The genomes sequenced for the neotropical stingless beesScaptotrigona bipunctata and S. depilisstrengthen the phylogenomics support for the taxonomy of socialbees

Bees

Different Nitidulid beetles can be found in social bee colonies (Ellis et al. 2008) andmay range from harmless associates (e.g.,Cychramus luteus , Neumann andRitter 2004) to damaging parasites (e.g., Aethina tumida , Neumann and Elzen 2004). Haptoncus luteolus (Coleoptera: Nitidulidae, Figure 1a, b) is a fruit pest (Yunus andHo 1980; Audisio et al. 1990) and has been reported as an associate of honeybee, Apis mellifera , colonies (Atkinson and Ellis 2011a, b). Here, we report for the first time its association with colonies of multiple stingless bee species. In July 2013, beetle larvae (N =6) were manually collected using entomological forceps from a thriving colony of the local stingless bee Trigona thoracica in Kelantan, Malaysia. The larvae were only found near and in multiple-punctured pollen pots in a bottom corner (Figure 1c). The pollen pots looked distorted oval and collapsed, which contrasts to the normal elongated oval shape in this stingless bee species (Michener 2013). No adult beetles were observed. In December 2013, adult beetles (N =21) were manually collected using entomological forceps from a thriving Heterotrigona itama , another local stingless bee. These beetles were hiding in the corners or under the brood comb of the colony. No beetle larvae was found. In January 2014, adult beetles (N >100), larvae (>100), and pupae (N >100) were manually collected using entomological forceps from the colony of a different local stingless bee, Tetragonula laeviceps . Several brood, honey, and pollen pots were infested by the larvae and adults (see Michener 2013 for respective details on the nesting biology of stingless bees). Interestingly, we observed distinct bulges on the surface of the nesting material. Careful dissections revealed that those apparently served as pupation sites (Figure 1d). This heavily infested T. laeviceps colony collapsed. All beetle samples were transported to the Biology Laboratory, Universiti Malaysia Kelantan. The adults were identified as H. luteolus (Coleoptera: Nitidulidae) [synonym—Epuraea luteola ] based on definitive morphological characteristics (see Gillogly 1982; Figure 1a). Then, a successful standard laboratoryrearing program for small hive beetles was established using plastic containers [18×12×6 cm, with a pin-holed lid (O=~1 mm), allowing for sufficient air circulation] and a diet consisting of pollen/honey/protein powder (Nutrilite®, Amway) in a 1:1:2 volume ratio (Neumann et al. 2013), thereby confirming that H. luteolus is able to successfully reproduce on bee products. Our observations show that H. luteolus can be associated with colonies of at least three different species of stingless bees. Taken together with earlier reports in honeybee, A. mellifera , colonies (Atkinson and Ellis 2011a, b), this suggests that this beetle species can exploit a variety of different social bee species. This is similar to the small hive beetle, which can infest colonies of honeybees (Neumann and Elzen 2004), bumblebees Bombus impatiens (Spiewok and Neumann 2006), Corresponding author: K. Krishnan, kumara_k2001@yahoo.com Manuscript editor: James Nieh Apidologie (2015) 46:262–264 Scientific note

Glyphosate, one of the most widely used herbicides globally, has been extensively applied in agriculture due to its efficacy in weed control. However, recent studies have raised concerns about its sublethal effects on non-target organisms, particularly social bee species such as honeybees (Apis mellifera), bumblebees (Bombus spp.), and stingless bees (e.g., Melipona, Trigona). While glyphosate's primary mechanism targets the shikimate pathway, which is absent in animals, emerging evidence suggests it can indirectly impact bees by altering their gut microbiota, immune responses, and behavior. Research shows that even at sublethal doses, glyphosate can impair navigation, learning, and foraging efficiency, leading to reduced colony growth and survival rates. Field and laboratory studies indicate that the impact is exacerbated when bees are exposed to formulated products containing surfactants, which increase glyphosate’s toxicity. Furthermore, the disruption of social behaviors within colonies, such as communication through the waggle dance in honeybees, has profound implications for hive health and productivity. Despite growing evidence, there remain significant gaps in our understanding of glyphosate’s long-term and chronic effects, especially across diverse ecosystems and bee species. Current research is limited by a lack of longitudinal field studies that assess the cumulative impact of low-dose exposure over multiple generations. Most studies have focused on honeybees, with less attention given to wild and native bee populations, which may respond differently to glyphosate. To address these challenges, future research must prioritize mechanistic studies, explore eco-friendly alternatives to glyphosate, and implement integrated pest management strategies to reduce agrochemical dependence. Collaboration among scientists, policymakers, and stakeholders is critical to developing evidence-based regulations that protect pollinator health. Given the essential role bees play in global food security through pollination, protecting these vital species from the sublethal effects of glyphosate is not only an ecological imperative but also a socioeconomic necessity. Immediate actions in research, policy reform, and sustainable agricultural practices are needed to mitigate the risks posed by glyphosate and safeguard the future of pollinators.

Abstract. Ibrahim YS, Rosazan MN, Mamat MII, Anuar ST, Azmi WA. 2025. Detection of microplastics in honey of stingless bee (Heterotrigona itama) and honey bee (Apis mellifera) from Malaysia. Biodiversitas 26: 1271-1278. The demand for stingless bee honey and European bee honey has increased rapidly due to its medicinal benefits. Honey of the Indo-Malaya stingless bee, Heterotrigona itama, and European honey bee, Apis mellifera, are among the most popular bee products that Malaysians commonly consume. It has been reported that the contamination of honey with microplastics (MPs) can occur at various stages of production, from bees collecting the contaminated floral sources to the harvesting and packaging processes. With the emerging plastics pollution in the environment and concerns about potential health risks, this study aims to investigate the presence of MPs in honey samples from stingless bees, H. itama, and honey bees, A. mellifera, from Malaysia. Microplastic particles were extracted from 10 g of honey and characterized under a stereomicroscope to determine their color, size, and type. Polymer types were further identified using FTIR analysis. All honey samples from both species were found to be 100% contaminated with microplastics. H. itama honey contained a higher concentration of microplastics (8.18 ± 2.57 MPs/g) compared to A. mellifera's honey (5.52 ± 1.13 MPs/g). The MPs found in honey from both species were predominantly fibers and fragments, mostly transparent in color, with sizes ranging from 0.7 to 1.8 mm. The findings of this preliminary study are intended to provide an awareness of MPs in honey, especially in the food safety aspect, which needs a better understanding of good practices of beekeeping and processing procedures to minimize the contamination of honey.

Bees can be exposed to pesticides when visiting crops or plants in adjacent areas affected by spray drift. Among pesticide categories, fungicides tend to be considered relatively safe, though they also can negatively affect pollinators. Most evidence of damage by fungicides to bees comes from laboratory tests; there is little information concerning contamination levels in the field. We examined exposure of honey bees (Apis mellifera L.) (Hymenoptera: Apidae) and a common Brazilian native species of social stingless bees (Scaptotrigona postica Latreille; Hymenoptera: Apidae), which is about a third the size of a honey bee, to a commercial fungicide (Fox Xpro), with three active ingredients (trifloxystrobin, bixafen, and prothioconazole), applied to crops they often visit according to label directions. A spraying apparatus mounted on tracks in a laboratory spray room was used to simulate field conditions. Soybean and cotton plants grown in pots were transferred to the spray room when the plants were in flower. Anaesthetized bees were attached with insect pins at the top and middle of the plants, on leaves and flowers. The fungicide was applied using fine or coarse droplets. The amounts of the individual active ingredients absorbed by bees were then quantified. Concentrations of trifloxystrobin were highest in both honey bees and stingless bees, followed by bixafen, and then prothioconazole, which was detected in the bees at much lower levels. Overall, bees at the top of the plants and those sprayed with fine droplets absorbed more pesticide. As a function of body mass, the stingless bees were more heavily contaminated than the larger honey bees. Tests using spraying systems that simulate field conditions can better estimate the actual doses that contaminate bees to help determine the impact of fungicides and other pesticides applied to crops.

Why do some social insects have sophisticated recruitment systems, while other species do not communicate about food source locations at all? To answer this question, it is necessary to identify the social or ecological factors that make recruitment adaptive and thus likely to evolve. We developed an individual-based model of honey bee foraging to quantify the benefits of recruitment under different spatial distributions of nondepleting resource patches and with different colony sizes. Benefits of recruitment were strongly dependent on resource patch quality, density, and variability. Communication was especially beneficial if patches were poor, few, and variable. A sensitivity analysis of the model showed that under conditions of high resource density recruitment could even become detrimental, especially if foraging duration was short, tendency to scout was high, or recruits needed a long time to find communicated locations. Colony size, a factor often suspected to influence recruitment evolution, had no significant effect. These results may explain the recent experimental findings that in honey bees, benefits of waggle dance recruitment seem to vary seasonally and with habitat. They may also explain why some, but not other, species of social bees have evolved a strategy to communicate food locations to nest mates. Key words: Apis mellifera, communication, foraging, individual-based model, social insects, waggle dance. [Behav Ecol]

Since the composition of honey varies with the species of bee as well as flowering and geographical aspects, this study aimed to evaluate the physicochemical and bioactive properties of Apisand stingless bees’honey from the Brazilian Caatinga. Samples of different species of Apis mellifera L.Meliponini (Melipona subnitida, Frieseomellita varia, Melipona mandacaia, Plebeia sp.) and Apis mellifera L.werecollected from honey producersin the state of Rio Grande do Norte. Honey from A. mellifera and stingless bees showed physicochemical differences in some parameters, especially in moisture, free acidity, HMF, water activity, sugars and electric conductivity. There were no differences in color between honeys from A. mellifera and stingless bees. Honeys fromPlebeia sp., F. varia and A. mellifera showed higher antioxidant capacity followed by honeys fromM. mandacaia and M. subnitida. Flavonoids had little influence on the differentiation of antioxidant activities of stingless bees, while the opposite occurred with the phenolic content, where honeys with the highest levels of phenolic also showed higher antioxidant capacity.

Although the importance of bees as the pollinators responsible for maintaining gene flow for many native and cultivated plants in ecosystems around the world is recognized, much of their biodiversity and behavior remains to be discovered. Stingless bees are considered key pollinators for several plant species in tropical and subtropical ecosystems and they also provide pollination services for economically important agricultural crops. Many countries are using the honey bee (Apis mellifera Linnaeus, 1758, Hymenoptera: Apidae) as a surrogate to evaluate the risk of pesticides to all species of bees. However, there is uncertainty regarding the extent to which honey bees can serve as surrogates for non-Apis bee species in the risk assessment for pesticides. This paper provides a short overview of the life history traits relevant in risk assessment of stingless bees. It summarizes what is known about stingless bee exposure to pesticides compared to that of honey bees and presents criteria for potential candidate species from Brazil for use in pesticide risk assessment in tropical environments. This paper also identifies gaps in knowledge of bee biology and pesticide exposure routes not covered by the current honey bee exposure assessment paradigm. Based on these gaps, research is needed on life history traits, estimates of nectar and pollen consumption, mud, resin, and water collection and available protocols to adequately assess toxic effects of pesticides to stingless bees. This review is part of a series of papers on the risk of exposure of non-Apis bees to pesticides.

Pollen loads gathered by honeybees and pot-pollen stored by stingless bees are functional foods with commercial potential. The botanical origin influences the physicochemical composition of the corbicular pollen, Apis mellifera’s bee bread, and the stingless bees’ pot-pollen processed in their nests. Thus, it is vital to determine the factors affecting the botanical composition of these apicultural products. As no prior research on the subject has been conducted, this study examines seasonality and bee species’ impact on the phytogeographic source of honeybee pollen loads and stingless bees’ pot-pollen through the use of palynological analysis, Network Analysis, and Generalized Linear Models (GLM). Due to seasonal variations and abiotic factors, Melipona quadrifasciata quadrifasciata, Melipona marginata, and Scaptotrigona bipunctata stingless bees produced monofloral pot-pollen from plant species within the Myrtaceae, Euphorbiaceae, and Fabaceae families. In contrast, A. mellifera and Tetragona clavipes bees exhibited a more generalist pollen foraging behavior, resulting in heterofloral pollen loads or pot-pollen in most seasons studied. The GLM results indicate that factors including ‘bee species’ and ‘season’ impacted the volume and varieties of pollen gathered by honeybees and stingless bees. It is vital to acknowledge the production area’s characteristics, such as climatic conditions, bee-friendly plants, flower types and quantity, and the behavioral patterns of the bee species regarding pollen foraging. These results highlight the significance of establishing best practices for producing monofloral and heterofloral pollen loads, or pot-pollen. This information may be utilized for commercial applications of A. mellifera pollen loads and stingless bee pot-pollen.

There is historical traditional use of stingless bee honey (SBH). However, no information concerning composition and antimicrobial properties of SBHs from Sudan was reported. Thus, the paper was aimed to focus on the components of the nests and honey characteristics of three species of stingless bee. Proximate analysis showed similar composition, except protein and ash were significantly (p > 0.01) different. Meliplebeia beccarii honey contained more protein 5.31% and slightly more ash 1.01% than the other species. Antibacterial activity was determined by cup-plate agar diffusion; Hypotrigona squamuligera and M. beccarii were more active against Staphylococcus aureus. None of the Studied SBHs showed activity against Escherichia coli, Apergillus niger, and Candida albicans. Anti-parasite activity as determined by sub-culture method showed that “almost all” SBHs have similar activities against trophozoites of Giardia lamblia and Entamoeba histolytica with statistically significant differences when compared to the untreated control. The amount of honey produced by M. beccarii varied from 100 to 150 ml per nest while honey produced by Hypotrigona ruspolii and H. squamuligera varied from 500 to 1000 ml/colony/year. Results also showed different nest architecture and components (entrance, brood areas, honey pots, and pollen pots) as indication for existence of many taxa of Meliponini in Sudan.

Summary To know basic information about the stingless bee, Trigona minangkabau , and the European honey bee, Apis mellifera , as pollinator of strawberry, we set three greenhouse areas: the honey bee introduced area, the stingless bee introduced area and the control area. Foraging and pollination efficiencies of the two bee species were studied comparatively. During the experimental period (10 days), the stingless bee foraged well and the nest weight did not change, though the honey bee often foraged inefficiently and the nest weight decreased by 2 kg. The average nectar volume of a flower was lower in the honey bee area (0.02 μl) and nearly the same in the other two areas (0.1 μl). We make a numerical model to describe pollination and fertilization process. This model shows that one visit of the honey bee pollinated 11% of achenes and one visit of the stingless bee did 4.7% on average and that 11 visits of the honey bee or 30 visits of the stingless bee are required per flower to attain normal berry (fertilization rate, 87%). In this study, the rate of deformed berries in the stingless bee area (73%) was lower than that of the control area (90%), but higher than that of the honey bee area (51%). From our numerical model, we conclude the stingless bee could pollinate strawberry as well as the honey bee if we introduced 1.8 times of bees used in this experiment.

Pollinating bees are stressed by highly variable environmental conditions, malnutrition, parasites and pathogens, but may also by getting in contact with microorganisms or entomopathogenic nematodes that are used to control plant pests and diseases. While foraging for water, food, or nest material social as well as solitary bees have direct contact or even consume the plant protection product with its active substance (e.g., viruses, bacteria, fungi, etc.). Here, we summarize the results of cage, microcolony, observation hive assays, semi-field and field studies using full-size queen-right colonies. By now, some species and subspecies of the Western and Eastern honey bee (Apis mellifera, A. cerana), few species of bumble bees, very few stingless bee species and only a single species of leafcutter bees have been studied as non-target host organisms. Survival and reproduction are the major criteria that have been evaluated. Especially sublethal effects on the bees' physiology, immune response and metabolisms will be targets of future investigations. By studying infectivity and pathogenic mechanisms, individual strains of the microorganism and impact on different bee species are future challenges, especially under field conditions. Overall, it became evident that honey bees, bumble bees and few stingless bee species may not be suitable surrogate species to make general conclusions for biological mechanisms of bee-microorganism interactions of other social bee species. Solitary bees have been studied on leafcutter bees (Megachile rotundata) only, which shows that this huge group of bees (∼20,000 species worldwide) is right at the beginning to get an insight into the interaction of wild pollinators and microbial plant protection organisms.

The price of stingless bee (Heterotrigona itama) honey is almost twice higher than the one produced by a non-stingless bee (Apis mellifera) due to its limited production and high content of polyphenol and flavonoids compounds. However, in the market, the authentication of stingless bees and non-stingless bees honey becomes difficult due to the similarity in the color of honey and its sugar content. In this research, we propose a simple analytical approach by combining ultraviolet (UV) spectroscopy and hierarchical cluster analysis (HCA) for discrimination between stingless and non-stingless bee honey. Fifty samples of monofloral stingless bee (Heterotrigona itama) honey and non-stingless bee (Apis mellifera) honey from Acacia mangium was used. The samples were diluted with a distilled water at a proportion of 1:30 (volume/volume). The spectral data were acquired in the range of 190-1100 nm using a benchtop UV-Vis spectrometer with fast scanning mode. The HCA was applied for selected preprocessed spectral data in the range of 230-400 nm. The result shows that HCA could be effective to discriminate between stingless and non-stingless bee honey. Shortly, it is expected to realize the simple and quick analytical method to authenticate stingless bee honey based on UV spectroscopy and the HCA method.

The demand for natural products that promote sustainable production chains is growing worldwide. Honey, while not a staple for food security due to its limited nutritional contribution, represents a valuable resource to foster dietary diversity, responsible consumption, and forest conservation. Its production depends directly on access to diverse flora with low pesticide exposure and requires minimal soil transformation or external inputs, making it a low-impact practice. Beekeeping also contributes to the maintenance of pollinator populations, which are key for biodiversity and sustainable agriculture. Consumer interest in the origin and authenticity of honey has grown in recent years, especially for Apis mellifera products (Escuredo and Seijo 2024, Mascarello et al. 2024). Less known is the diversity and economic potential of stingless bees (Meliponini). Across tropical and neotropical regions, nearly 500 species have been described (Grüter 2020), many of which produce honey and act as key pollinators. In Colombia, about 34 species are reportedly used in meliponiculture (Nates-Parra and Rosso-Londoño 2013). This practice is expanding in rural areas, where it complements agriculture and is compatible with family-centered management. Traditional knowledge attributes medicinal properties to stingless bee products (Sgariglia et al. 2010, Rosales 2012). Although clinical validation remains limited, biochemical studies of their honeys have identified distinctive compositions rich in phenolic compounds and other bioactive molecules, providing preliminary evidence of antioxidant and anti-inflammatory properties that warrant further research (Gomes et al. 2022). These properties are linked to microbial communities naturally present in stingless bee honeys. Fermentative yeasts and lactic acid bacteria shape these products, creating unique ecological niches with potential probiotic and bioactive value (da Silva et al. 2024). Microbial assessment is also essential to detect possible pathogens, as not all stingless bee species are recommended for meliponiculture: Trigona species, for example, may incorporate feces or dead animal tissue into nests, raising sanitary concerns (Gómez et al. 2023) Developing markets for stingless bee products requires robust methodologies for authentication and safety. We propose environmental DNA (eDNA) as a tool to address three dimensions of traceability and diversity assessment: Entomological origin of honey. Despite vast bee diversity, taxonomic expertise has declined, limiting our ability to monitor wild bees. DNA-based approaches, particularly metabarcoding, enable verification of the bee taxa involved in honey production—critical in regions with limited expertise. Challenges include primer selection for non- Apis bees, expanding reference databases, and training in molecular methods. Incorporating eDNA and metabarcoding into product traceability could democratize biodiversity monitoring and inform public policy. Botanical foraging spectrum. DNA from bee pollen and honey reveals floral visitation patterns, offering a proxy for ecosystem health. Progress is constrained by incomplete plant reference libraries. Diversity indices—borrowed from microbiome studies—can help characterize floral communities, though interpretation must account for variable foraging behavior across bee species. Microbial diversity and bioprospecting. Stingless bee honeys act as natural “filters,” concentrating microbial communities with industrial, probiotic, or antibiotic potential. Although generally safe, rigorous biosafety evaluation is needed to balance opportunities with risks. Reliable isolation and cultivation methods will be key to characterize diversity, establish reference libraries, and develop pipelines for future bioprospecting. Entomological origin of honey. Despite vast bee diversity, taxonomic expertise has declined, limiting our ability to monitor wild bees. DNA-based approaches, particularly metabarcoding, enable verification of the bee taxa involved in honey production—critical in regions with limited expertise. Challenges include primer selection for non- Apis bees, expanding reference databases, and training in molecular methods. Incorporating eDNA and metabarcoding into product traceability could democratize biodiversity monitoring and inform public policy. Botanical foraging spectrum. DNA from bee pollen and honey reveals floral visitation patterns, offering a proxy for ecosystem health. Progress is constrained by incomplete plant reference libraries. Diversity indices—borrowed from microbiome studies—can help characterize floral communities, though interpretation must account for variable foraging behavior across bee species. Microbial diversity and bioprospecting. Stingless bee honeys act as natural “filters,” concentrating microbial communities with industrial, probiotic, or antibiotic potential. Although generally safe, rigorous biosafety evaluation is needed to balance opportunities with risks. Reliable isolation and cultivation methods will be key to characterize diversity, establish reference libraries, and develop pipelines for future bioprospecting. Our pilot project, in the Magdalena Medio region of Colombia, applies eDNA analyses to bees, honey, pollen, and associated microorganisms. The study focuses on three genera of stingless bees maintained in boxes derived from colonies collected in situ in the El Silencio nature reserve of Fundación Biodiversa Colombia, ensuring foraging in a pesticide-free environment with high floral diversity. As one of the first efforts to apply DNA-based tools to stingless bee biodiversity surveys and product authentication, this initiative highlights the need for stronger collaboration, standardized data frameworks, and investment to unlock the potential of eDNA for conservation and bioeconomic development.

The study was conducted to characterize the physicochemical properties of honey produced from underground nesting stingless bees (Meliponula beccarii) in the Dandi and Meta Robi districts of the West Shewa zone, Ethiopia. A total of 27 honey samples, including 24 samples collected through careful investigation and excavation of natural nests and 3 samples purchased from the market, were analyzed in the laboratory. The evaluated physicochemical properties showed an overall mean of 306.64±87.95 meq./kg free acidity, 28.05±3.52% moisture content, 1.31±0.44 mS/cm electrical conductivity, 3.29±0.16 pH, 0.89±1.14 mg/kg HMF, 0.63±0.24% mineral (ash), 9.39±4.26% glucose, 0.24±0.01 g/100g sucrose, 10.81±4.95 g/100g maltose, and 16.57±2.55 g/100g fructose, turanose (0.20 ± 0.00 g/100g). The differences between the two district's honey samples were significant (p > 0.05) for fructose value and considerable for free acidity, moisture content, and pH values. The honey samples purchased from the market showed similar physicochemical properties to the honey from the feral nests, with a mean of 314.33±88.72 meq./kg free acidity, 27.73±2.52% moisture content, 1.43±0.41 mS/cm electrical conductivity, 3.26±0.13 pH, 0.95±1.23 mg/kg HMF, 0.59±0.19% mineral (ash), 10.11±4.11% glucose, 0.25±0.02 g/100g sucrose, 11.23±4.52 g/100g maltose, and 16.33±2.41 g/100g fructose. The study found that the stingless bee honey from the study areas had distinctive low HMF, high free acidity, and low pH values, which may indicate the honey's potential medicinal properties. The high free acidity in the Meliponula beccarii honey appear unusually elevated compared to other stingless bee honey and the Codex Alimentarius standards for Apis honey, suggesting increased fermentation that can originate from the bee species, plant and geographical origins, improper handling, and high moisture content. This study demonstrated that the honey in the study areas has distinctive physicochemical properties from A. mellifera-produced honey, which may support its traditional medicinal uses. Further detailed studies on ground-nesting and other stingless species' honey medicinal values are recommended to provide scientific evidence.