Brazilian stingless bee honey: A pioneer study on the in vitro bioaccessibility of phenolic compounds.

In this study, the effect of in vitro gastrointestinal digestion of phenolic compounds, the total phenolic content, and the antioxidant potential of stingless bee honey were investigated. Among the 33 phenolic compounds investigated, 25 were quantified, and only eight were not bioaccessible (p-aminobenzoic acid, sinapic acid, pinobanksin, isorhamnetin, quercetin-3-glucoside, syringaldehyde, coumarin, and coniferaldehyde). Benzoic acid was predominant in most undigested samples (21.3 to 2414 µg 100 g-1), but its bioaccessibility varied widely (2.5 to 534%). Rutin, a glycosylated flavonoid, was quantified in all samples and might have been deglycosylated during digestion, increasing the bioaccessibility of quercetin in a few samples. Overall, the concentration of phenolic compounds prior digestion and their bioaccessibility varied greatly among samples. Nevertheless, higher concentrations before digestion were not correlated to greater bioaccessibility. This study is the first to assess the in vitro bioaccessibility of phenolic compounds in SBH, providing novel insights into SBH research.

Pollen analysis of honey and pollen stored by Melipona (Melikerria) fasciculata Smith, 1854 (Apidae Meliponini), in an Amazon and Cerrado transition area, Maranhão, Brazil.

The identification of meliponicultural flora is fundamental for the preservation of bees, as well as enabling the development of pollinator management and reforestation programs. In this context, this study aimed to investigate the types of pollen found in the honey and pollen pots of the Melipona fasciculata bee in Arari, State of Maranhão, between August 2022 and July 2023. 40 types of pollen were identified, distributed among 18 botanical families, with Fabaceae standing out (46.15% in pollen pots and 37.86% in honey pots), Myrtaceae (11.53% in pollen and 13.51% in honey) and Rubiaceae (7.69% in pollen and 5.40% in honey). Sapindaceae, in the honey pots, accounted for 5.40%. The other families showed lower percentages, 3.84% in the pollen pots and 2.70% in the honey pots. In terms of similarity, two distinct groups were observed in both the pollen and honey pots. In the pollen pots, group A (May-June ~ 97%) and group B (Feb-Mar ~ 99%) stood out, while in the honey pots, group A (Mar-Apr ~ 98%) and group B (Jun-Jul-Sept ~ 98%) showed the highest percentages. These findings highlight the wide range of resources used by the M. fasciculata species, as well asits preference for Fabaceae and Myrtaceae, due to the diversity and availability of trophic resources. An understanding of the meliponicultural flora is essential to support effective conservation strategies, which aim not only to guarantee the survival of the bees, but also to ensure the continued production of honey, a resource of great importance to local communities.

Preparative Fractionation of Brazilian Red Propolis Extract Using Step-Gradient Counter-Current Chromatography.

Propolis is a resinous bee product with a very complex composition, which is dependent upon the plant sources that bees visit. Due to the promising antimicrobial activities of red Brazilian propolis, it is paramount to identify the compounds responsible for it, which, in most of the cases, are not commercially available. The aim of this study was to develop a quick and clean preparative-scale methodology for preparing fractions of red propolis directly from a complex crude ethanol extract by combining the extractive capacity of counter-current chromatography (CCC) with preparative HPLC. The CCC method development included step gradient elution for the removal of waxes (which can bind to and block HPLC columns), sample injection in a single solvent to improve stationary phase stability, and a change in the mobile phase flow pattern, resulting in the loading of 2.5 g of the Brazilian red propolis crude extract on a 912.5 mL Midi CCC column. Three compounds were subsequently isolated from the concentrated fractions by preparative HPLC and identified by NMR and high-resolution MS: red pigment, retusapurpurin A; the isoflavan 3(R)-7-O-methylvestitol; and the prenylated benzophenone isomers xanthochymol/isoxanthochymol. These compounds are markers of red propolis that contribute to its therapeutic properties, and the amount isolated allows for further biological activities testing and for their use as chromatographic standards.

Extraction Kinetics of Brazilian Green Propolis and Chemical Characterization of its Volatiles.

Propolis is a natural resinous product produced by Apis mellifera bees from the exudates of various plants. The color of propolis (green) is a consequence of its botanical origin, as bees collect young tissues and leaves of Baccaris dracunculifolia. This study evaluated the chemical composition and extraction kinetics of essential oils obtained from Brazilian green propolis by hydrodistillation. Hydrodistillation was performed for 360 min and analyzed at different times (30, 60, 120, 240, and 360 min), allowing the calculation of the accumulated content (% w/w) and the identification of the essential oil chemical profile. The GC/FID and GC/MS analysis led to the annotation of 60 compounds with estragole (13.30 %), benzyl propanoate (14.59 %), and (E)-nerolidol (13.57 %) as the main compounds. The optimum conditions for extraction of phenylpropanoids (PP), hydrocarbons (HD), monoterpenes (MT), and oxygenated monoterpenes (OMT) are between 30 and 120 min. In comparison, sesquiterpenes (ST) and oxygenated sesquiterpenes (OST) are extracted more efficiently between 240 and 360 min. The optimal extraction speed determination is essential for industrial-scale processing to obtain components such as sesquiterpenes, which have a high economic value in the cosmetic/perfumery and pharmaceutical industries.

Floral Pollen Bioactive Properties and Their Synergy in Honeybee Pollen.

Honeybee pollen (HBP) is a mixture of floral pollen collected by honeybees near the hive. It is characterized by a composition rich in phenolic compounds, carotenoids and vitamins that act as free radicals scavengers, conferring antioxidant and antibacterial capacity to the matrix. These bioactive properties are related to the botanical origin of the honeybee pollen. Honeybee pollen samples were collected from different geographical locations in central Chile, and their total carotenoid content, polyphenols profile by HPLC/MS/MS, DPPH radical scavenging capacity, and antimicrobial capacity against S. pyogenes, E. coli, S. aureus, and P. auriginosa strains were evaluated. Our results showed a good carotenoids content and polyphenols composition, while antioxidant capacity presented values between 0-95 % for the scavenging effect related to the botanical origin of the samples. Inhibition diameter for the different strains presented less variability among the samples, Furthermore, binary mixtures representing the two most abundant species in each HBP were prepared to assess the synergy effect of the floral pollen (FP) present in the samples. Data shows an antagonist effect was observed when assessing the carotenoid content, and a synergy effect often presents for antimicrobial and antioxidant capacity for bee pollen samples. The bioactive capacities of the honeybee pollen and their synergy effect could apply to develop new functional ingredients for the food industry.

Chemical Composition of Volatile Compounds in Apis mellifera Propolis from the Northeast Region of Pará State, Brazil.

Propolis is a balsamic product obtained from vegetable resins by exotic Africanized bees Apis mellifera L., transported and processed by them, originating from the activity that explores and maintains these individuals. Because of its vegetable and natural origins, propolis is a complex mixture of different compound classes; among them are the volatile compounds present in the aroma. In this sense, in the present study we evaluated the volatile fraction of propolis present in the aroma obtained by distillation and simultaneous extraction, and its chemical composition was determined using coupled gas chromatography, mass spectrometry, and flame ionization detection. The majority of compounds were sesquiterpene and hydrocarbons, comprising 8.2-22.19% α-copaene and 6.2-21.7% ß-caryophyllene, with additional compounds identified in greater concentrations. Multivariate analysis showed that samples collected from one region may have different chemical compositions, which may be related to the location of the resin's production. This may be related to other bee products.

Stingless Bee-Collected Pollen (Bee Bread): Chemical and Microbiology Properties and Health Benefits.

Stingless bee-collected pollen (bee bread) is a mixture of bee pollen, bee salivary enzymes, and regurgitated honey, fermented by indigenous microbes during storage in the cerumen pot. Current literature data for bee bread is overshadowed by bee pollen, particularly of honeybee Apis. In regions such as South America, Australia, and Southeast Asia, information on stingless bee bee bread is mainly sought to promote the meliponiculture industry for socioeconomic development. This review aims to highlight the physicochemical properties and health benefits of bee bread from the stingless bee. In addition, it describes the current progress on identification of beneficial microbes associated with bee bread and its relation to the bee gut. This review provides the basis for promoting research on stingless bee bee bread, its nutrients, and microbes for application in the food and pharmaceutical industries.

Nano-selenium and nano-zinc oxide supplementation in syrup on laying area, population size and hsp gene expression of honey bees in hot climate

This study was designed to examine the protective effects of nano-selenium and nano-zinc oxide on queen and workers performance under heat stress condition and gene expression of heat shock protein 70 (hsp70) as an index of heat tolerance. Sixty colonies were randomly assigned to five treatments with 12 replicates from June until early September. Sugar syrup (50%) containing no supplement or nano-selenium at levels of 50 and 100 µg L-1 or nano-zinc at levels of 100 and 200 µg L-1 was fed to colonies. Nano-selenium supplementations had no effect, but nano-zinc at level of 100 µg L-1 significantly decreased body malondialdehyde concentration. The highest bee population was seen in nano-zinc at level of 100 µg L-1 and the lowest one in control group. The lowest and the highest body weight, fat and protein deposition was found in group received nano-zinc at level of 100 µg L-1 and control, respectively. The highest gene expression was for group received nano-zinc at level of 100 µg L-1 In group received nano-zinc at level of 100 µg L-1, an increase in hsp70 gene expression was found. In conclusion, nano-zinc oxide at level of 100 µg L-1 could increase queen and worker performance and heat resistance of bees in the hot climate condition.(AU)

Analysis of a dose-response assay in Scaptotrigona bipunctatabees, Lepeletier, 1836 (Hymenoptera: Apidae) using the logistic regression model under the Bayesian approach

This paper shows the results of a dose-response study in Scaptotrigona bipunctatabees, Lepeletier, 1836 (Hymenoptera: Apidae) exposed to the insecticide Fastac Duo. The aim was to evaluate the lethal concentration that causes the death of 50% of bees (LC50) and investigate the odd of mortality after exposure to different concentrations, using the logistic regression model under the Bayesian approach. In this approach, it is possible to incorporate a prior information and gives more accurate inferential results. Three independent dose-response experiments were analyzed, dissimilar in their lead time according to guidelines from the Organisation for Economic Co-operation and Development (OECD), in which each assay contained four replicates at the concentration levels investigated, including control. Observing exposure to the agrochemical, it was identified that the higher the concentration, the greater the odd of mortality. Regarding the estimated lethal concentrations for each experiment, the following values were found, 0.03 g a.i. L-1, for 24hours, 0.04 g a.i. L-1, for 48hoursand 0.06 g a.i. L-1for 72hours, showing that in experiments with longer exposure times there was an increase in LC50. Concluding, the study showed an alternative approach to classical methods for dose-response studies in Scaptotrigona bipunctatabees exposed to the insecticide Fastac Duo.(AU)

Nano-selenium and nano-zinc oxide supplementation in syrup on laying area, population size and hsp gene expression of honey bees in hot climate

This study was designed to examine the protective effects of nano-selenium and nano-zinc oxide on queen and workers performance under heat stress condition and gene expression of heat shock protein 70 (hsp70) as an index of heat tolerance. Sixty colonies were randomly assigned to five treatments with 12 replicates from June until early September. Sugar syrup (50%) containing no supplement or nano-selenium at levels of 50 and 100 µg L-1 or nano-zinc at levels of 100 and 200 µg L-1 was fed to colonies. Nano-selenium supplementations had no effect, but nano-zinc at level of 100 µg L-1 significantly decreased body malondialdehyde concentration. The highest bee population was seen in nano-zinc at level of 100 µg L-1 and the lowest one in control group. The lowest and the highest body weight, fat and protein deposition was found in group received nano-zinc at level of 100 µg L-1 and control, respectively. The highest gene expression was for group received nano-zinc at level of 100 µg L-1 In group received nano-zinc at level of 100 µg L-1, an increase in hsp70 gene expression was found. In conclusion, nano-zinc oxide at level of 100 µg L-1 could increase queen and worker performance and heat resistance of bees in the hot climate condition.

Orchid bees in riparian and terra-firme forest fragments in an urban matrix in southwestern Brazilian Amazonia / Abelhas das orquídeas em fragmentos de floresta ripária e de terra-firme em uma matriz urbana no sudoeste da Amazônia brasileira

Riparian forests are important ecosystems that support an enormous biodiversity in Brazil. Despite being protected under Brazilian legislation, these forests suffer great impact from the fragmentation of habitats. Orchid bees are a key group of pollinators in the Neotropical region, yet few data are available on the assamblage structure of these bees in riparian forests. We evaluated the role of fragments of riparian and terra-firme forest on the conservation and maintenance of orchid bees in an urban landscape in the southwestern Amazon basin. Specifically, we evaluated whether bee assemblages in riparian and terra-firme forests differed significantly in abundance, species richness and composition. We also evaluated whether species richness and abundance of bees vary with the size of the forest fragment. Male bees were attracted using odoriferous baits and collected with entomological nets in 10 forest fragments. There was no significant difference between riparian and terra-firme fragments in species abundance, richness and composition, but there was a positive correlation between fragment size and species richness and abundance. Our results suggest that, in an urban landscape, the remaining riparian and terra-firme forest fragments still could maintain 62.7% of the orchid bee species known to occur in the region, reinforcing the conservation value of these forest remnants. Our findings indicate that these fragments provide a potentially important habitat for the maintenance of local bee populations in the landscape.(AU)

As florestas ripárias no Brasil são importantes ecossistemas que sustentam uma enorme biodiversidade. Apesar de protegidas pela legislação brasileira, elas têm sofrido grandes impactos decorrentes da fragmentação florestal. As abelhas das orquídeas constituem um grupo chave de polinizadores na região Neotropical, porém, pouco se sabe sobre suas assembleias em florestas ripárias. Nós avaliamos o papel de fragmentos de floresta ripária e de terra-firme na conservação e manutenção da fauna de abelhas das orquídeas em uma paisagem urbana no sudoeste da Amazônia. Especificamente, avaliamos se assembleias de abelhas de fragmentos ripários e de terra-firme diferem significativamente em abundância, riqueza e composição de espécies. Também avaliamos se a abundância e a riqueza de espécies variam em função do tamanho do fragmento. Machos de abelhas foram atraídos por iscas odoríferas e coletados com redes entomológicas em 10 fragmentos florestais. Não houve diferença significativa entre fragmentos ripários e de terra-firme quanto à abundância, riqueza e composição de espécies, mas houve uma correlação positiva entre o tamanho do fragmento e a riqueza e abundância de espécies. Nossos resultados sugerem que, em uma paisagem urbana, os fragmentos de floresta ripária e de terra firme ainda podem manter 62,7% do número de espécies de abelhas das orquídeas conhecido para a região, reforçando o valor da conservação desses remanescentes florestais. Nossos dados indicam que esses fragmentos fornecem um habitat potencialmente importante para a manutenção das populações locais de abelhas na paisagem.(AU)

Analysis of a dose-response assay in Scaptotrigona bipunctatabees, Lepeletier, 1836 (Hymenoptera: Apidae) using the logistic regression model under the Bayesian approach

This paper shows the results of a dose-response study in Scaptotrigona bipunctatabees, Lepeletier, 1836 (Hymenoptera: Apidae) exposed to the insecticide Fastac Duo. The aim was to evaluate the lethal concentration that causes the death of 50% of bees (LC50) and investigate the odd of mortality after exposure to different concentrations, using the logistic regression model under the Bayesian approach. In this approach, it is possible to incorporate a prior information and gives more accurate inferential results. Three independent dose-response experiments were analyzed, dissimilar in their lead time according to guidelines from the Organisation for Economic Co-operation and Development (OECD), in which each assay contained four replicates at the concentration levels investigated, including control. Observing exposure to the agrochemical, it was identified that the higher the concentration, the greater the odd of mortality. Regarding the estimated lethal concentrations for each experiment, the following values were found, 0.03 g a.i. L-1, for 24hours, 0.04 g a.i. L-1, for 48hoursand 0.06 g a.i. L-1for 72hours, showing that in experiments with longer exposure times there was an increase in LC50. Concluding, the study showed an alternative approach to classical methods for dose-response studies in Scaptotrigona bipunctatabees exposed to the insecticide Fastac Duo.

Development of alginate beads with encapsulated jabuticaba peel and propolis extracts to achieve a new natural colorant antioxidant additive.

This work aims to encapsulate anthocyanins and phenolic compounds extracted from a native Brazilian fruit peel - jabuticaba (Plinia cauliflora (Mart.) Kausel) and propolis from Tubuna (Scaptotrigona bipunctata) stingless bees, with great potential benefits for human health. The alginate encapsulation was conducted by the ionotropic gelation through the dripping into the CaCl2 solution. Both raw extracts were characterized by TPC - total phenolic content (Folin-Ciocalteu), AA -antioxidant activity (DPPH and ABTS assays), and TMAC - total monomeric anthocyanin concentration (pH differential method); as well as their resultant mixture (2:1 jabuticaba/propolis). The obtained beads presented highly efficient encapsulation of total polyphenols (~98%) and monomeric anthocyanins (~89%), with spherical morphology and smooth surface obtaining a mean diameter between 200 and 250 µm. In vitro release study showed that JPE/alginate beads were completely disintegrated at pH 7.4 (intestinal pH), but they were resistant to gastric pH (1.2) presenting a slow release of about 40% in 240 min. This is the first report that encapsulates the mixture of jabuticaba and propolis extracts and may contribute to the utilization of a great source of bioactive compounds besides the potential pigment of anthocyanins, an alternative to natural and healthy food/beverage colorants.

Occurrence of virus, microsporidia, and pesticide residues in three species of stingless bees (Apidae: Meliponini) in the field.

Bees are important pollinators whose population has declined due to several factors, including infections by parasites and pathogens. Resource sharing may play a role in the dispersal dynamics of pathogens among bees. This study evaluated the occurrence of viruses (DWV, BQCV, ABPV, IAPV, KBV, and CBPV) and microsporidia (Nosema ceranae and Nosema apis) that infect Apis mellifera, as well as pesticide residues in the stingless bees Nannotrigona testaceicornis, Tetragonisca angustula, and Tetragona elongata sharing the same foraging area with A. mellifera. Stingless bees were obtained from 10 nests (two of N. testaceicornis, five of T. angustula, and three of T. elongata) which were kept in the field for 1 year and analyzed for the occurrence of pathogens. Spores of N. ceranae were detected in stingless bees but were not found in their midgut, which indicates that these bees are not affected, but may be vectors of the microsporidium. Viruses were found in 23.4% of stingless bees samples. APBV was the most prevalent virus (10.8%) followed by DWV and BQCV (both in 5.1% of samples). We detected glyphosate and its metabolites in small amounts in all samples. The highest occurrence of N. ceranae spores and viruses was found in autumn-winter and may be related to both the higher frequency of bee defecation into the colony and the low food resources available in the field, which increases the sharing of plant species among the stingless bees and honey bees. This study shows the simultaneous occurrence of viruses and spores of the microsporidium N. ceranae in asymptomatic stingless bees, which suggest that these bees may be vectors of pathogens.

Active phloem sap collection by Trigona spinipes (Fabricius) (Hymenoptera, Apidae)in Artemisia annuaLinn (Asteraceae)

Trigona spinipes Fabricius (Apidae: Meliponini), is a common stingless bee widely seen in urban and rural areas in Brazil, popularly known as irapuá, arapuá or bee-dog. Although these bees are considered pollinators of some cultivated plants, they are better known for the damage they cause in different crops. During experimental agroecological cultivation of Artemisia (Artemisia annuaLinn, Asteraceae), in Jaguariúna (SP, Brazil), stingless bees Trigona spinipes (Fabricius) were observed sucking phloem sap directly from the plant, a phenomenon not yet described in scientific literature. This study aimed to register and describe the phloem sap-sucking behavior performed by T. spinipesfor the first time, as well as to assess the potential impact of this behavior on A. annuacultivation. The behavior and the material collected by bees and the severity of attack were also analyzed. The aging and premature death of observed A. annuaspecimens occurred because of extensive lesions caused by T. spinipes, confirming the negative consequence of sap-sucking attacks of T. spinipesbees on the plants. Factors that could induce this unusual behavior were presented, pointing out the need for future studies on the development of strategies to protect plants, without causing damage to the T. spinipesbee populations, which are elements of Brazilian bee fauna and, therefore, protected by law.

Royal jelly production with queens produced by single and double grafting in Africanized honeybee colonies

The objective of this research was to evaluate royal jelly production from Africanized honeybee queens of different lineages (lineage selected for honey production, lineage selected for royal jelly production and unselected) produced by single and double grafting and to compare royal jelly production among their offspring. Data were tested by double factorial analysis of variance and the means were compared by Tukey test at 5%. The parameters evaluated were: queen weight at emergence, percentage of larvae acceptance in the upper and lower bars, royal jelly per cup (mg) and royal jelly per colony/collection (g). Queens selected for honey presented greater weight at emergence, while the unselected queens were the lightest. Double grafting was better than simple grafting, since the queens were born 2.38% heavier. There was no difference (p > 0.05) in relation to royal jelly production according to neither the lineage nor even comparing the method by which the queens were produced. The cost of labor to produce queens by double grafting was much higher, as by the simple grafting about 170% more queens were born. The potential of Africanized honeybees with adequate production management and favorable environmental conditions favors the production of royal jelly.(AU)

Royal jelly production with queens produced by single and double grafting in Africanized honeybee colonies

The objective of this research was to evaluate royal jelly production from Africanized honeybee queens of different lineages (lineage selected for honey production, lineage selected for royal jelly production and unselected) produced by single and double grafting and to compare royal jelly production among their offspring. Data were tested by double factorial analysis of variance and the means were compared by Tukey test at 5%. The parameters evaluated were: queen weight at emergence, percentage of larvae acceptance in the upper and lower bars, royal jelly per cup (mg) and royal jelly per colony/collection (g). Queens selected for honey presented greater weight at emergence, while the unselected queens were the lightest. Double grafting was better than simple grafting, since the queens were born 2.38% heavier. There was no difference (p > 0.05) in relation to royal jelly production according to neither the lineage nor even comparing the method by which the queens were produced. The cost of labor to produce queens by double grafting was much higher, as by the simple grafting about 170% more queens were born. The potential of Africanized honeybees with adequate production management and favorable environmental conditions favors the production of royal jelly.

Matrix solid-phase dispersion associated to gas chromatography for the assessment in honey bee of a group of pesticides of concern in the apicultural field.

A method based on matrix solid-phase dispersion (MSPD) associated to gas chromatography-flame photometric detection (GC-FPD), GC-electron capture detection (GC-ECD) and GC-mass spectrometry (GC-MS) for confirmation purposes, was developed for the determination of a representative group of twelve pesticides in honeybee with particular concern in the apicultural field (fipronil, thiamethoxam, acetamiprid, acrinathrin, metamidophos, dimetoathe, diazinon, chlorpyrifos, methidathion, profenophos, azinphos methyl and coumaphos). Factors influencing the extraction efficiency of MSPD were investigated and optimized through response surface method. The use of octadecylsilyl (C18) sorbent combined with a florisil clean-up and acetonitrile-methanol (99:1) elution was the optimal condition for the extraction of the selected pesticides. Under this condition the recovery of pesticides at the limit of quantification of the method (0.007 to 0.050 µg g-1) ranged from 68 to 102% with RSDs for within-laboratory reproducibility ≤20%. The proposed method was applied to the analysis of honeybees collected in 68 field hives from areas of great apicultural and agricultural development in central Chile. In 65% of these samples eight different pesticides were detected. Pesticides most frequently found were chlorpyrifos (34% of the samples, <0.017-0.067 µg g-1), acrinathrin (32% of the samples, <0.020-0.026 µg g-1) and diazinon (10% of the samples at values <0.015 µg g-1). The incidence of these pesticides in bees can be related to their high employ in central Chile, use to combat the varroosis in hives and hydrophobicity.

Antioxidant, antihyperglycemic, and antidiabetic activity of Apis mellifera bee tea.

Diabetes has emerged as one of the largest global epidemics; it is estimated that by 2035, there will be 592 million diabetic people in the world. Brazilian biodiversity and the knowledge of traditional peoples have contributed to the treatment of several diseases, including diabetes. Apis mellifera bee tea is used by indigenous Brazilians to treat diabetes, and this traditional knowledge needs to be recorded and studied.The objective of this study was to record the use and to evaluate the antioxidant, antihyperglycemic, and antidiabetic activity of Apis mellifera bee tea, which is used by the Guarani and Kaiowá indigenous people for the treatment of diabetes. Semi-structured interviews were performed with Guarani and Kaiowá ethnic indigenous people from the State of Mato Grosso do Sul, Brazil, seeking to identify the animal species used for medicinal purposes. For the experimental procedures, tea prepared with macerated Apis mellifera bees was used. In vitro assays were performed to evaluate antioxidant activity; direct free radical scavenging, protection against oxidative hemolysis, lipid peroxidation were evaluated in human erythrocytes and potential in inhibiting the formation of advanced glycation end products (AGEs). In vivo, normoglycemic Swiss male mice treated with Apis mellifera tea (AmT) were subjected to the oral glucose tolerance test and compared with control and metformin-treated groups. Diet-induced diabetic mice were treated for 21 days with AmT and evaluated for glycemia and malondialdehyde levels in the blood, liver, nervous system, and eyes. During interviews, the indigenous people described the use of Apis mellifera bee tea for the treatment of diabetes. In in vitro assays, AmT showed direct antioxidant activity and reduced oxidative hemolysis and malondialdehyde generation in human erythrocytes. The AmT inhibited the formation of AGEs by albumin-fructose pathways and methylglyoxal products. In vivo, after oral glucose overload, normoglycemic mice treated with AmT had reduced hyperglycemia at all times evaluated up to 180 min. AmT also reduced hyperglycemia and malondialdehyde levels in the blood, liver, nervous system, and eyes of diabetic mice to similar levels as those in metformin-treated mice and normoglycemic controls. In summary, Apis mellifera bee tea showed antioxidant, antihyperglycemic, and antidiabetic activity, which provides support for the therapeutic application of Guarani and Kaiowá indigenous knowledge.

MALDI Imaging Analysis of Neuropeptides in Africanized Honeybee ( Apis mellifera) Brain: Effect of Aggressiveness.

Aggressiveness in honeybees seems to be regulated by multiple genes, under the influence of different factors, such as polyethism of workers, environmental factors, and response to alarm pheromones, creating a series of behavioral responses. It is suspected that neuropeptides seem to be involved with the regulation of the aggressive behavior. The role of allatostatin and tachykinin-related neuropeptides in honeybee brain during the aggressive behavior is unknown, and thus worker honeybees were stimulated to attack and to sting leather targets hung in front of the colonies. The aggressive individuals were collected and immediately frozen in liquid nitrogen; the heads were removed and sliced at sagittal plan. The brain slices were submitted to MALDI spectral imaging analysis, and the results of the present study reported the processing of the precursors proteins into mature forms of the neuropeptides AmAST A (59-76) (AYTYVSEYKRLPVYNFGL-NH2), AmAST A (69-76) (LPVYNFGL-NH2), AmTRP (88-96) (APMGFQGMR-NH2), and AmTRP (254-262) (ARMGFHGMR-NH2), which apparently acted in different neuropils of the honeybee brain during the aggressive behavior, possibly taking part in the neuromodulation of different aspects of this complex behavior. These results were biologically validated by performing aggressiveness-related behavioral assays using young honeybee workers that received 1 ng of AmAST A (69-76) or AmTRP (88-96) via hemocele. The young workers that were not expected to be aggressive individuals presented a complete series of aggressive behaviors in the presence of the neuropeptides, corroborating the hypothesis that correlates the presence of mature AmASTs A and AmTRPs in the honeybee brain with the aggressiveness of this insect.