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.

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

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.

Morphometric differences and fluctuating asymmetry in Melipona subnitida Ducke 1910 (Hymenoptera: Apidae) in different types of housing / Diferenças morfométricas e assimetria flutuante em Melipona subnitida Ducke 1910 (Hymenoptera: Apidae) em diferentes tipos de habitação

Abstract A geometric morphometrics approach was applied to evaluate differences in forewing patterns of the Jandaira bee (Melipona subnitida Ducke). For this, we studied the presence of fluctuating asymmetry (FA) in forewing shape and size of colonies kept in either rational hive boxes or natural tree trunks. We detected significant FA for wing size as well as wing shape independent of the type of housing (rational box or tree trunks), indicating the overall presence of stress during the development of the studied specimens. FA was also significant (p < 0.01) between rational boxes, possibly related to the use of various models of rational boxes used for keeping stingless bees. In addition, a Principal Component Analysis indicated morphometric variation between bee colonies kept in either rational hive boxes or in tree trunks, that may be related to the different origins of the bees: tree trunk colonies were relocated natural colonies while rational box colonies originated from multiplying other colonies. We conclude that adequate measures should be taken to reduce the amount of stress during bee handling by using standard models of rational boxes that cause the least disruption.

Resumo A abordagem da morfometria geométrica foi aplicada para avaliar as diferenças nos padrões das asas anteriores da abelha Jandaíra (Melipona subnitida Ducke). Para isso, estudou-se a presença de assimetria flutuante (AF) na forma das asas anteriores e tamanho das colônias mantidas tanto em caixas de colméia racional ou troncos de árvores naturais. Foi detectado AF significativa para o tamanho da asa, bem como a forma da asa independente do tipo de habitação (caixa racional ou cortiço), indicando a presença global de estresse durante o desenvolvimento dos espécimes estudados. AF também foi significativa (p < 0,01) entre as caixas racionais, possivelmente relacionados com a utilização de vários modelos de caixas racionais utilizados para a conservação de abelhas sem ferrão. Além disso, a Análise de Componentes Principais indicou variações morfométricas entre as colônias de abelhas mantidos em caixas racionais ou em cortiços, que podem estar relacionados com as diferentes origens das abelhas: os cortiços foram colônias naturais realocados, enquanto as colônias das caixas racionais foram originadas da multiplicação outras colônias. Conclui-se que devem ser tomadas medidas adequadas para reduzir a quantidade de estresse durante o manuseio abelha usando modelos padrão de caixas racionais que causam a menor perturbação.

Morphometric differences and fluctuating asymmetry in Melipona subnitida Ducke 1910 (Hymenoptera: Apidae) in different types of housing.

A geometric morphometrics approach was applied to evaluate differences in forewing patterns of the Jandaira bee (Melipona subnitida Ducke). For this, we studied the presence of fluctuating asymmetry (FA) in forewing shape and size of colonies kept in either rational hive boxes or natural tree trunks. We detected significant FA for wing size as well as wing shape independent of the type of housing (rational box or tree trunks), indicating the overall presence of stress during the development of the studied specimens. FA was also significant (p < 0.01) between rational boxes, possibly related to the use of various models of rational boxes used for keeping stingless bees. In addition, a Principal Component Analysis indicated morphometric variation between bee colonies kept in either rational hive boxes or in tree trunks, that may be related to the different origins of the bees: tree trunk colonies were relocated natural colonies while rational box colonies originated from multiplying other colonies. We conclude that adequate measures should be taken to reduce the amount of stress during bee handling by using standard models of rational boxes that cause the least disruption.

Parâmetros genéticos para características produtivas e biométricas em abelha Melipona quadrifasciata anthidioides LEPELETIER / Genetic parameters for production and biometrics traits in honey bee Melipona quadrifasciata anthidioides LEPELETIER

Os objetivos deste trabalho foram estimar componentes de variância genética aditiva, fenotípica e residual e a herdabilidade para características relacionadas com a produção de mel e com a estrutura do ninho de abelhas Melipona quadrifasciata anthidioides. Sessenta colônias de diferentes regiões da Bahia foram transladadas para caixas padronizadas modelo INPA e divididas, originando as gerações parentais G1 e G2. Foram medidas as características: estimativa da produção de mel; número, largura, volume e altura dos potes de mel; número, altura e diâmetro dos potes de pólen; peso; número, largura e diâmetro dos discos de cria e estimativa da população da colônia. As medidas foram corrigidas para o efeito fixo de mês de mensuração. Os componentes de variância e herdabilidade foram estimados por meio do método de semelhança entre parentes, utilizando-se abordagem Bayesiana. As médias e os desvios padrão variaram de 2,01±0,70cm para diâmetro de potes de pólen a 2.333,0±384,1kg para o peso das caixas. Houve indicação de convergência para todas as cadeias obtidas. As estimativas de variância genética aditiva variaram de 0,02cm para as características largura dos potes de mel a 38.587,72kg para o peso. Para as estimativas de variâncias fenotípicas, os valores variaram de 0,05 para a altura dos potes de pólen a 95.136,43kg para o peso; e para as variâncias residuais, os valores encontrados variaram de 0,02 para a variável largura dos potes de mel a 56.548,71kg para o peso. As estimativas de herdabilidade variaram de 0,35 a 0,53. Os resultados demonstraram que as características avaliadas possuem variação genética aditiva que garante boa resposta à seleção.

The aim of this study was to estimate components of genetic variance, phenotypic and residual and heritability for traits related to the production of honey and the nest structure of bee Melipona quadrifasciata anthidioides. Sixty colonies from different regions of Bahia were transferred to standard INPA model boxes and divided, creating the parental generations G1 and G2. The following characteristics were measured: estimated production of honey, number, width, height and volume of the honey pots, number, height and diameter of pollen storage pots, weight, number, length and diameter of the brood combs and estimate the population of the colony. The measurements were corrected for the fixed effects of month of measurement. The variance components and heritability were estimated by the method of similarity between relatives using the Bayesian approach. The mean and standard deviations ranged from 2.01±0.70cm diameter pots for pollen to 2333.0±384.1kg to the weight of the boxes. There was indication of convergence for all the chains obtained. Estimates of the additive genetic variance ranged from 0.02cm to the width characteristics of honey pots to 38587.72kg for weight. For the estimates of phenotypic variance the values ranged from 0.05 for the height of the pollen pots to 95136.43kg for weight; and the residual variance the values varied from 0.02 for the variable width of the honey pots to 56548.71kg for weight. The results showed that the characteristics assessed have additive genetic variation that ensures good response to selection.

Size and shape in Melipona quadrifasciata anthidioides Lepeletier, 1836 (Hymenoptera; Meliponini) / Tamanho e forma em Melipona quadrifasciata anthidioides Lepeletier, 1836 (Hymenoptera; Meliponini)

This study aimed to identify differences in wing shape among populations of Melipona quadrifasciata anthidioides obtained in 23 locations in the semi-arid region of Bahia state (Brazil). Analysis of the Procrustes distances among mean wing shapes indicated that population structure did not determine shape variation. Instead, populations were structured geographically according to wing size. The Partial Mantel Test between morphometric (shape and size) distance matrices and altitude, taking geographic distances into account, was used for a more detailed understanding of size and shape determinants. A partial Mantel test between morphometris (shape and size) variation and altitude, taking geographic distances into account, revealed that size (but not shape) is largely influenced by altitude (r = 0.54 p 0.01). These results indicate greater evolutionary constraints for the shape variation, which must be directly associated with aerodynamic issues in this structure. The size, however, indicates that the bees tend to have larger wings in populations located at higher altitudes.

Este trabalho avaliou a divergência de forma entre populações de Melipona quadrifasciata anthidioides, utilizando caracteres morfométricos em 23 localidades da região semi-árida do estado da Bahia (Brasil). As análises das distâncias de Procrustes entre as formas médias das asas indicaram que não há estruturação populacional para a variação dessa estrutura. Entretanto, nossas análises demonstraram que as populações estavam estruturadas geograficamente pelo tamanho das asas. O teste parcial de Mantel entre matrizes de distâncias morfométricas (forma e tamanho) e altitude, levando em conta as distâncias geográficas, foi utilizado para uma compreensão mais detalhada dos determinantes de tamanho e forma. O teste de Mantel entre as variações morfométricas (forma e tamanho) e altitude, tendo em conta as distâncias geográficas, revelou que o tamanho (mas não a forma) é amplamente influenciado pela altitude (r = 0,54 p 0,01). Tais resultados indicam maiores restrições evolutivas para a variação de forma, o que deve estar diretamente associado às questões aerodinâmicas dessa estrutura. O tamanho, por outro lado, indica que as abelhas estudadas tendem a apresentar asas maiores nas populações localizadas em regiões de maior altitude.

Genetic variability in Melipona scutellaris from Recôncavo, Bahia, Brazil.

Bees play a key role in pollination and thereby help maintain plant diversity. The stingless bee Melipona scutellaris is an important pollinator in northeastern Brazil because it is endemic to this region. Both deforestation and timber harvesting have reduced the nesting sites for this species, thus reducing its population and range. Genetic studies may help reverse this process by providing important tools for their proper management with a view to conservation of this species. Microsatellite markers have proven to be ideal for mapping genes and population genetic studies. Our aim was to study, using microsatellite markers, the interpopulation genetic variability of M. scutellaris in different parts of the Recôncavo region in Bahia State, Brazil. In all, 95 adult workers from 11 localities in Recôncavo Baiano (Amargosa, Cabaceiras do Paraguaçu, Conceição da Feira, Conceição do Almeida, Domingos Macedo Costa, Governador Mangabeira, Jaguaripe, Jiquiriça, Maragojipe, São Felipe, and Vera Cruz) were analyzed using 10 pairs of microsatellite primers developed for different Meliponini species. The total number of alleles, allele richness, and genetic diversity ranged from 2 to 7 per locus (average = 4.4), 1.00 to 4.88, and 0.0 to 0.850, respectively. The expected and observed heterozygosities varied from 0.0 to 0.76 and 0.0 to 0.84, respectively. No locus showed deviation from the expected frequencies in the chi-square test or linkage disequilibrium. The fixation index, analysis of molecular variance, and unweighted pair-group method using the arithmetic average revealed the effects of human activities on the populations of M. scutellaris, as little genetic structure was detected.

Size and shape in Melipona quadrifasciata anthidioides Lepeletier, 1836 (Hymenoptera; Meliponini).

This study aimed to identify differences in wing shape among populations of Melipona quadrifasciata anthidioides obtained in 23 locations in the semi-arid region of Bahia state (Brazil). Analysis of the Procrustes distances among mean wing shapes indicated that population structure did not determine shape variation. Instead, populations were structured geographically according to wing size. The Partial Mantel Test between morphometric (shape and size) distance matrices and altitude, taking geographic distances into account, was used for a more detailed understanding of size and shape determinants. A partial Mantel test between morphometris (shape and size) variation and altitude, taking geographic distances into account, revealed that size (but not shape) is largely influenced by altitude (r = 0.54 p < 0.01). These results indicate greater evolutionary constraints for the shape variation, which must be directly associated with aerodynamic issues in this structure. The size, however, indicates that the bees tend to have larger wings in populations located at higher altitudes.

Transferability of microsatellite primers developed for stingless bees to four other species of the genus Melipona.

Microsatellite markers are a useful tool for ecological monitoring of natural and managed populations. A technical limitation is the necessity for investment in the development of primers. Heterologous primers can provide an alternative to searching for new loci. In bees, these markers have been used in populational and intracolonial genetic analyses. The genus Melipona has the largest number of species among bee genera, about 70, occurring throughout the Neotropical region. However, only five species of the genus Melipona have specific microsatellite markers. Given the great diversity of this genus, this number is not representative. We analyzed the transferability of 49 microsatellite loci to four other species of the genus Melipona (M. scutellaris, M. mondury, M. mandacaia, and M. quadrifasciata). Four individuals of each species, from different localities, were used in amplification tests. Primer pairs described for five Melipona species and for Trigona carbonaria were tested. Among the 49 loci, 22 gave amplification products for all four species, while three gave nonspecific bands and five showed no amplification products. The remaining loci varied in the pattern of amplification, according to the species examined. The number of alleles ranged from 1 to 6. The results demonstrate the possibility of using these heterologous markers in other Melipona species, increasing the number of loci that can be analyzed and contributing to further genetic analyses of intra- and intercolonial structure, which is required for conservation measure planning, genetic improvement and resolution of taxonomic problems.

Flight activity of Melipona asilvai Moure (Hymenoptera: Apidae).

Many stingless bee species are specific to their areas of occurrence. Even when adapted to their local climate and flora conditions, they are subject to modifications in the environment, directly influencing flight activity. The aim of this work is to obtain information about the flight activity of the stingless bee Melipona asilvai Moure, thus contributing to the knowledge of this species. The flow of bees entering and leaving the colony was evaluated, and the type of transported material was identified. This information was correlated with climatic data collected at the time of observations, performed between June 2002 and March 2003. It can be proved that temperature was the factor with the greatest influence on the external activity of this species, showing a significant positive correlation with the entry of bees into the colony and pollen collection. Mud collecting showed a significant positive correlation with a humidity increase. Flight activity began at a temperature of 21.0 degrees C and humidity of 84.5%, peaking at 27.4 degrees C and 60.6% RH, respectively.

Flight activity of Melipona asilvai Moure (Hymenoptera: Apidae)

Muitas espécies de meliponíneos são específicas de suas regiões de ocorrência. Mesmo sendo adaptadas às condições locais de clima e flora, estão sujeitas às modificações do ambiente externo, influenciando diretamente a sua atividade de vôo. Este trabalho teve como objetivo obter informações sobre a atividade de vôo de Melipona asilvai Moure, contribuindo para o conhecimento desta espécie. Foi avaliado o fluxo de abelhas, entrando e saindo da colônia, e a identificação do tipo de material transportado. Estas informações foram correlacionadas com dados climáticos coletados no momento das observações, realizadas entre junho de 2002 e março de 2003. A temperatura foi o fator de maior influência sobre a atividade de vôo desta espécie, apresentando correlação positiva significativa com a entrada de abelhas na colônia e com a coleta de pólen. A coleta de barro apresentou correlação positiva significativa com o aumento da umidade. A atividade de vôo foi iniciada à temperatura de 21,0 °C e umidade de 84,5%, apresentando pico com 27,4 °C e 60,6%, respectivamente.

Pollen spectrum of honey of "uruçu" bee (Melipona scutellaris Latreille, 1811)

In spite of the importance of the "uruçu" bee as honey producer of excellent quality, as well, potential pollinator both in agricultural and natural ecosystems, mainly in North-eastern Brazil, just some information is found in literature about sources that such bees utilize to collect nectar and pollen. The identification of the plants visited by Melipona scutellaris was accomplished with base on the analysis of pollen types found in the honey samples collected every two months, from March 1997 to February 1998, in 15 colonies located in Catu, State of Bahia, Brazil (12§21'00"S, 38§22'40"W, 76 m of altitude). Qualitative and quantitative analysis of the pollen types were carried out determining the percentage and occurrence classes. Twenty-eight pollen types were found, being considered dominant pollen, the Eucalyptus spp. and Psidium sp. types and secondary pollen, Bauhinia sp., Caesalpinia sp. and Mimosa verrucata types. It was verified dominant pollen of Eucalyptus spp. in honeys produced in November/December 1997 and January/February 1998. The families Caesalpiniaceae (14 percent), Mimosaceae (25 percent) and Myrtaceae (56 percent) were the most represented in the pollen spectrum