Effects of croton urucurana extracts and crude resin on Anagasta kuehniella (Lepidoptera: Pyralidae)

Hundreds of plant species have been studied in order to find out the active ingredient responsible for their insecticidal activity against the pests of economic importance. To verify the insecticidal activity in the husk of stem of Croton urucurana Baillon 1864 (Euphorbiaceae) against Anagasta kuehniella Zeller 1879 (Lepidoptera: Pyralidae), the methanolic (EMeOH) extract, dichloromethane fraction (FDM), ethyl acetate fraction (FAE) and crude resin, incorporated into an artificial diet were evaluated. EMeOH (0.5, 1.0 and 2.0%) and crude resin (2.0%) interfered with neither the weight nor the survival of fourth instar larvae and other analyzed parameters. FDM (2.0%) fraction caused mortality of 65%, and the artificial diet containing 2.0, 1.0 and 0.5% FAE caused 100, 55 and 68% mortality respectively when compared with the control, confirming the least efficiency rates of food conversion for FDM(2.0%) and FAE(1.0%). The tryptic analysis performed with the midgut fluid of fourth-instar larvae demonstrated that tryptic and chymiotryptic activities for the larvae fed artificial diet containing EMeOH and crude resin were not different.

Encontro de imaturos de Anopheles cruzii em bromélias de área urbana, litoral de São Paulo

Primeiro relato da ocorrência de larvas de Anopheles (Kerteszia) cruzii, mosquito essencialmente silvestre, em bromélias de solo em área urbana do município de Ilhabela, litoral norte do estado de São Paulo. De março de 1998 a julho de 1999 foram capturadas 312 formas imaturas de An. cruzii, sendo 8,6% em bromélias do ambiente urbano, 40,1% em bromélias do periurbano e 51,3% na mata. O número médio de bromélias com An. cruzii foi de 4,0% dentre o total de pesquisadas, com valores próximos de positividade para ambiente periurbano e mata. A presença de An. cruzii no ambiente urbano provavelmente é resultante da sua ocorrência prévia na mata, aliada à elevada presença desse criadouro na área urbana, de fonte alimentar e abrigos disponíveis. Alerta-se para a possibilidade de transferência de infecções entre esses ambientes

The Anopheles albitarsis complex with the recognition of Anopheles oryzalimnetes Wilkerson and Motoki, n. sp. and Anopheles janconnae Wilkerson and Sallum, n. sp. (Diptera: Culicidae)

The Anopheles (Nyssorhynchus) albitarsis complex includes six species: An. albitarsis, Anopheles oryzalymnetes Wilkerson and Motoki, n. sp., Anopheles marajoara, Anopheles dencorum, Anopheles janconnae Wilkerson and Sallum, n. sp., and An. albitarsis F. Except for An. deancorum, species of the complex are indistinguishable when only using morphology. The problematic distinction among species of the complex has made study of malaria transmission and ecology of An. albitarsis s.l. difficult. Consequently, involvement of species of the An. albitarsis complex in human Plasmodium transmission is not clear throughout its distribution range. With the aim of clarifying the taxonomy of the above species, with the exception of An. albitarsis F, we present comparative morphological and morphometric analyses, morphological redescriptions of three species and descriptions of two new species using individuals from populations in Brazil, Paraguay, Argentina and Venezuela. The study included characters from adult females, males, fourth-instar larvae, pupae and male genitalia of An. albitarsis, An. deaneorum and An. oryzalimnetes n. sp. For An. janconnae n. sp. only characters of the female, male and male genitalia were analysed. Fourth-instar larvae and pupae and male genitalia characteristics of all five species are illustrated. Bionomics and distribution data are given based on published literature records

Insight into Anopheles (Nyssorhynchus) (Diptera: Culicidae) species from Brazil full access

Anopheles (Nyssorhynchus) benarrochi s.l., Anopheles (Nyssorhynchus) oswaldoi s.l., and Anopheles (Nyssorhynchus) konderi s.l. collected in Acrelândia, state of Acre, Brazil, were identified based on morphological characters of the male genitalia, fourth-instar larvae, and pupae. Morphological variation was observed in the male genitalia of these species in comparison with specimens from other localities in Brazil. DNA sequence from the nuclear ribosomal second internal transcribed spacer of individuals identified as An. benarrochi s.l. by using male genitalia characteristics showed that the various morphological forms are conspecific but are distinct from An. benarrochi B from Colombia. Anopheles konderi s.l. and An. oswaldoi s.l. both misidentified as An. oswaldoi s.s. (Peryassú) throughout Brazil, may actually comprise at least two undescribed species. Diagnostic morphological characteristics of the male genitalia are provided to distinguish Anopheles benarrochi s.l., Anopheles oswaldoi s.l., and Anopheles konderi s.l. from morphologically similar species. Incrimination of An. oswaldoi s.s. in malaria transmission in Brazil needs further investigation because other undescribed species from Acre may have been confounded with this taxon

Redescription of Culex (Culex) dolosus (Lynch Arribálzaga)(Diptera: Culicidae), based on specimens from Pico do Itapeva, Serra da Mantiqueira, São Paulo, Brazil

Culex (Culex) dolosus (Lych Arribálzaga, 1891) is re-described and compared with Cx. eduardoi Casal & Garcia (1968) based on morphological characteristics. Both species are morphologically similar, and they have been largely misidentified throughout Brazil. Adult male and female, fourth instar larvae, and pupae of Cx. dolosus were examined, based on specimens from Pico do Itapeva, Pindamonhangaba Municipality, Serra da Mantiqueira, São Paulo State, southern Brazil. Male genitalia, larvae and pupae are illustrated. Geographical distribution is summarized from published records, and information on bionomics is based on literature and field data

Redescription of Anopheles (Nyssorhynchus) antunesi Galvão & Amaral and description of a new species of the Myzorhynchella Section (Diptera: Culicidae) from Serra da Mantiqueira, Brazil

Anopheles (Nyssorhynchus) pristinus Nagaki & Sallum, n. sp. of the Myzorhynchella Section is described based on morphological characters of adult females, males, fourth-instar larvae, pupae and male genitalia. Anopheles (Nyssorhynchus) antunesi Galvão & Amaral is characterized to fix its identity and distinguish it from An. pristinus. The eggs of An. antunesi are described for the first time. Molecular characterization employing sequences of the COI mitochondrial gene and the ITS2 region of ribosomal DNA are provided for each species. An. antunesi and An. pristinus are compared with morphologically similar species of the Myzorhynchella Section. The results of the present study suggest that the new species has been misidentified as both An. antunesi and Anopheles lutzii Cruz. An. antunesi and An. pristinus are sympatric, occurring at high altitudes in Serra da Mantiqueira, southeastern Brazil

Primeiro registro de Anopheles (Kertezia) homunculus Komp (Diptera, Culicidae) no Estado do Espírito Santo, Brasil

Em pesquisa de campo realizada no interior do Estado do Espírito Santo, Brasil, em janeiro de 2006, como parte de projeto sobre a transmissão de Plasmodium, foram coletadas larvas de anofelinos em bromélias. Os imaturos foram mantidos no laboratório até a obtenção dos adultos machos e fêmeas associados com as exúvias das larvas e das pupas, para serem identificados. Conseqüentemente, verificou-se que dois espécimes pertenciam a Anopheles (Kerteszia) homunculus Komp, 1937. Este é o primeiro registro dessa espécie de Kerteszia no Espírito Santo. O encontro evidencia a importância de estudos adicionais de modo a estabelecer a distribuição geográfica do An. homunculus, bem como o status taxonômico e a importância epidemiológica da espécie na dinâmica da transmissão da malária em áreas de Mata Atlântica

Embryonic, Larval, and Juvenile Development of the Sea Biscuit Clypeaster subdepressus (Echinodermata: Clypeasteroida)

Sea biscuits and sand dollars diverged from other irregular echinoids approximately 55 million years ago and rapidly dispersed to oceans worldwide. A series of morphological changes were associated with the occupation of sand beds such as flattening of the body, shortening of primary spines, multiplication of podia, and retention of the lantern of Aristotle into adulthood. To investigate the developmental basis of such morphological changes we documented the ontogeny of Clypeaster subdepressus. We obtained gametes from adult specimens by KCl injection and raised the embryos at 26 degrees C. Ciliated blastulae hatched 7.5 h after sperm entry. During gastrulation the archenteron elongated continuously while ectodermal red-pigmented cells migrated synchronously to the apical plate. Pluteus larvae began to feed in 3 d and were similar to 20 d old at metamorphosis; starved larvae died 17 d after fertilization. Postlarval juveniles had neither mouth nor anus nor plates on the aboral side, except for the remnants of larval spicules, but their bilateral symmetry became evident after the resorption of larval tissues. Ossicles of the lantern were present and organized in 5 groups. Each group had 1 tooth, 2 demipyramids, and 2 epiphyses with a rotula in between. Early appendages consisted of 15 spines, 15 podia (2 types), and 5 sphaeridia. Podial types were distributed in accordance to Loven's rule and the first podium of each ambulacrum was not encircled by the skeleton. Seven days after metamorphosis juveniles began to feed by rasping sand grains with the lantern. Juveniles survived in laboratory cultures for similar to 9 months and died with <500 mu m wide, a single open sphaeridium per ambulacrum, aboral anus, and no differentiated food grooves or petaloids. Tracking the morphogenesis of early juveniles is a necessary step to elucidate the developmental mechanisms of echinoid growth and important groundwork to clarify homologies between irregular urchins.

Description of the immatures of the ant, Myrmelachista catharinae

The Neotropical ant genus Myrmelachista Roger comprises 69 described species and subspecies, and still is a poorly studied group. Larvae play a paramount role in colony nutrition in social hymenopterans and bear considerable value in the reconstruction of group phylogenies, however, they are generally neglected. Larvae of different instars of Myrmelachista catharinae Mayr (Hymenoptera: Formicidae) are herein described in detail by light and scanning electron microscopy. The number of larval instars was estimated as three based on the frequency distribution of maximum head capsule widths. The described larvae confirmed some traits typical of the genus: general shape of body and mandibles, general aspect and distribution of body hairs, and the number of sensilla on the palps and galea. Differently from other Myrmelachista larvae previously described, M. catharinae presented two distinct kinds of second instars, some additional types of body hairs, different number of antennal sensilla, and a distinct labrum shape. M. catharinae presented ten pairs of spiracles, which is the first record for this genus.

DESCRIPTION OF THE IMMATURES OF WORKERS OF THE ANT CAMPONOTUS VITTATUS (HYMENOPTERA: FORMICIDAE)

Camponotus vittatus Forel is a poorly studied Neotropical ant, which is very common in Brazil. Larval descriptions are useful to systematics, as larval characters aid with genus-level differentiation, and ant larvae lie at the basis of ant social organization. This study presents the first description of the immatures of C. vittatus with the aid of light and scanning electron microscopy. There are three instars based on the frequency distribution of larval head widths. The larvae had some characteristics typical of Camponotus, specifically, a 'pogono-myrmecoid' body shape, 10 pairs of spiracles, antennae with 3 sensilla, mature larvae with pronounced labial pseudopalps, and conspicuous 'chiloscleres' on the labrum. Unique characteristics found would include the greatest diversity of body hair types recorded in an ant larva and 'camponotoid' mandibles with 6 medial denticles over the blade. The number of antennal sensilla proved variable.

The four hexamerin genes in the honey bee: structure, molecular evolution and function deduced from expression patterns in queens, workers and drones

Background: Hexamerins are hemocyanin-derived proteins that have lost the ability to bind copper ions and transport oxygen; instead, they became storage proteins. The current study aimed to broaden our knowledge on the hexamerin genes found in the honey bee genome by exploring their structural characteristics, expression profiles, evolution, and functions in the life cycle of workers, drones and queens. Results: The hexamerin genes of the honey bee (hex 70a, hex 70b, hex 70c and hex 110) diverge considerably in structure, so that the overall amino acid identity shared among their deduced protein subunits varies from 30 to 42%. Bioinformatics search for motifs in the respective upstream control regions (UCRs) revealed six overrepresented motifs including a potential binding site for Ultraspiracle (Usp), a target of juvenile hormone (JH). The expression of these genes was induced by topical application of JH on worker larvae. The four genes are highly transcribed by the larval fat body, although with significant differences in transcript levels, but only hex 110 and hex 70a are re-induced in the adult fat body in a caste-and sex-specific fashion, workers showing the highest expression. Transcripts for hex 110, hex 70a and hex70b were detected in developing ovaries and testes, and hex 110 was highly transcribed in the ovaries of egg-laying queens. A phylogenetic analysis revealed that HEX 110 is located at the most basal position among the holometabola hexamerins, and like HEX 70a and HEX 70c, it shares potential orthology relationship with hexamerins from other hymenopteran species. Conclusions: Striking differences were found in the structure and developmental expression of the four hexamerin genes in the honey bee. The presence of a potential binding site for Usp in the respective 5' UCRs, and the results of experiments on JH level manipulation in vivo support the hypothesis of regulation by JH. Transcript levels and patterns in the fat body and gonads suggest that, in addition to their primary role in supplying amino acids for metamorphosis, hexamerins serve as storage proteins for gonad development, egg production, and to support foraging activity. A phylogenetic analysis including the four deduced hexamerins and related proteins revealed a complex pattern of evolution, with independent radiation in insect orders.

Reproductive aspects of Meloetyphlus fuscatus a meloid beetle cleptoparasite of the bee Eulaema nigrita (Hymenoptera, Apidae, Euglossini)

This study investigated the reproductive biology of the meloid beetle Meloetyphlus fuscatus Waterhouse, a cleptoparasite of Eulaema nigrita nests. New E. nigrita nests had rates of cell parasitism by meloids ranging from 3.7% to 15.8%, while in re-used nests the rate of cell parasitism ranged from 1.4% to 18.7%. The adult parasites were never observed trying to leave the host nests. Both sexes mated more than once. Females had a high fecundity (more than 8,000 eggs), and in most cases, deposited their eggs into the empty, old cells of the host. The triungulins (the first larval instars) hatched from eggs 18-20 days after oviposition and dispersed from the host nest by attaching themselves to males as they emerged. The triungulins most likely transfer to female bees during mating and are transported to the nests of their hosts. Within an attacked cell, the triungulin consumes the bee egg and completes its development by consuming the larval food stored in the cell.

Rearing Africanized honey bee (Apis mellifera L.) brood under laboratory conditions

We developed a method for rearing larvae of Africanized bees under laboratory conditions to determine the amount of diet needed during larval development to obtain a worker bee. We started with larvae 18-24 h old, which were transferred to polyethylene cell cups and fed for five days. We found that the amount of diet needed for successful larval development was: 4, 15, 25, 50, and 70 mu L during the first to fifth days, respectively. The survival rate to the adult stage was 88.6% when the larvae received the daily amount of diet divided into two feedings, and 80% when they received only one feeding per day. The adult weight obtained in the laboratory, when the larvae received the daily amount of diet in a single dose, did not differ from those that were developed under field conditions (our control). All adults that we obtained in laboratory appeared to be normal. This technique has the potential to facilitate studies on brood pathogens, resistance mechanisms to diseases and also might be useful to test the impacts of transgenic products on honey bee brood.

Functional characterization of the sciarid BhC4-1 core promoter in transgenic Drosophila

Background: Core promoters are cis-regulatory modules to which bind the basal transcriptional machinery and which participate in the regulation of transcription initiation. Although core promoters have not been extensively investigated through functional assays in a chromosomal context, the available data suggested that the response of a given core promoter might vary depending on the promoter context. Previous studies suggest that a (-57/+40) fragment constitutes the core promoter of the BhC4-1 gene which is located in DNA puff C4 of the sciarid fly Bradysia hygida. Here we tested this (-57/+40) fragment in distinct regulatory contexts in order to verify if promoter context affects its core promoter activity. Results: Consistent with the activity of a core promoter, we showed that in the absence of upstream regulatory sequences the (-57/+40) fragment drives low levels of reporter gene mRNA expression throughout development in transgenic Drosophila. By assaying the (-57/+40) fragment in two distinct regulatory contexts, either downstream of the previously characterized Fbp1 enhancer or downstream of the UAS element, we showed that the BhC4-1 core promoter drives regulated transcription in both the germline and in various tissues throughout development. Furthermore, the use of the BhC4-1 core promoter in a UAS construct significantly reduced salivary gland ectopic expression in third instar larvae, which was previously described to occur in the context of the GAL4/UAS system. Conclusions: Our results from functional analysis in transgenic Drosophila show that the BhC4-1 core promoter drives gene expression regardless of the promoter context that was assayed. New insights into the functioning of the GAL4/UAS system in Drosophila were obtained, indicating that the presence of the SV40 sequence in the 3' UTR of a UAS construct does not preclude expression in the germline. Furthermore, our analysis indicated that ectopic salivary gland expression in the GAL4/UAS system does not depend only on sequences present in the GAL4 construct, but can also be affected by the core promoter sequences in the UAS construct. In this context, we propose that the sciarid BhC4-1 core promoter constitutes a valuable core promoter which can be employed in functional assays in insects.

Notes on Population Dynamics of Amblyomma Ticks (Acari: Ixodidae) in Brazil

Previous population dynamics data, generated for Amblyomma parvum Aragao and Aniblyomma cajennense (Fabricius) in Argentina and southeastern Brazil, have indicated that these ticks complete I generation per year, with larvae predominating in autumn, nymphs in winter, and mostly adults during spring and summer. The present study reports population dynamics data for free-living Amblyomma spp. ticks in northern Brazil (Amazon forest, latitude 10 degrees S, 63 degrees W), and for Amblyomma spp. ticks collected oil birds in Southeastern Brazil (latitude 23 degrees S, 45 degrees W). In northern Brazil, adult ticks predominated from mid-spring to mid-autumn, larvae predominated in early winter, and nymphs from mid-winter to mid-spring. Seven Amblyomma spp. were identified, although A. cajannense predominated in I of the 2 sites sampled. In southeastern Brazil, larval infestations on birds peaked in autumn, followed by a nymphal infestation peak in late winter. At least 32% and 75% of these larvae and nymphs, respectively, were identified as Amblyomma longirostre (Koch). Similar to previous work, the present study showed that Amblyomma spp. larvae and nymphs predominated during autumn-winter months, and mostly adults during spring-summer months, a pattern compatible with 1 genration/yr, even at latitude 10 degrees S in northern Brazil.