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Made available in DSpace on 2022-04-28T19:47:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Satellite DNAs (satDNAs) and transposable elements (TEs) are among the main components of constitutive heterochromatin (c-heterochromatin) and are related to their functionality, dynamics, and evolution. A peculiar case regarding the quantity and distribution of c-heterochromatin is observed in the genus of bees, Melipona, with species having a low amount of heterochromatin and species with high amount occupying almost all chromosomes. By combining low-pass genome sequencing and chromosomal analysis, we characterized the satDNAs and TEs of Melipona quadrifasciata (low c-heterochromatin) and Melipona scutellaris (high low c-heterochromatin) to understand c-heterochromatin composition and evolution. We identified 15 satDNA families and 20 TEs for both species. Significant variations in the repeat landscapes were observed between the species. In M. quadrifasciata, the repetitive fraction corresponded to only 3.78% of the genome library studied, whereas in M. scutellaris, it represented 54.95%. Massive quantitative and qualitative changes contributed to the differential amplification of c-heterochromatin, mainly due to the amplification of exclusive repetitions in M. scutellaris, as the satDNA MscuSat01-195 and the TE LTR/Gypsy_1 that represent 38.20 and 14.4% of its genome, respectively. The amplification of these two repeats is evident at the chromosomal level, with observation of their occurrence on most c-heterochromatin. Moreover, we detected repeats shared between species, revealing that they experienced mainly quantitative variations and varied in the organization on chromosomes and evolutionary patterns. Together, our data allow the discussion of patterns of evolution of repetitive DNAs and c-heterochromatin that occurred in a short period of time, after separation of the Michmelia and Melipona subgenera. Laboratório de Citogenética de Insetos Departamento de Biologia Geral Universidade Federal de Viçosa, Campus Viçosa, Av. P.H. Rolfs s/n Departamento de Biologia Geral E Aplicada Instituto de Biociências/IB UNESP–Universidade Estadual Paulista Departamento de Biologia Geral E Aplicada Instituto de Biociências/IB UNESP–Universidade Estadual Paulista

Bee populations are declining globally due to different environmental stressors, such as pathogens, malnutrition, and agrochemicals. Brazil is the home of hundreds of stingless bee species, some of...

Mites can establish ecologically mutualistic and/or commensal relationships with stingless bees. In the present study, we evaluated the acarine diversity associated with colonies of three stingless bee species (Melipona quadrifasciata quadrifasciata, Scaptotrigona bipunctata and Tetragonisca fiebrigi) in seven localities of southern Brazil. A total of 1458 mites from 18 species/morphospecies were sampled. The colonies of Melipona q. quadrifasciata showed the highest richness of mites, but T. fiebrigi was the bee species with the highest acari diversity among the evaluated species. This information suggests a close relationship between mites and stingless bees, which has direct implications for the transport of hives and non-naturally occurring species to different regions.

Stingless bees are responsible for pollinating up to 90% of Brazilian flora, so their study is of commercial and ecological importance. Stingless bees mix plant resins with wax and soil, forming geopropolis. Studies of the variability in composition and activity due to species, place and season of collection are lacking. Yield, total phenolic and flavonoid content, and antioxidant activity (via DPPH) of the extracts of fifteen M. quadrifasciata (Mandacaia) geopropolis samples collected over a year in the State of Sao Paulo, Brazil, as well as two samples from different regions of the state of Minas Gerais in Brazil were compared. Composition was determined by ultra-high-resolution liquid chromatography and mass spectrometry, identifying 16 compounds. The month and region of collection affected the available plant resins and influenced their yield, composition and antioxidant capacity. Depending on the place of collection, M. quadrifasciata geopropolis is a promising natural source of antioxidant phenolic compounds.

O objetivo do trabalho foi determinar as espécies vegetais utilizadas por abelhas da espécie Melipona quadrifasciata Lepeletier, 1836, para a coleta de pólen. O estudo foi realizado por meio da análise melissopalinológica das cargas de pólen coletadas nas corbículas das abelhas campeiras durante o regresso aos seus ninhos. A comparação do material coletado com os grãos de pólen constituintes da palinoteca, da Universidade Federal do Piauí, permitiu a identificação de seis espécies vegetais associadas aos tipos polínicos mais representativos nas análises: Anadenanthera macrocarpa, Caesalpinia pyramidalis, Mimosa arenosa, Mimosa verrucosa, Spondias tuberosa, Waltheria rotudifolia. Dentre estas, quatro são pertencentes à família Fabaceae, com espécies vegetais da subfamília Mimosoideae. Observou-se que o mês de novembro apresentou maior quantidade e diversidade de pólen coletado. Embora tenha sido observada uma diversidade de espécies vegetais em florescimento na região de estudo, as abelhas apresentaram preferências bastante específicas no que diz respeito à coleta de pólen para a alimentação e manutenção das colônias. Dessa maneira, concluiu-se que as abelhas Melipona quadrifasciata L., no presente estudo, apresentaram preferência pelo pólen proveniente de plantas nativas da Caatinga, especialmente de espécies arbóreas.

International audience; We conducted several tests on the stingless bee Melipona quadrifasciata aiming to determine the impact of the glue used for applying radio frequency identification (RFID) tags on this bee species. The study was organized in three experimental sets, in which we evaluated the effects of a synthetic glue, a natural glue (shellac), and the effects of the bee manipulation alone (control group). We performed (i) an in vitro experiment (bioassay), in which we tested five different experimental treatments in triplicate: chip plus synthetic glue, chip plus shellac, synthetic glue, shellac, and control, totaling 150 bees (n = 30 per treatment); (ii) field experiments, in which we tested the RFID tracking system composed of RFID tags, reading units, antennas, and circuit boards; and (iii) the morphological and histochemical analyses of the flight muscles of bees collected from each experimental treatment (n = 5 per treatment) at 48 h after the beginning of bioassay. Use of the natural glue, as opposed to the synthetic glue, promoted an increase of the bees’ longevity while inhibiting detrimental impacts on their foraging activities, as observed by both the bioassay and field experiments. We found negative responses to the synthetic glue treatment combined with the electronic tags, showing that the natural glue induces less morphological damage to the flight muscles of tagged stingless bees.

The presence of Brazilian native bees can improve tomato production by increasing pollination effectiveness. However, the extensive use of pesticides in tomato cultures may be harmful to bees. Imidacloprid-based insecticides are used in tomato plantations because of its high efficiency against tomato pests. This study investigated the effects of oral intake of field-realistic concentrations of imidacloprid by M. quadrifasciata workers, a stingless native bee from Brazil and effective pollinators of tomato crops. The oral intake of sucrose syrup added with 10, 35, or 70 ppb of imidacloprid did not increase the mortality rate when compared with the control group. However, we observed a reduction in the workers’ motility and food consumption. We also treated M. quadrifasciata workers with sucrose syrup mixed with an imidacloprid-based insecticide (Evidence 700 WG®, Bayer), with the final concentration of 250 ppb of imidacloprid. This treatment did not cause visible alterations of the intestine absorptive cells of the bees’ midgut and did not increase DNA damage. Therefore, the observed reduction of food consumption and locomotion behavior of M. quadrifasciata workers may contribute to the global effort to understand the contribution of neonicotinoids on bees’ population decline process.

International audience; AbstractSex determination is one of the major developmental events in higher metazoans, where complex molecular mechanisms define two physiologically and behaviorally distinct organisms still genetically compatible. In honey bees, the sex determination cascade is initiated by the allelic composition of complementary sex determiner (csd) gene: males develop from hemi or homozygous embryos, whereas females develop from heterozygous embryos. In females, different alleles of csd lead to the formation of female-specific variants of feminizer (fem) and doublesex (dsx). In males, male-specific variants of fem and dsx are formed by default. In this paper, we investigated the genes of sex determination in the stingless bees Melipona quadrifasciata, Scaptotrigona postica, and Frieseomelitta varia. Our results revealed that the architecture of fem and dsx transcripts is highly conserved among the three stingless bee species, and also with honey bees.

Rio de Janeiro city presents one of the biggest urban forests today comprising several types of vegetation, highlighting the Atlantic tropical rainforest. Maintenance of these ecosystems requires protection and strong and efficient pollination. The stingless bees are between its main pollinators. The present investigation aims to recognize and to compare the pollen source preference and competition between Melipona rufiventris and Melipona quadrifasciata anthidioides in the Parque Nacional da Tijuca (PNT) urban forest at Rio de Janeiro. Bee pollen pellets were collected and prepared without the use of acetolysis. Evaluation intended the comparison of the taxon percentages of pollen grains collected by the bees in the same morning time. Dominant plant species were of Alchornea, Eucalyptus, Mimosa caesalpiniifolia, Mimosa scabrella, Melastomataceae, Myrcia and Solanum. The favorite sources of the two bee species in collecting pollen in the rainforest of the PNT were partially distinct among them. Therefore, an overlapping of harvesting sources was not so frequent during a year. In this respect, the forest needs several species of bees for pollination and perpetuation.

Bees represent efficient bioindicators of environmental contamination, including elements that can accumulate in both biomes and bee-derived products. Therefore, the goal of this study was to evaluate the presence of metallic elements over the body surface of Melipona quadrifasciata anthidioides workers from rural and urban areas from Caatinga in Brazil. Using SEM/XEDS (scanning electron microscope/energy dispersive X-ray spectroscopy) identified magnesium (Mg), aluminum (Al), calcium (Ca), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), molybdenum (Mo), cadmium (Cd), barium (Ba), and lead (Pb) in the corbiculae of bees. The areas had similar profiles, but As was characterized as a "trace element" in the agricultural area and "minor element" in the urban area. The presence of toxic elements such as As, Cd, and Pb indicates that both areas may be contaminated, revealing the importance of metallic elements microanalysis in native bee species for environmental biomonitoring.

Machine learning (ML) is a branch of artificial intelligence (AI) that enables the analysis of complex multivariate data. ML has significant potential in risk assessments of non-target insects for modeling the multiple factors affecting insect health, including the adverse effects of agrochemicals. Here, the potential of ML for risk assessments of glyphosate (herbicide; formulation) and imidacloprid (insecticide, neonicotinoid; formulation) on the stingless bee Melipona quadrifasciata was explored. The collective behavior of forager bees was analyzed after in vitro exposure to agrochemicals. ML algorithms were applied to identify the agrochemicals that the bees have been exposed to based on multivariate behavioral features. Changes in the in situ detection of different proteins in the midgut were also studied. Imidacloprid exposure leads to the greatest changes in behavior. The ML algorithms achieved a higher accuracy (up to 91%) in identifying agrochemical contamination. The two agrochemicals altered the detection of cells positive for different proteins, which can be detrimental to midgut physiology. This study provides a holistic assessment of the sublethal effects of glyphosate and imidacloprid on a key pollinator. The procedures used here can be applied in future studies to monitor and predict multiple environmental factors affecting insect health in the field.

Meliponiculture - the management of stingless bee colonies - is an expanding activity in Brazil with economic, social and environmental potential. However, unlike in apiculture, the pathogens that impact on meliponiculture remain largely unknown. In southern Brazil, every year at the end of the summer, managed colonies of the stingless bee Melipona quadrifasciata manifest a syndrome that eventually leads to collapse. Here we characterize the M. quadrifasciata virome using high-throughput sequencing, with the aim of identifying potentially pathogenic viruses, and test whether they are related to the syndrome outbreaks. Two paired viromes are explored, one from healthy bees and another from unhealthy ones. Each virome is built from metagenomes assembled from sequencing reads derived either from RNA or DNA. A total of 40 621 reads map to viral contigs of the unhealthy bees' metagenomes, whereas only 11 reads map to contigs identified as viruses of healthy bees. The viruses showing the largest copy numbers in the virome of unhealthy bees belong to the family Dicistroviridae - common pathogenic honeybee viruses - as well as Parvoviridae and Circoviridae, which have never been reported as being pathogenic in insects. Our analyses indicate that they represent seven novel viruses associated with stingless bees. PCR-based detection of these viruses in individual bees (healthy or unhealthy) from three different localities revealed a statistically significant association between viral infection and symptom manifestation in one meliponary. We conclude that although viral infections may contribute to colony collapses in the annual syndrome in some meliponaries, viruses spread opportunistically during the outbreak, perhaps due to colony weakness.

Meliponine is a honey with particular characteristics; it has a high percentage of humidity, which can favor the development of microorganisms, thereby causing changes in the quality of the product. The objective of this study was to evaluate the microbiological characteristics of meliponine honey produced by different species and marketed in the State of Paraná-Brazil. The sample unit was composed of honey of Tetragonisca angustula (n = 15), Scaptotrigona bipunctata (n = 05), Melipona quadrifasciata quadrifasciata (n = 05) and Melipona bicolor schencki (n = 01). The following microbiological parameters were evaluated: counts of total and thermotolerant coliforms, fungi count and detection of Salmonella spp. Principal component analysis (PCA) was used to evaluate the possible relationships among species, locality and type of microorganism. For microorganisms of the total coliform group, 15.38% of the analyzed samples had values >3 MPN/g. The presence of thermotolerant coliforms was observed in 7.69% of the samples, all of which were T. angustula samples. For fungi counts, 100.00% of the analyzed samples had values within the established range created by the Agricultural Defense Agency of Paraná. The presence of Salmonella spp. was not observed in any of the samples evaluated. The amount of water in the honey is related to the location of the apiary and edaphoclimatic factors. Adoption of good handling practices by beekeepers is indispensable for the safety of the final product. La meliponíne produce una miel diferenciada con características particulares, presenta un alto porcentaje de humedad, lo que puede favorecer el desarrollo de microorganismos provocando cambios en la calidad del producto. El objetivo de este estudio fue evaluar las características microbiológicas de la miel de meliponíne producida por diferentes especies y comercializada en el Estado de Paraná-Brazil. La unidad de muestra estuvo compuesta por miel de Tetragonisca angustula (n = 15), Scaptotrigona bipunctata (n = 05), Melipona quadrifasciata quadrifasciata (n = 05) y Melipona bicolor schencki (n = 01). Se evaluaron parámetros microbiológicos: recuento de coliformes totales y termotolerantes, recuento de hongos y detección de Salmonella spp. Análisis de componentes principales: se utilizó ACP para evaluar las posibles relaciones entre especies, localidad y tipo de microorganismo. Para los microorganismos del grupo de coliformes totales, el 15,38% de las muestras analizadas presentaron valores >3 NMP/g. En el 7.69% de las muestras se observó la presencia de coliformes termotolerantes, siendo todas muestras de T. angustula. Para contar hongos, el 100,00% de las muestras analizadas presentaron valores dentro de lo establecido por la Agencia de Defensa Agropecuaria de Paraná. No se observó presencia de Salmonella spp en ninguna de las muestras evaluadas. Algunas muestras fueron similares según la especie productora y la cantidad de agua en la miel está relacionada con la ubicación del meliponario, debido a factores edafoclimáticos. La adopción de buenas prácticas de manipulación por parte de los productores de miel es fundamental para garantizar la seguridad alimentaria del producto final. Os meliponíneos elaboram um mel diferenciado com características particulares, apresenta elevado percentual de umidade o que pode favorecer o desenvolvimento de microrganismo ocasionando alterações na qualidade do produto. O objetivo deste estudo foi avaliar as características microbiológicas do mel de meliponíneos produzido por diferentes espécies e comercializado no Estado do Paraná-Brasil. A unidade amostral foi composta de mel de Tetragonisca angustula (n=15), Scaptotrigona bipunctata (n=05), Melipona quadrifasciata quadrifasciata (n=05) e Melipona bicolor schencki (n=01). Parâmetros microbiológicos foram avaliados: contagem de coliformes totais e termotolerantes, contagem de fungos e detecção de Salmonella spp. A análise de componentes principais - ACP foi utilizada para avaliar as possíveis relações entre espécie, localidade e tipo de microrganismo. Para microrganismos do grupo coliformes totais, 15,38% das amostras analisadas apresentaram valores >3 NMP/g. Em 7,69% das amostras foi observado a presença de coliformes termotolerantes, sendo todas amostras de T. angustula. Para contagem de fungos, 100,00% das amostras analisadas apresentaram valores dentro do estabelecido pela Agência de Defesa Agropecuária do Paraná. Não foi observado a presença de Salmonella spp em nenhuma das amostras avaliadas. Algumas amostras se assemelharam segundo a espécie produtora e a quantidade de água no mel é relacionada com a localização do meliponário, devido a fatores edafoclimáticos. Adoção de boas práticas de manipulação pelos meliponicultores se tornam indispensáveis para conferir segurança alimentar ao produto final.

Little information about the contribution of plants to the production of honey by bees exists for southern Brazil. Information on this dynamic is crucial to maintaining the health of bees, the pollination of forests and assisting in the management of apiaries and meliponaries. The objective of the study was to elucidate which plants the bees visit for the production of honey in Rio Grande do Sul (RS), southern Brazil. 43 honey samples were selected from Apis mellifera, Tetragonisca angustula, Melipona quadrifasciata, Scaptotrigona bipunctata, Plebeia remota and Plebeia droryana. The honey samples were submitted to acetolysis. 110 pollen types were found in all honey samples from bees of Apis mellifera (97 pollen types), Melipona quadrifasciata quadrifasciata (14 pollen types), Tetragonisca angustula (46 pollen types), Scaptotrigona bipunctata (8 pollen types), Plebeia remota (17 pollen types) and Plebeia droryana (8 pollen types). The high pollen percentage of tree species in the samples stands out, ranging from 16.8% to 99.8%. The samples were grouped into seven groups by CONISS, called Melis-1, Melis-2, Melis-3, Melis-4, Melis-5, Melis-6 and Melis 7, and these are related to the floral resources of the studied bees. Some pollen types showed high percentages in honeys from Apis mellifera (pollen types of Eucalyptus sp. Myrcia type, Casearia sylvestris, Hovenia dulcis, Mimosa scabrella, Lamanonia ternata, Clethra scabra, Baccharis type and Weinmannia paulliniifolia), Tetragonisca angustula (pollen types of Eryngium sp./Petroselinum crispum, Handroanthus sp., Sorocea bonplandii and Parapiptadenia rigida), Melipona quadrifasciata quadrifasciata (pollen types of Myrcia type, Eucalyptus sp. and Mimosa bimucronata), Scaptotrigona bipunctata (pollen types of Allophylus edulis, Butia type and Parapiptadenia rigida), Plebeia remota (pollen types of Myrcia type, Butia type and Sebastiania sp.) and Plebeia droryana (pollen types of Myrcia type). The different types of vegetation in RS accounts for the different pollen spectra in honeys from southern Brazil.

International audience; Exposure to agrochemicals and pathogens has been indicated as critical factors of declining pollinator populations, including species of native tropical bees. In the present study, we investigated the sublethal effects of acute oral exposure to azadirachtin and spinosad bioinsecticides on the expression of immunity-related genes and the number of hemocytes in uninfected and Escherichia coli–infected workers of the stingless bee Melipona quadrifasciata. Expression of the immune-related gene vitellogenin decreased and the number of circulating hemocytes increased in bioinsecticide-exposed uninfected bees and in unexposed E. coli–infected bees as compared with unexposed uninfected bees. Our results show that though bioinsecticides are considered safe, contamination by sublethal doses of azadirachtin and spinosad can interfere with the immune system of bees.

Microbes have been identified as fundamental for the good health of bees, acting as pathogens, protective agent against infection/inorganic toxic compounds, degradation of recalcitrant secondary plant metabolites, definition of social group membership, carbohydrate metabolism, honey and bee pollen production. However, study of microbiota associated with bees have been largely confined to the honeybees and solitary bees. Here, I characterized the microbiota of indoor surface nest of four brazilian stingless bee species (Apidae: Meliponini) with different construction behaviors and populations. Bees that use predominantly plant material to build the nest (Frieseomelitta varia and Tetragonisca angustula) have a microbiome dominated by bacteria found in the phylloplane and flowers such as Pseudomonas sp. and Sphingomonas sp. Species that use mud and feces (Trigona spinipes) possess a microbiome dominated by coliforms such as Escherichia coli and Alcaligenes faecalis. Melipona quadrifasciata, which uses both mud / feces and plant resin, showed a hybrid microbiome with microbes found in soil, feces and plant material. These findings indicate that indoor surface microbiome varies widely among bees and reflects the materials used in the construction of the nests.

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

A Bacia do Rio Doce é uma das principais bacias hidrográficas do estado de Minas Gerais, cujos recursos naturais e minerais têm sido explorados desde o processo de ocupação e desenvolvimento da região. No dia 05 de novembro de 2015, a Bacia do Rio Doce sofreu o que foi considerada a maior tragédia ambiental do país, no qual a Barragem de Fundão, pertencente à Samarco Mineração S.A. se rompeu, lançando cerca de 45 milhões de m3 de rejeitos de mineração no meio ambiente. A lama de rejeitos, canalizada pelo Rio Doce, causou a morte de 19 pessoas, poluiu as águas desde o local do rompimento, devastou matas ciliares remanescentes, soterrou formas vegetais de pequeno porte e bancos de sementes, dificultando assim processos de recuperação natural e sucessão das áreas atingidas. Sabendo da importância dos serviços de polinização realizada por abelhas sem ferrão para a regeneração da vegetação nativa, este trabalho teve como objetivo selecionar genótipos prioritários para a formação de populações de duas espécies de abelhas sem ferrão, Tetragonisca angustula e Melipona quadrifasciata. Popularmente conhecidas como Jataí e Mandaçaia, respectivamente, a reintrodução e aumento populacional destas abelhas nas áreas atingidas pelo rompimento da Barragem de Fundão, podem contribuir com a restauração ativa das áreas degradadas. Abelhas de ambas as espécies foram amostradas de populações nativas e de meliponários, em pontos localizados na região metropolitana de Belo Horizonte e na região de Mariana. Fragmentos do gene COI (citocromo oxidase I) foram utilizados para análise de estruturação populacional. Os resultados obtidos mostraram que existe uma estruturação populacional de T. angustula, sendo que, abelhas coletadas na região de Mariana formam grupos geneticamente diferentes de outras localidades amostrais. Para M. quadrifasciata não houve estruturação populacional, no entanto, haplótipos coletados em Mariana são exclusivos desta região. Desta forma, estratégias de reprodução de colônias e reintrodução na natureza devem considerar como prioritários os genótipos manejados localmente ou capturados diretamente na natureza por meio de ninhos-iscas na região de Mariana-MG. Os resultados obtidos reforçam a importância do estudo populacional previamente à tomada de decisões sobre reintrodução de colônias, para que esta seja feita de modo a respeitar a genética de populações locais, resguardando a saúde de colônias que serão deslocadas e garantido a sobrevivência e o crescimento populacional destes polinizadores. Palavras-chave: Abelhas sem ferrão. Reintrodução de colônias. Restauração ativa. Meliponicultura. Genotipagem. Tetragonisca angustula. Melipona quadrifasciata. The Rio Doce Basin is one of the main hydrographic basins in the state of Minas Gerais, whose natural and mineral resources have been exploited since the occupation and development of the region. On November 5, 2015, the Rio Doce Basin suffered what was considered the greatest environmental tragedy in the country, in which the Fundão Dam, belonging to Samarco Mineração SA broke, launching about 45 million m3 of mining tailings in the environment. The tailings mud, channeled through the Rio Doce, caused the death of 19 people, polluted the waters since the site of the breach, devastated remaining riparian forests, buried small vegetable forms and seed banks, thus hindering natural recovery and succession processes affected areas. Knowing the importance of pollination services performed by stingless bees for the regeneration of native vegetation, this study aimed to select priority genotypes for the formation of populations of two stingless bee species, Tetragonisca angustula and Melipona quadrifasciata. Popularly known as Jataí and Mandaçaia, respectively, the reintroduction and population increase of these bees in the areas affected by the Fundão Dam breach, can contribute to the active restoration of degraded areas. Bees of both species were sampled from native populations and meliponaries, at points located in the metropolitan region of Belo Horizonte and in the region of Mariana. Fragments of the COI gene (cytochrome oxidase I) were used for population structure analysis. The results obtained showed that there is a population structure of T. angustula, and bees collected in the Mariana region form groups that are genetically different from other sampling locations. For M. quadrifasciata there was no population structure, however, haplotypes collected in Mariana are exclusive to this region. Thus, strategies for colony reproduction and reintroduction into the wild must consider as a priority the genotypes managed locally or captured directly in the wild through bait nests in the Mariana-MG region. The results obtained reinforce the importance of the population study prior to making decisions on the reintroduction of colonies, so that it is done in a way that respects the genetics of local populations, safeguarding the health of colonies that will be displaced and ensuring population survival and growth of these pollinators. Keywords: Stingless bees. Colony reintroduction. Active restoration. Meliponiculture. Genotyping. Tetragonisca angustula. Melipona quadrifasciata.

Bee populations are declining globally due to different environmental stressors, such as pathogens, malnutrition, and agrochemicals. Brazil is the home of hundreds of stingless bee species, some of them now considered endangered, though very little is known about the impact of disease on native bees. In Southern Brazil the endangered stingless bee Melipona quadrifasciata is affected by an annual syndrome that causes sudden death of workers, eventually leading colonies to collapse. Although novel viruses were found in foragers from diseased colonies, none has been consistently implicated in the outbreak. Here we used transcriptomics in combination with an integrative longitudinal survey on managed colonies to identify predictors for preventing M. quadrifasciata colony failures. We found that key genes related to xenobiotic metabolization, nutrition and immune responses are downregulated in foragers from colonies that became diseased three months later. The period that preceded the outbreak was marked by pronounced forager weight loss as well as behavioural changes. Our findings support the proposition that worldwide bee mortality is influenced by a combination of diverse sublethal factors, and increase awareness of the long-term effects of genetic diversity erosion in stingless bee species, which enhances their vulnerability to environmental stressors.

Melipona quadrifasciata anthidioides is a species of stingless bee popularly known in Brazil as "mandacaia". Products derived from bees for food and therapeutic uses, have stimulated the evaluation of the chemical composition of geopropolis. Concentrations of 24 essential and potentially toxic elements were determined in geopropolis samples, using a sequential optical emission spectrometer with inductively coupled plasma (ICP OES) after microwave-assisted acid digestion. Principal component analysis (PCA) and Hierarchical cluster analysis (HCA) were used to carry out an exploratory analysis of the samples. The following elements were quantified (in mg Kg-1): Al (320,414.40-36,911.1), As (

Even though meliponines are among the greatest pollinating agents in Neotropical ecosystems, the populations of these bees are in decline due to anthropic actions, such as the indiscriminate use of pesticides. Neonicotinoids, such as Imidacloprid, are used in many crops and have been shown to be one of the pesticides groups with the most damaging effects on the health of bee colonies. The goals of this work were (1) to evaluate the ability of Melipona quadrifasciata, a native Brazilian stingless bee species, to detect the presence of Imidacloprid in artificial diets, and (2) to evaluate how this insecticide may affect their behavior. For this purpose, behavioral experiments were carried out in observation arenas where artificial flowers containing different concentrations of Imidacloprid (0%, 1%, 5%, 10%, and 20%) were made available to forager bees and their choice and behavior were registered. Our results showed that bee choices were not affected by the different concentrations of Imidacloprid, even at extremely high doses in the flowers (20%). It was also found that the presence of this insecticide interfered with the communication between individuals as a result of reduced antennation events. Both findings, the apparent inability of bees to detect the presence of the insecticide and the change in behavior caused by the exposition to the volatilized product, may rapidly lead to colony losses in nature.

Many deceptive orchids present variation in floral color and fragrance. This might be advantageous for the plant, as it can disturb the associative avoidance learning of pollinators, promoting more visits to the flowers. Some studies have shown that color and fragrance can be correlated in polymorphic deceptive orchids, but these studies employed color traits based on the human visual system and not the visual perception of pollinators. Thus, we investigated the composition of the floral fragrance of Ionopsis utricularioides (Sw.) Lindl., a polymorphic deceptive orchid, and analyzed possible correlations with the floral color as seen by bees, Apis mellifera L. and Melipona quadrifasciata Lepeletier, using the color hexagon model. We found high color and fragrance intraspecific variation, as expected for deceptive species. However, we found no color-fragrance association in individuals, either by comparing fragrance profiles with the color variable saturation or by comparing them with the placement of individuals in the color hexagon for both bee species. This lack of correlation contradicts the biochemical pathway hypothesis, which proposes that associations between floral color and scent in polymorphic flowers arise from shared biochemical pathways. However, a complete absence of correlation between floral signals is consistent with selection arising through pollinator cognitive ecology. Lack of correlation would increase the floral variability perceived by bees, given their multimodal learning, and this variability could disrupt avoidance learning of deceptive flowers, thus enhancing the efficacy of the plant's deceptive pollination mechanism.

The use of pesticides is considered one of the most important threats to pollinators, especially since they are widely used in agriculture for pest control. In the last years, several studies have reported severe secondary effects on various bee species, including exotic and native bees. In this study, lethal (mortality) and sublethal (locomotor activity) effects of insecticides and acaricides used in strawberries in Brazil (abamectin, novaluron, spinetoram, and thiamethoxam) were evaluated on the native stingless bees Melipona quadrifasciata and Tetragonisca fiebrigi. The results showed that the effects varied significantly according to the pesticide, type of exposure (oral or topical), and bee species. Through oral exposure, M. quadrifasciata was more susceptible to all insecticides except for abamectin, while in topical exposure, T. fiebrigi was more sensitive. Thiamethoxam followed by spinetoram and abamectin were the most lethal, regardless of species or exposure route; novaluron was not harmful at the highest tested dose. The locomotor activity of bees was altered in the presence of sublethal doses (LC10 and LC50) of all insecticides. Spinetoram and abamectin can be as much as toxic as thiamethoxam against M. quadrifasciata and T. fiebrigi in laboratory experiments. These findings should be confirmed in field experiments to define possibilities to combine pest control and pollinator management. In crops like strawberries, the selectivity of native pollinators should be considered.

Pesticides effects on pollinators and representativeness of native species in ecological risk assessment and studies are an increasing concern. Stingless bees are especially important in Brazil, a major agricultural producer. Given the lack of colony-level data, a feeding study methodology for Melipona quadrifasciata was proposed and tested with the toxic standard dimethoate. Free foraging colonies at five sites were offered sugar syrup containing 0 (C), 75 (T1), 100 (T2) and 200 (T3) μg dimethoate/Kg and assessed weekly up to five weeks after exposure. Dimethoate concentration was analytically verified in the artificial syrup and honey from colonies. Pollen from pots was identified throughout the experiment. Actual concentrations in the syrup were 75-117% of nominal levels and low dissipation/dilution occurred in honey pots during the study. Mean numbers of foragers, brood, honey pots, proportions of brood mortality and incomplete honey pots, behavior and colony survival were significantly affected in T3 colonies, compared to control. Transient effects on brood mortality and incomplete pots (T1) or on number of new cells (T2) were observed at the lowest concentrations, but were rapidly recovered and may not be treatment related. Pollen diversity varied between sites, but no clear treatment differences were observed. The proposed methodology allowed identification of suitable parameters for colony-level studies using M. quadrifasciata, and of the no-observed effect concentration (nominal: 100 μg/Kg) Efeitos de pesticidas em polinizadores e a representatividade de espécies nativas em estudos e avaliações de risco ecológicas são uma preocupação crescente. As abelhas sem ferrão são especialmente importantes no Brasil, um dos maiores produtores agrícolas do mundo. Dada a falta de dados no nível de colônias, uma metodologia de estudo de alimentação para Melipona quadrifasciata foi proposta e testada com o padrão tóxico dimetoato. Colônias livres para forragear foram expostas por seis semanas a 0 (C), 75 (T1), 100 (T2) e 200 (T3) μg de dimetoato/Kg de xarope de açúcar em cinco locais, e avaliadas semanalmente até cinco semanas após a exposição. Foi realizada uma verificação analítica do dimetoato no xarope artificial e no mel das colônias. O pólen dos potes foi identificado ao longo do experimento. As concentrações medidas no xarope foram 75-117% dos níveis nominais e baixa dissipação/diluição ocorreu nos potes de mel durante o estudo. Os números médios de forrageiras, cria, potes de néctar, proporções de mortalidade da cria e de potes incompletos de néctar, comportamento e sobrevivência das colônias foram significativamente afetados em colônias T3, em comparação com o controle. Efeitos transitórios na mortalidade de crias e potes incompletos (T1) ou no número de novas células (T2) foram observados nas concentrações mais baixas, mas foram recuperados e podem não estar relacionados ao tratamento. A diversidade de pólen variou entre os locais, mas não foram observadas diferenças claras entre tratamentos. A metodologia proposta permitiu a identificação de parâmetros adequados para estudos em nível de colônia com Melipona quadrifasciata e identificação da concentração de efeito adverso não observado para dimetoato (nominal: 100 μg/Kg)

International audience; The species-specific responses to climate change can result in discordant range shifts, leading to changes in associations among species. In this study, we tested whether climate change can affect the spatial association between the plant Mimosa scabrella and the stingless bees Melipona quadrifasciata anthidioides and M. quadrifasciata quadrifasciata. Ecological niche models were used to predict current and future species distributions using intermediate (RCP 4.5) and high (RCP 8.5) greenhouse gas emission scenarios. Our models projected a pronounced reduction in climatically suitable habitats available for the studied species and identified a species-specific response to climate change. As a result, the models estimated a drastic reduction in the spatial association among species of up to 94% in the coming years, revealing that climate change may affect species interactions.

Human disturbances are one of the main reasons for the recent deficit of pollinators, which can cause severe habitat loss, decreasing food availability and nesting resources for pollinators. However, few studies show how landscapes composition and configuration can impact the pollen foraging of bees. We aimed to evaluate how landscape structure and local characteristics can influence the pollen foraging patterns of the stingless bee Melipona quadrifasciata anthidioides in agricultural landscapes of Chapada Diamantina, a region of the Bahia state in Brazil. Pollen samples were collected from fourteen M. quadrifasciata anthidioides colonies to evaluate how pollen collection varies according to different proportions of human land-use, landscape heterogeneity, plant richness and distance from crops. To explain the richness and diversity of pollen collected by bees, we used generalized linear models, and linear models, compared through the Akaike information criteria corrected for small samples (AICc). Pollen richness can be explained by the distance from crops, plant richness and human land-use proportion. Bees will collect pollen from more plant species when they are in a natural and diverse landscape. Pollinator friendly crop areas must be planned to be surrounded by diverse natural nearby environments, guaranteeing yearlong pollen resources available for bees’ maintenance.

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

The pollination of tomatoes in greenhouses using bees (especially Bombus species) has increased worldwide, providing increased productivity. However, no concrete initiative has been carried out with bees for tomato pollination on a commercial scale, in greenhouses in Brazil. Thus, the aim of this study was to evaluate Melipona quadrifasciata efficiency in pollination and consequent production of tomatoes of different growth habits in greenhouse environment. Two experiments were conducted. The pollen load of the creeping tomato (determined growth) flowers and the fruit quality was quantified in the first experiment, in the greenhouse cultivation treatments with bees, without bees, and in the open environment. In the second experiment, the treatments were carried out with tomato of indeterminate growth in greenhouse with bees and without bees. The bees presented foraging behavior and visitation to tomato flowers. The pollen load for the determined tomato was 433% larger in a closed environment with bees whe...

Honey and bee bread samples from storage pots of Melipona quadrifasciata anthidioides were collected monthly from April 2015 to May 2016 in the Mogi Guacu Biological Reserve (22o 10ʹ S, 47o...

Bees are major pollinators of both native flora and cultured crops. Nonetheless, despite their key functional role in ecosystems and agriculture, bee populations have been affected worldwide by deforestation and contamination by insecticides. Conversely, little is known about the effects of pesticides on morphogenetic development of neotropical stingless bees. We compared the fluctuating asymmetry (FA) in newly emerged bees and foragers of Melipona quadrifasciata anthidioides exposed to pesticides (experimental greenhouse and cultivated field). In addition, visitation behavior of foragers was inferred from pollen analyses and direct observation. A significant increase of FA (P

Pollen loads collected by Melipona quadrifasciata anthidioides were examined in order to detect the botanical origin of pollen and to recognise the pollination action of these bees in an urban Atlantic secondary forest. Pollen analysis followed standard methodology and without the use of acetolysis. The results revealed a great contribution of monofloral (more than 90% of a single pollen type or more than 60% if no accessory pollen was present). Quantitatively, Myrtaceae pollen samples followed Melastomataceae ones. Bifloral or heterofloral samples combine several pollen types. Besides Myrcia and Eucalyptus, the most frequent pollen types were Solanum, Mimosa caesalpiniifolia and Alchornea. The current study was related to the apiaries' surrounding vegetation and reflected both the resources available and the preference for native plant species by the bees. Furthermore, these results are relevant to the management of primary and secondary forests in order to preserve the environments.

Objective. The objective of the present paper was to test the in vitro antibacterial activity of the Mandaçaia (Melipona quadrifaciata) bee’s geopropolis. Materials and methods. The experiment was carried out with secretion samples collected from animals with ear diseases evaluated at Unifeso’s Clinic School of Veterinary Medicine, where in vitro tests took place, antibiograns with alcoholic extract of geopropolis were made 10 test tubes (two tubes for each concentration of 50%, 60%, 70%, 80%, 90%). Results. Were not identified microorganisms at concentrations of 70, 80 and 90%. Conclusions. The geopropolis of Melipona quadrifasciata showed antibacterial activity in vitro against microorganisms from ear secretion of dogs with otitis externa., Objetivo. El objetivo del presente trabajo fue probar la actividad antibacteriana in vitro de la geopropólea de la abeja Mandaçaia (Melipona quadrifasciata). Materiales y métodos. Se realizó el experimento con muestras de secreciones obtenidas de animales con enfermedades del oído evaluados en la Clínica Escuela de Medicina Veterinaria de Unifeso, donde tuvieron lugar las pruebas in vitro, antibiograns con extracto alcohólico de geopropolis fueron realizados 10 tubos de ensayo (dos tubos para cada concentración del 50%, 60%, 70%, 80%, 90%). Resultados. No se identificaron los microorganismos a concentraciones de 70, 80% y 90%. Conclusiones. La geopropolis de Melipona quadrifasciata mostró actividad antibacteriana, in vitro, frente a microorganismos de la secreción del oído de los perros con otitis externa.

International audience; AbstractHygienic behavior in stingless bees is a trait of workers that confers colony-level resistance against some brood diseases. Workers of hygienic colonies detect, uncap and remove dead or diseased brood from the nest cells. We examined the hygienic behavior in stingless bees (Melipona quadrifasciata anthidioides) from freeze-killed brood assay using liquid nitrogen. Responses were measured at 14 times (3, 6, 12, 24, 48, 72, 96, 120, 144, 168, 192, 216, 240 and 264 h after freeze-killing of the brood). Workers were estimated to remove on average 65% of larvae and 34% of dead pupae within 48 h of freezing. Workers removed dead brood rapidly after uncapping the cells. Strong colonies showed a greater removal of dead pupae, while the size of the population did not influence the removal of dead larvae. These findings report for the first time the hygienic behavior in M. q. anthidioides and confirm that workers have more difficulty removing pupae compared with larvae from the combs.