Mollicute Anti-Adhesive and Growth Inhibition Properties of the Methanolic Extract of Propolis from the Brazilian Native Bee Melipona quadrifasciata.

Hydroalcoholic propolis extracts from the bee species Melipona quadrifasciata have been shown to possess antimicrobial activity against different mollicute strains, but a methanolic extract (ME) could contain an increased diversity of nonpolar bioactive components with a potentially higher antimicrobial activity. The ME obtained by maceration of the propolis sample was fractionated with solvents of different polarities and then, purified by silica gel column chromatography through biomonitoring of its antimicrobial activity against mollicute strains. Analysis by gas chromatography-mass spectrometry (GC/MS) enabled the identification of compounds using the NIST library. Minimum inhibitory concentrations (MICs) of the samples were determined by broth microdilution. Anti-adhesive assays were performed with Mycoplasma pneumoniae cells. The hexane (MIC=62.5 mg/L) and dichloromethane (MIC=125 mg/L) fractions presented the most promising results against M. pneumoniae. They were fractionated into 74 subfractions, and even the best ones did not show better results (MIC>250 mg/L) than their original fractions, likely due to the loss of terpene compounds that seem to act in synergy. The dichloromethane subfraction FD4 was highlighted in the anti-adhesive assay with an inhibitory activity of 21.6 %. A synergistic effect of the nonpolar compounds in M. quadrifasciata propolis may be responsible for its antibacterial activity, but several purified components can improve its anti-adhesive properties.

Dataset about Southern-Brazilian geopropolis: Physical and chemical perspectives.

The dataset showed in this manuscript belongs to the investigation of the Southern-Brazilian geopropolis of stingless bees. Stingless bees are native species of insects from tropical areas; they produce honey, pollen and geopropolis that is composed of a mix of vegetal extracts, digestive enzymes, and mostly by soil. Used in folk medicine as antiseptic, antioxidant and antimicrobial agent, the composition is due to bee species, climate changes, local flora, and soil type. Moreover, the complex chemical content gives to the geopropolis a bioactive potential, with scavenging characteristics that is important to avoid free radical damages in the human health. Regarding the importance of exploring new natural matrices sources with bioactive potential, the first approach of chemical characterization of geopropolis is indispensable. Thus, ten samples of Southern-Brazilian geopropolis were analyzed and the bioactive responses obtained were discussed in the accompanying article titled "Southern-Brazilian geopropolis: A potential source of polyphenolic compounds and assessment of mineral composition". Furthermore, the physicochemical analysis of moisture and ash content, the yield of extraction, the reducing activity and free radical scavenging potential of ethanolic extracts, the antimicrobial activity, and the analysis of HPLC-ESI-MS/MS chromatograms are the main data presented in brief. The data can guide scientists in order to know methods and data for these samples.

Simple discrimination in stingless bees (Melipona quadrifasciata): Probing for select- and reject-stimulus control.

Simple and conditional discrimination training may produce various types of controlling relations. Responses may be controlled primarily by the positive stimulus (select-control relation) or by the negative stimulus (reject-control relation; the subject excludes the negative stimulus and chooses the positive). Bees learn to respond in simple and conditional discriminations. However, no study has searched for reject-control responding in Melipona bees. We trained Melipona quadrifasciata on a simple discrimination task (S+ vs. S-; e.g., blue vs. yellow) and then probed for stimulus control with two types of probe trials, S+ versus a new stimulus (Select-control probes) and S- versus a new stimulus (Reject-control probes). For Group Different, a new-stimulus color (e.g., white) was used in one type of probe and another color (e.g., black) was used in the other type. For Group Same, a single new-stimulus color was used in both types of probes. On Select probes, the bees always preferred S+ to the new stimulus. On Reject probes, results were mixed. Depending on the colors used in training and probing, bees responded to both stimuli, and even preferred the S-. The data suggest no control by the negative function of the S- and support the select-stimulus control hypothesis of responding.

Changes in social behavior are induced by pesticide ingestion in a Neotropical stingless bee.

Throughout evolutionary history bees have developed complex communication systems. For social bees, communication is important for both the individual and the development of the colony. Successful communication helps bees to recognize relatives, defend the colony, and promote recruitment to optimize foraging of floral resources. Bees' contribution to pollination is of broad environmental and economic importance. However, studies have reported that anthropogenic actions, such as the use of pesticides, negatively affect bee survival and behavior. We tested the effect of a commercially available pesticide mix containing two pesticide classes, a neonicotinoid and a pyrethroid, on the social behavior of the stingless bee, Melipona quadrifasciata (Lepeletier, 1863). After determining a sublethal dose of the pesticides, we tested the effect of an acute dose on antennation and trophallaxis behaviors of worker bees. Our results showed a drastic reduction in the communication and social interactions of bees.

Cytotoxicity mechanisms in melanoma cells and UPLC-QTOF/MS2 chemical characterization of two Brazilian stingless bee propolis: Uncommon presence of piperidinic alkaloids.

The present study characterized propolis extracts produced by Scaptotrigona bipunctata (Tubuna) and Melipona quadrifasciata (Mandaçaia) by LC-MS/MS; their cytotoxicity as well as the mechanism of action in a melanoma cellular model were also assessed. The chemical characterization performed by UPLC-ESI-QTOF/MS2 analysis revealed uncommon presence of piperidinic alkaloids in Tubuna's propolis extract together with C-glycopyranoside flavonoids. Mandaçaia's propolis collected in the same area rather presented terpenoids and flavonoids. Regarding the mechanism of cytotoxicity, propolis extracts increased the accumulation of reactive oxygen species (ROS), reduced the potential of mitochondrial membrane, induced a decrease in the proteins Bcl-2 and AKT-3 levels, and decreased melanoma cells' migration and invasion. Both propolis extracts induced apoptosis while only Mandaçaia's propolis extract induced cell cycle arrest in G2/M.

Sequence and expression pattern of the germ line marker vasa in honey bees and stingless bees.

Queens and workers of social insects differ in the rates of egg laying. Using genomic information we determined the sequence of vasa, a highly conserved gene specific to the germ line of metazoans, for the honey bee and four stingless bees. The vasa sequence of social bees differed from that of other insects in two motifs. By RT-PCR we confirmed the germ line specificity of Amvasa expression in honey bees. In situ hybridization on ovarioles showed that Amvasa is expressed throughout the germarium, except for the transition zone beneath the terminal filament. A diffuse vasa signal was also seen in terminal filaments suggesting the presence of germ line cells. Oocytes showed elevated levels of Amvasa transcripts in the lower germarium and after follicles became segregated. In previtellogenic follicles, Amvasa transcription was detected in the trophocytes, which appear to supply its mRNA to the growing oocyte. A similar picture was obtained for ovarioles of the stingless bee Melipona quadrifasciata, except that Amvasa expression was higher in the oocytes of previtellogenic follicles. The social bees differ in this respect from Drosophila, the model system for insect oogenesis, suggesting that changes in the sequence and expression pattern of vasa may have occurred during social evolution.

Sequence and expression pattern of the germ line marker vasa in honey bees and stingless bees

Queens and workers of social insects differ in the rates of egg laying. Using genomic information we determined the sequence of vasa, a highly conserved gene specific to the germ line of metazoans, for the honey bee and four stingless bees. The vasa sequence of social bees differed from that of other insects in two motifs. By RT-PCR we confirmed the germ line specificity of Amvasa expression in honey bees. In situ hybridization on ovarioles showed that Amvasa is expressed throughout the germarium, except for the transition zone beneath the terminal filament. A diffuse vasa signal was also seen in terminal filaments suggesting the presence of germ line cells. Oocytes showed elevated levels of Amvasa transcripts in the lower germarium and after follicles became segregated. In previtellogenic follicles, Amvasa transcription was detected in the trophocytes, which appear to supply its mRNA to the growing oocyte. A similar picture was obtained for ovarioles of the stingless bee Melipona quadrifasciata, except that Amvasa expression was higher in the oocytes of previtellogenic follicles. The social bees differ in this respect from Drosophila, the model system for insect oogenesis, suggesting that changes in the sequence and expression pattern of vasa may have occurred during social evolution.

Differentiation of Melipona quadrifasciata L. (Hymenoptera, Apidae, Meliponini) subspecies using cytochrome b PCR-RFLP patterns

Melipona quadrifasciata quadrifasciata and M. quadrifasciata anthidioides are subspecies of M. quadrifasciata, a stingless bee species common in coastal Brazil. These subspecies are discriminated by the yellow stripe pattern of the abdominal tergites. We found Vsp I restriction patterns in the cytochrome b region closely associated to each subspecies in 155 M. quadrifasciata colonies of different geographical origin. This mitochondrial DNA molecular marker facilitates diagnosis of M. quadrifasciata subspecies matrilines and can be used to establish their natural distribution and identify hybrid colonies.

Effect of juvenile hormone III on the ultrastructure of the corpora allata in Melipona quadrifasciata (Hymenoptera,. Apidae, Meliponýný)

Changes in hormonal levels can produce alternative phenotypes. Juvenile hormone III plays an importantrole in the regulation of metamorphosis, caste determination and age in bees. In this work, we examined theultrastructure of corpora allata cells from stingless bees (Melipona quadrifasciata) treated with juvenilehormone during development. The corpora allata cells of M. quadrifasciata queens showed greater activitythan those of workers. The topical application of juvenile hormone III altered the cellular ultrastructureand either delayed development (as shown by fewer mitochondria and greater chromatin condensation) orenhanced development (looser chromatin and numerous mitochondria) when compared to untreated (control)bees. Our results show that corpora allata cells differ in their ultrastructural characteristics and that thecessation of juvenile hormone production by these cells in M. quadrifasciata is not synchronous.

Genetic analysis of Melipona quadrifasciata Lep. (Hymenoptera: Apidae, Meliponinae) with RAPD markers

Melipona quadrifasciata ("mandaçaia") can be subdivided into two subspecies: M. q. anthidioides and M. q. quadrifasciata. In the present study we used RAPD markers to estimate intercolonial genetic variation among 69 colonies of Melipona quadrifasciata. Ten workers per colony were analyzed. The intercolony genetic distances based on RAPD markers ranged from 29.5% (colonies collected in the State of São Paulo vs colonies from the State of Minas Gerais) to 34.2% (São Paulo vs Santa Catarina). These results indicate a high genetic similarity among the colonies analyzed.According to the genetic distances two different groups could be distinguished. The first containing the samples from Santa Catarina region and the second, samples from Paraná, São Paulo, Minas Gerais, and Espírito Santo. Based on the molecular analysis, bees belonging to the different subspecies M. q. quadrifasciata (from Santa Catarina) and M. q. anthidioides (from the other regions) were distinguished.

A abelha Melipona quadrifasciata Lep., conhecida popularmente como "mandaçaia", apresenta duas subespécies: Melipona quadrifasciata anthidioides e Melipona quadrifasciata quadrifasciata. Utilizando-se marcadores RAPD, foram calculadas as distâncias genéticas entre 69 colônias de Melipona quadrifasciata. Foram coletadas 10 operárias de cada colônia. As distâncias genéticas entre as colônias dessas regiões variaram de 29,5% (entre São Paulo e Minas Gerais) a 34,2% (entre São Paulo e Santa Catarina), indicando alto grau de similaridade genética entre as abelhas provenientes de diferentes regiões. De acordo com a distância genética, dois grupos podem ser distinguidos, um pertencente à subespécie M. q. quadrifasciata (região de Santa Catarina) e outro pertencente à subespécie M. q. anthidioides (demais regiões amostradas).

Genetic analysis of Melipona quadrifasciata Lep. (Hymenoptera: Apidae, Meliponinae) with RAPD markers

Melipona quadrifasciata ("mandaçaia") can be subdivided into two subspecies: M. q. anthidioides and M. q. quadrifasciata. In the present study we used RAPD markers to estimate intercolonial genetic variation among 69 colonies of Melipona quadrifasciata. Ten workers per colony were analyzed. The intercolony genetic distances based on RAPD markers ranged from 29.5% (colonies collected in the State of São Paulo vs colonies from the State of Minas Gerais) to 34.2% (São Paulo vs Santa Catarina). These results indicate a high genetic similarity among the colonies analyzed.According to the genetic distances two different groups could be distinguished. The first containing the samples from Santa Catarina region and the second, samples from Paraná, São Paulo, Minas Gerais, and Espírito Santo. Based on the molecular analysis, bees belonging to the different subspecies M. q. quadrifasciata (from Santa Catarina) and M. q. anthidioides (from the other regions) were distinguished.

A abelha Melipona quadrifasciata Lep., conhecida popularmente como "mandaçaia", apresenta duas subespécies: Melipona quadrifasciata anthidioides e Melipona quadrifasciata quadrifasciata. Utilizando-se marcadores RAPD, foram calculadas as distâncias genéticas entre 69 colônias de Melipona quadrifasciata. Foram coletadas 10 operárias de cada colônia. As distâncias genéticas entre as colônias dessas regiões variaram de 29,5% (entre São Paulo e Minas Gerais) a 34,2% (entre São Paulo e Santa Catarina), indicando alto grau de similaridade genética entre as abelhas provenientes de diferentes regiões. De acordo com a distância genética, dois grupos podem ser distinguidos, um pertencente à subespécie M. q. quadrifasciata (região de Santa Catarina) e outro pertencente à subespécie M. q. anthidioides (demais regiões amostradas).