Ferulic acid as major antioxidant phenolic compound of the Tetragonisca angustula honey collected in Vera Cruz - Itaparica Island, Bahia, Brazil / Ácido ferúlico como principal composto fenólico antioxidante do mel de Tetragonisca angustula coletado em Vera Cruz - Ilha de Itaparica, Bahia, Brasil

Abstract The antioxidant activity of Tetragonisca angustula honey (TAH) and its ethanolic extract (TAEE) were investigated. The total levels of phenolic (TPC) and flavonoids (TFC) were also evaluated. The results for TPC were 19.91 ± 0.38 and 29.37 ± 1.82 mg GAE g-1 and for TFC 0.20 ± 0.02 and 0.14 ± 0.01 mg QE g-1 of TAH and TAEE, respectively. Antioxidant activities were 73.29 ± 0.49% and 93.36 ± 0.27% in the DPPH● assay and 71.73 ± 4.07% and 97.86 ± 0.35% in ABTS●+ for TAH and TAEE, respectively. The total reducing activity was determined by the method of reducing power (PR) and iron ion (Fe III) and the results varied in PR from 151.7 ± 25.7 and 230.7 ± 25.2 mg GAE L-1, for TAH and TAEE respectively and for (Fe III) in EC50 0.284 in TAEE and 0.687 in TAH. Chemical analysis by HPLC-DAD of the ethanolic extract (TAEE) revealed the presence of ferulic acid as majority phenolic component in the extract. The 1H NMR analysis confirmed this structure and showed the also presence of glucose, citric acid, succinic acid, proline and hydrocarbon derivatives. In addition, the botanical origin was also investigated and showed a multifloral characteristic, having found 19 pollen types with a botanical predominance of the Anacardiaceae family, with Tapirira pollen occurring as predominant (42.6%) and Schinus as secondary (25.7%). The results showed that T. angustula honey is an interesting source of antioxidant phenolic compounds due to its floral origin and can act as a protector of human health when consumed.

Resumo A atividade antioxidante do mel de Tetragonisca angustula (TAH) e seu extrato etanólico (TAEE) foram investigados. Os níveis totais de fenólicos (TPC) e flavonóides (TFC) também foram avaliados. Os resultados para TPC foram 19,91 ± 0,38 e 29,37 ± 1,82 mg GAE g-1 e para TFC 0,20 ± 0,02 e 0,14 ± 0,01 mg QE g-1 de TAH e TAEE, respectivamente. As atividades antioxidantes foram 73,29 ± 0,49% e 93,36 ± 0,27% no ensaio DPPH● e 71,73 ± 4,07% e 97,86 ± 0,35% no ABTS●+ para TAH e TAEE, respectivamente. A atividade redutora total foi determinada pelo método de poder redutor (PR) e íon ferrico (Fe III) e os resultados variaram em PR de 151,7 ± 25,7 e 230,7 ± 25,2 mg GAE L-1, para TAH e TAEE respectivamente e para (Fe III) em EC50 0,284 em TAEE e 0,687 em TAH. A análise química por HPLC-DAD do extrato etanólico (TAEE) revelou a presença de ácido ferúlico como componente majoritário no extrato. A análise de RMN 1H confirmou esta estrutura e mostrou a presença de glicose, ácido cítrico, ácido succínico, prolina e derivados de hidrocarbonetos no TAEE. Além disso, a origem botânica também foi investigada e apresentou característica multifloral, tendo encontrado 19 tipos polínicos com predomínio botânico da família Anacardiaceae, sendo o pólen Tapirira predominante (42,6%) e o Schinus secundário (25,7%). Os resultados mostraram que o mel de T. angustula é uma interessante fonte de compostos fenólicos antioxidantes devido à sua origem floral e pode atuar como protetor da saúde humana quando consumido.

Ferulic acid as major antioxidant phenolic compound of the Tetragonisca angustula honey collected in Vera Cruz - Itaparica Island, Bahia, Brazil

Abstract The antioxidant activity of Tetragonisca angustula honey (TAH) and its ethanolic extract (TAEE) were investigated. The total levels of phenolic (TPC) and flavonoids (TFC) were also evaluated. The results for TPC were 19.91 ± 0.38 and 29.37 ± 1.82 mg GAE g-1 and for TFC 0.20 ± 0.02 and 0.14 ± 0.01 mg QE g-1 of TAH and TAEE, respectively. Antioxidant activities were 73.29 ± 0.49% and 93.36 ± 0.27% in the DPPH assay and 71.73 ± 4.07% and 97.86 ± 0.35% in ABTS+ for TAH and TAEE, respectively. The total reducing activity was determined by the method of reducing power (PR) and iron ion (Fe III) and the results varied in PR from 151.7 ± 25.7 and 230.7 ± 25.2 mg GAE L-1, for TAH and TAEE respectively and for (Fe III) in EC50 0.284 in TAEE and 0.687 in TAH. Chemical analysis by HPLC-DAD of the ethanolic extract (TAEE) revealed the presence of ferulic acid as majority phenolic component in the extract. The 1H NMR analysis confirmed this structure and showed the also presence of glucose, citric acid, succinic acid, proline and hydrocarbon derivatives. In addition, the botanical origin was also investigated and showed a multifloral characteristic, having found 19 pollen types with a botanical predominance of the Anacardiaceae family, with Tapirira pollen occurring as predominant (42.6%) and Schinus as secondary (25.7%). The results showed that T. angustula honey is an interesting source of antioxidant phenolic compounds due to its floral origin and can act as a protector of human health when consumed.

Resumo A atividade antioxidante do mel de Tetragonisca angustula (TAH) e seu extrato etanólico (TAEE) foram investigados. Os níveis totais de fenólicos (TPC) e flavonóides (TFC) também foram avaliados. Os resultados para TPC foram 19,91 ± 0,38 e 29,37 ± 1,82 mg GAE g-1 e para TFC 0,20 ± 0,02 e 0,14 ± 0,01 mg QE g-1 de TAH e TAEE, respectivamente. As atividades antioxidantes foram 73,29 ± 0,49% e 93,36 ± 0,27% no ensaio DPPH e 71,73 ± 4,07% e 97,86 ± 0,35% no ABTS+ para TAH e TAEE, respectivamente. A atividade redutora total foi determinada pelo método de poder redutor (PR) e íon ferrico (Fe III) e os resultados variaram em PR de 151,7 ± 25,7 e 230,7 ± 25,2 mg GAE L-1, para TAH e TAEE respectivamente e para (Fe III) em EC50 0,284 em TAEE e 0,687 em TAH. A análise química por HPLC-DAD do extrato etanólico (TAEE) revelou a presença de ácido ferúlico como componente majoritário no extrato. A análise de RMN 1H confirmou esta estrutura e mostrou a presença de glicose, ácido cítrico, ácido succínico, prolina e derivados de hidrocarbonetos no TAEE. Além disso, a origem botânica também foi investigada e apresentou característica multifloral, tendo encontrado 19 tipos polínicos com predomínio botânico da família Anacardiaceae, sendo o pólen Tapirira predominante (42,6%) e o Schinus secundário (25,7%). Os resultados mostraram que o mel de T. angustula é uma interessante fonte de compostos fenólicos antioxidantes devido à sua origem floral e pode atuar como protetor da saúde humana quando consumido.

The characteristics of stingless bee nests (Apidae: Meliponini) in the Cycloop Mountain Nature Reserve, Indonesia

Introduction: Stingless bees are widespread in tropical and subtropical regions. In Indonesia, the distribution of stingless bees are grouped in three regions, namely Indo-Malayan, Wallacea, and Indo-Australian. Ten species of stingless bees have been recorded in Papua, seven of which are endemic. The Cycloop Mountains Nature Reserve (CMNR) is one of the conservation area in Papua, Indonesia, for flora and fauna. Unfortunately, the study of the diversity of stingless bees in Papua has been limited. Objective: To measure the diversity, nesting sites, nest entrance characteristics and nest architecture of stingless bees. Methods: Observation of the stingless bee nests in the nature reserves and in the residential areas used a road sampling method and information from local people, respectively. A total of 22 colonies were studied. Results: Two species of stingless bee were found, namely Tetragonula sapiens (Cockerell, 1911) and Heterotrigona (Platytrigona) planifrons (Smith, 1865). The current study showed new distribution records for T. sapiens and H. planifrons in the CMNR. The nesting site of T. sapiens was commonly found in house foundation, while that of H. planifrons was in coconut palm cavities. The nest entrance of T. sapiens varied, i.e., elliptical, oval, rounded, irregular, horizontally or vertically elongated. Meanwhile, the nest entrance of H. planifrons was vertically elongated. The brood cells of T. sapiens varied, i.e., vertical, horizontal, or semi-clusters, while in H. planifrons was layered vertically. Conclusions: Two species of stingless bees found, T. sapiens and H. planifrons, showed a new distribution records and T. sapiens was a dominant species in Papua. The nest entrance of the species varied in shape, color, and texture.

Introducción: Las abejas sin aguijón están muy extendidas en las regiones tropicales y subtropicales. En Indonesia, la distribución de las abejas sin aguijón se agrupa en tres regiones: Indo-Malayan, Wallacea e Indo-Australian. Se han registrado diez especies de abejas sin aguijón en Papua, siete de las cuales son endémicas. La Reserva Natural de las Montañas Cycloop (CMNR) es una de las áreas para la conservación de flora y la fauna en Papua, Indonesia. Desafortunadamente, el estudio de la diversidad de abejas sin aguijón en Papua ha sido limitado. Objetivo: Medir la diversidad, los sitios de anidación y describir la arquitectura y características de entrada al nido de las abejas sin aguijón. Métodos: Se observaron los nidos de abejas sin aguijón en reservas naturales y áreas residenciales, mediante el método de muestreo de caminos e información de la población local, respectivamente. Se estudiaron 22 colonias. Resultados: Se encontraron dos especies de abejas sin aguijón, Tetragonula sapiens (Cockerell, 1911) y Heterotrigona (Platytrigona) planifrons (Smith, 1865). Este estudio mostró nuevos registros de distribución de T. sapiens y H. planifrons en el CMNR. El sitio de anidación de T. sapiens se encontró comúnmente en los cimientos de las casas, mientras que el de H. planifrons estaba en las cavidades de las palmas de coco. La forma de la entrada al nido de T. sapiens varió: elíptica, ovalada, redondeada, irregular, alargada horizontal o verticalmente. Mientras tanto, la entrada al de H. planifrons se alarga verticalmente. Las celdas de cría de T. sapiens variaron, entre verticales, horizontales o semi-racimos, mientras que en H. planifrons eran verticales. Conclusiones: Se encontraron dos especies de abejas sin aguijón, T. sapiens y H. planifrons, que mostraron nuevos registros de distribución y T. sapiens fue una especie dominante en Papúa. La entrada al nido de las especies varió en forma, color y textura.

In Vitro Investigation of Antioxidant and Antidiabetic Properties of Phenolic-Rich Extract from Stingless Bee Honey (Heterotrigona itama)

@#Introduction: The increase of Type 2 diabetes mellitus has prompted numerous research toward finding an alternative to manage the disease through the oxidant-antioxidant balance, mainly through bioactive compounds in natural products. This study explored the antioxidant and antidiabetic properties of phenolic-rich extract (PRE) from Stingless bee honey (SBH) (Heterotrigona itama) as therapeutic agent to restore the redox balance. Methods: The total phenolic content (TPC), total flavonoid content (TFC) and antioxidant assays of PRE and SBH, were determined to provide preliminary insight into the sample’s antioxidant properties, followed by high-performance liquid chromatography analysis of PRE. The antidiabetic potential of PRE and SBH were determined based on their inhibition against α-amylase and α-glucosidase enzymes. The cytotoxicity analysis of PRE was conducted on 3T3-L1 adipocytes and L6 muscle cells before the glucose uptake and cellular antioxidant analyses were performed on both cell lines, respectively. Results: PRE yielded higher TPC, TFC and antioxidant activities than SBH. The phytochemical profile of PRE comprises gallic acid, myricetin, kaempferol, epicatechin, chlorogenic acid, quercetin, syringic acid, and cinnamic acid. The results from carbohydrate enzymatic inhibitory assays collectively suggested that PRE exhibited more robust antidiabetic activities than SBH. PRE showed good glucose uptake stimulating and reactive oxygen species scavenging effects in those cell lines. Conclusion: Overall, PRE from SBH showed higher carbohydrate enzymatic inhibition, glucose uptake, and protection against intracellular oxidative stress, primarily due to its high antioxidant content and may serve as an alternative therapeutic agent for managing T2DM.

Effect of symbiotic bee fungus on survival of stingless bee Heterotrigona itama larvae

Aims@#Symbiotic bee fungus associated with the stingless bee larval food has been revealed to play a major role in performing a mutual relationship with the host. The fungus is believed capable to produce crucial nutrients that are required for larval pupation. However, detailed information on the fungus identification isolated from the larval food of our native Indo-Malaya stingless bee, Heterotrigona itama (Hymenoptera: Apidae), is poorly understood. Hence, this study was conducted to determine the effect of fungus isolated from the stingless bee larval food on the survival of H. itama larvae and identify the isolated fungus using both morphological and molecular analyses.@*Methodology and results@#Elisa plates designed with F-bottom type were used as artificial brood cells in a controlled condition. The eggs transferred to Elisa plates were kept in the incubator with 75%-100% humidity and 25 ± 2.5 °C of temperature. This study carried out in three different treatments: (1) microbes-free larval food, (2) fresh larval food and (3) microbes-free larval food with fungus supplement. Results showed that the survival of H. itama larvae depends on the presence of beneficial fungus with the highest survival rate (Treatment 2 = 85.71%). The absence of the beneficial fungus in the fluid food of H. itama showed the lowest survival rate (Treatment 1 = 37.14%). Microscopy analysis showed that the fungus had various forms, including unconjugated asci, round-shaped ascospore and pseudohyphae. The molecular characterisation of the isolated fungi was performed using 23S rRNA gene sequencing using universal primers ITS1 and ITS4. DNA barcoding of three isolated fungi confirmed all isolated fungi matched Panus lecomtei (Basidiomycetes; Poriales; Polyporaceae) with 99.70% of similarity.@*Conclusion, significance and impact of study@#This study provides information on the importance of whitish fungus in appearance that existed in the brood cell, which is proposed to be the crucial component of in-vitro stingless bee queen rearing protocol.

Stingless bee propolis, metformin, and their combination alleviate diabetic cardiomyopathy

Abstract Background and aim: Stingless bee propolis, a resinous compound processed by mandibular secretion of stingless bees, is used for maintenance of hygiene and stability of beehives. Research on stingless bee propolis shows therapeutic properties attributed to polyphenols exhibiting antioxidative, antihyperglycemic and antiischemic effect. However, the cardioprotective effect of stingless bee propolis on diabetic cardiomyopathy is unknown. Methods: Adult male Sprague Dawley rats were randomised to five groups: normal group, diabetic group, diabetic given metformin (DM+M), diabetic given propolis (DM+P) and diabetic given combination therapy (DM+M+P) and treated for four weeks. Body weight, fasting blood glucose, food and water intake were taken weekly. At the end of experiment, biomarkers of oxidative damage were measured in serum and heart tissue. Antioxidants in heart tissue were quantified. Part of left ventricle of heart was processed for histological staining including Haematoxylin and Eosin (H&E) stain for myocyte size and Masson's Trichrome (MT) stain for heart fibrosis and perivascular fibrosis. Results: Propolis alleviated features of diabetic cardiomyopathy such as myocyte hypertrophy, heart fibrosis and perivascular fibrosis associated with improvement in antioxidative status. Conclusion: This study reports beneficial effect of propolis and combination with metformin in alleviating histopathological feature of diabetic cardiomyopathy by modulating antioxidants, making propolis an emerging complementary therapy.

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.

Diversity and foraging patterns of bees on flowers of Cucurbita pepo (Cucurbitaceae) in Costa Rica

Abstract Introduction: The species and functional diversity of pollinators are important components for the reproduction of cultivated plants. More information is necessary about this diversity and its geographical variation in crops such as Cucurbita pepo, an important crop in global agriculture. Objective: To describe the taxonomic diversity, geographic variation and foraging patterns of the community of bees that visit C. pepo crops in Costa Rica. Methods: Squash fields were visited at 11 locations within three geographic regions of the country, where the groups of bees and their relative frequency were determined. Through video recordings, information was obtained on their behavior at two locations. Results: A minimum of 27 species belonging to 19 genera and 2 families of bees were found. Three species were dominant in 10 localities (Eucera limitaris, Apis mellifera and Trigona corvina). Altitude reduces bee diversity due to the dominance of Bombus ephipiatus in high regions. Two genera of halictids (Megalopta and Caenaugochlora) that are rarely reported in this crop were frequently observed. Trigona bees dominated among the flowers later in the morning, lacerating nectary holes to facilitate nectar collection. Conclusions: Squash fields in Costa Rica are visited by a highly diverse bee community, which may ensure pollination via complementarity in the face of spatial or seasonal changes in environmental conditions.

Resumen Introducción: Dada la importancia del componente diversidad para la polinización de plantas cultivadas, es necesario obtener más información de esta diversidad y su variación geográfica en cultivos como Cucurbita pepo, uno de los cultivos más importantes de la agricultura centroamericana. Objetivo: Describir la diversidad y la variación geográfica de la comunidad de abejas que visitan este cultivo en Costa Rica, y algunos aspectos de sus patrones de búsqueda de alimento. Métodos: Se visitaron cultivos de C. pepo en 11 localidades dentro de tres regiones geográficas de Costa Rica, donde se determinó los grupos de abejas y su frecuencia relativa. Por medio de grabaciones de video se registró el comportamiento de cada grupo en dos localidades. Resultados: Fueron encontradas un mínimo de 27 especies pertenecientes a 19 géneros y 2 familias de abejas. Tres especies son dominantes en 10 localidades (Eucera limitaris, Apis mellifera y Trigona corvina). La altitud reduce la diversidad de abejas debido a la dominancia de Bombus ephipiatus en regiones altas. Se observaron dos géneros de halíctidos (Megalopta y Caenaugochlora) no previamente reportados en este cultivo. Abejas Trigona dominan las flores en horarios más tardíos de la mañana, donde algunas veces muerden los orificios de los nectarios para facilitar la recolecta de néctar. Conclusiones: La diversidad de abejas que visitan C. pepo en Costa Rica parece asegurar su polinización ante cambios espaciales o estacionales en condiciones ambientales.

Foraging preferences of the native stingless bee Melipona seminigra pernigra (Apidae: Meliponini) in campo rupestre on canga of Serra dos Carajás, southeastern Amazonia / Preferências de forrageamento da abelha nativa sem ferrão Melipona seminigra pernigra (Apidae: Meliponini) em campo rupestre na canga da Serra dos Carajás, sudeste da Amazônia

Abstract: Honey pollen samples of Melipona seminigra pernigraMoure & Kerr 1950 sampled between 2017 and 2019 from experimental apiaries installed in campo rupestre on canga (CRC) vegetation of the Serra dos Carajás aimed to evaluated seasonal floral availability of undisturbed and mining-influenced areas. Around one hundred pollen types were identified mainly belonging to Fabaceae, Myrtaceae and Euphorbiaceae (31, 6 and 5 species, respectively). Mining area presented the highest pollen richness, almost twice those identified in the undisturbed areas. 80% of the pollen types are rare with concentrations ≤ 2,000 pollen grains/10 g, while the remaining were the most abundant, frequent and the primary bee sources. These latter correspond mostly to native plants species such as Tapirira guianensis Aubl., Protium spp., Aparisthmium cordatum (A.Juss.) Baill., Mimosa acutistipula var. ferrea Barneby, Periandra mediterranea (Vell.) Taub., Miconia spp., Pleroma carajasense K.Rocha, Myrcia splendens (Sw.) DC., Serjania spp. and Solanum crinitum Lam. All pollen types were identified during both seasons, but higher concentration values are related to the dry period (June-September). The statistical analysis of the pollen data indicated that there was no significant difference between undisturbed and mining-influenced areas, since primary bee sources of this study are widespread used in revegetation of mined areas.

Resumo: O conteúdo polínico de amostras de mel coletadas nos anos de 2017 e 2019 de apiários experimentais de Melipona seminigra pernigraMoure & Kerr 1950, instalado dentro de uma vegetação de campo rupestre em um afloramento de canga na Serra dos Carajás, sudeste da Amazônia, foi analisado para entender a variabilidade local dos recursos florais em áreas naturais e perturbadas. Aproximadamente 100% dos tipos polínicos foram identificados e pertencem principalmente às famílias Fabaceae, Myrtaceae e Euphorbiaceae (31, 6 e 5 espécies, respectivamente). Áreas de mineração apresentaram a maior riqueza de pólen, quase o dobro daquelas identificadas em áreas perturbadas. 80% dos tipos de pólen são raros com concentrações ≤ 2.000 grãos de pólen/10g, enquanto que os restantes foram os mais abundantes, frequentes e fontes primárias para as abelhas. Este últimos correspondem principalmente a plantas nativas como Tapirira guianensis Aubl., Protium spp., Aparisthmium cordatum (A.Juss.) Baill., Mimosa acutistipula var. ferrea Barneby, Periandra mediterrânea (Vell.) Taub., Miconia spp., Pleroma carajasense K.Rocha, Myrcia splendens (Sw.) DC., Serjania spp. e Solanum crinitum Lam. Todos os tipos polínicos foram identificados durante ambas as estações, mas altas concentrações estão relacionadas ao período seco (junho-setembro). A análise estatística indicou que não houve diferença significativa nos dados de pólen de mel entre áreas naturais e áreas anteriormente degradadas, uma vez que as fontes primárias das abelhas deste estudo são amplamente utilizadas na revegetação de áreas mineradas.

Effects of biopesticides in Tetragonisca angustula latreille (Hymenoptera: Meliponinae) pollinators / Efeitos de biopesticidas em polinizadores Tetragonisca angustula latreille (Hymenoptera: Meliponinae) / Efectos de los bioplaguicidas sobre los polinizadores Tetragonisca angustula latreille (Hymenoptera: Meliponinae)

Stingless bees Tetragonisca angustula (Latreille) (Hymenoptera: Meliponinae) are pollinators of native and cultivated plants and are therefore in contact with areas contaminated by pesticides. These native bees were evaluated for changes in gene expression of esterase isoenzymes (EST) and peptides after contamination by contact with growth regulators from insecticides Gallaxy® EC 100, Natuneem and Azamax after 48, 120, 168 hours, 30 and 60 days. EST-4 presented an increase in relative activity after contamination with Gallaxy® EC 100 at 6.2 × 10-2 g a.i./mL; Natuneem at 7.5 × 10-5 g a.i./mL; and Azamax at 1.2 × 10-3 g a.i/mL after 60 days, 48 h, and 60 days, respectively. Inhibition of the relative activity of EST-4 was detected after contamination by Natuneem at 1.5 × 10-5 g a.i./mL and Azamax at 1.2 × 10-3 g a.i./mL after 48 h and 30 days, respectively. The insecticide growth regulators promoted changes in protein synthesis of T. angustula adult workers resulting in an increase or decrease in the relative intensity of bands, and the appearance of new peptides when compared with controls. Changes in protein synthesis have been identified mainly after long period of contamination, 120 and 168 h with the IGRs Gallaxy® EC 100 (at 0.78 and 1.25 g a.i./mL), Azamax (at 1.2 × 10-3 and 6 × 10-3 g a.i./mL), and Natuneem (at 7.5 × 10-5 and 3 × 10-3 g a.i./mL), and at 60 days with Natuneem (at 1.5 × 10-5 g a.i./mL).(AU)

Abelhas sem ferrão Tetragonisca angustula (Latreille) (Hymenoptera: Meliponinae) são polinizadores de plantas nativas e cultivadas e, portanto, estão em contato com áreas contaminadas por biopesticidas. Essas abelhas nativas foram avaliadas quanto a alterações na expressão gênica de isoenzimas esterases (EST) e peptídeos após contaminação por contato com reguladores de crescimento de inseticidas Gallaxy® EC 100, Natuneem e Azamax após 48, 120, 168 horas, 30 e 60 dias. A EST-4 apresentou um aumento na atividade relativa após a contaminação com Gallaxy® 100 EC em 6,2 × 10-2 g i.a./mL, Natuneem em 7,5 × 10-5 g i.a./mL e Azamax em 1,2 × 10-3 g i.a./mL após 60 dias, 48 h e 60 dias, respectivamente. A inibição da atividade relativa de EST-4 foi detectada após contaminação pelo Natuneem a 1,5 × 10-5 g i.a./mL e Azamax a 1,2 × 10-3 g i.a./mL após 48 he 30 dias, respectivamente. Os reguladores de crescimento de inseticidas promoveram alterações na síntese protéica de trabalhadores adultos de T. angustula, resultando em um aumento ou diminuição da intensidade relativa das bandas e no aparecimento de novos peptídeos em comparação com os controles. Alterações na síntese de proteínas foram identificadas principalmente após um longo período de contaminação, 120 e 168 h com o IGRs Gallaxy® EC 100 (0,78 e 1,25 g i.a./mL), Azamax (1,2 × 10-3 e 6 × 10-3 g i.a./mL) e Natuneem (7,5 × 10-5 e 3 × 10-3 g i.a./mL) e 60 dias com Natuneem (1,5 × 10-5 g i.a./mL).(AU)

Las abejas sin aguijón Tetragonisca angustula (Latreille) (Hymenoptera: Meliponinae) son polinizadores de plantas nativas y cultivadas y, por lo tanto, están en contacto con áreas contaminadas por bioplaguicidas. Estas abejas nativas fueron evaluadas para detectar cambios en la expresión génica de isoenzimas esterasa (EST) y péptidos después de la contaminación por contacto con los reguladores del crecimiento insecticidas Gallaxy® EC 100, Natuneem y Azamax después de 48, 120, 168 horas, 30 y 60 días. EST-4 mostró un aumento en la actividad relativa después de la contaminación con Gallaxy® 100 EC a 6.2 × 10-2 g i.a./mL, Natuneem a 7.5 × 10-5 g i.a./mL y Azamax a 1.2 × 10-3 g i.a./mL después de 60 días, 48 hy 60 días, respectivamente. La inhibición de la actividad relativa de EST-4 se detectó después de la contaminación por Natuneem a 1.5 × 10-5 g i.a./mL y Azamax a 1.2 × 10-3 g i.a./mL después de 48 hy 30 días. respectivamente. Los insecticidas reguladores del crecimiento promovieron cambios en la síntesis de proteínas de trabajadores adultos de T. angustula, resultando en un aumento o disminución de la intensidad relativa de las bandas y en la aparición de nuevos péptidos en relación a los controles. Los cambios en la síntesis de proteínas se identificaron principalmente después de un largo período de contaminación, 120 y 168 h con IGRs Gallaxy® EC 100 (0.78 y 1.25 g i.a./mL), Azamax (1.2 × 10-3 y 6 × 10-3 g i.a./mL) y Natuneem (7.5 × 10-5 y 3 × 10-3 g i.a./mL) y 60 días con Natuneem (1.5 × 10-5 g i.a./mL).(AU)

Pollination service of Nannotrigona testaceicornis stingless bees in strawberry / Serviço de polinização da abelha Nannotrigona testaceicornis no morangueiro

Strawberry presents moderate dependence on bee pollination service, and pollination is related to the production and quality of fruits. The aim of this study was to evaluate the pollination service provided by Nannotrigona testaceicornis stingless bees in strawberry. Primary flowers of cultivar San Andreas were used in a completely randomized experimental design, with twelve replicates and five types of pollination: one N. testaceicornis visit (1V); two N. testaceicornis visits (2V); three N. testaceicornis visits (3V); natural pollination (NP); self-pollination (SP). In flowers visited by N. testaceicornis, movements and visit time were observed. The following fruit characteristics were evaluated: fruit biometry, fecundation rate of achenes, postharvest fruit quality, contribution of pollination agents and mechanisms on average fruit weight. In fruit biometry, pollination service contributed only in longitudinal length, which was higher in NP and 3V, compared to SP. In the different types of pollination, the fertilization rate of achenes did not differ and showed no effect on fresh fruit weight. In post-harvest fruit quality, 1V, 3V and NP showed better results regarding degree of deformation and marketability. Nannotrigona testaceicornis stingless bees and natural pollination contributed to the average fresh weight of strawberry fruits. Nannotrigona testaceicornis stingless bees and natural pollination provided improvement in quality and added value of fruits. Nannotrigona testaceicornis stingless bees were effective strawberry pollinators.(AU)

O morangueiro apresenta dependência moderada quanto ao serviço de polinização por abelhas, e sua polinização está relacionada com a produção e a qualidade das frutas. Diante disso, o objetivo deste estudo foi avaliar o serviço de polinização da abelha Nannotrigona testaceicornis no morangueiro. Utilizaram-se flores primárias do cultivar San Andreas no delineamento experimental inteiramente casualizado, com doze repetições e cinco tipos de polinização: uma visita de N. testaceicornis (1V); duas visitas de N. testaceicornis (2V); três visitas de N. testaceicornis (3V); polinização natural (PN) e autopolinização (AP). Os movimentos e o tempo de visita nas flores foram observados nas visitas de N. testaceicornis. Nas frutas, avaliaram-se as características: biometria das frutas, taxa de fecundação dos aquênios, qualidade pós-colheita, contribuição de agentes e mecanismos de polinização na massa média das frutas. Na biometria das frutas, o serviço de polinização contribuiu no comprimento longitudinal, que se apresentou mais elevado na PN e 3V, em relação à AP. Nos diferentes tipos de polinização, a taxa de fecundação dos aquênios não diferiu e não apresentou efeito sobre a massa fresca das frutas. Na qualidadepós-colheita, 1V, 3V e PN apresentaram melhores resultados no grau de deformação e frutas comercializáveis. A abelha N. testaceicornis e a polinização natural contribuíram na massa fresca média das frutas de morangueiro. Os serviços de polinização natural e de N. testaceicornis proporcionaram melhoria na qualidade e agregação de valor das frutas. A abelha N. testaceicornis apresentou-se como polinizadora efetiva do morangueiro.(AU)

Introducing a Technique in Sustaining Adult Worker Bees of Heterotrigona itama (Cockerell, 1918) in Laboratory to Facilitate Future Health Research on Kelulut Honey

@#Introduction: The study is introducing a laboratory technique to sustain the longevity of Heterotrigona itama stingless bees collected from the farm in order to facilitate future health research on Kelulut honey. Methods: The worker bees were held in laboratory at 26 ± 2 oC, 57 ± 8 % relative humidity (RH) and 12:12 hours (light:dark period) in a cup covered with meshed cloth and installed with an inverted-wick system consists of a drinking straw with the bottom end loosely plugged with cotton wool. The artificial diet was pipetted into the straw to wet the cotton wool. The bees were divided into five diet groups, namely a) unfed - control, b) distilled water, c) purified tap water, d) non-carbonated isotonic drink or e) 5% honey solution. Feeding activity and survival of worker bees were observed daily. Results: The worker bees are seen to frequent and lick the cotton wool wetted with artificial diets. Comparison between the artificial diets, Kaplan-Meier statistical analysis showed that the 5% honey solution and non-carbonated isotonic drink have significantly (P < 0.05) extended the longevity of the worker bees with 50% survival probability at least 8 days. When the similar holding and feeding technique used for the bees from commercial farms, the 50% survival probability was extended to 14 days. Conclusion: The inverted-wick system with the use of 5% honey solution or isotonic drink as the artificial diet is capable to hold the H. itama worker bees at least for a week with survival above 50% for laboratory experiments.

Management practices and honey characteristics of Melipona eburnea in the Peruvian Amazon / Práticas de manejo e características do mel de Melipona eburnea Friese na Amazônia peruana

ABSTRACT: Meliponiculture and associated honey production are activities with a great potential for general use and commercialization in the Peruvian Amazon. Lack of management techniques and limited knowledge of honey quality are two factors which may hinder the implementation of meliponiculture. The present study was conducted in three communities in the Peruvian Amazon where the production and physicochemical and microbiological characteristics of honey from Melipona eburnea were evaluated at 90 days following transfer from natural to rational hives. Honey from M. eburnea was also compared with the 90-day-old honey from Melipona grandis, Melipona illota, and Melipona titania. In addition, 180-day-old honey from M. eburnea was sampled and compared with the younger 90-day-old honey. The production of honey varied from 900 to 1400 mL/colony/3 months for M. eburnea. When moisture and total sugars among the different species of Melipona were compared, no significant differences were observed. Neither were there significant differences between the M. eburnea honey at 90 and 180 days of maturation. Microbiological analyses for both treatments had values <0.3/g most probable number (MPN) of coliforms and organisms of fecal origin. Stingless bee honey can therefore be harvested after 3 months, by which time its physicochemical characteristics are stable and it is microbiologically suitable for human consumption.

RESUMO: A meliponicultura e a produção associada de mel é uma atividade com grande potencial de uso e comercialização na Amazônia peruana, contudo, as poucas técnicas de manejo e o pouco conhecimento da qualidade do mel oferecido limitam seu desenvolvimento. O presente estudo foi realizado em três comunidades da Amazônia peruana, onde a produção e as características físico-químicas e microbiológicas do mel de Melipona eburnea foram avaliadas 90 dias após a instalação em colmeias racionais. O mel de M. eburnea também foi comparado ao mel de 90 dias de M. grandis, M. illota e M. titania. Além disso, o mel de M. eburnea foi amostrado aos 180 dias. A produção de mel variou de 900 a 1400 ml/colônia/três meses para M. eburnea. Quando a umidade e os açúcares totais foram comparados entre as diferentes espécies, não foram encontradas diferenças significativas. Também não foram encontradas diferenças significativas para o mel de M. eburnea aos 90 e 180 dias de maturação. As análises microbiológicas para os dois tratamentos apresentaram valores <0,3 / g MPN (número mais provável) de coliformes e organismos de origem fecal. O mel de abelha sem ferrão pode, portanto, ser colhido após três meses, pois mantém suas características físico-químicas e é microbiologicamente adequado para consumo humano.

Determination of phenolic profile by HPLC-ESI-MS/MS, antioxidant activity, in vitro cytotoxicity and anti-herpetic activity of propolis from the Brazilian native bee Melipona quadrifasciata

ABSTRACT Propolis is a rich source of bioactive molecules made by bees by collecting an infinite variety of secondary metabolites from plants. This study determined the chromatographic and mass spectrometry profile along with the biological activity of propolis produced by the native Brazilian bee Melipona quadrifasciata. A hydroalcoholic crude extract was prepared and partitioned in solvents of different polarities, generating aqueous, dichloromethane, ethyl acetate, butanol and insoluble fractions. The phenolic and flavonoid content was also determined in crude aqueous and methanolic extracts as well. The antioxidant activity was determined by three different methods and correlated with the phenolic and flavonoid content. Extracts and fractions were tested against the herpes simplex virus type 1, and the cytotoxicity was evaluated in fibroblast L929 cells. The chromatographic and mass spectrometry analysis revealed the presence of catechin, epicatechin, aromadendrin, naringenin, pinocembrin and p-coumaric acid. The methanolic extract, ethyl acetate fraction and insoluble fraction possess higher phenolic and flavonoid content along with better antioxidant activities. The ethyl acetate and butanol fractions, along with the FM14 and FM45 subfractions demonstrated better viral inhibition, with 50% inhibitory concentrations of 90.5, 294, 188 and 58.5 µg ml−1, respectively. Among these, the ones presenting the best selectivity index were the ethyl acetate and FM45 fractions, with a pronounced virucidal effect. The methanolic extract stood out in the antioxidant activities and showed no cytotoxicity bellow 125 µg ml−1, exhibiting the most promising biological activity. Although this propolis seems to act in the early stages of herpetic infections, it is still difficult to correlate this activity with a single constituent of this complex mixture, suggesting a synergism in the extract components. To our knowledge, this is the first study of its kind with this propolis type.

Brazilian stingless bee propolis and geopropolis: promising sources of biologically active compounds

ABSTRACT Stingless bee products such as honey, pollen, propolis, and geopropolis have been used for centuries in traditional medicine for the treatment of several illnesses. Investigation of the biological activity of stingless bee products, especially propolis and geopropolis, has revealed promising therapeutic properties. About 20% of total Neotropical stingless bees can be found in Brazil. Despite the species diversity, studies on their biological activity are scarce. The present review focuses on the antioxidant and antimicrobial activities of propolis and geopropolis from Brazilian stingless bees. In addition, the toxicity of these natural products was addressed. In order to provide new evidences for the toxic potential of propolis and geopropolis components, an in silico analysis was performed using the ADMET PredictorTM software. We observed that most of studies evaluated only crude ethanol extracts of a limited number of stingless bees species. Propolis and geopropolis displayed antioxidant capacity and antimicrobial activity. Concerning the toxic potential, the extracts of stingless bees propolis and geopropolis were considered safe. Nonetheless, in vitro and in vivo toxicological studies are still necessary.

Partial characterization of ethanolic extract of Melipona beecheii propolis and in vitro evaluation of its antifungal activity

ABSTRACT The ethanolic extract of Melipona beecheii Bennett 1831, propolis from Yucatán Mexico was evaluated in vitro for the determination of its phenolic compound content, antioxidant capacity and antifungal activity. The results were compared against those of the ethanolic extract of Apis mellifera propolis. The total phenolic content, flavonoid content and flavanones-dihydroflavonols content were assessed by colorimetric methods. The antifungal activity was assessed in vitro against Candida albicans. For the ethanolic extract of M. beecheii propolis; total phenolic content, was 263.25 ± 8.78 µg/ml, total flavonoid content was 768 ± 204 µg/ml and flavanones-dihydroflavonols content was 335.42 ± 15.08 µg/ml. For antioxidant activity assessed as DPPH scavenging and iron reducing power ethanolic extract of M. beecheii propolis reported IC50 of 32.47 and 1.60 µg/ml of gallic acid equivalent respectively. Regarding antifungal activity against C. albicans, the minimal inhibitory concentration and minimal fungicidal concentration for ethanolic extract of M. beecheii propolis were 1.62 ± 0.33 and 2.50 ± 0.22 µg/ml of dry extract; For both minimal inhibitory concentration and minimal fungicidal concentration, ethanolic extract of M. beecheii propolis required 30% less concentration of dry extract than ethanol extract of A. mellifera propolis to exert the same antifungal actions against C. albicans. To the best of our knowledge, this is the first report about the flavonoid and flavanones-dihydroflavonols content of M. beecheii propolis. Due to the lack of information available about the stingless bee's honeycomb products, the study and conservation of endemic honeybees should remain as an active focus of research.

Pentacyclic triterpenes and other constituents in propolis extract from Melipona beecheii collected in Yucatan, México

ABSTRACT Thirteen pentacyclic triterpenes, methyl 3-oxours-12-en-23-oate, marsformosanone, taraxerone, β-amyrenone, α-amyrenone, lupenone, 24-methylencycloartan-3-one, moretenol acetate, β-amyrin acetate, germanicol acetate, 24-methylencycloartanyl acetate, β-amyrin, and α-amyrin were identified in a chloroform-methanol propolis extract from Melipona beecheii. Additionally, were identified in this propolis, hexadecanoic acid, methyl ester, octadecanoic acid, methyl ester and 1-triacontanol. The purification of the propolis extract was carried out using different chromatographic techniques, including vacuum liquid chromatography, gravity column chromatography and gel filtration chromatography Sephadex LH-20. The identification of the metabolites was performed using mass spectrometry.

Development and validation of a high-performance thin layer chromatography method for the simultaneous quantitation of α- and γ-mangostins in Thai stingless bee propolis

ABSTRACT Stingless bees (Apoidea) are widely distributed and commercially cultivated in artificial hives in fruit gardens. Their propolis are commonly used in traditional medicine to treat various diseases (e.g., abscesses, inflammations, and toothaches) and as a constituent of numerous health products. Thus, this study aimed to (i) develop and validate a high-performance thin layer chromatography method for the quantitation of major active constituents (α- and γ-mangostins) in propolis produced by five stingless bee species (Tetragonula fuscobalteata Cameron, T. laeviceps Smith, T. pagdeni Schwarz, Lepidotrigona terminata Smith, and L. ventralis Smith) cultivated in Thai mangosteen orchards and (ii) determine an optimal extraction solvent. Separation was performed on a silica gel 60 F254 plate using toluene/ethyl acetate/formic acid (8:2:0.1, v/v/v) as a mobile phase, and the developed method was validated to assure its linearity, precision, accuracy, and limits of detection/quantitation. Propolis extract from T. fuscobalteata exhibited the highest mangostin content, and acetone was shown to be more a more effective extraction solvent than dichloromethane, ethanol, or methanol. Thus, the simplicity and reliability of the developed method make it well suited for the routine analysis (e.g., for quality control) of commercial products containing stingless bee propolis.

Nest architecture and colony characteristics of Meliponula (Axestotrigona) ferruginea (Hymenoptera, Apidae, Meliponini) in Cameroon

Stingless bees have evolved adaptive nest constructions strategies which have resulted in sophisticated nest architecture inmany species while others lack certain structural components. However, no information exists on the nest biology andecology on the genus Meliponula in Cameroon. This study aims to contribute to knowledge on the nest architecture andcolony characteristics of Meliponula (Axestotrigona) ferruginea. Meliponula ferruginea belongs to the genus Meliponulaand subgenus Axestotrigona. This species was first described by Le Peletier De Saini-Farrgeau (1836) and Michener (Thebees of the world, The Johns Hopkins University Press, Baltimore and London, 2000) recently. In Cameroon the specieshas been collected in the northern parts of the country, but there has been no attempt to describe the nest architecture. Ourstingless bee survey from the Bamenda highlands afromontane forests of Cameroon reveals that this species can either nestin tree trunks or in abandoned traditional hollow hives. Interestingly, 50% of colonies studied nested in traditional hollowhives originally baited for honeybees (Apis mellifera adansonii). The nest entrance of Meliponula (Axestotrigona) ferruginea ranged from 1 to 1.5 cm (1.2±0.0.24 cm) in diameter, while the external entrance tube extended up to 1.6±0.0.4cm and the nest entrance tube can go up to 5 cm inward in depending on the thickness of the tree trunk. The nest is made upof sticky brown 5–7 (6±0.98) involucra sheets. The size of brood area depends on the age and condition of the nest. Thelength of the nest could be between 5.5 and 7.6 cm (6.8±0.98 cm) and up to 13± 4.16 cm combs with a comb diameter ofabout 7.5±1.29 cm. The cells and storage pots are oval in shape but the storage pots are much larger, about 3 times the sizeof cell. Generally, this study shows a lot of similarities with other previously studied species in terms of the nestcharacteristics and measurements. However, the above characteristics can be very useful in taxonomical, phylogenetic andecological studies of M. ferruginea.

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

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