Number of Malpighian Tubules in Larvae and Adults of Stingless Bees (Hymenoptera: Apidae) from Amazonia

The number of Malpighian tubules in larvae and adults of bees is variable. Larvae of Apis mellifera L. have four Malpighian tubules, while adults have 100 tubules. In stingless bees, this number varies from four to eight. The objectives of this study were to provide characteristics of the Malpighian tubules as well as to quantify their number in larvae and adults of six species of Meliponinae, Melipona seminigra merrillae Cockerell, Melipona compressipes manaosensis Schwarz, Melipona rufiventris Lepeletier, Scaptotrigona Moure, Frieseomelitta Ihering, and Trigona williana Friese. Malpighian tubules were dissected from larvae and adults, measured, quantified, and maintained in microtubes with Dietrich??s solution. The numbers of Malpighian tubules were constant only for larvae of M. rufiventris (four and eight) and Scaptotrigona sp. (four). The most frequent number of tubules in the Melipona group was seven and eight in larvae, and 70 and 90 in adults. In the Trigona group were four and 20 to 40, for larvae and adults, respectively. The results showed differences in the number of Malpighian tubules among the species analyzed and also between the larvae and adults of the same species. Despite the variation observed, species of the group Melipona always have a larger number and longer Malpighian tubules in both larvae and adults as compared to the Trigona group, which may indicate an evolutionary trend of differentiation between these groups.     

An overview of cytogenetics of the tribe Meliponini (Hymenoptera: Apidae)

The present study provides a comprehensive review of cytogenetic data on Meliponini and their chromosomal evolution. The compiled data show that only 104 species of stingless bees, representing 32 of the 54 living genera have been studied cytogenetically and that among these species, it is possible to recognize three main groups with n?=?9, 15 and 17, respectively. The first group comprises the species of the genus Melipona, whereas karyotypes with n?=?15 and n?=?17 have been detected in species from different genera. Karyotypes with n?=?17 are the most common among the Meliponini studied to date. Cytogenetic information on Meliponini also shows that although chromosome number, in general, is conserved among species of a certain genus, other aspects, such as chromosome morphology, quantity, distribution and composition of heterochromatin, may vary between them. This reinforces the fact that the variations observed in the karyotypes of different Meliponini groups cannot be explained by a single theory or a single type of structural change. In addition, we present a discussion about how these karyotype variations are related to the phylogenetic relationships among the different genera of this tribe.     

Genetic structure of Partamona helleri (Apidae, Meliponini) from Neotropical Atlantic rainforest

Stingless bees of the genus Partamona are distributed from southern Mexico to southern Brazil. This genus has been subject to different approaches to solve questions concerning general biology, taxonomy, systematics and biogeography, but population studies applying molecular techniques are inexistent. We analyzed the genetic structure of P. helleri across its geographic distribution along the coastal Atlantic tropical rainforest in Brazil. Ten mtDNA haplotypes were observed in 47 colonies of P. helleri of which some were exclusive and others shared among geographic sub-groups. Statistical analysis showed high genetic differentiation between geographic areas sampled. Fragmentation of the Atlantic forest during Pleistocene glaciations is discussed as a possible cause of the present haplotype distribution and frequency.     

Olfactory learning in the stingless bee Tetragonisca angustula (Hymenoptera, Apidae, Meliponini)

Tetragonisca angustula stingless bees are considered as solitary foragers that lack specific communication strategies. In their orientation towards a food source, these social bees use chemical cues left by co-specifics and the information obtained in previous foraging trips by the association of visual stimuli with the food reward. Here, we investigated their ability to learn the association between odors and reward (sugar solution) and the effect on learning of previous encounters with scented food either inside the hive or during foraging. During food choice experiments, when the odor associated with the food was encountered at the feeding site, the bees’ choice is biased to the same odor afterwards. The same was not the case when scented food was placed inside the nest. We also performed a differential olfactory conditioning of proboscis extension response with this species for the first time. Inexperienced bees did not show significant discrimination levels. However, when they had had already interacted with scented food inside the hive, they were able to learn the association with a specific odor. Possible olfactory information circulation inside the hive and its use in their foraging strategies is discussed.     

Pollen Harvest by Stingless Bee Foragers (Hymenoptera,Apidae, Meliponinae)

Data on pollen load capacity and flower constancy are discussed for nine stingless bee species. The foragers present high levels of flower constancy and often visit only one flower type (on average 97% of bee foragers), rarely a few flower types (on average 3% of bee foragers), during the same foraging trip. The latter foragers exhibit no tendencies for choosing similar sources, related either to flower type or to pollen type. Pollen load capacity (the ratio between pollen load weight/worker weight) decreases as forager body weight increases, so it is larger in smaller stingless bees species and smaller in larger ones. Nevertheless, it seems that specific load capacity also depends on pollen types.     

Communities of Social Bees (Apidae: Meliponini) in Trap-Nests: the Spatial Dynamics of Reproduction in an Area of Atlantic Forest

As most stingless bee species depend on preexisting cavities, principally tree hollows, nesting site availability may represent an important restriction in the structuring of their forest communities. The present study examined the spatial dynamics of stingless bee communities in an area of Atlantic Forest by evaluating their swarming to trap-nests. The field work was performed in the Michelin Ecological Reserve (MER) on the southeastern coast of the state of Bahia, Brazil. Seven hundred and twenty trap-nests were distributed within two forest habitats in advanced and initial stages of regeneration. The trap-nests were monitored between September 2009 and March 2011. Twenty-five trap-nests were occupied by five bee species, resulting in a capture ratio of 0.035 swarms/trap (approximately 0.14 swarms/ha), corresponding to 10 swarms/year (0.056 swarms/ha/year). According to previous study at MER, the most abundant species in natural nests were also the most common in trap-nests in the two forest habitats examined, with the exception of Melipona scutellaris Latreille. Swarms of higher numbers of species were captured in initial regeneration stage forests than in advanced regeneration stage areas, and differences in species compositions were significant between both habitats (p?=?0.03); these apparent differences were not consistent, however, when considering richness (p?=?0.14) and total abundance (p?=?0.08). The present study suggests the existence of a minimum cavity size threshold of approximately 1 L for most local species of stingless bees and sustains the hypothesis of a mass effect of Tetragonisca angustula Latreille populations from surrounding disturbed habitats on the MER forest community in terms of propagule (swarm) pressure. Examining swarm densities with trap-nests can be a promising technique for comparative analyses of the carrying capacities of forest habitats for stingless bee colonies, as long as size thresholds of cavities for nesting are taken into consideration.     

Cytogenetic characterization of two species of Frieseomelitta Ihering, 1912 (Hymenoptera, Apidae, Meliponini)

The cytogenetic analysis of Frieseomelitta dispar and F. francoi revealed the chromosome numbers 2n = 30 and n = 15 and a karyotypic formula 2K = 4M+2M(t)+4A+20A(M). The number of chromosomes observed was consistent with those reported for other Frieseomelitta species. The occurrence of the M(t) chromosome and other features of the karyotype formulae suggest a close relationship between F. dispar, F. francoi and F. varia. Nevertheless, it was possible to differentiate the karyotypes of the species by DAPI/CMA(3) staining, which revealed GC-rich regions on two chromosome pairs of F. dispar: one acrocentric and one pseudoacrocentric. In F. francoi, the same kinds of regions were observed on a pair of metacentrics and on a pair of acrocentrics. Our analysis also confirmed the chromosome number conservation in Frieseomelitta and suggests that infrequent pericentric inversion could constitute a synapomorphy for the group including F. dispar, F. francoi, and F. varia.     

Variation the oviposition behavior by the stingless bee,Heterotrigona itama (Hymenoptera,Apidae, Meliponini)

A study to determine the variation of ovipositioning behavior of stingless bees, Heterotrigona itama (Cockerell, 1918) was conducted on three colonies on June 2015. A digital single-lens reflex (DSLR) camera with a macro lens attached was used to record every movement of H. itama in its colonies for 20 min hour between 0800 h and 2000 h for seven days and seven month. The daily egg laying rate and time for laying eggs in colony-B and colony-C were significantly (P ? 0.05) higher than the colony-A. However, time to close the brood was not significantly different among colonies. The fastest egg oviposition time was 4 s by the colony-B and the slowest was 6 s by the colony-A. In addition there are no significant trends on brood produced per day, laying time of eggs, and the closing time of the brood after the oviposition process from June to December 2016. This result is useful for understanding the behavior of egg laying process by the queen bees and necessary to deal with problems of management and reproduction in the near future.     

Production of Sexuals and Mating Frequency in the Stingless Bee Tetragonisca angustula (Latreille) (Hymenoptera, Apidae)

Queen, worker, and male production was studied for 1 year in three queenright colonies of Tetragonisca angustula (Latreille). We sampled brood combs monthly and noticed that the number of brood cells and production of individuals were similar among colonies. Although the production of queens did not vary significantly, the frequencies of workers, males, and the number of cells among the combs varied over time. The production of males was highly seasonal, occurring mostly from February to April, coinciding with the period of intense brood cells production, when colonies produced more males and less workers, resulting in a negative correlation. Although the frequency of queens has not varied in time, the seasonal availability of males affected the mating frequency and the time spent since emergence until fertilization of queens. In the T. angustula colonies studied, the population dynamics was highly seasonal and the mating success depended of male production, according to the season.     

The Panama microplate,island studies and relictual species of Melipona (Melikerria) (Hymenoptera: Apidae: Meliponini)

The endemic stingless honey‐making bee Melipona (Melikerria) insularis sp.n. on Coiba and Ranchería Islands in Pacific Panama is described, together with the proposed sister species, M. ambigua sp.n. from northeast Colombia. The Coiba Island group and Panama mainland were surveyed, yielding one meliponine endemic (M. insularis sp.n. ) and six meliponine genera and species. The poor Coiba fauna of amphibians and birds corresponds to the poor social bee fauna and suggests habitat barriers generally precluded recolonization from the mainland during glacial periods. Many animals became extinct, yet some remain as relicts. Melipona insularis sp.n. was isolated on accreted terranes of Coiba rainforest in the Panama microplate. Morphology suggests that M. insularis sp.n. is not a direct descendant of the San Blas‐E. Panama endemic Melikerria, M. triplaridis. A phylogenetic hypothesis corroborates disjunct distributions. Rainforest endemics such as Peltogyne purpurea (Fabaceae) and Ptilotrigona occidentalis (Apidae, Meliponini) also occur as relictual, disjunct populations in Central and South America. These may have been isolated before accelerated biotic exchange began 2.4 Ma. Our work supports the geological findings of both a volcanic arc and the San Blas massif providing a substantial bridge for Melikerria from Colombia and Panama in Eocene to Miocene times. We suggest there have been taxon cycles permitting recolonization during glaciations, whereby colonies of M. insularis sp.n. were able to recolonize Ranchería, a 250 ha island, 2 km from Coiba. However, rafting colonies nesting in trees, carried on vegetation mats, may have produced founding populations of Melipona in Central America and on oceanic islands such as Coiba.     

Alelle number and heterozigosity for microsatellite loci in different stingless bee species (Hymenoptera: Apidae, Meliponini)

In the present study we compare genetic characteristics (allele diversity and observed heterozygosity) of microsatellite loci, from three stingless bee species (Plebeia remota Holmberg, Partamona mulata Moure In Camargo and Partamona helleri Friese), amplified by using heterospecific primers originally designed for Melipona bicolor Lepeletier and Scaptotrigona postica Latreille. We analyzed 360 individuals of P. remota from 72 nests, 58 individuals of R. mulata from 58 nests, and 47 individuals of P. helleri from 47 nests. The three species studied showed low level of polymorphism for the loci amplified with primers derived from M. bicolor. However, for the loci amplified with primers derived from S. postica, only P. remota presented low level of polymorphism.     

The identity of the Neotropical stingless bee Frieseomelitta meadewaldoi (Cockerell, 1915) (Hymenoptera, Apidae)

A new study of a surviving syntype of Trigona meadewaldoi Cockerell, 1915, was undertaken and several widely employed names for Neotropical stingless bees recognized as junior synonyms. A lectotype is designated for Trigona meadewaldoi and the following new synonymies established: Tetragona francoi Moure, 1946, and Trigona (Frieseomelitta) freiremaiai Moure, 1963. These nomenclatural matters are here settled and the species thoroughly characterized in advance of a forthcoming phylogenetic consideration of the genus Frieseomelitta von Ihering, 1912.     

Pollen sources used by species of Meliponini (Hymenoptera: Apidae) along the Rio Negro channel in Amazonas,Brazil

This work analyses the pollen stored by Meliponini bees whose nests were sampled along the Rio Negro channel, Amazonas, Brazil, during the final phase of the rainy season, which is also the time of pollen scarcity. From a total of 51 samples studied, 78 pollen types were identified, of which 39 were determined to species level. The pollen types were from 70 genera and 42 families. Based on the pollen records, a clustering analysis was performed, as well as the calculation of the trophic overlap index. The study demonstrates that the plants used by the bees do not depend on them for pollination in most cases; it rather suggests scarcity of pollen resources from the plants adapted to the pollination of Meliponini, and emphasises the ability of such bees to switch to new available resources. This work indicates a list of important plants for meliponiculture suitable for cultures near meliponaries.     

Gene Variation, Population Differentiation, and Sociogenetic Structure of Nests of Partamona seridoensis (Hymenoptera: Apidae, Meliponini)

Gene variation and the differentiation of two populations of Partamona seridoensis (Hymenoptera: Apidae: Meliponini) from the Caatinga biome, a semiarid ecosystem unique to Brazil, were estimated through allozymic and microsatellite analyses. These populations exhibited similar low degrees of enzyme gene variation. Observed genotype frequencies at the allozyme and microsatellite loci were in accordance with Hardy-Weinberg equilibrium in the two populations. Both markers demonstrated that the two populations are not genetically homogeneous and must be considered distinct populations. The occurrence of private alleles at the allozyme and microsatellite loci corroborates this differentiation, sustaining the hypothesis of a low level of interpopulation gene flow. The phenotypic segregations clearly demonstrated that the progeny inside each nest were the result of mating between the queen of the colony and only one male.