Scaptotrigona Depilis Research Articles

The inheritance of alternative nest architectural traditions in stingless bees

The transmission of complex behavior and culture in humans has long been attributed to advanced forms of social learning,1,2 which play a crucial role in our technological advancement.3 While similar phenomena of behavioral traditions and cultural inheritance have been observed in animals,1,2,4,5,6 including in primates,7 whales,8 birds,9 and even insects,10 the underlying mechanisms enabling the persistence of such animal traditions, particularly in insects, are less well understood. This study introduces pioneering evidence of enduring architectural traditions in the stingless bee Scaptotrigona depilis, which are maintained without any evidence for social learning. We demonstrate that S.depilis exhibits two distinct nest architectures, comprising either helicoidal or flat, stacked horizontal combs, which are transmitted across generations through stigmergy11,12,13,14,15,16,17-an environmental feedback mechanism whereby the presence of the existing comb structures guides subsequent construction behaviors-thereby leading to a form of environmental inheritance.18,19,20 Cross-fostering experiments further show that genetic factors or prior experience does not drive the observed variation in nest architecture. Moreover, the experimental introduction of corkscrew dislocations within the combs prompted helicoidal building, confirming the use of stigmergic building rules. At a theoretical level, we establish that the long-term equilibrium of building in the helicoidal pattern fits with the expectations of a two-state Markov chain model. Overall, our findings provide compelling evidence for the persistence of behavioral traditions in an insect, based on a simple mechanism of environmental inheritance and stigmergic interactions, without requiring any sophisticated learning mechanism, thereby expanding our understanding of how traditions can be maintained in non-human species.

Antibacterial and Toxic Activity of Geopropolis Extracts from Melipona subnitida (Ducke, 1910) (Hymenoptera: Apidae) and Scaptotrigona depilis (Moure, 1942) (Hymenoptera: Apidae).

Bacteria are associated with many infections that affect humans and present antibiotic resistance mechanisms, causing problems in health organisations and increased mortality rates. Therefore, it is necessary to find new antibacterial agents that can be used in the treatment of these microorganisms. Geopropolis is a natural product from stingless bees, formed by a mixture of plant resins, salivary secretions, wax and soil particles, the chemical composition of this natural product is diverse. Thus, this study aimed to evaluate antibacterial activity, antibiotic modulation and the toxicity of geopropolis extracts from the stingless bees, Melipona subnitida (Ducke, 1910) and Scaptotrigona depilis (Moure, 1942) against standard and multi-resistant Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa bacteria. Geopropolis samples were collected in a meliponary located in Camaragibe, Pernambuco, Brazil. To determine the Minimum Inhibitory Concentration (MIC) and antibiotic modulation we performed broth microdilution tests. Mortality tests were used to verify extract toxicity in the model Drosophila melanogaster. The microbiological tests showing that the M. subnitida extracts had better inhibitory effects compared to S. depilis, presenting direct antibacterial activity against standard and multi-resistant strains. The extracts potentialized antibiotic effects, suggesting possible synergy and did not present toxicity in the model used. The information obtained in this study highlights extracts as promising antibacterial agents and is the first study to evaluate bacterial activity in these extracts, in addition to verifying their modulating effects and determining toxicity in the model used.

Further evidences of an emerging stingless bee-yeast symbiosis.

Symbiotic interactions between microorganisms and social insects have been described as crucial for the maintenance of these multitrophic systems, as observed for the stingless bee Scaptotrigona depilis and the yeast Zygosaccharomyces sp. SDBC30G1. The larvae of S. depilis ingest fungal filaments of Zygosaccharomyces sp. SDBC30G1 to obtain ergosterol, which is the precursor for the biosynthesis of ecdysteroids that modulate insect metamorphosis. In this work, we find a similar insect-microbe interaction in other species of stingless bees. We analyzed brood cell samples from 19 species of stingless bees collected in Brazil. The osmophilic yeast Zygosaccharomyces spp. was isolated from eight bee species, namely Scaptotrigona bipunctata, S. postica, S. tubiba, Tetragona clavipes, Melipona quadrifasciata, M. fasciculata, M. bicolor, and Partamona helleri. These yeasts form pseudohyphae and also accumulate ergosterol in lipid droplets, similar to the pattern observed for S. depilis. The phylogenetic analyses including various Zygosaccharomyces revealed that strains isolated from the brood cells formed a branch separated from the previously described Zygosaccharomyces species, suggesting that they are new species of this genus and reinforcing the symbiotic interaction with the host insects.

Laboratory Risk Assessment of Three Entomopathogenic Fungi Used for Pest Control toward Social Bee Pollinators.

The use of fungal-based biopesticides to reduce pest damage and protect crop quality is often considered a low-risk control strategy. Nevertheless, risk assessment of mycopesticides is still needed since pests and beneficial insects, such as pollinators, co-exist in the same agroecosystem where mass use of this strategy occurs. In this context, we evaluated the effect of five concentrations of three commercial entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae, and Cordyceps fumosorosea, by direct contact and ingestion, on the tropical stingless bees Scaptotrigona depilis and Tetragonisca angustula, temperate bee species, the honey bee Apis mellifera, and the bumble bee Bombus terrestris, at the individual level. Furthermore, we studied the potential of two infection routes, either by direct contact or ingestion. In general, all three fungi caused considerable mortalities in the four bee species, which differed in their response to the different fungal species. Scaptotrigona depilis and B. terrestris were more susceptible to B. bassiana than the other fungi when exposed topically, and B. terrestris and A. mellifera were more susceptible to M. anisopliae when exposed orally. Interestingly, increased positive concentration responses were not observed for all fungal species and application methods. For example, B. terrestris mortalities were similar at the lowest and highest fungal concentrations for both exposure methods. This study demonstrates that under laboratory conditions, the three fungal species can potentially reduce the survival of social bees at the individual level. However, further colony and field studies are needed to elucidate the susceptibility of these fungi towards social bees to fully assess the ecological risks.

Molecular network‐guided chemical profile and mass spectrometry, volatile compounds, and antimicrobial activity of Scaptotrigona depilis propolis

RationalePropolis has a great diversity in its composition due to numerous factors; therefore, each study is an important contribution to the knoFwledge of its composition and biological action. The objective of this study was to determine the chemical profile and biological activity of propolis produced by Scaptotrigona depilis.MethodsExtracts with 70% ethanol (EPE70) and with cereal alcohol (CAPE) were elaborated, and then characterized using UHPLC‐ESI(+)‐MS/MS. Volatile compounds were extracted and then characterized using gas chromatography mass spectrometry (GC–MS). In addition, antimicrobial activities were verified against resistant strains.ResultsThe volatile compounds of propolis predominantly consist of sesquiterpenes. Using the exploratory metabolomic approach, compounds of different classes were putatively identified in the ethanolic extracts, of which the most representative were terpenes, and some of the sesquiterpenes identified among the volatiles were also detected. The extracts were shown to be active against Escherichia coli and Staphylococcus aureus bacteria with a minimum inhibitory concentration (MIC) of 0.5 and 1.0 mg mL−1, respectively.ConclusionsThe molecular network approach proved to be determining the chemical profile of S. depilis propolis rapidly and accurately, and led to the identification of lipophilic compounds. The identification of compounds using GC–MS and UHPLC‐ESI(+)‐MS/MS is complementary and useful for the characterization of propolis.

Bioactive compounds and antiradical activity of propolis from Scaptotrigona bipunctata and Scaptotrigona depilis

Propolis is a viscous substance produced and collected from plant shoots or exudates by worker bees. This study aimed to identify phenolic compounds, carotenoids, volatile compounds, and antiradical activity of propolis obtained from two stingless bees (SB), Scaptotrigona bipunctata (Tubuna) and Scaptotrigona depilis (Canudo), all native from Rio Grande do Sul. The two propolis showed a similar qualitative composition, but with quantitative differences. Catechin was the major phenolic compound found in propolis from S. bipuncata and S. depilis. The propolis from S. depilis showed a higher concentration of carotenoids, mainly lutein. The α-pinene was the volatile compound found in greater concentration in the two propolis.

Evidences of High Genetic Differentiation among Populations of the Stingless Bee Scaptotrigona depilis (Moure, 1942) in Piauí, Brazil

Bees of the Meliponini tribe (Hymenoptera, Apidae), known as stingless bees, comprise one of the most important groups of pollinators of the tropical flora (Michener, 2007). Its ecological importan...

Pollen resources partitioning of stingless bees (Hymenoptera: Apidae) from the southern Atlantic forest

Meliponinae eusocial bees are among the most important pollinators in the Neotropics and their beekeeping has been growing as both recreational and economic activity. Information on the pollen preferences and niche overlap among species coexisting in the same location is still scarce. This study focused on the use of pollen resources by three native species of Meliponinae in the Atlantic Forest, Melipona quadrifasciata quadrifasciata (Lepeletier 1836), Scaptotrigona depilis (Moure 1942), and Tetragonisca angustula (Latreille 1811), at two different sites. The use of pollen resources was evaluated from reserves inside hives collected monthly between February 2015 and January 2016. It was also estimated the Mean Minimal Distance (Dm) traveled by the bees to obtain pollen. The bees differed regarding the diversity of pollen types, Dm, and proportion of pollen from different plant life-form. Local factor promoted differences only for Dm. M. q. quadrifasciata foraged a low diversity of pollen species and traveled greater distances with a higher proportion of trees plus palm species. Tetragonisca angustula foraged a higher diversity of pollen and flew shorter distances with a lower proportion of trees and palms. The species factor predominates over the local factor in the clustering pattern, based on pollen profile similarity.

Queen Execution, Diploid Males, and Selection For and Against Polyandry in the Brazilian Stingless Bee Scaptotrigona depilis.

Female mating frequency varies. Determining the causes of this variation is an active research area. We tested the hypothesis that in stingless bees, Meliponini, single mating is due to the execution of queens that make a matched mating at the complementary sex determination locus and have diploid male offspring. We studied the Brazilian species Scaptotrigona depilis. We made up 70 test colonies so that 50% (single matched mating), 25% (double mating), 12.5% (quadruple mating), or 0% (single nonmatched mating) of the emerging brood were diploid males. Queen execution following diploid male emergence was equal and high in colonies producing 50% (77% executed) and 25% (75%) diploid males versus equal and low in colonies producing 12.5% (7%) and 0% (0%) diploid males. These results show that queens that mate with two males with similar paternity suffer an increased chance of being executed, which selects against double mating. However, double mating with unequal paternity (e.g., 25∶75), which occasionally occurs in S. depilis, is selectively neutral. Single mating and double mating with unequal paternity form one adaptive peak. The results show a second adaptive peak at quadruple mating. However, this is inaccessible via gradual evolutionary change in a selective landscape with reduced fitness at double mating.

Chemical Diversity in a Stingless Bee-Plant Symbiosis.

Bees are essential pollinators on earth, supporting forest equilibrium and human agriculture. The chemistry of the stingless bee–plant symbiosis is a complex and not completely understood phenomenon. Here, we combined untargeted tandem mass spectrometry, molecular networking, and multivariate statistical analysis to investigate the chemical diversity in colonies of the stingless bee Scaptotrigona depilis. Flavonoids were the most representative and diverse group of plant metabolites detected, indicating the importance of these biologically active natural products to the bees. We unveiled the metabolome, mapped the distribution of plant metabolites in stingless bee colonies, and digitized the chemical data into a public database.

Microbial community modulates growth of symbiotic fungus required for stingless bee metamorphosis

The Brazilian stingless bee Scaptotrigona depilis requires the brood cells-associated fungus Zygosaccharomyces sp. as steroid source for metamorphosis. Besides the presence of Zygosaccharomyces sp., other fungi inhabit S. depilis brood cells, but their biological functions are unknown. Here we show that Candida sp. and Monascus ruber, isolated from cerumen of S. depilis brood provisions, interact with Zygosaccharomyces sp. and modulate its growth. Candida sp. produces volatile organic compounds (VOCs) that stimulate Zygosacchromyces sp. development. Monascus ruber inhibits Zygosacchromyces sp. growth by producing lovastatin, which blocks steroid biosynthesis. We also observed that in co-cultures M. ruber inhibits Candida sp. through the production of monascin. The modulation of Zygosaccharomyces sp. growth by brood cell-associated fungi suggests their involvement in S. depilis larval development. This tripartite fungal community opens new perspectives in the research of microbial interactions with bees.

Unique nest entrance structure of Partamona helleri stingless bees leads to remarkable \u2018crash-landing\u2019 behaviour

Partamona helleri stingless bees construct a unique, funnel-shaped entrance that resolves an evolutionary conflict between foraging efficiency and defence. The large outer entrance allows many foragers to pass while the narrow inner entrance requires few guards to defend. This structure has given rise to a remarkable behaviour in returning foragers, which appear to approach the nest entrance at high speed and ‘crash’ head first into the entrance. We compared P. helleri landing behaviour with two related species with architecturally different entrances that land conventionally using their legs: Melipona scutellaris, whose narrow entrance allows only a single bee to pass, and Scaptotrigona depilis, which has a wide entrance tube. All three species initially decelerated on their approach to the nest entrance. However, 0.2 m from the entrance P. helleri began accelerating, whereas the other species continued to decelerate. Partamona helleri entered its nest at 1.14 ms−1, double the velocity of the other species. Despite its greater velocity, P. helleri made no fewer errors than the other species when attempting to enter its nest, probably by virtue of the large target provided by the outer entrance. We then used a bioassay that suggests that this behaviour is a defence against ambush predators at the nest entrance. Finally, we use a scaling argument to show that the crash impact should not cause any damage to a small animal such as a bee, such that no morphological adaptation is required.

Quality of Meliponinae honey: Pesticides residues, pollen identity, and microbiological profiles

AbstractThis study aims at evaluating the quality of honey of Melipona quadrifasciata quadrifasciata Tetragonisca angustula and Scaptotrigona depilis. Pollen analysis was performed to identify the floral resources used for the production of honey, and 19 pollen types were identified. Microbiological analyses identified the presence of coliforms, molds, and yeasts, but Clostridium botulinum was not found. Using the modified “Quick, Easy, Cheap, Effective, Rugged, and Safe” method followed by ultra‐high performance liquid chromatography‐tandem mass spectrometry determination, eight pesticides were detected in the honey of T. angustula and only two in the honey from S. depilis. The honey of M. q. quadrifasciata did not present pesticide residues. The data pointed out the relevance of good practices to avoid the contamination of honey with sources of microbiological contaminants, which may impair the product's safety for human consumption.

Stingless Bee Larvae Require Fungal Steroid to Pupate

The larval stage of the stingless bee Scaptotrigona depilis must consume a specific brood cell fungus in order to continue development. Here we show that this fungus is a member of the genus Zygosaccharomyces and provides essential steroid precursors to the developing bee. Insect pupation requires ecdysteroid hormones, and as insects cannot synthesize sterols de novo, they must obtain steroids in their diet. Larval in vitro culturing assays demonstrated that consuming ergosterol recapitulates the developmental effects on S. depilis as ingestion of Zygosaccharomyces sp. cells. Thus, we determined the molecular underpinning of this intimate mutualistic symbiosis. Phylogenetic analyses showed that similar cases of bee-Zygosaccharomyces symbiosis may exist. This unprecedented case of bee-fungus symbiosis driven by steroid requirement brings new perspectives regarding pollinator-microbiota interaction and preservation.

Diploid Male Production Results in Queen Death in the Stingless Bee Scaptotrigona depilis.

As in most Hymenoptera, the eusocial stingless bees (Meliponini) have a complementary sex determination (CSD) system. When a queen makes a "matched mating" with a male that shares a CSD allele with her, half of their diploid offspring are diploid males rather than females. Matched mating imposes a cost, since diploid male production reduces the colony workforce. Hence, adaptations preventing the occurrence or attenuating its effects are likely to arise. Here we provide clear evidence that in the stingless bee Scaptotrigona depilis, the emergence of diploid males induces queen death, and this usually occurs within 10-20days of the emergence of diploid male offspring from their pupae. Queens that have not made a matched mating die when introduced into a colony in which diploid males are emerging. This shows that the adult diploid males, and not the queen that has made a matched mating herself, are the proximate cause of queen death. Analysis of the cuticular hydrocarbon profiles of adult haploid and diploid males shows six compounds with significant differences. Moreover, the diploid and haploid males only acquire distinct cuticular hydrocarbon profiles 10days after emergence. Our data shows that the timing of queen death occurs when the cuticular hydrocarbons of haploid and diploid males differ significantly, suggesting that these chemical differences could be used as cues or signals to trigger queen death.

Hygienic behaviour in Brazilian stingless bees.

ABSTRACTSocial insects have many defence mechanisms against pests and pathogens. One of these is hygienic behaviour, which has been studied in detail in the honey bee, Apis mellifera. Hygienic honey bee workers remove dead and diseased larvae and pupae from sealed brood cells, thereby reducing disease transfer within the colony. Stingless bees, Meliponini, also rear broods in sealed cells. We investigated hygienic behaviour in three species of Brazilian stingless bees (Melipona scutellaris, Scaptotrigona depilis, Tetragonisca angustula) in response to freeze-killed brood. All three species had high mean levels of freeze-killed brood removal after 48 h ∼99% in M. scutellaris, 80% in S. depilis and 62% in T. angustula (N=8 colonies per species; three trials per colony). These levels are greater than in unselected honey bee populations, ∼46%. In S. depilis there was also considerable intercolony variation, ranging from 27% to 100% removal after 2 days. Interestingly, in the S. depilis colony with the slowest removal of freeze-killed brood, 15% of the adult bees emerging from their cells had shrivelled wings indicating a disease or disorder, which is as yet unidentified. Although the gross symptoms resembled the effects of deformed wing virus in the honey bee, this virus was not detected in the samples. When brood comb from the diseased colony was introduced to the other S. depilis colonies, there was a significant negative correlation between freeze-killed brood removal and the emergence of deformed worker bees (P=0.001), and a positive correlation with the cleaning out of brood cells (P=0.0008). This shows that the more hygienic colonies were detecting and removing unhealthy brood prior to adult emergence. Our results indicate that hygienic behaviour may play an important role in colony health in stingless bees. The low levels of disease normally seen in stingless bees may be because they have effective mechanisms of disease management, not because they lack diseases.

Whole-Genome Sequence of Bacillus sp. SDLI1, Isolated from the Social Bee Scaptotrigona depilis.

We announce the complete genome sequence of Bacillus sp. strain SDLI1, isolated from larval gut of the stingless bee Scaptotrigona depilis. The 4.13-Mb circular chromosome harbors biosynthetic gene clusters for the production of antimicrobial compounds.

The origin and evolution of queen and fertility signals in Corbiculate bees.

BackgroundIn social Hymenoptera (ants, bees and wasps), various chemical compounds present on the cuticle have been shown to act as fertility signals. In addition, specific queen-characteristic hydrocarbons have been implicated as sterility-inducing queen signals in ants, wasps and bumblebees. In Corbiculate bees, however, the chemical nature of queen-characteristic and fertility-linked compounds appears to be more diverse than in ants and wasps. Moreover, it remains unknown how queen signals evolved across this group and how they might have been co-opted from fertility signals in solitary ancestors.ResultsHere, we perform a phylogenetic analysis of fertility-linked compounds across 16 species of solitary and eusocial bee species, comprising both literature data as well as new primary data from a key solitary outgroup species, the oil-collecting bee Centris analis, and the highly eusocial stingless bee Scaptotrigona depilis. Our results demonstrate the presence of fertility-linked compounds belonging to 12 different chemical classes. In addition, we find that some classes of compounds (linear and branched alkanes, alkenes, esters and fatty acids) were already present as fertility-linked signals in the solitary ancestors of Corbiculate bees, while others appear to be specific to certain species.ConclusionOverall, our results suggest that queen signals in Corbiculate bees are likely derived from ancestral fertility-linked compounds present in solitary bees that lacked reproductive castes. These original fertility-linked cues or signals could have been produced either as a by-product of ovarian activation or could have served other communicative purposes, such as in mate recognition or the regulation of egg-laying.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-015-0509-8) contains supplementary material, which is available to authorized users.

A Brazilian Social Bee Must Cultivate Fungus to Survive

The nests of social insects provide suitable microenvironments for many microorganisms as they offer stable environmental conditions and a rich source of food [1-4]. Microorganisms in turn may provide several benefits to their hosts, such as nutrients and protection against pathogens [1, 4-6]. Several examples of symbiosis between social insects and microorganisms have been found in ants and termites. These symbioses have driven the evolution of complex behaviors and nest structures associated with the culturing of the symbiotic microorganisms [5, 7, 8]. However, while much is known about these relationships in many species of ants and termites, symbiotic relationships between microorganisms and social bees have been poorly explored [3,4, 9, 10]. Here, we report the first case of an obligatory relationship between the Brazilian stingless bee Scaptotrigona depilis and a fungus of the genus Monascus (Ascomycotina). Fungal mycelia growing on the provisioned food inside the brood cell are eaten by the larva. Larvae reared invitro on sterilized larval food supplemented with fungal mycelia had a much higher survival rate (76%) compared to larvae reared under identical conditions but without fungal mycelia (8% survival). The fungus was found to originate from the material from which the brood cells are made. Since the bees recycle and transport this material between nests, fungus would be transferred to newly built cells and also to newly founded nests. This is the first report of a fungus cultivation mutualism in a social bee.

Co-culture between insect-associated microorganisms stimulates polyketide production

Social insects have been associated with microorganisms that can produce antimicrobial compounds in order to protect their colonies against pathogens. Natural products biosynthesis can be stimulated using co-culture techniques, once in natural conditions microorganisms are competing for nutrients and space. Monascus ruber SdFCP01 and Candida sp. SdFCP02, both isolated from brood cells of the Brazilian stingless bee Scaptotrigona depilis, were co-cultured and the production of the polyketide monascuspiloin by M. ruber was elicited. The ecological roles that M. ruber and Candida sp. play in S. depilis colony have been investigated.