Treatment of an Aedes aegypti colony with the Cry11Aa toxin for 54 generations results in the development of resistance

To study the potential for the emergence of resistance in Aedes aegypti populations, a wild colony was subjected to selective pressure with Cry11Aa, one of four endotoxins that compose the Bacillus thuringiensis serovar israelensis toxin. This bacterium is the base component of the most important biopesticide used in the control of mosquitoes worldwide. After 54 generations of selection, significant resistance levels were observed. At the beginning of the selection experiment, the half lethal concentration was 26.3 ng/mL and had risen to 345.6 ng/mL by generation 54. The highest rate of resistance, 13.1, was detected in the 54th generation. Because digestive proteases play a key role in the processing and activation of B. thuringiensis toxin, we analysed the involvement of insect gut proteases in resistance to the Cry11Aa B. thuringiensis serovar israelensis toxin. The protease activity from larval gut extracts from the Cry11Aa resistant population was lower than that of the B. thuringiensisserovar israelensis susceptible colony. We suggest that differences in protoxin proteolysis could contribute to the resistance of this Ae. aegypti colony.

Terbinafine inhibits Cryptococcus neoformans growth and modulates fungal morphology

Cryptococcus neoformans is an encapsulated fungus that causes cryptococcosis. Central nervous system infection is the most common clinical presentation followed by pulmonary, skin and eye manifestations. Cryptococcosis is primarily treated with amphotericin B (AMB), fluconazole (FLC) and itraconazole (ITC). In the present work, we evaluated the in vitro effect of terbinafine (TRB), an antifungal not commonly used to treat cryptococcosis. We specifically examined the effects of TRB, either alone or in conjunction with AMB, FLC and ITC, on clinical C. neoformans isolates, including some isolates resistant to AMB and ITC. Broth microdilution assays showed that TRB was the most effective drug in vitro. Antifungal combinations demonstrated synergism of TRB with AMB, FLC and ITC. The drug concentrations used for the combination formulations were as much as 32 and 16-fold lower than the minimum inhibitory concentration (MIC) values of FLC and AMB alone, respectively. In addition, calcofluor white staining revealed the presence of true septa in hyphae structures that were generated after drug treatment. Ultrastructural analyses demonstrated several alterations in response to drug treatment, such as cell wall alterations, plasma membrane detachment, presence of several cytoplasmic vacuoles and mitochondrial swelling. Therefore, we believe that the use of TRB alone or in combination with AMB and azoles should be explored as an alternative treatment for cryptococcosis patients who do not respond to standard therapies.

Revalidation and redescription of Triatoma brasiliensis macromelasoma Galvão, 1956 and an identification key for the Triatoma brasiliensis complex (Hemiptera: Reduviidae: Triatominae)

Triatoma brasiliensis macromelasoma is revalidated based on the results of previous multidisciplinary studies on the Triatoma brasiliensis complex, consisting of crossing experiments and morphological, biological, ecological and molecular analyses. These taxonomic tools showed the closest relationship between T. b. macromelasoma and Triatoma brasiliensis brasiliensis. T. b. macromelasoma is redescribed based on specimens collected in the type locality and specimens from a F1 colony. The complex now comprises T. b. brasiliensis, T. b. macromelasoma, Triatoma melanica, Triatoma juazeirensis and Triatoma sherlocki. An identification key for all members of the complex is presented. This detailed comparative study of the morphological features of T. b. macromelasoma and the remaining members of the complex corroborates results from multidisciplinary analyses, suggesting that the subspecific status is applicable. This subspecies can be distinguished by the following combination of features: a pronotum with 1+1 narrow brownish-yellow stripes on the submedian carinae, not attaining its apex, hemelytra with membrane cells darkened on the central portion and legs with an incomplete brownish-yellow ring on the apical half of the femora. Because the T. brasiliensis complex is of distinct epidemiological importance throughout its geographic distribution, a precise identification of its five members is important for monitoring and controlling actions against Chagas disease transmission.

Morphology of the larvae, male genitalia and DNA sequences of Anopheles (Kerteszia) pholidotus (Diptera: Culicidae) from Colombia

Since 1984, Anopheles (Kerteszia) lepidotus has been considered a mosquito species that is involved in the transmission of malaria in Colombia, after having been incriminated as such with epidemiological evidence from a malaria outbreak in Cunday-Villarrica, Tolima. Subsequent morphological analyses of females captured in the same place and at the time of the outbreak showed that the species responsible for the transmission was not An. lepidotus, but rather Anopheles pholidotus. However, the associated morphological stages and DNA sequences of An. pholidotus from the foci of Cunday-Villarrica had not been analysed. Using samples that were caught recently from the outbreak region, the purpose of this study was to provide updated and additional information by analysing the morphology of female mosquitoes, the genitalia of male mosquitoes and fourth instar larvae of An. pholidotus, which was confirmed with DNA sequences of cytochrome oxidase I and rDNA internal transcribed spacer. A total of 1,596 adult females were collected in addition to 37 larval collections in bromeliads. Furthermore, 141 adult females, which were captured from the same area in the years 1981-1982, were analysed morphologically. Ninety-five DNA sequences were analysed for this study. Morphological and molecular analyses showed that the species present in this region corresponds to An. pholidotus. Given the absence of An. lepidotus, even in recent years, we consider that the species of mosquitoes that was previously incriminated as the malaria vector during the outbreak was indeed An. pholidotus, thus ending the controversy.

Co-administration of plasmid-encoded granulocyte-macrophage colony-stimulating factor increases human immunodeficiency virus-1 DNA vaccine-induced polyfunctional CD4+ T-cell responses

T-cell based vaccines against human immunodeficiency virus (HIV) generate specific responses that may limit both transmission and disease progression by controlling viral load. Broad, polyfunctional, and cytotoxic CD4+T-cell responses have been associated with control of simian immunodeficiency virus/HIV-1 replication, supporting the inclusion of CD4+ T-cell epitopes in vaccine formulations. Plasmid-encoded granulocyte-macrophage colony-stimulating factor (pGM-CSF) co-administration has been shown to induce potent CD4+ T-cell responses and to promote accelerated priming and increased migration of antigen-specific CD4+ T-cells. However, no study has shown whether co-immunisation with pGM-CSF enhances the number of vaccine-induced polyfunctional CD4+ T-cells. Our group has previously developed a DNA vaccine encoding conserved, multiple human leukocyte antigen (HLA)-DR binding HIV-1 subtype B peptides, which elicited broad, polyfunctional and long-lived CD4+ T-cell responses. Here, we show that pGM-CSF co-immunisation improved both magnitude and quality of vaccine-induced T-cell responses, particularly by increasing proliferating CD4+ T-cells that produce simultaneously interferon-γ, tumour necrosis factor-α and interleukin-2. Thus, we believe that the use of pGM-CSF may be helpful for vaccine strategies focused on the activation of anti-HIV CD4+ T-cell immunity.

Occurrence, morphology and ultrastructure of the Dufour gland in Melipona bicolor Lepeletier (Hymenoptera, Meliponini)

The occurrence, morphology and ultrastructure of the Dufour gland in Melipona bicolor Lepeletier, 1836 are presented. The Dufour gland is not present in workers. In virgin queens the gland cells show characteristics of low activity, which are described in the text. In physogastric queens the gland epithelium is higher and the cells more active than in virgin queens, showing numerous basal plasmic membrane invaginations impregnated by an electrondense material, increased apical invaginations and accumulation of substances that will be released to the gland lumen in the subcuticular space. Therefore, the data show that the Dufour gland is more developed in physogastric than in virgin queens, indicating a possible involvement of the Dufour gland in the reproduction of this species.

Taxonomy and morphology of Apsil Malloch (Diptera, Muscidae, Coenosiinae, Coenosiini) with new records, description of a new species and a key to identification

Taxonomy and morphology of Apsil Malloch (Diptera, Muscidae, Coenosiinae, Coenosiini) with new records, description of a new species and a key to identification. Apsil Malloch (Diptera, Muscidae, Coenosiinae, Coenosiini) includes 10 species, most of them described from Chile, but some also from Argentina. Five of them and one new species were found at the California Academy of Sciences collection (San Francisco, California) during the course of a project developed in that institution. Almost all material studied was collected in Chile, mainly by M. E. Irwin, during the year of 1966. Brief diagnosis of the known studied species (A. apicata Malloch, A. atripes Malloch, A. dilata Malloch, A. maculiventris Malloch (female described for the first time) and A. spatulata, Malloch), the description of A. mallochi, sp. nov. and a key for the identification of all known species are given. Color illustrations of some morphological characters make easier the recognition of the species. New geographic records were assigned.

External morphology of the immature stages of Neotropical heliconians: VIII. Philaethria wernickei (Röber) (Lepidoptera, Nymphalidae, Heliconiinae)

The external features of egg, larva, and pupa of Philaethria wernickei (Röber, 1906) are described and illustrated, based upon light and scanning electron microscopy.

Colony cycle of the social wasp Mischocyttarus consimilis Zikán (Hymenoptera, Vespidae)

Colony cycle of the social wasp Mischocyttarus consimilis Zikán (Hymenoptera, Vespidae). This study describes some aspects of the colony cycle of the Neotropical social wasp Mischocyttarus consimilis, from data obtained under field conditions. Our results showed that the colony cycle in M. consimilis is annual and asynchronous in relation to the months of the year. The colonies remained active for approximately eight months. Most of the abandonments were associated with natural causes, and were most frequent in the pre-emergence stage. The nests were constructed preferentially in man-made structures, especially in sites protected from direct sunlight and rain. Colony foundation was either by haplometrosis or pleometrosis, being the first form predominant.

Pineus boerneri Annand, 1928 (Hemiptera, Adelgidae) - a new species to Brazil: morphology of eggs, nymphs and adults

Pineus boerneri Annand, 1928 (Hemiptera, Adelgidae) _ a new species to Brazil: morphology of eggs, nymphs and adults. Pineus boerneri represents the first Adelgidae species recorded in Brazil. This aphid species forms extensive colonies on branches and trunk of Pinus spp., with apterous oviparous females, eggs and nymphs covered with white wax. The aim of this research is to study the morphology of eggs, nymphs, and adults to provide useful data for species identification in order to solve taxonomic issues. The study was based on morphometric data and optical and scanning electron microscopy images. First instar nymphs are active and can be easily distinguished from the others by their elongate minute yellowish body; well developed legs bearing a pair of sensorial setae at the apex of the second tarsomere; and antenna with three segments with a large rhinarium and distinct apical setae on the last segment. From the second to the fourth instar, nymphs are sessile, with round red body; they loose the third antennal segment and its sensorial structures, as well as the setae on the second tarsomere. The oviparous female is reddish-brown, with round body with about 0.76 mm diameter; legs reduced; antennae one-segmented; ovipositor distinct; numerous wax glands are present, mainly on the head. Accurate characterization of the species and distinction of the nymphal instars of P. boerneri were made possible by canonical analysis of morphometric data and morphological characters.

Comparative morphology of the mandibles of female polistine social wasps (Hymenoptera, Vespidae, Polistinae)

Comparative morphology of the mandibles of female polistine social wasps (Hymenoptera, Vespidae, Polistinae). Diversity of mandibular forms in female polistines is explored and compared among 116 species of all polistine genera. Inferences about function are made and discussed based on observed form differences. Mandible length and width measurements are analyzed for a subset of polistine species plus two vespines and two eumenines. A variable expressing the ratio between these variables is also considered in morphometric analyses. The following mechanical interactions among mandible structural elements are highlighted: opposition and crossing of mandibles' apical teeth at the middle of the closing trajectory; shearing action of the apical teeth against mesial denticles of the opposite mandible; shearing action of the mandible anterior margin against the ventral margin of the clypeus. In the genera Agelaia and Angiopolybia, exceptionally developed mesial mandibular structures may be related to necrophagy. In some epiponine genera, poorly developed mesial denticles and strong torsion of mandibles may be partially related to use of short wood fibers in nest construction as advanced by Sarmiento (2004). The considerable morphological variation found across the subfamily Polistinae is certain to be important in taxonomic and phylogenetic studies at the genus and species levels.

Comparative morphology of the spermathecae of some species of Chrysomya Robineau-Desvoidy and Cochliomyia Townsend (Diptera, Calliphoridae)

Comparative morphology of the spermathecae of some species of Chrysomya Robineau-Desvoidy and Cochliomyia Townsend (Diptera, Calliphoridae). Little is known about the morphology of the chitinized structures of the spermathecae of the Calliphoridae. In this work, the spermathecae of Chrysomya albiceps Wiedemann, 1819, C. megacephala Fabricius, 1794, Cochliomyia macellaria Fabricius, 1775 and C. hominivorax Coquerel, 1858 are described and illustrated. The occurrence in one species of four spermathecae, an atypical form for blow flies, was recorded for the first time. The analysis of these structures will allow a better understanding of this group as well as provide taxonomic characters for future phylogenetic studies.

Emigration of a colony of the leaf-cutting ant Acromyrmex heyeri Forel (Hymenoptera, Formicidae)

Emigration of a colony of the leaf-cutting ant Acromyrmex heyeri Forel (Hymenoptera, Formicidae). Colony migration is a poorly studied phenomenon in leaf-cutting ants. Here we report on the emigration of a colony of the leaf-cutting ant A. heyeri in Brazil. The colony emigrated to a new location 47.4 m away from the original nest site, possibly because it had undergone considerable stress due to competitive interactions with a colony of Acromyrmex crassispinus.

External morphology of the adult of Dynamine postverta (Cramer) (Lepidoptera, Nymphalidae, Biblidinae) and patterns of morphological similarity among species from eight tribes of Nymphalidae

External morphology of the adult of Dynamine postverta (Cramer) (Lepidoptera, Nymphalidae, Biblidinae) and patterns of morphological similarity among species from eight tribes of Nymphalidae. The external structure of the integument of Dynamine postverta postverta (Cramer, 1779) is based on detailed morphological drawings and scanning electron microscopy. The data are compared with other species belonging to eight tribes of Nymphalidae, to assist future studies on the taxonomy and systematics of Neotropical Biblidinae.

Lagochile emarginata (Gyllenhal): morphology of immature and imago, and biological records (Coleoptera, Scarabaeidae, Rutelinae)

Lagochile emarginata (Gyllenhal): morphology of immature and imago, and biological records (Coleoptera, Scarabaeidae, Rutelinae). The last larval instar and pupa of Lagochile emarginata are described. Pupa of the genus Lagochile Hoffmannsegg, 1817 is described for the first time. Redescription of the imago, clarifications on the morphology of immature Scarabaeoidea and biological notes are presented.