotaphytidae). Main compounds were steroids (mainly two triunsaturated steroids and cholesterol), carboxylic acids (mainly hexadecanoic acid), waxy esters of long chain fatty acids, alcohols (mainly hexadecanol), aldehydes and other minor compounds. We compared these compounds with those found in other lizard species and discussed the potential signaling function of some compounds and how the xeric habitat of this lizard could have conditioned the composition of secretions.     

Wolbachia在熊蜂中的双重感染

Wolbachia是一类广泛存在于节肢动物体内细胞质遗传的细菌,它可以通过诱导产雌孤雌生殖、引起细胞质不亲和、遗传雄性的雌性化、雄性致死和增强生殖力等作用方式引起其寄主生殖行为的改变.本文以16S rDNA为标记检测了3种熊蜂不同组织(头,胸,足,卵巢或雄外生殖器)的Wolbachia感染.其中明亮熊蜂Bombus l...     

Selection of reference genes for real-time polymerase chain reaction analysis in tissues from Bombus terrestris and Bombus lucorum of different ages

Quantitative real-time polymerase chain reaction (PCR) is an accurate and sensitive technique for gene expression analysis. However, it requires data normalization using reference genes. Here we assessed the stability of eight reference genes in the labial gland and fat body of the bumblebees Bombus terrestris and Bombus lucorum of different ages. To date, no reference genes have been identified for these species. Our data show that arginine kinase (AK) and phospholipase A2 (PLA2) are the most stable genes in both tissues of B. terrestris. The most stable genes for the labial gland and fat body of B. lucorum were found to be elongation factor 1α (EEF1A) and PLA2.