Vitellogenin content in fat body and ovary homogenates of workers and queens of Melipona quadrifasciata anthidioides during vitellogenesis

Abstract Vitellogenin (Vg) is an egg yolk protein that is produced primarily in the fat body of most female insects. In the advanced social structure of eusocial honeybees, the presence of the queen inhibits egg maturation in the workers’ ovaries. However in the stingless bee Melipona quadrifasciata, the workers always develop ovaries and lay a certain amount of eggs while provisioning the brood cells with larval food during what is known as the worker nurse phase. The present work is a comparative study of the presence of Vg in homogenates of the fat bodies and ovaries of the nurse workers, and the virgin and physogastric queens of M. quadrifasciata. The presence of Vg was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting using Apis mellifera anti‐egg antibody. Vg was not detected in the fat bodies or ovaries of the workers, but it was found in the ovaries of virgin and physogastric queens and in the fat body of physogastric queens. The results are discussed, taking into account the reproductive state of the individuals and the other possible roles of Vg, such as a storage protein for metoabolism of other organs.     

Ecdysteroid titers in pupae of highly social bees relate to distinct modes of caste development

Modifications in endocrine programs are common mechanisms that generate alternative phenotypes. In order to understand how such changes may have evolved, we analyzed the pupal ecdysteroid titers in two closely related, highly social bees: the honey bee, Apis mellifera, and a stingless bee, Melipona quadrifasciata. In both species, the ecdysteroid titers in queens reached their peak levels earlier than in workers. Titer levels at peak maxima did not differ for the honey bee castes, but in Melipona they were twofold higher in queens than in workers. During the second half of pupal development, when the ecdysteroid titers decrease and the cuticle progressively melanizes, the titer in honey bee queens remained higher than in workers, while the reverse situation was observed in Melipona. Application of the juvenile hormone analog Pyriproxyfen^® to spinning-stage larvae of Melipona induced queen development. Endocrinologically this was manifest in a queen-like profile of the pupal ecdysteroid titer. Comparing these data with previous results on preimaginal hormone titers in another stingless bee, we conclude that the timing and height of the pupal ecdysteroid peak may depend on the nature of the specific stimuli that initially trigger diverging queen/worker development. In contrast, the interspecific differences in the late pupal ecdysteroid titer profiles mainly seem to be related to caste-specific programs in tissue differentiation, including cuticle pigmentation.     

Do army ant queens re-mate later in life?

Queens of eusocial Hymenoptera are inseminated only during a brief period before they start to lay eggs. This has probably been kin-selected because repeated insemination of old queens would normally be against the inclusive fitness interest of their daughter workers. Army ants have been considered to be the only possible exception to this rule due to their idiosyncratic life-history. We studied two distantly related species of army ants, the African Dorylus (Anomma) molestus and the Neotropical Eciton burchellii and present data from microsatellite genotyping, behavioural observations and sperm counts.We also describe the copulation behaviour of African army ants for the first time. Our results strongly suggest that, contradictory to earlier contentions, army ant queens do not mate repeatedly throughout their life and thus do not constitute an exception among the eusocial Hymenoptera in this respect. Sperm counts for males and queens of both species show that army ant queens have to mate with several males to become fully inseminated. However, sperm limitation by queens is unlikely to have been the prime reason for the evolution of high queen-mating frequencies in this group. Received 5 July 2006; revised 26 September 2006; accepted 11 October 2006.     

Conflict over male parentage in stingless bees

Summary Stingless bees usually have one, singly-mated queen. This can lead to a genetic conflict of interest between the queen and the non-laying workers over who should produce the males. In many stingless bee species workers have developed ovaries and can produce male-destined eggs. In this study we compile the available data on who produces the males in stingless bees. Worker reproduction is common but less frequent than expected from predictions built on relatedness-based preferences of non-laying workers. We tested whether the pattern in worker reproduction can be explained best by queen control, by an arms race between workers and their queen, by the costs of losing workers to reproductive competition, or by phylogenetic constraints. The data are consistent with the view that there is ongoing conflict over male production that is resolved differently depending on the specific dynamics of costs and benefits of worker reproduction. There was also a role for phylogeny; Melipona workers often reproduced while Plebeia and Australian stingless bee workers seldom or never did. The high worker reproduction in Melipona may reflect low costs, because many of the replaced queen-laid eggs would become excess queens.Received 17 April 2003; revised 2 September 2003; accepted 5 September 2003.     

Smelling like resin: terpenoids account for species-specific cuticular profiles in Southeast-Asian stingless bees

Insects may be unique in having a cuticle with a species-specific chemical profile. In social insects, colony survival depends not only on species-specific but also on colony-specific cuticular compounds with hydrocarbons playing an important role in the communication systems of ants, termites, wasps and bees. We investigated inter- and intraspecific differences in the composition of compounds found on the body surface of seven paleotropical stingless bee species (Apidae: Meliponini) at two different sites in Borneo (Sabah, Malaysia). Besides hydrocarbons, the body surface of all seven stingless bee species comprised terpenoid compounds, a substance class that has not been reported for chemical profiles of any social insect so far. Moreover, the chemical profile of some species differed fundamentally in the composition of terpenoids with one group (e.g. sesquiterpenes) being present in one species, but missing in another. Chemical profiles of different colonies from the same species showed the same hydrocarbon- and terpenoid compounds over different regions, as tested for Tetragonilla collina and Tetragonula melanocephala. However, chemical profiles differed quantitatively between the different colonies especially in T. melanocephala. It is likely that the terpenoids are derived from plant resins because stingless bees are known to collect and use large amounts of resins for nest construction and defence, suggesting an environmental origin of the terpenoids in the chemical profile of paleotropical stingless bees.     

Secretions of stingless bees: cephalic secretions of two Frieseomelitta species

Gas chromatographic and mass spectrometric analysis of the volatile compounds in the mandibular glands of Frieseomelitta varia and the heads of Frieseomelitta silvestrii have revealed relatively simple or more complex mixtures of volatile oxygenated compounds. 2-Alkanols were found to be important components for both species. In F. varia the composition was essentially the same in samples from two widely separated areas and there were small differences between callows and mature, foraging bees. The first electroantennographic studies on stingless bees, using hexane extracts of heads of both species, have demonstrated a response in the workers’ antennae to these, as well as to the pure compounds 2-heptanol and 2-nonanol .     

Ergonomics in stingless bees: changes in intranidal behavior after partial removal of storage pots and honey inMelipona favosa (Hym. Apidae,Meliponinae)

Summary The ergonomics of task allocation in stingless bees was examined in two laboratory colonies ofMelipona favosa containing individually marked workers. Performance of intranidal behavior patterns, brood cell production and the processing and storage of nectar, were studied during a control and a wax-deprived period. Experimental wax deprivation consisted of regular controlled removal of storage pots and their contents. Stored carbohydrate concentration and storage pot mass declined during the wax-deprived period. Behavior was measured by sampling for a complete ethogram at seven intranidal areas.In comparison withApis mellifera, M. favosa appeared to be more sensitive to stress, with a distinct reallocation of effort between tasks. The significant increase under stress of brood production and brood-rearing behavior patterns observed was previously also found inApis. Self-oriented behavior patterns declined under stressed circumstances.Involucrum construction declined in wax-deprivedMelipona colonies. Relatively low rates of walking behavior in general inMelipona suggest a low mean free distance between tasks. These results are of particular interest in relation to the basically different nest structure of stingless bees, e.g. very distinct areas for brood production and the processing and storage of food. This architecture of stingless bee nests appeared to influence strongly their specific responses to wax deprivation.     

Worker policing in the common wasp Vespula vulgaris is not aimed at improving colony hygiene

In insect societies, eggs laid by workers are frequently killed by other workers – a behaviour known as “worker policing”. The traditional explanation of worker policing is that it is a mechanism to resolve intracolony conflict, and maintain the reproductive monopoly of the queen. Recently, Pirk et al. (2004) proposed that worker policing instead is aimed at removing unviable worker-laid eggs and is ultimately just another example of hygienic behaviour. Here we test this hypothesis for the common wasp Vespula vulgaris, a species with highly effective worker policing. We show that worker-laid eggs from queenless colonies have a lower hatch rate (68%) than queen-laid eggs (82%). Analysis of egg laying rates of queens and workers, however, shows that the difference is not big enough to explain the apparent absence of adult worker-derived males in this species. Received 30 January 2006; revised 2 May 2006; accepted 5 May 2006.     

Several workers lay eggs in the same brood cell in queenless honey bee (Apis mellifera) colonies

Queenless honey bee (Apis mellifera) colonies are often characterized by the presence of multiple eggs in brood cells. This is surprising because only one egg can be reared to maturity per cell. Moreover, worker honey bees cannot produce many eggs per day. There are several reasons that could explain the presence of multiple eggs in single cells: a) workers cannot control how many eggs they release; in this case we would expect all eggs to be from the same mother; b) excess eggs could be provided as food for the first larva to hatch in the absence of adequate brood care and this would again result in all eggs in one cell sharing the same mother; c) the number of cells available for oviposition may be limiting, obliging workers to lay eggs in cells that already contain eggs, resulting in eggs of mixed maternity. Here we show that the majority of brood cells in queenless colonies contain eggs from multiple mothers. Therefore our results suggest that the presence of multiple eggs in brood cells arises from a limitation on the number of suitable cells available for oviposition. Received 29 September 2008; revised 21 November 2008; accepted 24 November 2008.     

How to fool a virgin: Artificial dealation triggers oviposition in virgin Lasius niger queens

Once inseminated, ant queens rapidly shed their wings and start to lay eggs. Here we test whether there is a causal link between dealation and oviposition in the ant Lasius niger. We show that artificially dealated virgin queens start to lay eggs shortly after wing removal, whereas winged virgin queens hardly ever lay eggs. Dealate virgins do, however, produce fewer eggs than mated queens. These findings indicate that dealation does induce egg-laying, and that other factors, such as mating and/or insemination, further stimulate oviposition under natural conditions. Received 2 January 2006; revised 8 February 2006; accepted 4 April 2006.     

Karyotype and Chromosomal Location of 18S–28S and 5S Ribosomal DNA in the Scallops Pecten maximus and Mimachlamys varia (Bivalvia: Pectinidae)

This work describes the karyotype and chromosomal location of the ribosomal DNA (rDNA) of Pecten maximus and Mimachlamys varia, two commercial scallop species from Europe. According to the chromosome centromeric index values found, the karyotype of P. maximus is composed of 1 metacentric, 2 metacentric–submetacentric, 1 telocentric–subtelocentric and 15 telocentric pairs, and that of M. varia of 4 metacentric, 2 subtelocentric–submetacentric, 9 subtelocentric, 3 subtelocentric–telocentric and 1 telocentric–subtelocentric pairs. In P. maximus, 18S-28S rDNA was located by FISH on a metacentric–submetacentric pair, and in M. varia on a subtelocentric–submetacentric pair using both silver staining and FISH. PCR amplification of the 5S rDNA unit yielded a single product of about 460 bp (P. maximus) and 450 bp (M. varia), that used as probe revealed a 5S rDNA site on a telocentric pair in P. maximus and a subtelocentric pair in M. varia. Two-color FISH or sequential silver staining of 5S rDNA-FISH-metaphases corroborated that the two gene families are located on different chromosomes in both species. A comparative analysis of the data allowed the inference of karyotypic relationships within scallops.     

Parasitism ofLygus lineolaris eggs onCoronilla varia,solanum tuberosum,and three host weeds in southeastern Québec

Parasitism ofLygus lineolaris (Palisot de Beauvois) eggs by three species ofMymaridae, Anaphes iole Girault,Erythmelus miridiphagus Dozier andPolynema pratensiphagum Walley and one species ofScelionidae, Telenomus sp., was investigated at Ste-Anne-de-Bellevue, Québec. The maximum level of field parasitism ofL. lineolaris eggs by individual species was 15.4, 53.8, 70.0 and 16,7%, respectively. Parasitoids were retrieved from eggs ofL. lineolaris inserted in stems and branches ofAmaranthus retroflexus L.,Chenopodium album L.,Coronilla varia L.,Rumex obtusifolius L. andSolanum tuberosum L. Amaranthus retroflexus andS. tuberosum sustained a large population ofL. lineolaris and egg parasitism was high enough to consider it as a potential control factor.      

Evidence that workers recognize unfertilized queen-laid eggs for male production in stingless bees

The behavioural variation in bee workers raises questions about the reproduction in eusocial bees, as well as possible triggers for ovarian activation and oviposition. However, there are no data on possible differences between queen-laid eggs found alone or together with worker-laid eggs in a brood cell. This study evaluated the ploidy of eggs from queens found together with worker-laid eggs in the brood cells of the stingless bee Scaptotrigona xanthotricha. In the brood cells with two eggs, there was a difference in size between them, with the smaller one being a similar size to those found in brood cells with only one egg. The frequency of diploid eggs in brood cells with one egg was 96.6%, whereas, in those with two or more, it was 39.3%. These results suggest that egg fertilization can be identified by workers, because when there is a worker-laid egg, those from the queen are often haploid, supporting a prediction from Hamilton's theory of kin selection.     

Subtle visits despite guards: Theft from nest of stingless bee (Meliponini) by orchid bee (Euglossini)

The theft of food items and building materials from nests of stingless bees (Hymenoptera: Apidae: Meliponini) is most commonly carried out by other Meliponini bee species, especially by those related to genus Lestrimelitta Friese, 1903. In this note, we recorded the theft of cerumen and propolis made by a species of the orchid bee, Euglossa annectans Dressler, 1982 (Apidae: Euglossini) from the nest of the stingless bee Tetragonisca angustula (Latreille, 1811). We noticed that the guard workers of T. angustula did not attack the thief bee even though it did not come sufficiently close to the entrance tube of the nest.     

Behavioral suites mediate group-level foraging dynamics in communities of tropical stingless bees

Competition for floral resources is a key force shaping pollinator communities, particularly among social bees. The ability of social bees to recruit nestmates for group foraging is hypothesized to be a major factor in their ability to dominate rich resources such as mass-flowering trees. We tested the role of group foraging in attaining dominance by stingless bees, eusocial tropical pollinators that exhibit high diversity in foraging strategies. We provide the first experimental evidence that meliponine group foraging strategies, large colony sizes and aggressive behavior form a suite of traits that enable colonies to improve dominance of rich resources. Using a diverse assemblage of Brazilian stingless bee species and an array of artificial “flowers” that provided a sucrose reward, we compared species’ dominance and visitation under unrestricted foraging conditions and with experimental removal of group-foraging species. Dominance does not vary with individual body size, but rather with foraging group size. Species that recruit larger numbers of nestmates (Scaptotrigona aff. depilis, Trigona hyalinata, Trigona spinipes) dominated both numerically (high local abundance) and behaviorally (controlling feeders). Removal of group-foraging species increased feeding opportunities for solitary foragers (Frieseomelitta varia, Melipona quadrifasciata and Nannotrigona testaceicornis). Trigona hyalinata always dominated under unrestricted conditions. When this species was removed, T. spinipes or S. aff. depilis controlled feeders and limited visitation by solitary-foraging species. Because bee foraging patterns determine plant pollination success, understanding the forces that shape these patterns is crucial to ensuring pollination of both crops and natural areas in the face of current pollinator declines.     

Isolation and characterization of ten microsatellite loci in stingless bee Trigona spinipes (Apidae: Meliponini)

Stingless bees are the most abundant pollinators of Brazilian tropical flora. Trigona spinipes has some of the largest colonies of any stingless bee species found in several types of environment. This work describes the isolation and characterization of microsatellite loci for this species. A microsatellite‐enriched genomic library was constructed and ten primer pairs were designed for T. spinipes. The primers were tested in 20 unrelated individuals. The mean number of alleles was 8.10 and mean observed and expected heterozygosity were 0.655 and 0.680, respectively. Primers were also tested in cross‐species amplification and five loci were successfully amplified in Trigona chanchamayoensis, Trigona hyalinata, Tetragonisca angustula, Partamona mulata and Frieseomelitta varia. The microsatellite primers described herein will be useful for evaluating genetic variability and gaining a better understanding of the population structure of T. spinipes as well as other species of stingless bees.     

Seed-coat microsculpturing and its systematic application in Isatis (Brassicaceae) and allied genera in Iran

In order to examine the systematic application of seed-coat microsculpturing in Isatis, seed surfaces of 23 species (41 populations) in four genera of tribe Isatideae were examined using scanning electron microscopy (SEM). Eight types of basic ornamentation patterns were recognized among the studied specimens. Of these, the reticulate–areolate type was the most common and was found in the genera Isatis, Pachypterygium, Samerari and Tauscheria and 15 species (e.g., I. cappodocica, I. kotschyana and I. tinctoria). The reticulate type, the second most frequent, occurred in 7 species while other types each were represented by only one or 2 species. Although different populations of a given species show similar seed-surface sculpturing in most cases, in some polymorphic species like I. cappadocica and I. kotschyana these patterns were variable among populations. To some extent the variation corresponds to infraspecific taxa for some species, but the differences are not significant enough to be useful in the delimitation of the subspecies recognized by previous workers. Moreover, seed-coat characters do not support the separation of genera Isatis, Pachypterygium, Sameraria and Tauscheria.     

Larval Sociality in Three Species of Central-place Foraging Lappet Moths (Lepidoptera: Lasiocampidae): A Comparative Survey

We studied three species of Lasiocampidae with social, tent-building caterpillars in Northern Bavaria, viz. Eriogaster lanestris, Eriogaster catax, and Malacosoma neustria. We used key life-history data (number of larval instars, sizes and weights of eggs, caterpillars, and moths, size of egg clutches) as well as behavioral data (activity patterns, tent-building behavior, trail following behavior) for a comparative study. Although larvae of all three species are active only in spring, show overlapping habitat requirements, and use the same major host-plant (Prunus spinosa) with only minor differences in phenology, they show markedly different life-history and behavioral strategies.E. catax lays comparatively few but large eggs while E. lanestris lays more but smaller eggs. M. neustria lays the smallest eggs but large clusters. E. lanestris caterpillars build a large tent with an accessible interior while those of E. catax build a small tent that is only used as a resting and molting platform. M. neustria shows a flexible behavior, may abandon the primary tent and build a new one several times. M. neustria colonies also subdivide and reunite regularly while Eriogaster colonies stay together until larvae become solitary. In E. lanestris all tentmates of a colony are highly synchronized while foraging or resting. Instead, in E. catax small subgroups leave the tent for foraging while at every time the majority rests on the tent. M. neustria caterpillars forage more or less individually and only synchronize by night. Results are discussed in relation to other species of the genera Eriogaster and Malacosoma and with regard to the evolution and diversification of caterpillar sociality.     

Suitability of pollen sources for the development and reproduction of Coleomegilla maculata (Coleoptera: Coccinellidae) under simulated drought conditions

Laboratory experiments compared the nutritive value of various pollen sources for the development of Coleomegilla maculata DeGeer under conditions of continuous water availability and simulated drought. When water was continuously available, larval survival was not different from 100% on diets of frozen eggs of Ephestia kuehniella Zeller, corn pollen, sorghum pollen, or pulverized bee pollen, whereas survival of larvae was significantly reduced on the latter three diets in the simulated drought treatment. Pollen of cultivated sunflower, Helianthus annus L., proved fatal to both larvae and adults; its surface structure caused clumping and accumulation on the insect cuticle that led to death from exhaustion/desiccation in petri dishes. The Ephestia egg diet yielded shorter developmental times and heavier adult weights than any pollen diet in both treatments. The drought treatment increased developmental time on all diets with a significant treatment–diet interaction. Drought reduced the adult weight of females on the sorghum pollen diet, and that of both sexes on the bee pollen diet, again with a significant treatment–diet interaction. Initial water content was highest in corn pollen (36.8%), followed by Ephestia eggs (29.2%), sorghum pollen (25.3%), sunflower pollen (8.7%), and bee pollen (4.6%), but did not appear correlated with C. maculata larval survival on pollen sources under drought conditions. Reproductive adult females that received corn or sorghum pollen as a supplement to Ephestia eggs did not differ in fecundity or fertility from those fed only Ephestia eggs.     

The ectoparasitic mite Tropilaelaps mercedesae (Acari, Laelapidae) as a vector of honeybee viruses

The ectoparasitic mites Varroa destructor and Tropilaelaps mercedesae share life history traits and both infect honeybee colonies, Apis mellifera. Since V. destructor is a biological vector of several honeybee viruses, we here test whether T. mercedesae can also be infected and enable virus replication. In Kunming (China), workers and T. mercedesae mites were sampled from three A. mellifera colonies, where workers were exhibiting clinical symptoms of deformed wing virus (DWV). We analysed a pooled bee sample (15 workers) and 29 mites for the presence of Deformed wing virus (DWV), Black queen cell virus (BQCV), Sacbrood virus (SBV), Kashmir bee virus (KBV), Acute bee paralysis virus (ABPV), and Chronic bee paralysis virus (CBPV). Virus positive samples were analysed with a qPCR. Only DWV +RNA was found but with a high titre of up to 10⁸ equivalent virus copies per mite and 10⁶ per bee. Moreover, in all DWV positive mites (N= 12) and in the bee sample virus–RNA was also detected using RT-PCR and tagged RT-PCR, strongly suggesting virus replication. Our data show for the first time that T. mercedesae may be a biological vector of DWV, which would open a novel route of virus spread in A. mellifera. Received 6 June 2008; revised 14 August 2008; accepted 10 September 2008.