A new scenario of bioprospecting of Hymenoptera venoms through proteomic approach

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

Mastoparan effects in skeletal muscle damage: An ultrastructural view until now concealed

Animal venoms have been valuable sources for development of new drugs and important tools to understand cellular functioning in health and disease. The venom of Polybia paulista, a neotropical social wasp belonging to the subfamily Polistinae, has been sampled by headspace solid phase microextraction and analyzed by gas chromatography-mass spectrometry. Recent study has shown that mastoparan, a major basic peptide isolated from the venom, reproduces the myotoxic effect of the whole venom. In this study, Polybia-MPII mastoparan was synthesized and studies using transmission electron microscopy were carried out in mice tibial anterior muscle to identify the subcellular targets of its myotoxic action. The effects were followed at 3 and 24 h, 3, 7, and 21 days after mastoparan (0.25 mu g/mu L) intramuscular injection. The peptide caused disruption of the sarcolemma and collapse of myofibril arrangement in myofibers. As a consequence, fibers presented heteromorphic amorphous masses of agglutinated myofilaments very often intermingled with denuded sarcoplasmic areas sometimes only surrounded by a persistent basal lamina. To a lesser extent, a number of fibers apparently did not present sarcolemma rupture but instead appeared with multiple small vacuoles. The results showed that sarcolemma, sarcoplasmic reticulum (SR), and mitochondria were the main targets for mastoparan. In addition, a number of fibers showed apoptotic-like nuclei suggesting that the peptide causes death both by necrosis and apoptosis. This study presents a hitherto unexplored view of the effects of mastoparan in skeletal muscle and contributes to discuss how the known pharmacology of the peptide is reflected in the sarcolemma, SR, mitochondria, and nucleus of muscle fibers, apparently its subcellular targets.

The neurotoxicological effects of mastoparan Polybia-MPII at the murine neuromuscular junction: an ultrastructural and immunocytochemical study

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Temporal patterns of parasitism in Diatraea saccharalis Fabr. (Lep., Crambidae) populations at different spatial scales in sugarcane fields in Brazil

Biological control of Diatraea saccharalis is regarded as one of the best examples of successful classical biological control in Brazil. Since the introduction of the exotic parasitoid, Cotesia flavipes, from Pakistan at the beginning of the 1970s, decrease in D. saccharalis infestation in sugarcane fields has been attributed to the effectiveness of this agent. Recently, the native Tachinidae fly parasitoids (Lydella minense and Paratheresia claripalpis) have also been implicated in this success. However, quantitative data confirming the actual contribution of these agents to the control of D. saccharalis are rather limited. The purpose of this study was to investigate the dynamics of the interactions between D. saccharalis and its parasitoids, emphasizing the temporal patterns of parasitism. To investigate this question, a large data set comprising information collected from two sugarcane mills located in the state of São Paulo, Brazil (Barra and Sao Joao sugarcane mills), was analysed. Basically, the data set contained monthly information about the number of D. saccharalis larvae and their parasitoids in each sample (man-hour per sample), the sugarcane varieties cultivated, the age of the sugarcane plants (only at the Sao Joao sugarcane mill) as well as the sugarcane cut at sampling time. The data were collected from March 1984 to March 1997 and from May 1982 to December 1996 for the Barra and Sao Joao sugarcane mills, respectively. Temporal inverse density-dependent parasitism was predominant for both parasitoid species with respect to all spatial scales. Although the temporal pattern of parasitism was not directly density dependent, it was evident that the tachinids and C. flavipes presented positive numerical responses according to variations in D. saccharalis densities through time.

Spatial and temporal population interactions between the parasitoids Cotesia flavipes and Tachinidae flies: considerations on the adverse effects of biological control practice

Biological control of Diatraea saccharalis is regarded as one of the best examples of successful classical biological control in Brazil. Since the introduction of the exotic parasitoid Cotesia flavipes, the decrease of D. saccharalis infestation in sugarcane fields has been attributed to the effectiveness of this agent. Recently, the native tachinid fly parasitoids (Lydella minense and Paratheresia claripalpis) have also been implicated in the success. Here, we investigated the spatial and temporal population interactions between C. flavipes and the tachinid flies, and provide a critical analysis of the biological control practice, focusing on the undesirable effects of introductions of exotic natural enemies. To investigate these questions, a large data set comprising information from two sugarcane mills located in the state of São Paulo, Brazil (Barra and Sao Joao Mills), was analysed. Analysis of the correlation between C. flavipes and tachinid fly population densities through time revealed that such populations were inversely correlated in the Sao Joao Mill and not correlated in the Barra Mill. Logistic regressions were computed to investigate the proportion of sites occupied by the parasitoid species at both mills as a function of time. An increasing trend in the proportion of sites occupied by C. flavipes was observed, with a concomitant decrease of the sites occupied by tachinid flies. This effect was more intense in the Sao Joao Mill. Thus, there is a convincing possibility that constant releases of C. flavipes decreased the tachinid fly populations, resulting in an undesirable effect of biological control practice.

Relationship between number of ovipositions of Cotesia flavipes (Cam.) and number of descendants emerged from its host Diatraea saccharalis (Fabr.)

Fêmeas de Cotesia flavipes (Cam.) foram introduzidas, individualmente, em placas de Petri contendo uma larva de seu hospedeiro Diatraea saccharalis (Fabr.). Permitiu-se que cada fêmea de C. flavipes realizasse posturas uma ou duas vezes em uma mesma larva. O período de emergência das larvas e dos adultos, a razão sexual, o número de larvas e pupas inviáveis e a longevidade dos adultos de C. flavipes registrados nas duas condições experimentais (uma ou duas posturas) foram comparados. Os resultados revelaram que duas oviposições sucessivas no hospedeiro não alteraram a razão sexual na descendência, mas aumentaram o número de parasitóides e também o número de larvas e pupas inviáveis.

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

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

Influência de variedades de cana-de-açúcar, incorporadas em dieta artificial, no desenvolvimento de Diatraea saccharalis (Fabr.) e no seu parasitismo por Cotesia flavipes (Cam.)

Verificou-se a influência de duas variedades de cana-de-açúcar (Saccharum spp.), incorparados em dieta artificial, na forma de colmos triturados, no desenvolvimento de Diatraea saccharalis (Fabr.) e no parasitismo de Cotesia flavipes (Cam.). A variedade SP71-1081 foi menos adequada ao desenvolvimento de D. saccharalis comparada à SP71-3146, aumentando os períodos larval e pupal (dias) e, diminuindo o peso larval; não houve diferenças para as viabilidades larval e pupal e peso de pupa. Lagartas oriundas das dietas artificiais e lagartas alojadas no interior dos toletes das variedades de cana, foram igualmente parasitadas por C. flavipes. Lagartas criadas em dieta artificial contendo colmos triturados das variedades testadas, promoveram desenvolvimento semelhante de C. flavipes. A percentagem de parasitismo foi semelhante nas larvas criadas em dietas artificiais contendo colmos das variedades SP 71 - 1081 (84,0%) e SP 71-3146 (81,3%).

The effect of acidic residues and amphipathicity on the lytic activities of mastoparan peptides studied by fluorescence and CD spectroscopy

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

Influence of the Bilayer Composition on the Binding and Membrane Disrupting Effect of Polybia-MP1, an Antimicrobial Mastoparan Peptide with Leukemic T-Lymphocyte Cell Selectivity

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Selectivity in the mechanism of action of antimicrobial mastoparan peptide Polybia-MP1

Many potent antimicrobial peptides also present hemolytic activity, an undesired collateral effect for the therapeutic application. Unlike other mastoparan peptides, Polybia-MP1 (IDWKKLLDAAKQIL), obtained from the venom of the social wasp Polybia paulista, is highly selective of bacterial cells. The study of its mechanism of action demonstrated that it permeates vesicles at a greater rate of leakage on the anionic over the zwitterionic, impaired by the presence of cholesterol or cardiolipin; its lytic activity is characterized by a threshold peptide to lipid molar ratio that depends on the phospholipid composition of the vesicles. At these particular threshold concentrations, the apparent average pore number is distinctive between anionic and zwitterionic vesicles, suggesting that pores are similarly formed depending on the ionic character of the bilayer. To prospect the molecular reasons for the strengthened selectivity in Polybia-MP1 and its absence in Mastoparan-X, MD simulations were carried out. Both peptides presented amphipathic alpha-helical structures, as previously observed in Circular Dichroism spectra, with important differences in the extension and stability of the helix; their backbone solvation analysis also indicate a different profile, suggesting that the selectivity of Polybia-MP1 is a consequence of the distribution of the charged and polar residues along the peptide helix, and on how the solvent molecules orient themselves according to these electrostatic interactions. We suggest that the lack of hemolytic activity of Polybia-MP1 is due to the presence and position of Asp residues that enable the equilibrium of electrostatic interactions and favor the preference for the more hydrophilic environment.

Study of the mechanism of action of anoplin, a helical antimicrobial decapeptide with ion channel-like activity, and the role of the amidated C-terminus

Anoplin, an antimicrobial, helical decapeptide from wasp venom, looses its biological activities by mere deamidation of its C-terminus. Secondary structure determination, by circular dichroism spectroscopy in amphipathic environments, and lytic activity in zwitterionic and anionic vesicles showed quite similar results for the amidated and the carboxylated forms of the peptide. The deamidation of the C-terminus introduced a negative charge at an all-positive charged peptide, causing a loss of amphipathicity, as indicated by molecular dynamics simulations in TFE/water mixtures and this subtle modification in a peptide's primary structure disturbed the interaction with bilayers and biological membranes. Although being poorly lytic, the amidated form, but not the carboxylated, presented ion channel-like activity on anionic bilayers with a well-defined conductance step; at approximately the same concentration it showed antimicrobial activity. The pores remain open at trans-negative potentials, preferentially conducting cations, and this situation is equivalent to the interaction of the peptide with bacterial membranes that also maintain a high negative potential inside. Copyright (C) 2007 European Peptide Society and John Wiley & Sons, Ltd.

Cladistic analysis of Charterginus Fox, 1898 (Hymenoptera, Vespidae, Epiponini). A neotropical genus of social wasps

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

An improved bait method for collecting Hymenoptera, especially social wasps (Vespidae: Polistinae)

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