Declines in pollinator colonies represent a worldwide concern. The widespread use of agricultural pesticides is recognized as a potential cause of these declines. Previous studies have examined the effects of neonicotinoid insecticides such as imidacloprid on pollinator colonies, but these investigations have mainly focused on adult honey bees. Native stingless bees (Hymenoptera: Apidae: Meliponinae) are key pollinators in neotropical areas and are threatened with extinction due to deforestation and pesticide use. Few studies have directly investigated the effects of pesticides on these pollinators. Furthermore, the existing impact studies did not address the issue of larval ingestion of contaminated pollen and nectar, which could potentially have dire consequences for the colony. Here, we assessed the effects of imidacloprid ingestion by stingless bee larvae on their survival, development, neuromorphology and adult walking behavior. Increasing doses of imidacloprid were added to the diet provided to individual worker larvae of the stingless bee Melipona quadrifasciata anthidioides throughout their development. Survival rates above 50% were only observed at insecticide doses lower than 0.0056 µg active ingredient (a.i.)/bee. No sublethal effect on body mass or developmental time was observed in the surviving insects, but the pesticide treatment negatively affected the development of mushroom bodies in the brain and impaired the walking behavior of newly emerged adult workers. Therefore, stingless bee larvae are particularly susceptible to imidacloprid, as it caused both high mortality and sublethal effects that impaired brain development and compromised mobility at the young adult stage. These findings demonstrate the lethal effects of imidacloprid on native stingless bees and provide evidence of novel serious sublethal effects that may compromise colony survival. The ecological and economic importance of neotropical stingless bees as pollinators, their susceptibility to insecticides and the vulnerability of their larvae to insecticide exposure emphasize the importance of studying these species.

The midgut epithelium of bees is formed by the digestive cells, responsible for enzyme secretion and nutrient absorption and for small regenerative cells that are placed in nests scattered among the digestive cells. During metamorphosis, the larval midgut epithelium degenerates and a new adult midgut epithelium is built during larval differentiation of regenerative cells. The present work focuses on the midgut epithelial modifications during the post-embryonic development of the stingless bee Melipona quadrifasciata anthidioides worker and the occurrence of regenerative cell proliferation during midgut metamorphosis in order to test the hypothesis that adult midgut epithelium of worker bees results from regenerative cell proliferation during the pupal stage. Regenerative cell proliferation was detected during larval lifespan. Larval aging is followed by an increase in the number and the size of the nests of regenerative cells. Larval epithelium degeneration begins 2 days after the start of defecation process and in this period the nests of regenerative cells are in contact by means of cytoplasmic extension which have many septate desmosomes and gap junctions. The BrdU immunoreactive regenerative cells were found in the prepupae 12 h after BrdU injection, suggesting that regenerative cell population increase during this larval period. Regenerative cell proliferation results in the increase of the regenerative cell population and not in the formation of new digestive cells because the proliferation of regenerative cells would not be enough to reestablish the nests of regenerative cells and at the same time form new adult digestive cells. In this sense the hypothesis that digestive adult cells originate from regenerative cell proliferation during midgut metamorphosis in M. quadrifasciata anthidioides was rejected.

2010Phylogeography and historical demography of the neotropical stingless bee Melipona quadrifasciata (Hymenoptera, Apidae):incongruence between morphology and mitochondrial DNA APIDOLOGIE, v.41, n.5, p.534-547, 2010 http://producao.usp.br/handle/BDPI/15811 Abstract -The stingless bees are among the most abundant and ecologically important social invertebrates in tropical communities. The Neotropical stingless bee Melipona quadrifasciata has two subspecies: M. quadrifasciata quadrifasciata and M. quadrifasciata anthidioides. The main difference between subspecies are the yellow metassomal stripes, which are continuous in M. q. quadrifasciata and discontinuous in M. q. anthidioides. Recently, two populations were described with continuous stripes and inhabiting clearly disjunct areas in relation to M. q. quadrifasciata. We sequenced 852 bp of the mtDNA COI gene from 145 colonies from 56 localities, and for the first time performed a detailed phylogeographic study of a neotropical stingless bee. Phylogenetic analyses revealed the existence of two clades exhibiting a south to north distribution: southern populations comprise the subspecies M. q. quadrifasciata, and northern populations are composed of M. q. anthidioides and two disjunct populations with continuous stripes. The divergence time of these two phylogroups was estimated between 0.233 and 0.840 million years ago in the Pleistocene, a period of climatic changes and geomorphological alterations in the Neotropical region. No evidence of genetic structure in relation to the tergal stripes was found, indicating that the morphological trait regarding the pattern of stripes on tergites is not an accurate diagnostic for the subspecies of M. quadrifasciata. biogeography / coalescence / tergal stripes / Melipona quadrifasciata / subspecies

Euglossine spermatozoa are the longest described to date for the Hymenoptera. This cell includes a head and a flagellar region. In transverse sections, the acrosome is circular at the tip but has an oval contour along most of its length. The perforatorium penetrates into a deep cavity in the nuclear tip. The flagellum consists in an axoneme, a pair of mitochondrial derivatives, a centriolar adjunct and a pair of accessory bodies. The axoneme has a 9+9+2 microtubule pattern which becomes gradually disorganized in the final portion, with the central microtubules and the nine doublets terminating simultaneously, followed by the accessory microtubules. The mitochondrial derivatives are asymmetric both in length and diameter. Sectioned transversally, the derivatives are ellipsoidal or have a pear shape. The larger one has a more obvious paracrystalline region. The centriolar adjunct begins at the nuclear base and extends parallel to the axoneme until it encounters the smaller mitochondrial derivative, on which it fits, making a concave groove. In addition to these consistent euglossine features, species-specific differences that might be useful in phylogenetic work on the group are also noted.

The present research represents the first description of a Vespidae species spermatozoon. In Agelaia vicina, the sperm head is composed of an acrosome and a nucleus. The acrosome consists of an acrosomal vesicle, oval shaped in cross sections, and a perforatorium, that projects into the anterior nuclear region. The nucleus is elongated and oval shaped in cross sections. The nucleus-flagellum transition region includes the centriolar adjunct, the nucleus base and the anterior portions of the axoneme and of the large mitochondrial derivative. The centriolar adjunct begins as a thin layer between the nucleus and the large mitochondrial derivative, while just below it acquires a lateral extension partially surrounding the large mitochondrial derivative. In its medial portion, it is triangular shaped, situated laterally to the axoneme and the large mitochondrial derivative, and it finishes in contact with the tip of the small mitochondrial derivative. The flagellum is composed of a 9+9+2 axoneme, two mitochondrial derivatives and two accessory bodies. The large mitochondrial derivative begins laterally to the nuclear base, and the small one below the centriolar adjunct. Only the large mitochondrial derivative presents a paracrystalline core, situated distally in relation to the axoneme. In the posterior flagellar region, the small mitochondrial derivative finishes first, followed by the large one and both accessory bodies. Although Vespidae belong to the superfamily Vespoidea, as Formicidae, the sperm ultrastructure of A. vicina is phylogenetically more closely related to Apoidea, and curiously very different from that observed for ants. This study supports the use of insect sperm morphology as a tool for phylogenetic analysis, especially in the Hymenoptera order.