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The stingless bees Melipona interrupta and Melipona seminigra have great ecological importance as pollinators and economical importance for its honey production. They are associated with a great diversity of entomofauna and microbiota. To understand more about the microbiota associated with these species, this study carried out the morphological and molecular identification of fungi present in the internal hive environment and the brood cells of larvae and pupae (queen and workers) of M. interrupta and M. seminigra, during rainy and dry seasons. Isolated fungi were identified by two methods: analysis of morphological structures and sequencing of the rDNA ITS1-5.8S-ITS2 region. A total of 1.542 isolates were obtained. The filamentous fungi found were: Aspergillus sp., Aspergillus fumigatus, Aspergillus niger, Aspergillus nomius, Aspergillus ochraceus, Cladosporium cladosporioides, Monascus ruber and Penicillium citrinum. The yeasts found were: Cystobasidium minutum, Debaryomyces hansenii, Starmerella apicola, Starmerella etchellsii and Wickerhamiella versatilis. Yeasts were found only in brood cells. The greatest diversity of fungi was observed in the rainy season for M. seminigra, and in the dry season for M. interrupta. There was a high overlap of fungi species associated with both species, but there were also specific associations. Starmerella etchellsii was found only in worker cells of M. interrupta, while Cystobasidium minutum in worker cells of M. seminigra. These findings raise questions about the different types of relationships that microbial communities have with bees in their different microenvironments, as well as with their castes, opening new and intriguing avenues of investigation.

MeliponaIlliger, 1806 is represented by 74 known species of stingless bees, distributed throughout the Neotropical region. Cytogenetically it is the most studied stingless bee genus of the tribe Meliponini. Member species are divided in two groups based on the volume of heterochromatin. This study aim was to analyze the composition and organization of chromatin of the stingless bee subspeciesMelipona seminigra merrillaeCockerell, 1919 using classical and molecular cytogenetic techniques, so contributing to a better understanding of the processes of chromosomal changes within the genus. We confirm thatM. seminigra merrillaehas a chromosome number of 2n = 22 and n = 11, results that differ from those reported for the genus in the absence of B chromosomes. The heterochromatic pattern revealed a karyotype composed of chromosomes with a high heterochromatin content, which makes it difficult to visualize the centromere. Silver nitrate impregnation (Ag-NOR) showed transcriptionally active sites on the second chromosomal pair. Staining of base-specific fluorophores DAPI-CMA3indicated a homogeneous distribution of intensely DAPI-stained heterochromatin, while CMA3markings appeared on those terminal portions of the chromosomes corresponding to euchromatin. Similar to Ag-NOR, fluorescence in situ hybridization (FISH) with 18S ribosomal DNA probe revealed distinct signals on the second pair of chromosomes. Microsatellite mapping (GA)15showed markings distributed in euchromatic regions, while mapping with (CA)15showed marking patterns in heterochromatic regions, together with a fully marked chromosome pair. Microsatellite hybridization, both in heterochromatic and euchromatic regions, may be related to the activity of transposable elements. These are capable of forming new microsatellites that can be dispersed and amplified in different regions of the genome, demonstrating that repetitive sequences can evolve rapidly, thus resulting in within-genus diversification.

Background Pot-pollen is a fermented product stored by stingless bees in cerumen pots and traditionally used as food or medicine by natives in tropical regions. The knowledge of pot-pollen composition from the Amazon region is important to strengthen the breeding of native bees and consequently contribute to sustainable development in this region. The aim of the present study was to determine the chemical composition of Melipona seminigra pot-pollen from Amazonas, Brazil. Results We report the identification of 21 phenolic compounds using UPLC-ESI-QToF-MS. Overall, the predominant constituents of pot-pollen were protein (333.8-470.7 g kg-1 ) and dietary fibers (247.6-353.3 g kg-1 ). The predominant fatty acids were polyunsaturated (44.95-59.57% of total fatty acid content) and the samples contained all essential amino acids. Total carotenoids content (TCC) ranged from 3.2-48.0 μg g-1 , total flavonoids content (TFC) from 1.9-4.5 mg CAE g-1 , total reducing capacity (TRC) from 7.0-15.0 mg GAE g-1 , ferric-reducing antioxidant power (FRAP) from 2.8-8.9 μmol TE g-1 and oxygen radical absorbance capacity (ORAC) from 224.9-1117.0 μmol TE g-1 . Conclusions This study reports for the first time a comprehensive chemical characterization of M. seminigra pot-pollen. The samples presented antioxidant activity, high values of nutrients and bioactive compounds such as polyphenols, carotenoids, insoluble dietary fibers, polyunsaturated fatty acids, and essential amino acids, comparable to other health foods. This article is protected by copyright. All rights reserved.

The Melipona fasciculata is a stingless bee and her honey has properties and flavor appreciated by different consumers. Its production provides a source of income for poor communities in Maranhão, Brazil. However, the absence of Brazilian legislation for this type of honey, as it exists for Apis mellifera, prevents the expansion of the commercialization of M. fasciculata honey. This study was carried out to identify microbiological and physicochemical patterns of honey in seven important locations in the Baixada Maranhense, in the north of Brazil. The honeys showed good hygienic-sanitary quality, in line with Brazilian legislation for A. mellifera honey and the proposal for regulation of stingless bee (SLB) honey. However, 16% of samples belonging to five locations showed contamination by mesophilic bacteria. The physicochemical parameters were variable, mainly moisture indexes (%), with means between 19.35 to 21.22. The HMF content of São Bento (70.70 mg.kg−1) was above current Brazilian legislation. Total acidity showed higher averages in half of São Bento, Viana and Palmeirândia when compared to legislation and the SLB (Max: 50 mEq.kg−1). As for the principal component analysis, HMF and TA showed a positive correlation. With regards to reducing sugar content, only the mean in Peri-Mirim city reached values registered with the Apis legislation and the proposed SLB. In general, the honeys in this study showed rates compatible with those of other SLB, however, in disagreement with parameters required by the Brazilian regulation of Apis, motivating the establishment of specific legislation for honey in Maranhão.

Transposable elements have driven genome evolution and plasticity in many ways across a range of organisms. Different types of biotic and abiotic stresses can stimulate the expression or transposition of these mobile elements. Here, we cytogenetically analyzed natural fish populations of the same species living under different environmental conditions to test the influence and organization of transposable elements in their genome. Differential behavior was observed for the markers Rex 1, Rex 3, and Rex 6 in the chromosomes of individuals of the same species but coming from different environments (polluted and unpolluted). An increase in the number of Rex transposable elements in the chromosomes and their influence on the genome of populations living in a polluted environment indicates that they must be under constant adaptive evolution.

Social organization in highly eusocial bees relies upon two important processes: caste differentiation in female larvae, and age polyethism in adult workers. Juvenile Hormone (JH) is a key regulator of both processes. Here we investigated the expression of two genes involved in JH metabolism - mfe (biosynthesis) and jhe (degradation) - in the context of social organization in the stingless bee Melipona interrupta. We found evidence that the expression of mfe and jhe genes is related to changes in JH levels during late larval development, where caste determination occurs. Also, both mfe and jhe were upregulated when workers engage in intranidal tasks, but only jhe expression was downregulated at the transition from nursing to foraging activities. This relation is different than expected, considering recent reports of lower JH levels in foragers than nurses in the closely related species Melipona scutellaris. Our findings suggest that highly eusocial bees have different mechanisms to regulate JH and, thus, to maintain their level of social organization.

As rainhas virgens, também chamadas de “princesas”, de Melipona são uma das castas físicas essenciais que compõem a sociedade das abelhas-sem-ferrão. Na colônia ou colmeia, são reprodutoras potenciais que, em conjunto com os machos, têm a função primordial de perpetuar a espécie nos ambientes onde habitam. Melipona eburnea apresenta ampla distribuição geográfica abrangendo Brasil, Bolívia, Colômbia, Equador, Peru e Venezuela. No Brasil (estados de Acre y Amazonas) sendo apontada como muito promissora para a meliponicultura pela produtividade e facilidade da sua criação e manejo. Assim, o objetivo deste trabalho foi realizar um estudo de caso observacional para conhecer, na estação chuvosa, em condições de laboratoriais, aspectos biológicos e comportamentais de três rainhas virgens desta espécie. Para isso, a colônia original foi adaptada à caixa de observação e em seguida, um disco de cria nascente foi retirado da mesma e colocado em condições ideais de temperatura e umidade relativa. Enseguida ao nascimento, três rainhas virgens foram marcadas no tórax, com etiquetas coloridas e numeradas, usando-se cola atóxica e colocadas na colônia para serem aceitas pelas outras abelhas. As observações foram feitas entre fevereiro e maio de 2005, pela manhã, entre as 07:00 - 09:00h e entre as 10:00 - 12:00h. Registramos que a longevidade ou sobrevivência máxima na colônia, sem serem mortas ou expulsas, foi de 10 dias, variando de 8-10 dias. Constatamos que, uma vez que deixavam a colônia, nunca mais retornavam. As atividades comportamentais realizadas durante esse período foram classificadas em três categorias: autolimpeza, mobilidade e imobilidade. Destas, a mais representativa foi a mobilidade, seguida da imobilidade e autolimpeza.

Males can control female reproduction using genital plugs to impede access by rivals. In social bees, ants, and wasps, plugging may involve traumatic mating, with females being harmed. In stingless bees, chances are that plugs may promote ovarian activan, and are thought to ensure single mating?a general tendency among the social Hymenoptera. However, understanding on relationships between mating plugs, traumatic mating, and mating systems in stingless bees remains limited. To address this, we (1) compared mated queens of 7 Neotropical species to understand the patterns of copulatory marks in females and (2) compared pre- and post-mating genitalia of males and females in Melipona fasciculata to depict plug functional morphology. Data revealed an unprecedented consequence of mating in stingless bees: the characteristic marks left by mating plugs on female abdomens and the inferences that can be made from them. To our surprise, in 1 species M. fasciculata we found that queens retain the plug long after mating, and may carry it for the rest of their lives. All the other 6 species retained the plug for only a short period. Remated queens were only found in M. seminigra, whose multiple copulatory marks match previous findings of polyandry in this species. Our study shows that queens can remate, and suggests that male genital morphology may determine in part the time persistence of plugs. We conclude that traumatic mating plugs do not fully prevent remating in stingless bees and that mating systems are not uniform in this group. Nonetheless, exceptional cases of facultative polyandry in social insects?for example, when mating plugs fail?may confirm a general tendency for single mating in close link with efficient mating plugs. Made available in DSpace on 2022-03-12T02:25:33Z (GMT). No. of bitstreams: 1 Menezes-Queens-remate-2021.pdf: 832682 bytes, checksum: 5f2af29141433e0774e54690deb690bb (MD5) Previous issue date: 2021

A meliponicultura cresce no Brasil, e meliponicultores se organizam coletivamente com o intuito de fortalecimento. Esse trabalho avaliou a meliponicultura em dois grupos de produtores do Amazonas. Foram entrevistadas 18 pessoas, 8 em MaraA£ e 10 em Boa Vista do Ramos (BVR). Abordou-se desde a razA£o do produtor em iniciar na atividade, atA© o retorno e dificuldades em relaA§A£o A meliponicultura. Os nA­veis organizacionais dos produtores diferem entre os municA­pios, havendo uma cooperativa em BVR e produtores individuais em MaraA£. A geraA§A£o de renda foi um dos principais motivos para o inA­cio na atividade. Observou-se diferenA§as relevantes quanto as dificuldades e anseios apontados pelos grupos. A atividade tem o potencial real de geraA§A£o de renda e de fortalecimento da identidade cultural dos produtores da regiA£o amazA´nica. O estado do Amazonas deve usar esses pontos para elaborar programas de fortalecimento da meliponicultura e elaborar de uma legislaA§A£o especA­fica para a atividade.

Pollution is a growing environmental problem throughout the world, and the impact of human activities on biodiversity and the genetic variability of natural populations is increasingly preoccupying, given that adaptive processes depend on this variability, in particular that found in the repetitive DNA. In the present study, the mitochondrial DNA (COI) and the distribution of repetitive DNA sequences (18S and 5S rDNA) in the fish genome were analysed in fish populations inhabiting both polluted and unpolluted waters in the northern Amazon basin. The results indicate highly complex ribosomal sequences in the fish genome from the polluted environment because these sequences are involved primarily in the maintenance of genome integrity, mediated by a systematic increase in the number of copies of the ribosomal DNA in response to changes in environmental conditions.

Temperature is a major driver of biological phenomena, from metabolism to ecological interactions and rates of evolutionary diversification. However, species vary greatly in their thermal tolerance, as well as the temperature under which they perform best. This study aimed to investigate the effect of experimental manipulation of environmental temperatures on the individual mortality and phenotypic composition of colonies of Melipona interrupta. To fulfill these objectives, 30 colonies in equivalent developmental conditions were artificially subjected to different temperatures. Temperatures were monitored by thermo-hygrometers, and immature mortality and sex and caste ratios were observed in brood combs during 14 months. A strong effect of external temperature on immatures was detected on deviations from 28 to 30 °C (the natural average temperature inside the colony), causing an increase in mortality. Likewise, a significant effect of temperature on sex ratio was detected, with male:female ratio decreasing at temperatures below and above 28–30 °C. Lastly, there was no clear evidence for an effect of temperature on caste ratio, although queens appeared to become relatively more frequent at warmer temperatures. The results of this study allow us to conclude that anthropogenic changes, whose effect can be extrapolated to the similar natural changes, that modify the environmental temperatures to which M. interrupta colonies are exposed are likely to compromise their survival, mainly through individual mortality.

In highly eusocial insects, development of reproductive traits are regulated not only by sex determination pathway, but it also depends on caste fate. The molecular basis of both mechanisms in stingless bees and possible interaction with each other is still obscure. Here, we investigate sex determination in Melipona interrupta, focusing on characterization and expression analysis of the feminizer gene (Mi-fem), and its association to a major component of caste determination, the juvenile hormone (JH). We present evidence that Mi-fem mRNA is sex-specifically spliced in which only the female splice variant encodes the full length protein, following the same principle known for other bee species. We quantified Mi-fem expression among developmental stages, sexes and castes. Mi-fem expression varies considerably throughout development, with higher expression levels in embryos. Also, fem levels in pupae and newly emerged adults were significantly higher in queens than workers and males. Finally, we ectopically applied JH in cocoon spinning larvae, which correspond to the time window where queen/worker phenotypes diverge. We observed a significantly increase in Mi-fem expression compared to control groups. Since up to 100% of females turn into queens when treated with JH (while control groups are composed mainly of workers), we propose that fem might act to regulate queens' development. Our findings provide support for the conserved regulatory function of fem in Melipona bees and demonstrate a significant correlation between key elements of sex and caste determination pathways, opening the avenue to further investigate the molecular basis of these complex traits.

Made available in DSpace on 2018-11-26T17:41:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-12-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) FAPEMA (Fundagao de amparo a pesquisa do Maranhao) The occurrence of Aedes aegypti, Culex quinquefasciatus, and mosquitoes of the genus Anopheles potentiate the spread of several diseases, such as dengue, Zika, chikungunya, urban yellow fever, filariasis, and malaria, a situation currently existing in Brazil and in Latin America. Control of the disease vectors is the most effective tool for containing the transmission of the pathogens causing these diseases, and the bacterium Bacillus thuringiensis var. israelensis has been widely used and has shown efficacy over many years. However, new B. thuringiensis (Bt) strains with different gene combinations should be sought for use as an alternative to Bti and to prevent the resistant insects selected. Aiming to identify diversity in the Bt in different Brazilian ecosystems and to assess the pathogenicity of this bacterium to larvae of Ae. aegypti, C. quinquefasciatus, and Anopheles darlingi, Bt strains were obtained from the Amazon, Caatinga (semi-arid region), and Cerrado (Brazilian savanna) biomes and tested in pathogenicity bioassays in third-instar larvae of Ae. aegypti under controlled conditions in the laboratory. The isolates with larvicidal activity to larvae of Ae. aegypti were used in bioassays with the larvae of C. quinquefasciatus and An. darlingi and characterized according to the presence of 14 cry genes (cry1, cry2, cry4, cry10, cry11, cry24, cry32, cry44Aa, cry1Ab, cry4Aa, cry4Ba, ctyl0Aa, cry11Aa, and cry11Ba), six cyt genes (cyt1, cyt2, cyt1Aa, cyt1Ab, cyt2Aa and cyt2Ba), and the chi gene. Four hundred strains of Bt were isolated: 244 from insects, 85 from Amazon soil, and 71 from the Caatinga biome. These strains, in addition to the 153 strains isolated from Cerrado soil and obtained from the Entomopathogenic Bacillus Bank of Maranhao, were tested in bioassays with Ae. aegypti larvae. A total of 37 (6.7%) strains showed larvicidal activity, with positive amplification of the cry, cyt, and chi genes. The most frequently amplified genes were cry4Aa and cry4Ba, both occurring in 59.4% in these strains, followed by cyt1Aa and cyt2Aa, with 56.7% and 48% occurrence, respectively. Twelve (2.2%) strains that presented 100% mortality within 24 h were used in bioassays to estimate the median lethal concentration (LC50) for Ae. aegypti larvae. Two strains (BtMA-690 and BtMA-1114) showed toxicity equal to that of the Bti standard strain, and the same LC50 value (0.003 mg/L) was recorded for the three bacteria after 48 h of exposure. Detection of the presence of the Bt strains that showed pathogenicity for mosquito larvae in the three biomes studied was possible. Therefore, these strains are promising for the control of insect vectors, particularly the BtMA-1114 strain, which presents a gene profile different from that of Bti but with the same toxic effect. Univ Fed Maranhao, Curso Ciencias Nat, Campus 7,Ave Dr Jose Anselmo 2008, BR-65400000 Codo, Maranhao, Brazil Inst Natl Pesquisas Amazonia, Lab Malaria & Dengue, Programa Posgrad Entomol, Ave Andre Arauljo 2936, BR-69067375 Manaus, Amazonas, Brazil Univ Estadual Maranhao, Ctr Estudos Super Caxias, Dept Quim & Biol, Lab Entomol Med, Praca Duque Caxias S-N, BR-65604380 Caxias, Maranhao, Brazil Univ Estadual Paulista, Dept Fitossanidade, Lab Controls Microbiano Artropodes Praga, Via Acesso Paulo Donato Castelllane S-N, BR-14884900 Sao Paulo, Brazil Inst Nacl de Pesquisas da Amazonia, Grpo Pesquisas Abelhas, Programa Posgrad Entomol, Ave Andre Araujo 2936, BR-69067375 Manaus, Amazonas, Brazil Univ Estadual Paulista, Dept Fitossanidade, Lab Controls Microbiano Artropodes Praga, Via Acesso Paulo Donato Castelllane S-N, BR-14884900 Sao Paulo, Brazil

The HpaII/MspI-PCR is a method based on the sensitivity of restriction enzymes to methylation at their cleavage sites, which permits the comparison of the DNA methylation status of different organisms, based on the differential patterns of digestion. Prior to the present study, nothing was known of the methylation profile of specific DNA sequences in Amazonian fish based on enzymes sensitive to methylation. Given this, the present study investigated the methylation profile of the 18S rRNA gene of the armored catfish, Hoplosternum littorale, which inhabits relatively inhospitable environments in the Amazon basin. A differential methylation profile was found in the 18S rRNA sites, indicating that polluted environments may influence the methylation of DNA, and would have evolved in the species, initially, as a defense mechanism against the expression of transposable elements.

SummaryThe knowledge of digestive processes correlated with the species of plant used by stingless bees is important for development of Meliponiculture, or stingless bee keeping, which is a lucrative activity in the Brazilian Amazon. The number and state of digestion of pollen grains in the midgut and rectum, as well as the pollinic diversity and activity of some digestive enzymes, were analyzed in different life stages of the stingless bee Melipona seminigra to compare the difference of digestion process in larvae, adult workers (nurse and forager) and virgin queens of Amazon stingless bees. Pollen consumption was found to be similar for nurse workers and virgin queens, while larvae consume significantly more pollen than other stages. A survey of pollinic diversity identified eight plant families, with the majority of digested pollen collected from the Melastomataceae (97.85%), and most undigested pollen from the Fabaceae (51%). The activity of hydrolytic enzymes was analyzed in larval food, pollen store...

The purpose of this study was to determine the physicochemical characteristics of pollen collected by the Amazonian stingless bees Melipona seminigra and Melipona interrupta , in order to verify whether their characteristics meet the physicochemical requirements established by the Brazilian Technical Regulation for Identity and Quality of Bee Pollen. Physicochemical analyses were performed through official analytical methods. Results of pollen analyses collected by M. seminigra and M. interrupta were respectively as follows: moisture: 53.39 and 37.12%; protein: 37.63 and 24.00%; lipids: 10.81 and 6.47%; ash: 4.03 and 2.74%; crude fiber: 9.30 and 13.65%; carbohydrates: 25.66 and 44.27%; energy: 350.47 and 331.33kcal%; pH: 3.70 and 3.34; total solids: 46.60 and 62.87%, and water activity: 0.91 and 0.85. The percentages of moisture and pH in pollen collected by both studied bees are not in agreement with the Technical Regulation for bee pollen. Since some characteristics, which are inherent to the Melipona pollen, were not in conform to the current Regulation, we recommend that further studies should be conducted to better characterize it, and correct the Regulation, if necessary. RESUMO: O objetivo deste estudo foi determinar as características físico-químicas do pólen coletado pelas abelhas amazônicas sem ferrão, Melipona seminigra e Melipona interrupta , para verificar se suas características atendem aos requisitos físico-químicos estabelecidos no Regulamento Técnico Brasileiro de Identidade e Qualidade de Pólen Apícola. As análises físico-químicas foram realizadas através de métodos analíticos oficiais. Os resultados das análises do pólen coletado por M. seminigra e M. interrupta foram, respectivamente: umidade: 53,39 e 37,12%; proteínas: 37,63 e 24,00%; lipídeos: 10,81 e 6,47%; cinzas: 4,03 e 2,74%; fibra bruta: 9,30 e 13,65%; carboidratos: 25,66 e 44,27%; energia: 350,47 e 331,33kcal%; pH: 3,70 e 3,34; sólidos totais: 46,60 e 62,87%; e atividade de água: 0,91 e 0,85. O percentual de umidade e o pH do pólen coletado por ambas as abelhas estudadas está em desacordo com o Regulamento Técnico para pólen apícola. Recomendamos que sejam realizados mais estudos para melhor caracterizar este produto e atualizar o Regulamento vigente, se necessário, pois algumas características que não atendem ao Regulamento são inerentes ao pólen de Melipona.

Poliginia, caracterizada pela presença de mais de uma rainha poedeira, é considerada como uma condição temporária em colônias. Em abelhas sem ferrão isso é raramente observado. Este artigo registra o primeiro caso de poliginia natural em colônia de Melipona scutellaris, com cinco rainhas poedeiras. Polygyny, characterized by the presence of several egg-laying queens, is considered as a temporary colony status. In stingless bees it is rarely observed. This paper reports the first case of natural polygyny in Melipona scutellaris colony, with five egg-laying queens.

Melipona interrupta manaosensis is an important pollinator in the Neotropics. This stingless bee is reared by locals for honey production and also for conservation purposes. Eleven microsatellite markers were isolated from M. i. manaosensis and characterized in 32 individuals from the Meliponary of the Instituto Nacional de Pesquisas da Amazonia, Manaus, AM, Brazil. The number of alleles observed for each locus ranged from 2 to 5 (average 2.77). The polymorphism information content (PIC) varied between 0.08 to 0.62 (average 0.34) and the discriminating power (D) ranged from 0.61 to 0.99 (average 0.85). The observed heterozygosity (HO) and the expected heterozygosity (HE) ranged from 0.04 to 1.00 and 0.04 to 0.67, respectively. These microsatellites will be useful for future genetic studies of M. i. manaosensis populations and correlated species contributing with Amazon biodiversity preservation.

Melipona seminigra merrillae is one of the main pollinators of Central Amazon native flora. These stingless bees are reared as economic alternative and for forest conservation purposes. Nine polymorphic microsatellite markers were characterized in 44 genotypes of M. seminigra merrillae from the Meliponary of the Instituto Nacional de Pesquisas da Amazonia, INPA (3° 05.838′S 059° 59.103′W), Manaus, Brazil. The number of alleles observed for each locus ranged from 3 to 8, with an average of 3.7 alleles per locus. The polymorphism information content (PIC) varied between 0.21 and 0.81 (average 0.50) and the discriminating power (D) ranged from 0.59 to 0.98 (average 0.77). The observed heterozygosity (H o) and the expected heterozygosity (H e) were 0.04–0.68 and 0.19–0.84, respectively. A cross-amplification test in three Melipona species suggested potential transferability of these microsatellites. These microsatellite markers should be useful for characterizing natural populations and specie conservation.

Este trabalho relata detalhes da vida reprodutiva de duas espécies de abelhas sem ferrão. Rainhas velhas de Melipona compressipes fasciculata, no Maranhão, morrem e são substituídas com sucesso em todos os meses. Rainhas de Melipona scutellaris, trazidas de Lençóis (BA - nordeste do Brasil) para Uberlândia (MG, centro-sudeste do Brasil) morreram durante todos os meses e não mostraram a existência de trimestre preferencial para as novas rainhas iniciarem postura. Quarenta machos de M. scutellaris, após serem marcados no tórax e libertados em grupos de 10 a 100, 400, 800 e 1000 metros do meliponário, tiveram seus retornos observados. Todos os machos libertados a 100 e 400 metros regressaram ao meliponário, 7 de 10 machos e 2 de 10 machos retornaram de 800 e 1000 metros, respectivamente. Os machos esperam constantemente pela saída de uma rainha virgem, próximos às colônias órfãs, o que indica que a maioria das rainhas é inseminada próximo aos seus ninhos, portanto, a dispersão dos genes depende do vôo dos machos e da distância de enxameagem para ocupação de uma cavidade para o novo ninho. This work reports on the reproductive life of the stingless bees (Meliponini). Old queens of Melipona compressipes fasciculata in Maranhão State, Brazil die and are successfully substituted by new ones in all months. Queens of Melipona scutellaris from Lençois (BA - northeastern Brazil) brought to Uberlândia (Central South eastern Brazil) died during all months, and there are no months where new queens are more able to begin egg laying. Forty males of M. scutellaris were marked in the thorax and released in groups of 10 at 100, 400, 800 and 1000 meters from the meliponary and their return to it was observed. All males released at 100 to 400 meters returned, 7 out of 10 and 2 out of 10 males returned respectively from 800 and 1000 meters. Males are constantly waiting for a virgin queen next to orphan colonies, the majority of the queens is inseminated in near the hive, therefore, gene dispersion depends more upon the males and of the occupation of a new cavity for nest.

In countries containing a mega diversity of wildlife, such as Brazil, identifying and characterizing biological diversity is a continuous process for the scientific community, even in face of technological and scientific advances. This activity demands initiatives for the taxonomic identification of highly diverse groups, such as stingless bees, including molecular analysis strategies. This type of bee is distributed in all of the Brazilian states, with the highest species diversity being found in the State of Amazônia. However, the estimated number of species diverges among taxonomists. These bees are considered the main pollinators in the Amazon rainforest, in which they obtain food and shelter; however, their persistence is constantly threatened by deforestation pressure. Hence, it is important to classify the number and abundance of bee specie, to measure their decline and implement meaningful, priority conservation strategies. This study aims to maximize the implementation of more direct, economic and successful techniques for the taxonomic identification of stingless bees. Specifically, the genes 16S rRNA and COI from mitochondrial DNA were used as molecular markers to differentiate 9 species of Amazonian stingless bees based on DNA polymorphism, using the polymerase chain reaction-single-strand conformation polymorphism technique. We registered different, exclusive SSCP haplotypes for both genes in all species analyzed. These results demonstrate that SSCP is a simple and cost-effective technique that is applicable to the molecular identification of stingless bee species. © FUNPEC-RP.

Méis são descritos possuindo diferentes propriedades, incluindo a antimicrobiana. Muitos estudos têm apresentado essa atividade de méis produzidos por abelhas Apis mellifera, no entanto estudos incluindo atividades de méis de abelhas sem ferrão são escassos. O objetivo deste estudo foi comparar a atividade antimicrobiana de méis de Melipona compressipes, Melipona seminigra e A. mellifera, coletados no Estado do Amazonas, contra Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Chromobacterium violaceum, e Candida albicans. As concentrações inibitórias mínimas foram determinadas usando o método de diluição em ágar, com ágar Muller-Hinton (para bactérias) ou ágar Sabouraud (para a levedura). S. aureus e E. faecalis foram inibidos por todos os méis em concentrações inferiores a 12%, enquanto E. coli e C. violaceum foram inibidos por méis de abelhas sem ferrão em altas concentrações entre 10 e 20%. A. mellifera inibiu E. coli na concentração de 7% e C. violaceum em baixa concentração (0,7%). C. albicans foi inibida apenas em concentrações entre 30 e 40% dos méis. Assim, todas as variedades de mel testadas apresentaram atividade antimicrobiana sobre os patógenos testados, servindo assim como agente antimicrobiano potencial para diversas abordagens terapêuticas. Honeys are described possessing different properties including antimicrobial. Many studies have presented this activity of honeys produced by Apis mellifera bees, however studies including activities of stingless bees honeys are scarce. The aim of this study was to compare the antimicrobial activity of honeys collected in the Amazonas State from Melipona compressipes, Melipona seminigra and Apis mellifera against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Chromobacterium violaceum, and Candida albicans. Minimum inhibitory concentrations were determined using the agar dilution method with Müller-Hinton agar (for bacteria) or Saboraud agar (for yeast). Staphylococcus aureus and E. faecalis were inhibited by all honeys at concentrations below 12%, while E. coli and C. violaceum were inhibited by stingless bee honeys at concentrations between 10 and 20%. A. mellifera honey inhibited E. coli at a concentration of 7% and Candida violaceum at 0.7%. C. albicans were inhibited only with honey concentrations between 30 and 40%. All examined honey had antimicrobial activity against the tested pathogens, thus serving as potential antimicrobial agents for several therapeutic approaches.

We investigated the phenolic constituents and antioxidant activity of geopropolis from two species of stingless Amazonian bees, Melipona interrupta and Melipona seminigra. The chemical investigation of geopropolis from Melipona interrupta led to the isolation of 5,7,4'-trihydroxyflavonone, 3,5,6,7,4'-pentahydroxyflavonol, naringenine-4'-O-b-D-glucopyranoside and myricetin-3-O-b-D-glucopyranoside. Their structures were assigned based on spectroscopic analyses, including two-dimensional NMR techniques. Antioxidant activity of methanol and ethanol extracts of M. interrupta and M. seminigra were measured using the 1,2-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay. This is also the first work reporting the chemical investigation of stingless bee species from the Amazonian region.

In colonies of Melipona scutellaris Latreille, 1811 workers can be found with four ganglion nerve cells, a morphological characteristic of the queen. It is hypothesized that these workers, called intercastes, or phenocopies, are phenotypically-like workers, but genotypically identical to queens due to this specific trait. Workers with the same number of ganglion as queens seem to be intercastes between queens and workers. Our objective was to analyze the mRNA pro files of workers, queens, and intercastes of M. scutellaris through DDRT-PCR. Three hundred (300) pupae with white eyes were collected and externally identified according to the number of abdominal nerve ganglions: workers (5 ganglions), queens (4 ganglions) and intercastes (4 ganglions). The analysis identified differentially expressed transcripts that were present only in workers, but absent in intercastes and queens, confirming the hypothesis, by demonstrating the environmental effect on the queen genotype that generated phenotype-like workers.

Many factors have contributed to reductions in wild populations of stingless bees, such as: deforestation, displacement and destruction of nests by honey gatherers, as well as use of insecticides and other agrochemicals. All of these can potentially affect the populational structure of native species. We analyzed genetic variability and populational structure of Melipona scutellaris, based on five microsatellite loci, using heterologous primers of M. bicolor. Samples were taken from 43 meliponaries distributed among 30 sites of four northeastern states of Brazil (Pernambuco, Alagoas, Sergipe, and Bahia). Thirty-one alleles were found to be well distributed among the populations, with sizes ranging from 85 to 146 bp. In general, there was a variable distribution and frequency of alleles among populations, with either exclusive and/or fixed alleles at some sites. The population of Pernambuco was the most polymorphic, followed by Bahia, Alagoas and Sergipe. The heterozygosity was Ho = 0.36 on average, much lower than what has been reported for M. bicolor (Ho = 0.65). Most populations were not under Hardy-Weinberg equilibrium. We found a higher variation within rather than among populations, indicating no genetic structuring in those bees maintained in meliponaries. This apparent homogenization may be due to intense beekeeping activity, including exchange of genetic material among beekeepers. Based on our findings, we recommend more studies of meliponaries and of wild populations in order to help orient management and conservation of these native pollinators.

O conhecimento de substâncias repelentes para forídeos é um passo importante para a meliponicultura brasileira, pois esses insetos podem causar sérios danos às colméias de abelhas nativas. Os óleos de copaíba e andiroba, naturalmente encontrados na região amazônica, são muito utilizados pelos povos tradicionais da região como repelentes de insetos. Foi observado o efeito de dois óleos vegetais (andiroba e copaíba) sobre a postura de ovos por fêmeas de forídeos em condições de laboratório. A postura das fêmeas foi realizada preferencialmente no substrato pólen e diferiu estatisticamente dos substratos contendo óleo de andiroba ou copaíba, nos quais houve considerável diminuição (até nenhuma postura), e do substrato contendo mel. Esses óleos são uma boa alternativa no controle preventivo e curativo dessa praga em colônias de Meliponineos, devido ao seu efeito repelente, ao baixo custo e disponibilidade na Região Amazônica. The knowlegment of repellants substances to phoride flies is an important step to the Brazilian stinglessbee beekeeping due to the serious damage to the native bee hives they can do. The copaiba and andiroba oils, found commonly in the Amazon region, are very employed by the traditional people as insects' repellants. We observed the effect of these oils over the phoride's laying in laboratory conditions. The female phoride's eggs occurred preferentially in the pollen substrate and statistically differed from those containing andiroba and copaiba oils, which was strongly reduced (until none eggs were laid), and from the substrate containing honey. These oils showed to be quite good alternative for this plague's preventive and curative control in stingless bee hives due to its repellant effect, low cost, and availability to get it in the region.", 'en

Cytogenetic analysis of the Amazon stingless bee Melipona seminigra merrillae, by conventional Giemsa staining and C-banding, revealed a different chromosome number for Melipona: 2n = 22 for females and diploid drones while the haploid drones present n = 11. There is no evidence of B chromosomes. This result contrasts with previous studies, in which the chromosome number of 19 Melipona species was determined as 2n = 18 for females and n = 9 for haploid males. Based on cytogenetic information available for other Melipona species, we propose that M. s. merrillae has a more derived diploid number. This indicates that chromosome number is not a conservative characteristic within the genus as previously thought. Cytogenetic data for stingless bees are scarce, especially in Amazon region. Additional studies will be very important in order to promote Melipona karyoevolution discussion and consequently a taxonomy review.