Isolamento e caracterização de leveduras de Polybia ignobilis (Hymenoptera: Vespidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Termofisiolofia da vespa Polybia (Trichothorax) ignobilis (Haliday, 1836) (Hymenoptera, Vespidae, Epiponini) durante a atividade de forrageamento

Many social wasps are known to use thermogenesis to warm up their flight muscles and are therefore able to forage under a broad range of ambient temperatures. However it is uncertain whether there exists a possible relation between ambient temperature and thermogenic capacity for tropical species, as we lack studies focusing on these species. Therefore, we examined the use of this mechanism in the neotropical Epiponini wasp Polybia ignobilis. More specifically, we used a thermographic camera to obtain data of the surface temperatures of three body regions (head, thorax and abdomen) of wasps during foraging activities (pre-flight, flight and post-flight) in cold [initial pe- riod of foraging activity: TAM : 15 − 20◦C] and warm [final period of foraging activity: TPM : 30 − 35◦C] conditions. Thorax temperature (Tth) was always higher than head (Th) and abdomen temperature (Tabd). In general, the lowest body temperatures were observed during the pre-flight period, while the highest values occurred upon the return of the wasps from the foraging flight. Except for the pre-flight period, Tth was always higher than Tabd, indicating that heat generated at the thorax was preferentially directed to the cephalic region. Therefore we confirmed the use of thermogenesis by a neotropical social wasp, although its magnitude was found modest compared to temperate species, which suggests a link between thermal environment and thermogenic capacity. We also showed that P. ignobilis modulates heat production as a function of ambient temperature (TA), maintaining a greater temperature difference (Tbody − TA) at cooler temperatures. Finally, we identified the cephalic region of wasps as an important route for the dissipation of the heat generated during flight

High-performance gel filtration patterns of venoms from three species of wasps of the genus Polybia (hymenoptera, vespidae).

The profiles of high-performance gel filtration of venoms from Polybia paulista, Polybia ignobilis and Polybia occidentalis occidentalis showed 13 peaks distributed among the three species. These profiles presented similarities that permitted the chromatographic characterization of the genus Polybia and differences that permitted the identification of each species studied. Thus, the comparative analysis of chromatographic profiles of high-performance gel filtration of venoms may be used as an auxiliary tool in taxonomic studies of Polybia wasps.

Hyaluronidase from the venom of the social wasp Polybia paulista (Hymenoptera, Vespidae): Cloning, structural modeling, purification, and immunological analysis

In this study, we describe the cDNA cloning, sequencing, and 3-D structure of the allergen hyaluronidase from Polybia paulista venom (Pp-Hyal). Using a proteomic approach, the native form of Pp-Hyal was purified to homogeneity and used to produce a Pp-specific polyclonal antibody. The results revealed that Pp-Hyal can be classified as a glycosyl hydrolase and that the full-length Pp-Hyal cDNA (1315 bp; GI: 302201582) is similar (80-90%) to hyaluronidase from the venoms of endemic Northern wasp species. The isolated mature protein is comprised of 338 amino acids, with a theoretical pI of 8.77 and a molecular mass of 39,648.8 Da versus a pI of 8.13 and 43,277.0 Da indicated by MS. The Pp-Hyal 3D-structural model revealed a central core (α/β)7 barrel, two sulfide bonds (Cys 19-308 and Cys 185-197), and three putative glycosylation sites (Asn79, Asn187, and Asn325), two of which are also found in the rVes v 2 protein. Based on the model, residues Ser299, Asp107, and Glu109 interact with the substrate and potential epitopes (five conformational and seven linear) located at surface-exposed regions of the structure. Purified native Pp-Hyal showed high similarity (97%) with hyaluronidase from Polistes annularis venom (Q9U6V9). Immunoblotting analysis confirmed the specificity of the Pp-Hyal-specific antibody as it recognized the Pp-Hyal protein in both the purified fraction and P. paulista crude venom. No reaction was observed with the venoms of Apis mellifera, Solenopsis invicta, Agelaia pallipes pallipes, and Polistes lanio lanio, with the exception of immune cross-reactivity with venoms of the genus Polybia (sericea and ignobilis). Our results demonstrate cross-reactivity only between wasp venoms from the genus Polybia. The absence of cross-reactivity between the venoms of wasps and bees observed here is important because it allows identification of the insect responsible for sensitization, or at least of the phylogenetically closest insect, in order to facilitate effective immunotherapy in allergic patients. © 2013 Elsevier Ltd.

A comparative study of protein and enzymatic activity in venoms of some common wasps (Hymenoptera: Vespidae) from São Paulo State

The Hymenoptera Aculeata venoms, with few exceptions, have been poorly studied and characterized. Nevertheless, they have raised increasing interest due to their medical importance, since accidents with these insects are fairly frequent in Brazil and may cause severe allergic reactions. The objectives of the present work were the quantitative characterization of the main allergenic enzymes present in the venom of the species Polybia paulista, Polybia ignobilis, Polistes simillimus, and Agelaia pallipes pallipes through biochemical assays for the determination of total protein content, as well as the level of the enzymatic activity of phospholipase, hyaluronidase, acid phosphatase and esterase. These results, in addition to providing biochemical knowledge about the venom of the species in question, also supply studies that allow phylogenetic inferences among them.

Characterization of the social wasp guild (Hymenoptera: Vespidae) visiting flowers in the Caatinga (Itatim, Bahia, Brazil)

Thirteen species of flower-visiting social wasps were collected from 41 plant species. The number of wasp species did not vary significantly. On the other hand, the number of individuals varied significantly during the data collection period. Four of the wasp species (Mischocyttarus lanei, Polybia ignobilis, Polybia occidentalis, and Polybia sericea) showed changes in body size over the year. The total wasp biomass and the number of plants monthly visited by wasps had a positive significant correlation. The structure of this social wasp community is characterized by a small number of dominant species, a large number species that are not frequently present and several plant species visited by few wasps. Social wasp species are differently affected by seasonal changes in the 'caatinga'.

Estudo comparativo da diversidade de vespas sociais (Hymenoptera, Vespidae) em mata ciliar e cultura de cana-de-açúcar na região de Ipeúna - SP

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Foraging by Polybia (Trichothorax) ignobilis (Hymenoptera, Vespidae) on flies at animal carcasses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caste Asymmetries in the Neotropical Swarm-Founding Wasp Polybia (Trychothorax) ignobilis (Hymenoptera, Vespidae, Epiponini)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Variação morfofisiológica das castas da vespa enxameante neotropical Polybia (Trichothorax) ignobilis durante sua ontogenia colonial (Hymenoptera, Vespidae, Epiponini)

Pós-graduação em Ciências Biológicas (Zoologia) - IBRC

Renal- and calcium-dependent vascular effects of Polybia paulista wasp venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

External Morphology of the Immatures of Polybia paulista (Hymenoptera: Vespidae)

The immatures of Polybia paulista Ihering were described using light and scanning electron microscopy and the results are compared with previous descriptions within the same or related wasps. This study is based on 2 whole nests collected in the municipality of Rio Claro, São Paulo, in Brazil. We have detected the existence of 5 larval instars. The main morphological alterations over development occur in the relative size of structures, yet certain structures appear with subsequent instars and become more evident later in development: increasing density in the number of body spines and papillae; the appearance of body setae in fifth-instar larvae; opening of spiracles upon second-instar larvae; 2 body shapes in fifth-instar larvae; the appearance of a lateral tooth on the mandibles of fourth instar; presence of spines on the maxillae of fifth-instar larvae; altered shape of galea and palps upon third-instar larvae from a cluster of sensilla to a conical elevation; and the appearance of spines on postmentum upon fourth-instar larvae. This way, the present study presents a detailed description of the immatures of P. paulista, and we hope the presented information can be useful to morphological, taxonomic, and phylogenetic studies.

Survey of preys captured and used by Polybia platycephala (Richards) (Hymenoptera: Vespidae, Epiponini)

As vespas sociais são predadoras de várias espécies de insetos e, portanto, o estudo de suas presas pode revelar seu potencial para programas de controle biológico de pragas. Durante o período de setembro de 2000 a janeiro de 2002, foram realizadas 70h de coleta de presas capturadas em doze ninhos de Polybia platycephala Richards, localizados em áreas urbanas do município de Juiz de Fora, MG. As presas capturadas por P. platycephala compreenderam cinco ordens de insetos: Diptera (33,4%), Lepidoptera (28,6%), Hemiptera (12,0%), Hymenoptera (9,4%) e Coleoptera (7,2%). O peso médio da carga protéica transportada pelas vespas foi 1,9 ±1,6 mg (n = 34, 0,3 - 6,2 mg), e a taxa média de proteína transportada por dia foi 22,8 mg. de acordo com os resultados, pode-se estimar a captura de 4.380 presas por ano por uma única colônia de P. platycephala. Desta forma, a espécie pode ser utilizada em programas de manejo em ambientes urbanos contribuindo para o controle de insetos pragas como larvas de pernilongos, lagartas desfolhadoras de plantas de jardins, pulgões e formas aladas de formigas.

Mass spectrometric characterization of two novel inflammatory peptides from the venom of the social wasp Polybia paulista

The social wasp P. paulista is relatively common in southeast Brazil causing many medically important stinging incidents. The seriousness of these incidents is dependent on the amount of venom inoculated by the wasps into the victims, and the characteristic envenomation symptoms are strongly dependent on the types of peptides present in the venom. In order to identify some of these naturally occurring peptides available in very low amounts, an analytical protocol was developed that uses a combination of reversed-phase and normal-phase high-performance liquid chromatography (HPLC) of wasp venom for peptide purification, with matrix-assisted laser desorption/ionization time-of-flight post-source decay mass spectrometry (MALDI-Tof-PSD-MS) and low-energy collision-induced dissociation (CID) in a quadrupole time-of-flight tandem mass spectrometry (QTof-MS/MS) instrument for peptide sequencing at the sub-picomole level. The distinction between Leu and Ile was achieved both by observing d-type fragment ions obtained under CID conditions and by comparison of retention times of the natural peptides and their synthetic counterparts (with different combinations of I and/or L at N- and C-terminal positions). To distinguish the isobaric residues K and Q, acetylation of peptides was followed by Q-Tof-MS analysis. The primary sequences obtained were INWLKLGKMVIDAL-NH2 (MW 1611.98Da) and IDWLKLGKMVMDVL-NH2 (MW 1658.98Da). Micro-scale bioassay protocols characterized both peptides as presenting potent hemolytic action, mast cell degranulation, and chemotaxis of poly-morphonucleated leukocyte (PMNL) cells. The primary sequences and the bioassay results suggest that these toxins constitute members of a new sub-class of mastoparan toxins, directly involved in the occurrence of inflammatory processes after wasp stinging. Copyright (C) 2004 John Wiley Sons, Ltd.

Structural and functional characterization of two novel peptide toxins isolated from the venom of the social wasp Polybia paulista

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)