Analysis of the Hive Yield of Three Species of Bees for Honey Production in Eastern Honduras

Objectives: The analysis of the performance of honey production hives of three species of honeybees was carried out. Study Design: Three harvests per year of honey per hive of each species the first was on March-April, the second was on June-July, and the third was on November -December, with a sampling of ten hives per species, data were evaluated by ANOVA analysis, and Duncan mean separation test developed by SAS/STAT program. Place and Duration of the Study: This research was developed in the department of Para韘o, municipality of Danl�, Honduras, Central America, in Cerro El 羐uila, latitude 13�'59" N, and longitude 86�'59" W, the sampling was carried out in 2020. Methodology: The qualitative variables were evaluated for each species by In Situ observation that characterized the honeycombs of the three species, and the data were subjected to an analysis of variance and Duncan's mean separation, where the dependent variable is the average yield of Kg of honey/hive. Results: According to the ANOVA, there are significant differences in: the harvest of the three species of bees (p<0.0001), for the three annual harvests (p=0.0024), and by hives (p<0.0001) and in the analysis of separation of means the three averages of honey yield observed are not homogeneous (P>0.05), the Apis mellifera produces 63.1 kg/hive-year, the Tetragonisca angustula obtained an average of 0.679 kg / hive-year, and Melipona beecheii obtained 13.19 kg/hive-year. Conclusion: In the three species is observed that they have differences in the structures of the construction of their hives, their entrance, the construction of their cells, production and storage of honey and pollen, birth of queens, and method of defense; and when evaluating the average annual yield by species, by harvest, and by hives, is found that the three variables evaluated of bees are different averages of honey production.

Propolis from native Stingless Bees: ultrasound-assisted extraction / Propóleos de abejas nativas sin aguijón: extracción asistida por ultrasonido

BACKGROUND: Propolis has been considered a highly valuable material due to its therapeutic properties. However, in Colombia, the commercialization of propolis is limited not only by low production but also by the little knowledge about its efficient extraction. Therefore, finding an optimal and economical extraction method to obtain propolis is a necessity for beekeepers that would open new possibilities for industrial use and, therefore, for the market. OBJECTIVES:The objective of this study was to evaluate a conventional and ultrasound-assisted extraction method, seeking to obtain the highest yield and a high amount of content of bioactive compounds in propolis extracts. METHODS: The extraction was carried out for three crude propolis from different types of bees: Tetragoniscaangustula or Angelita (ANG), Meliponaeburnea or Melipona(MEL), and Scaptotrigonaspp (SCT). The extracts were characterized by color, pH, visual appearance, solid content, antioxidant capacity, total polyphenol content, and bacterial inhibition capacity. RESULTS: The highest extraction performance was obtained when the ultrasound-assisted method was used, especially for the ANG extract, which in addition to presenting inhibition for gram-negative (E. coli) and gram-positive (S. Aureus) bacteria, had the best antioxidant activity with a value of 545 mg GAE / 100 g of sample and total polyphenol content of 1,884 mg GAE / 100 g of sample. CONCLUSIONS: Ultrasound-assisted extraction can be considered a low-cost alternative to increase the extraction performance of crude propolis, together with its total polyphenol content and antioxidant capacity, without altering its physical properties

ANTECEDENTES: El propóleos ha sido considerado un material de alto valor por sus propiedades terapéuticas. Sin embargo, en Colombia la comercialización de propóleos está limitada no solo por la baja producción sino también por el incipiente conocimiento sobre la extracción eficiente de este. Por ello, encontrar un método de extracción óptimo y económico para la obtención de propóleos es una necesidad para los apicultores que abriría nuevas posibilidades para el uso industrial y por tanto para el mercado. OBJETIVOS: El objetivo de este estudio fue evaluar un método de extracción convencional y asistido por ultrasonido (US) buscando el mayor rendimiento y alto contenido de compuestos bioactivos en extractos de propóleos. MÉTODOS: La extracción se realizó para tres propóleos crudos de diferentes tipos de abejas Tetragonisca angustula o Angelita(ANG), Melipona eburnea o Melipona (MEL) y Scaptotrigona spp (SCT). Todos los extractos se caracterizaron por su color, pH, apariencia visual, contenido de sólidos, capacidad antioxidante, contenido total de polifenoles y capacidad de inhibición bacteriana. RESULTADOS: El mayor rendimiento de extracción se obtuvo cuando se usó el método asistido por ultrasonido y específicamente para el extracto ANG, que además de presentar inhibición para bacterias gram negativas (E. coli) y gram positivas (S. Aureus), tuvo la mejor actividad antioxidante con un valor de 545 mg GAE / 100 g de muestra y contenido total de polifenoles de 1884 mg GAE / 100 g de muestra. CONCLUSIONES: La extracción asistida por ultrasonido puede considerarse una alternativa de bajo costo para aumentar el rendimiento de extracción del propóleos crudo, así como su contenido total de polifenoles y capacidad antioxidante sin alterar sus propiedades físicas

Efficiency of two digestion methods in determining the presence of metals (Cd, Cu, Cr, Pb and Zn) in geopropolis produced by Melipona scutellaris / Eficacia de dos métodos de digestión para determinar metales (Cd, Cu, Cr, Pb y Zn) en geopropóleos producidos por Melipona scutellaris / Eficiência de dois métodos de digestão para determinar metais (Cd, Cu, Cr, Pb e Zn) na geoprópolis produzida por Melipona scutellaris

Abstract This study evaluates the efficiency of acid extraction and total digestion to determine the presence of metals in geopropolis produced by the stingless bee Melipona scutellaris. Geopropolis samples were collected at five meliponaries in the city and in the metropolitan region of Salvador, Bahia State, Brazil. The sample treatment methods comprised acid extraction and total digestion. The Inductively Coupled Plasma Optical Emission Spectrometry (ICP OES) technique was used to quantify Cd, Cu, Cr, Pb, and Zn. Geopropolis samples submitted to both digestion methods showed statistical differences. For both methods, Cr and Zn showed the highest concentrations, while those of Cd were the lowest. The Cr concentration for determination by total digestion was 37.53 mg/kg, while for acid extraction it was 32.90 mg/kg. For Zn, the concentration was 17.65 mg/kg and 8.85 mg/kg for total digestion and acid extraction, respectively. The total digestion method showed the highest concentrations of the metals evaluated; however, acid extraction (USEPA 3050b) is a more straightforward procedure for metal evaluation in geopropolis samples and presented values that support the use of geopropolis as a bioindicator. The acid extraction method USEPA 3050b, in combination with detection using ICP OES, showed efficiency in analyses carried out to determine metals in geopropolis.

Resumen El objetivo de este artículo fue evaluar la eficacia da extracción ácida y de la digestión total para determinación de metales absorbidos en geopropóleos producidos por Melipona scutellaris. Se colectaron muestras de geopropóleos en cinco meliponarios localizadas en la región metropolitana del Salvador, estado de Bahia, Brasil. Se utilizaron como métodos de tratamiento para muestras, la extracción ácida y la digestión total. Para determinar los metales Cd, Cu, Cr, Pb y Zn se utilizó Espectroscopia de Emisión Óptica con Plasma Acoplado Inductivamente (ICP OES). Se encontraron diferencias estadísticas en la concentración de metales hallados en las muestras de geopropóleos sometidas a los diferentes métodos de digestión. Para ambos métodos de digestión, las mayores concentraciones de metal fueron observadas para Cr y Zn. Entre los metales evaluados Cd presentó la menor concentración. La concentración de Cr para la determinación por digestión total fue de 37,53 mg/kg y para la extracción ácida fue de 32,90 mg/ kg. Para el Zn, la concentración fue de 17,65 mg/kg y 8,85 mg/kg para la digestión total y la extracción ácida, respectivamente. El método de digestión total mostró los mayores valores para concentraciones de los metales evaluados. Entretanto, la extracción ácida, USEPA 3050b, es un procedimiento más simple para la evaluación de metales en muestras de geopropóleos y también mostró valores que pueden satisfacer la necesidad de su uso en evaluaciones de colmenas como bio-indicador. El método de extracción ácida USEPA 3050b en combinación con la detección a través de ICP OES se mostró eficiente para el análisis de metales en geopropóleos.

Resumo O objetivo deste estudo foi avaliar a eficiência da extração ácida e da digestão total para determinação de metais adsorvidos na geoprópolis produzida por Melipona scutellaris. Foram coletadas amostras de geoprópolis em cinco meliponários, situados em Salvador, Estado da Bahia e região metropolitana. Foram utilizados como métodos de tratamento das amostras a extração ácida e a digestão total. Para determinação dos metais Cd, Cu, Cr, Pb e Zn utilizou-se a Espectrometria de Emissão Óptica com Plasma Indutivamente Acoplado (ICP OES). Houve diferença estatística na concentração de metais encontrados nas amostras de geoprópolis submetidas aos diferentes métodos de digestão. Para ambos os métodos de digestão as maiores concentrações de metais foi observada para o Cr e Zn. Dentre os metais avaliados o Cd apresentou a menor concentração. A concentração de Cr para determinação por digestão total foi de 37,53 mg/kg e para extração ácida de 32,90 mg/ kg. Para o Zn, a concentração foi de 17,65 mg/kg e 8,85 mg/kg para digestão total e extração ácida, respectivamente. O método de digestão total apresentou os maiores valores para concentrações dos metais avaliados. No entanto, a extração ácida, USEPA 3050b, é um procedimento mais simples para a avaliação dos metais em amostras de geoprópolis e também apresentou valores que podem satisfazer a necessidade de utilização na avaliação deste produto da colmeia como bioindicador. O método de extração ácida USEPA 3050b em combinação com a detecção por ICP OES mostrou eficiência para análise de metais em geoprópolis.

Evaluation of physic contamiants and contamination with coliforms, molds and yeasts of honey from the Northern Brazil / Avaliação de contaminantes físicos e contaminação com coliformes, bolores e leveduras de mel do Norte do Brasil

The Mercosur Technical Regulation for honey determines microbiological parameters for total coliforms and molds and/ yeasts, and absence of physical contaminants, of any nature. The purpose of this study was to verify the biological and physical contamin of Apis mellifera ligustica and Melipona fasciculata, commercialized in the northeastern region of the State of Pará, Brazil. The fourteen samples from informal trade of the municipalities of Bragança, Capanema, Nova Timboteua, São João de Pirabas, Salinas and Tracauateua were analyzed for the Most Probable Number of colifroms at 35ºC, Fungus and Yeast counts, besides the detection of dirt by Macro and Microscopy techniques. The results obtained from the microbiological analyzes were compared to the standard required by MERCOSUR, for coliforms. 57.14% of the samples were non-standard and ranged from <3.0 NMP/g to 20 NMP/g. The molds and/ yeast counts ranged from 1x10¹ CFU/g to 8.6x10² CFU/g, with atotal of 21.42% of the samples being in disagreement with the legislation. From the macroscopic analysis, it was observed that, although 50% of the samples did not present any apparent dirt, microscopically it was possible to observe dirt in 100% of the samples of honey analyzed, which is out of the standard required by the Brazilian legislation. This shows the low sanitary quality of honey commercialized in this region of the state of Pará.

O Regulamento Técnico do Mercosul para mel, determina parâmetros microbiológicos para coliformes totais e fungos e/leveduras, e ausência de contaminantes físicos, de qualquer natureza. A proposta do presente estudo foi verificar a contaminação biológica e física em méis de abelha das espécies Apis mellifera ligustica e Melipona fasciculata, comercializados na região Nordeste do Estado do Pará, Brasil. As amostras provenientes do comercio informal dos de municípios de, Bragança, Capanema, Nova Timboteua, São João de Pirabas, Salinas e Tracauateua, foram analisadas quanto a determinação do Número Mais Provável de coliformes a 35ºC, Contagem de Fungos e/leveduras, além da detecção de sujidades pelas técnicas de Macro e Microscopia. Os resultados obtidos a partir das análises microbiológicas foram comparadas ao padrão exigido pelo MERCOSUL, para coliformes, 57,14% das amostras encontravam-se fora do padrão variaram de <3,0 NMP/g a 20NMP/g. Já a contagem de fungos e/leveduras variou de 1x10¹ UFC/g a 8,6x10² UFC/g, no total de 21,42 % das amostras apresentaram-se em desacordo com a legislação. A partir da análise macroscópica observou-se que, embora 50% das amostras não apresentassem sujidades aparentes, porém microscopicamente foi possível observar a presença sujidades em 100% das amostras de méis analisadas encontrando-se fora do padrão exigido pela legislação Brasileira. Demostrando a baixa qualidade sanitária do mel comercializado nessa região do estado do Pará.

Geopropolis gel for the adjuvant treatment of candidiasis - formulation and in vitro release assay

ABSTRACT The geopropolis produced by the stingless bee Melipona subnitida (popularly called "jandaíra" in Brazil) is a mixture of resin, wax, and mud. This study analyzed the antifungal activity of the geopropolis extract from Candida spp., developed a gel formulation with this extract and analyzed the delivery of bioactives (kinetics release) in the formulation and their chemical profile by UHPLC-PDA-qTOF-MS/MS. Three different gels were prepared using the geopropolis extract, carbomer, propylene glycol, and water. Formulations with different amounts of propylene glycol were investigated. Physical, visual, pH, viscosity, adhesion, spreadability, leakage, and in vitro release tests were performed in the proposed formulations. Antifungal tests with the geopropolis ethanolic extract were carried out against six Candida species. The chemical profile of the geopropolis extract and compounds released from the formulations were analyzed after the release test. The formulations had a pH between 4.6 and 4.8 and viscosity between 535,600 and 920,400 cPs. The geopropolis extract presented excellent antifungal activity against the tested yeasts. The results of the release test in semipermeable cellulose membrane showed that all formulations containing 5%, 10% and 40% propylene glycol presented release of geopropolis extract. For adhesion and leakage tests, the gel formulation with 5% propylene glycol was more effective. Both geopropolis ethanolic extract and the liquid obtained in the release test showed the presence of flavonoids (flavonol/flavone, flavanone, and chalcones). Gel formulations with geopropolis extract that are rich in flavonoids are promising as an adjuvant treatment of vaginal candidiasis.

Caracterização microbiológica do mel produzido por melíponas no Estado de Alagoas / Microbiological characterization of honey produced by melíponas in the State of Alagoas

O mel de abelhas sem ferrão possui um grande valor agregado, chegando a ultrapassar o preço do mel das abelhas com ferrão (gênero Apis) em até quinze vezes. As melíponas são de suma importância na manutenção de seus ecossistemas de origem, contribuindo de forma para a conservação e a sobrevivência da flora e da fauna. A pesquisa objetivou caracterizar os méis de meliponídeos nativos do Estado de Alagoas. Foram coletadas e analisadas quatro amostras de méis, sendo todas elas obtidas da espécie Melipona scutellaris (mais conhecida como Uruçu) em diferentes municipios, condições e momentos. As amostras estavam próprias para consumo humano e de acordo com a legislação vigente verificou-se que os padrões adotados para o mel produzido por Apis melífera podem ser aplicados para o mel produzido por Meliponas.

Microbiological and physical-chemical characteristics of honeys from the bee Melipona fasciculata produced in two regions of Brazil / Características microbiológicas e físico-químicas de méis de abelha Melipona fasciculata produzidos em duas regiões do Brasil

ABSTRACT: The honey from Tiúba bees (Melipona fasciculata) is commercially important in the Brazilian state of Maranhão. However, the absence of any specific legislation for this kind of honey is an obstacle to its increased production and commercialization. Determining the microbiological and physical-chemical characteristics of different Tiúba honey can inform the elaboration of specific legislation. Thus, honey samples from two Water Catchment Areas in Maranhão (Munim and Pericumã, sample size 20 for each) were collected and submitted for microbiological analysis (total coliforms, thermotolerant coliforms, molds and yeasts, Clostridium sulfite reducers and Salmonella sp.) and physical-chemical analysis (moisture content, reducing and non-reducing sugars, insoluble and soluble solids, ash content, acidity and pH). Most of honeys sampled were suitable for human consumption, except for one sample from Pericumã. Honeys from the two water catchment areas presented differences in some physicochemical characteristics, which can be attributed to the botanic, soil and climatic diversity of the two areas. Still, these Tiúba honey samples did not fall within the Brazilian legislation for Apis mellifera honey for some of the physical-chemical characteristics, and so is not properly regulated, thus reinforcing the need for specific legislation for this type of honey.

RESUMO: O mel de Tiúba (Melipona fasciculata) tem importância comercial na região do estado do Maranhão, no entanto a ausência de uma legislação específica para este mel entrava a ampliação de sua produção e comercialização. Dessa forma, faz-se necessário determinar as características microbiológicas e físico-químicas de méis de Tiúba, com o objetivo de fornecer subsídios para a elaboração de uma legislação específica. Assim, amostras de méis provenientes de duas bacias hidrográficas do Maranhão (Munim n = 20, e Pericumã n = 20) foram coletadas e submetidas às análises microbiológicas (coliformes totais, coliformes termotolerantes, bolores e leveduras, Clostridium sulfito redutores e Salmonella sp.) e físico-químicas (umidade, açúcares redutores e não redutores, sólidos insolúveis e solúveis, cinzas, acidez e pH). A maioria dos méis apresentou-se adequado microbiologicamente ao consumo humano, exceto uma amostra de mel proveniente de Pericumã. Os méis provenientes das duas bacias hidrográficas apresentaram diferenças em algumas características físico-químicas, o que pode ser atribuído à diversidade botânica, edáfica (solo) e climática das duas bacias. Ainda, os méis de Tiúba não se adequaram à legislação brasileira de Apis mellifera para algumas características físico-químicas, o que não permite sua regulamentação e, assim, reforça a necessidade de legislação específica para este tipo de mel.

Foraging behavior, environmental parameters and nests development of Melipona colimana Ayala (Hymenoptera: Meliponini) in temperate climate of Jalisco, México / Comportamento de forragem, parâmetros ambientais e desenvolvimento de ninhos de Melipona colimana (Hymenoptera: Meliponini) em clima temperado de Jalisco, México

Abstract Melipona colimana Ayala is an endemic species inhabiting temperate forests of pine and oak of south of Jalisco in Mexico. During a year, it was recorded every 15 days foraging activity, environmental parameters and the development of colonies of M. colimana in its wild habitat. For five minutes every hour from 7:00 to 21:00, the bees that entered and left the hive and bringing pollen and resin were registered. Every hour the relative humidity, temperature, wind speed and light intensity was recorded and related to foraging activity. Additionally, the weight of the colonies recently transferred to wooden boxes, the number of brood combs, honey pots and pollen were registered. The time of beginning and ending of the foraging activity differs from the reports of stingless bees of tropical weather and the same happens with the pollen collection. The environmental parameters that affect other tropical stingless bees in the foraging activity also affect M. colimana in temperate climate. It was determined that the major activity season and the presence of more pollen pots in the colony is from November through February, for what it could be the best time of the year for the division and obtainance of new colonies, while the critical period of minor activity and pollen flow was during rainy season. These data may be useful for the future sustainable use of this species in temperate climate.

Resumo Melipona colimana Ayala é uma espécie endémica de clima temperado que habita nas florestas de pinho e de azinheira do sul de Jalisco, no México. Registrou-se durante um ano, todos os quinze dias, a atividade de forragem, os parâmetros ambientais e o desenvolvimento de colônias de M. colimana no seu habitat silvestre. Registrou-se durante cinco minutos, cada hora das 7:00 às 21:00, as abelhas que entraram e saíram da colmeia e as que traziam pólen e resinas. Registrou-se toda hora, a humidade relativa, temperatura, velocidade do vento e intensidade da luz e relacionaram-se à atividade de forragem. Além do mais, registrou-se o peso de colônias recém transferidas a gavetas de madeira e contabilizou-se o número de panais, potes de mel e pólen. A hora de início e de fim da atividade de forragem difere com os reportes das abelhas sem ferrão de clima tropical e o mesmo acontece com a recolecção de pólen. Os parâmetros ambientais que afetam outros meliponinos de clima tropical na forragem também afetam a M. colimana em clima temperado. Determinou-se que a temporada de maior atividade e a presença de mais potes de pólen na colônia é de Novembro a Fevereiro, razão pela qual pode ser a melhor temporada do ano para a divisão e obtenção de novas colônias, enquanto que o período crítico de menor atividade e fluxo de pólen foi durante a temporada de chuvas. Esses dados podem ser úteis para o futuro aproveitamento sustentável de esta espécie em clima temperado.

Melipona mondury produces a geopropolis with antioxidant, antibacterial and antiproliferative activities

ABSTRACT Geopropolis is a special type of propolis produced by stingless bees. Several pharmacological properties have been described for different types of geopropolis, but there have been no previous studies of the geopropolis from Melipona mondury. In this study, we investigated the antioxidant, antibacterial, and antiproliferative activities of M. mondury geopropolis, and determined its chemical profile. The antioxidant activity was determined using in vitro ABTS·+, ·DPPH, and β-carotene/linoleic acid co-oxidation methods. The antibacterial activity was determined using a microdilution method with Pseudomonas aeruginosa, Staphylococcus aureus, and methicillin-resistant S. aureus. The antiproliferative effect was determined in tumor cell lines using the Alamar Blue assay. The chemical profile was obtained using UHPLC-MS and UHPLC-MS/MS. The butanolic fraction had the highest concentration of phenolic compounds and more potent antioxidant properties in all assays. This fraction also had bacteriostatic and bactericidal effects against all bacterial strains at low concentrations, especially S. aureus. The hexane fraction had the highest antiproliferative potential, with IC50 values ranging from 24.2 to 46.6 µg/mL in HL-60 (human promyelocytic leukemia cell) and K562 (human chronic myelocytic leukemia cell), respectively. Preliminary chemical analysis indicates the presence of terpenes and gallic acid in the geopropolis. Our results indicate the therapeutic potential of geopropolis from M. mondury against inflammatory, oxidative, infectious, and neoplastic diseases.

Antimicrobial activity of honeys from two stingless honeybee species and Apis mellifera (Hymenoptera: Apidae) against pathogenic microorganisms / Atividade antimicrobiana de méis de duas espécies de abelhas sem ferrão e Apis mellifera (Hymenoptera, Apidae) contra micro-organismos patogênicos

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.

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.

Conociendo la miel de Melipona favosa en la Península de Paraguaná, estado Falcón, Venezuela / Knowing the Melipona favosa honey from Paraguaná Peninsula, Falcon state, Venezuela

Diversas especies de abejas sin aguijón producen miel de botija en Venezuela, la cual no está incluida en las normas venezolanas de calidad. Una de estas especies, la Meli po na favosa, vive en la Península de Paraguaná, donde se conoce como erica o maba. A fin de conocer las mieles de M. favosa, se realizó la determinación de su origen botánico, su caracterización físicoquímica según los métodos de la norma COVENIN 2136-84 para miel de abejas y contenido de nitrógeno por microkjeldahl, y de actividad biológica con métodos espectrofotométricos y de concentración inhibitoria mínima, en seis mieles. Son mieles claras de color ámbar entre 18 y 79 mm Pfund. La composición físicoquímica varió así: acidez libre 12,72-95,86 meq/kg, pH 3,53-4,44, humedad 25,40-32,00 g agua/100 g, 0,01-0.16 g ce ni zas/100g, 10,48-57,55 mgN/100 g, 62,60-69,50 g azúcares reductores/100 g, 0,60-5,10 g sacarosa aparente/100 g, flavonoides 0,10-8,15 mgEQ/100g miel, y polifenoles 51,50-217,19 mgEAG/100 g. La actividad antibac teriana (g miel/100 mL medio de cultivo) fue mayor contra E. coli (12.50-50,00) que contra S. aureus (50,00- 50,00). La actividad antioxidante se ubicó en un rango bajo a alto para mieles, con 45,91-227,92 mmoles equivalentes de Trolox/100 g. El análisis melisopalinológico indicó polen dominante del género Portulaca y la especie Carica papaya.

Diverse species of stingless bees produce por honey in Venezuela,which is not included in the Venezuelan regulations of honey quality. One of these species , the Melipona favosa, lives in the Paraguaná Peninsula, where it is known as erica or maba. In order to know the honeys of M. favosa, the botanical origin was determined, their physicochemical characterization was done according to the methods of the norm COVENIN 2136-84 for honey, the nitrogen content by microkjeldahl, and biological activity by spectrophotometric and minimum inhibitory concentration methods, in six ho neys. They are light honey amber color between 18 and 79 mm Pfund. The physicochemical composition varied as follows: free acidity 12.72- 95.86 meq/kg, pH 3.53-4.44, moisture 25.40-32.00 g water/100 g, 0.01-0.16 g ash/100g, 10.48-57.55 mgN/100 g, 62.60-69.50 g reducing su gars/100 g, 0.60-5.10 g apparent sucrose/100 g, flavonoids 0.10-8.15 mgQE/100g honey, and polyphenols 51.50-217.19 mgGAE/100 g. The antibacterial ac tivity (g ho ney/100 mLculture media) was higher against E. coli (12.50-50.00) than S. aureus (50.00-50.00). The antioxidant activity was in a low to high range for honeys, with 45.91- 227.92 mmoles Trolox equivalents/100 g.

Variación métrica inter e intraespecifica de Melipona beecheii, M. solani y M. yucatanica de Guatemala / Inter-and intraspecific variation metric beecheii Melipona, M. solani and M. yucatanica of Guatemala

El objetivo del estudio fue analizar la variación en caracteres métricos, a nivel interespecíficos e intraespecíficos, de tres especies de Meliponas provenientes de varios departamentos de Guatemala. El análisis se realizó mediante técnicas morfométricas basadas en estadística multivariada, aplicadas sobre distancias medidas sobre las cabezas y las alas anteriores derechas de especímenes de Melipona beecheii, M. solani y M. yucatanica. Esto con el fin de determinar si existía variación geográfica a nivel poblacional, y poder ampliar así los conocimientos sobre la biología y la dinámica poblacional de estas poblaciones y en última instancia contribuir a la correcta identificación taxonómica de las especies mencionadas.

Prediction of social structure and genetic relatedness in colonies of the facultative polygynous stingless bee Melipona bicolor (Hymenoptera, Apidae)

Stingless bee colonies typically consist of one single-mated mother queen and her worker offspring. The stingless bee Melipona bicolor (Hymenoptera: Apidae) shows facultative polygyny, which makes this species particularly suitable for testing theoretical expectations concerning social behavior. In this study, we investigated the social structure and genetic relatedness among workers from eight natural and six manipulated colonies of M. bicolor over a period of one year. The populations of M. bicolor contained monogynous and polygynous colonies. The estimated genetic relatedness among workers from monogynous and polygynous colonies was 0.75 ± 0.12 and 0.53 ± 0.16 (mean ± SEM), respectively. Although the parental genotypes had significant effects on genetic relatedness in monogynous and polygynous colonies, polygyny markedly decreased the relatedness among nestmate workers. Our findings also demonstrate that polygyny in M. bicolor may arise from the adoption of related or unrelated queens.

Comparative study on the use of specific and heterologous microsatellite primers in the stingless bees Melipona rufiventris and M. mondury (Hymenoptera, Apidae)

Due to their high degree of polymorphism, microsatellites are considered useful tools for studying population genetics. Nevertheless, studies of genetic diversity in stingless bees by means of these primers have revealed a low level of polymorphism, possibly the consequence of the heterologous primers used, since in most cases these were not specifically designed for the species under consideration. Herein we compared the number of polymorphic loci and alleles per locus, as well as observed heterozygosity in Melipona rufiventris and M. mondury populations, using specific and heterologous primers. The use of specific primers placed in evidence the greater frequency of polymorphic loci and alleles per locus, besides an expressive increase in observed heterozygosity in M. rufiventris and M. mondury, thereby reinforcing the idea that populational studies should be undertaken by preferably using species-specific microsatellite primers.

Variation and genetic structure of Melipona quadrifasciata Lepeletier (Hymenoptera, Apidae) populations based on ISSR pattern

For a study of diversity and genetic structuring in Melipona quadrifasciata, 61 colonies were collected in eight locations in the state of Minas Gerais, Brazil. By means of PCR analysis, 119 ISSR bands were obtained, 80 (68 percent) being polymorphic. He and H B were 0.20 and 0.16, respectively. Two large groups were obtained by the UPGMA method, one formed by individuals from Januária, Urucuia, Rio Vermelho and Caeté and the other by individuals from São João Del Rei, Barbacena, Ressaquinha and Cristiano Otoni. The Φst and θB values were 0.65 and 0.58, respectively, thereby indicating high population structuring. UPGMA grouping did not reveal genetic structuring of M. quadrifasciata in function of the tergite stripe pattern. The significant correlation between dissimilarity values and geographic distances (r = 0.3998; p < 0.05) implies possible geographic isolation. The genetic differentiation in population grouping was probably the result of an interruption in gene flow, brought about by geographic barriers between mutually close geographical locations. Our results also demonstrate the potential of ISSR markers in the study of Melipona quadrifasciata population structuring, possibly applicable to the studies of other bee species.

Chemical composition, botanical evaluation and screening of radical scavenging activity of collected pollen by the stingless bees Melipona rufiventris (Uruçu-amarela)

Stingless bees in Brazil are indigenous and found all over the country. Bee pollen is used for its nutritional value in the human diet. It is made up of natural flower pollen mixed with nectar and bee secretions. In order to evaluate the chemical composition, free radical scavenging activity, and botanical origin, sample of pollen loads from stingless bee, Melipona rufiventris (Uruçu amarela) was studied. The EtOAc extract of pollen of Melipona rufiventris yielded the following compounds: p-hydroxycinnamic acid, dihydroquercetin, isorhamnetin, isorhamnetin3-O-(6"-O-E-p-coumaroyl)-β-D-glucopyranoside, luteolin, and quercetin. This is the first report of the isolation of isorhamnetin3-O-(6"O-E-p-coumaroyl)β-D-glucopyranoside from pollen. The free radicalscavenging activities of different solvent extracts of pollen were determined using DPPH assay. This activity decreases in the order: EtOAc>EtOH>Hexane extract. It appears that the EtOAc extract of the pollen is a good scavenger of active oxygen species. The botanical evaluation of pollen loads showed the composition by two pollen types, with the dominant type (97.3 percent) being Scopariadulcis (L.) (Scrophulariaceae) and the minor one Senna obtusifolia (L.) Irwin & Barneby (Fabaceae). This suggests a specific foraging behavior in Melipona rufiventris bees, even in an environment with such a rich botanical diversity as the Northeastern Brazil.

As abelhas sem ferrão são espécies indígenas encontradas em todo o Brasil. Seu pólen é utilizado devido ao seu valor nutricional na dieta humana. É produzido a partir de pólen floral misturado com néctar e líquidos secretados pelas abelhas. Visando avaliar a composição química, a atividade sequestradora de radicais livres e a origem botânica foi estudado o pólencoletado pela abelha sem ferrão Melipona rufiventris (Uruçu-amarela). Do extrato acetato de etila foram isolados os compostos: ácido phidroxicinâmico, dihidroquercetina, isoramnetina, 3O(6"OEpcoumaroil)βDglicopiranosideoisoramnetina, luteolina e quercetina. Esta é a primeira vez que a 3O(6"OEpcoumaroil)βDglicopiranosideoisoramnetina é isolada de pólen apícola. A atividade sequestradora de radicais livres de vários extratos com solventes diferentes foi determinada pelo teste com DPPH (difenilpicrilhidrazida). A atividade mostrou a ordem decrescente para os extratos AcOEt>EtOH>Hexano. O extrato AcOEt apresenta melhor atividade sequestradora de radicais. A avaliação botânica palinológicamostrou que o pólen era composto de dois tipos, um majoritário(97.3 por cento) proveniente de Scoparia dulcis (L.) (Scrophulariaceae) e outro minoritário proveniente de Senna obtusifolia (L.) Irwin& Barneby (Fabaceae). Estes resultados sugerem o comportamento de forragem bastante específico exibido pela abelha Melipona rufiventris, mesmo em um ambiente tão rico em diversidade vegetal como o Nordeste do Brasil.

Analysis of the intercaste transcriptional profile of Melipona scutellaris Latreille, 1811 (Hymenoptera, Apidae, Meliponini) by mRNA differential display

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.

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.

Avaliação farmacognóstica de geoprópolis de Melipona fasciculata Smith da Baixada maranhense, Brasil / Pharmacognostic evaluation of geopropolis of Melipona fasciculata Smith from Baixada maranhense, Brazil

Melipona fasciculata Smith é uma abelha nativa, social e sem ferrão que produz cera, mel e geoprópolis. O presente trabalho objetivou caracterizar, do ponto de vista farmacognóstico, o geoprópolis de Melipona fasciculata, coletado em meliponários nos municípios de Arari, São Bento e São João Batista, da Baixada maranhense, no Estado do Maranhão, visando estabelecer dados para o controle de qualidade e padronização do produto. A metodologia utilizada constou de análises sensoriais, abordagem química, perfis cromatográficos e determinação dos teores de flavonóides. Os resultados demonstram que o geoprópolis apresenta características sensoriais similares às encontradas para própolis de Apis mellifera, presença de compostos fenólicos em maior concentração, além de substâncias da classe dos terpenos e saponinas e ausência de alcalóides. Os perfis cromatográficos indicaram que a composição e concentração das substâncias químicas são diferentes entre as amostras. Os teores de flavonóides variaram de 0,17 - 2,6%, os quais para a maioria das amostras apresentaram valores acima do mínimo exigido pela legislação brasileira para própolis de Apis mellifera. Os dados encontrados sugerem que as variações qualitativa e quantitativa de flavonóides e outros constituintes químicos no geoprópolis são fortemente afetados pela flora visitada pelas abelhas, região geográfica e fatores ambientais.

Melipona fasciculata Smith is a native, social, stingless bee species that produces wax, honey and geopropolis. This work aimed to do a pharmacological evaluation of Melipona fasciculata geopropolis, collected in beehives in the municipal districts of Arari, São Bento and São João Batista, in Baixada maranhense, in the state of Maranhão, Brazil, seeking to obtain basis for the quality control and standardization of the product. The methodology used included the accomplishment of sensorial analyses, chemical approach, chromatographic profiles and determination of the flavonoids contents. The results demonstrate that the geopropolis have organoleptic characteristics similar to the ones found for the propolis of Apis mellifera, presence of phenolic compounds in larger concentration, besides substances of the class of the terpenes and saponins and absence of alkaloids. The chromatographic profiles indicated that the composition and concentration of the chemical substances are different among the samples. The flavonoids contents ranged between 0.17 - 2.6%. The content for most of the samples presented values above the minimum demanded by the Brazilian legislation for propolis of Apis mellifera. The found data suggest that the qualitative and quantitative variations of flavonoids and other chemical substances in geopropolis are strongly affected for the flora visited by the bees, geographical area and environmental factors.

Caracterización fisicoquímica y antimicrobiana de la miel de nueve especies de abejas sin aguijón (Meliponini) de Guatemala / Physicochemical and antimicrobial characterization of nine stingless bees (Meliponini) honey from Guatemala / Caracterização fisicoquímica e antimicrobiana do mel de nove especies de abelhas sem ferrão (Meliponini) de Guatemala

La miel de meliponinos es reconocida popularmente en Latinoamérica por sus propiedades terapéuticas. En Guatemala, unas 13 especies son criadas artesanalmente para la obtención de miel. Debido al reciente interés en la utilización de la miel de meliponinos en Guatemala y a su comercialización, se hace necesario conocer las características fisicoquímicas y validar el uso terapéutico de la misma. En el presente trabajo se realizó una caracterización fisicoquímica y antibacteriana de la miel producida en Guatemala de nueve especies de meliponinos: Geotrigona acapulconis (Strand, 1919); Melipona beecheii Bennett, 1831; Melipona solani Cockerell, 1912; Melipona aff. yucatanica; Nannotrigona perilampoides Cresson, 1878; Plebeia sp.; Scaptotrigona mexicana Guérin, 1845; Scaptotrigona pectoralis Dalla Torre, 1896; y Tetragonisca angustula (Latreille, 1811). Los valores promedio obtenidos en 18 muestras fueron: humedad 21,55g/100g de miel; acidez 22,45meq/kg; cenizas 0,29g/100g; hidroximetilfurfural 0,23mg/kg; actividad de la diastasa 10,94DN; y pH 3,89. Se evaluó la actividad antibacteriana frente a ocho microorganismos, determinando una concentración inhibitoria mínima de 2,5 a 10% (v/v). Se realizó un análisis preliminar de tipos polínicos que permitió identificar la presencia de 21 familias vegetales visitadas por los meliponinos en Guatemala, donde la miel de T. angustula presentó la mayor diversidad. Este trabajo es el primer aporte al conocimiento sobre la composición fisicoquímica y antibacteriana de las mieles de nueve especies de meliponinos en este país.

The honey of stingless bees is popularly known in Latin America for its therapeutic properties. In Guatemala some 13 species are traditionally bred to obtain honey. Due to the recent interest and trading of meliponine honey, it is necessary to know the physicochemical properties and to validate its therapeutic use. The antimicrobial activity, the physicochemical composition and the botanical origin of the honey produced by nine species of stingless bees from Guatemala is reported. The species were Geotrigona acapulconis (Strand, 1919); Melipona beecheii Bennett, 1831; Melipona solani Cockerell, 1912; Melipona aff. yucatanica; Nannotrigona perilampoides Cresson, 1878; Plebeia sp.; Scaptotrigona mexicana Guérin, 1845; Scaptotrigona pectoralis Dalla Torre, 1896; and Tetragonisca angustula (Latreille, 1811). The average values obtained in 18 samples were: Moisture 21.55g for 100g of honey, acidity 22.45meq/100g, ash 0.29g/100g, hidroximetilfurfural 0.23mg/kg, diastase activity 10.94 N, and pH 3.89. The antibacterial activity of the honey against eight microorganisms was determined; the minimum inhibitory concentration was found to be of 2.5 to 10% (v/v). A preliminary palynological analysis permitted the identification of 21 vegetal families in the honey samples of Guatemala, among which the honey of T. angustula presented the greatest diversity. This work is the first contribution about the composition of honey from nine species of meliponini in the country.

O mel de meliponíneos é reconhecido popularmente na América Latina por suas propriedades terapêuticas. Na Guatemala, umas 13 espécies são criadas artesanalmente para a obtenção de mel. Devido ao recente interesse na utilização do mel de meliponíneos na Guatemala e a sua comercialização, se faz necessário conhecer as características físico-químicas e validar o uso terapêutico da mesma. No presente trabalho se realizou uma caracterização físico-química e antibacteriana do mel produzido na Guatemala de nove espécies de meliponíneos: Geotrigona acapulconis (Strand, 1919); Melipona beecheii Bennett, 1831; Melipona solani Cockerell, 1912; Melipona aff. yucatanica; Nannotrigona perilampoides Cresson, 1878; Plebeia sp.; Scaptotrigona mexicana Guérin, 1845; Scaptotrigona pectoralis Dalla Torre, 1896; e Tetragonisca angustula (Latreille, 1811). Os valores médios obtidos em 18 amostras foram: umidade 21,55g/100 g de mel; acidez 22,45meq/kg; cinzas 0,29g/100g; hidroximetilfurfural 0,23mg/kg; atividade da diastasa 10,94DN; y pH 3,89. Avaliou-se a atividade antibacteriana frente a oito microorganismos, determinando uma concentração inibitória mínima de 2,5 a 10% (v/v). A análise preliminar de tipos polínicos permitiu identificar a presença de 21 famílias vegetais visitadas pelos meliponíneos na Guatemala, onde o mel de T. angustula apresentou a maior diversidade. Este trabalho é o primeiro aporte ao conhecimento sobre a composição físico-química e antibacteriana do mel de noves espécies de meliponíneos neste país.