Genotypic and phenotypic diversity of a baculovirus population within an individual insect host

It is becoming increasingly apparent that many pathogen populations, including those of insects, show high levels of genotypic variation. Baculoviruses are known to be highly variable, with isolates collected from the same species in different geographical locations frequently showing genetic variation and differences in their biology. More recent Studies at smaller scales have also shown that virus DNA profiles from individual larvae can show polymorphisms within and between populations of the same species. Here, we investigate the genotypic and phenotypic variation of an insect baculovirus infection within a single insect host. Twenty four genotypically distinct nucleopolyhedrovirus (NPV) variants were isolated from an individual pine beauty moth, Panolis flammea, caterpillar by in vivo cloning techniques. No variant appeared to be dominant in the population. The Pafl NPV variants have been mapped using three restriction endonucleases and shown to contain three hypervariable regions containing insertions of 70-750 bp. Comparison of seven of these variants in an alternative host, Mamestra brassicae, demonstrated that the variants differed significantly in both pathogenicity and speed of kill. The generation and maintenance of pathogen heterogeneity are discussed. (c) 2005 Elsevier Inc. All rights reserved.

Galleria mellonella as an infection model for Campylobacter jejuni virulence

Larvae of Galleria mellonella (Greater Wax Moth) have been shown to be susceptible to Campylobacter jejuni infection and our study characterizes this infection model. Following infection with C. jejuni human isolates, bacteria were visible in the haemocoel and gut of challenged larvae, and there was extensive damage to the gut. Bacteria were found in the extracellular and cell-associated fraction in the haemocoel, and it was shown that C. jejuni can survive in insect cells. Finally, we have used the model to screen a further 67 C. jejuni isolates belonging to different MLST types. Isolates belonging to ST257 were the most virulent in the Galleria model, whereas those belonging to ST21 were the least virulent.

Insect infection model for campylobacter jejuni reveals that O-methyl phosphoramidate has insecticidal activity

Galleria mellonella (wax moth) larvae have elsewhere been shown to be susceptible to pathogens such as Francisella tularensis, Burkholderia mallei, and Pseudomonas aeruginosa. We report that the larvae are rapidly killed by Campylobacter jejuni at 37 degrees C. Three strains of C. jejuni tested, 11168H (human diarrheal isolate), G1 (human Guillain-Barre syndrome isolate), and 81-176 (human diarrheal isolate), were equally effective at killing G. mellonella larvae. A panel of defined mutants of C. jejuni 11168H, in known or putative virulence genes, showed different degrees of attenuation in G. mellonella larvae. A mutant lacking the O-methyl phosphoramidate (MeOPN) capsule side group was attenuated, clearly demonstrating that MeOPN has a role in virulence. This new model of C. jejuni infection should facilitate the identification of novel virulence genes.

Enhancement of buffer strips can improve provision of multiple ecosystem services

Farmland invertebrates play a pivotal role in the provision of ecosystem services, i.e. services that benefit humans. For example, bumblebees, solitary bees and honeybees, are crucial to the pollination of many of the world's crops and wildflowers, with over 70% of the world's major food crops dependent on the pollination services provided by these insects. The larvae of some butterfly species are considered to be pests; however, together with moth and sawfly larvae, they represent a key dietary component for many farmland birds. Spiders and ground beetles predate on crop pests including aphids, whilst soil macrofauna such as earthworms are vital for soil fertility services and nutrient recycling. Despite their importance, population declines of invertebrates have been observed during the last sixty years in the UK and NW Europe. For example, seven UK bumblebee species are in decline, and in the last 20 years, the species Bombus subterraneus (short-haired bumblebee) has become extinct, whilst there was a 54% decline in honeybee colony numbers in England from 1985 to 2005. Comparable trends have been documented for butterflies with a 23% decline in UK farmland species such as Anthocharis cardamines (orange tip) between 1990 and 2007. These declines have been widely attributed to the modern intensive arable management practices that have been developed to maximise crop yield. For example, loss and fragmentation of foraging and nesting habitats, including species-rich meadows and hedgerows, have been implicated in the decline of bees and butterflies. Increased use of herbicides and fertilisers has caused detrimental effects on many plant species with negative consequences for predatory invertebrates such as spiders and beetles which rely on plants for food and shelter.

Adding value to the urban lawnscape: insect abundance and diversity in grass-free lawns

Insect diversity may be declining even more rapidly than in plants and vertebrates, particularly in areas where indigenous habitats are replaced by an anthropogenic one. The most common component of anthropogenic greenspace is the ornamental lawn. Intensively managed and offering limited habitat opportunities for both plants and insects, lawns are biodiversity poor and ecologically insensitive. An alternative lawn format that positively influences biodiversity and reduces management requirements would be a useful tool in eco-friendly urban greenspace management. In investigating the potential for a forb-only alternative to the grass lawn we sampled both trial grass-free lawn formats and turf lawns to identify any influence that lawn composition and grass-free lawn specific mowing regimes might have on the abundance and diversity of insect families. In addition to the mowing regimes, both the composition and origin of lawn flora were found to significantly influence insect abundance and diversity and these factors rarely interacted. Native-only and mixed origin grass-free lawns hosted greater numbers of adult insects than found in turf and an equivalent diversity of insect families, however the mowing regime applied was distinct from traditional turf lawn management by being substantially less intensive and our results suggest that there is the potential for even greater abundance and diversity via the grass-free format that may offer additional resources to insectivorous garden species such as birds. When the composition of grass-free lawns included native forbs the diversity of insect families was found be sufficiently different from turf lawns to form distinct assemblages and in so doing contribute to beta diversity within urban greenspace. In sum, grass-free lawns may be a useful and aesthetically appropriate tool for adding value to the generally biodiversity poor urban lawnscape.

Analysis and manipulation of the structure of odor plumes from a piezo-electric release system and measurements of upwind flight of male Almond Moths, Cadra cautella, to pheromone plumes

We investigated the plume structure of a piezo-electric sprayer system, set up to release ethanol in a wind tunnel, using a fast response mini-photoionizaton detector. We recorded the plume structure of four different piezo-sprayer configurations: the sprayer alone; with a 1.6-mm steel mesh shield; with a 3.2-mm steel mesh shield; and with a 5 cm circular upwind baffle. We measured a 12 × 12-mm core at the center of the plume, and both a horizontal and vertical cross-section of the plume, all at 100-, 200-, and 400-mm downwind of the odor source. Significant differences in plume structure were found among all configurations in terms of conditional relative mean concentration, intermittency, ratio of peak concentration to conditional mean concentration, and cross-sectional area of the plume. We then measured the flight responses of the almond moth, Cadra cautella, to odor plumes generated with the sprayer alone, and with the upwind baffle piezo-sprayer configuration, releasing a 13:1 ratio of (9Z,12E)-tetradecadienyl acetate and (Z)-9-tetradecenyl acetate diluted in ethanol at release rates of 1, 10, 100, and 1,000 pg/min. For each configuration, differences in pheromone release rate resulted in significant differences in the proportions of moths performing oriented flight and landing behaviors. Additionally, there were apparent differences in the moths’ behaviors between the two sprayer configurations, although this requires confirmation with further experiments. This study provides evidence that both pheromone concentration and plume structure affect moth orientation behavior and demonstrates that care is needed when setting up experiments that use a piezo-electric release system to ensure the optimal conditions for behavioral observations.

Long-term changes to the frequency of occurrence of British moths are consistent with opposing and synergistic effects of climate and land-use changes

1. Species’ distributions are likely to be affected by a combination of environmental drivers. We used a data set of 11 million species occurrence records over the period 1970–2010 to assess changes in the frequency of occurrence of 673 macro-moth species in Great Britain. Groups of species with different predicted sensitivities showed divergent trends, which we interpret in the context of land-use and climatic changes. 2. A diversity of responses was revealed: 260 moth species declined significantly, whereas 160 increased significantly. Overall, frequencies of occurrence declined, mirroring trends in less species-rich, yet more intensively studied taxa. 3. Geographically widespread species, which were predicted to be more sensitive to land use than to climate change, declined significantly in southern Britain, where the cover of urban and arable land has increased. 4. Moths associated with low nitrogen and open environments (based on their larval host plant characteristics) declined most strongly, which is also consistent with a land-use change explanation. 5. Some moths that reach their northern (leading edge) range limit in southern Britain increased, whereas species restricted to northern Britain (trailing edge) declined significantly, consistent with a climate change explanation. 6. Not all species of a given type behaved similarly, suggesting that complex interactions between species’ attributes and different combinations of environmental drivers determine frequency of occurrence changes. 7. Synthesis and applications. Our findings are consistent with large-scale responses to climatic and land-use changes, with some species increasing and others decreasing. We suggest that land-use change (e.g. habitat loss, nitrogen deposition) and climate change are both major drivers of moth biodiversity change, acting independently and in combination. Importantly, the diverse responses revealed in this species-rich taxon show that multifaceted conservation strategies are needed to minimize negative biodiversity impacts of multiple environmental changes. We suggest that habitat protection, management and ecological restoration can mitigate combined impacts of land-use change and climate change by providing environments that are suitable for existing populations and also enable species to shift their ranges.

Three-dimensional Reconstruction of Ovaries of Leaf-cutting Ant (Atta sexdens rubropilosa) Queens (Hymenoptera: Formicidae)

In this study, the ovary morphology of newly emerged ant queens of Atta sexdens rubropilosa was studied in whole mount preparations by confocal microscopy. The ovaries are composed of approximately 40 ovarioles, showing non-synchronic oocyte maturation. The terminal filament with clusters of undifferentiated cells was found at the distal end of the ovarioles. Next to this region is the germarium, composed of several elongated cystocytes interconnected by cytoplasmic bridges. The nurse cells (23-28 cells) result from asymmetric mitosis. Cytoskeleton analysis showed F-actin concentrated at the muscle cells of the external tunica and in fusomes inside the ovarioles. Microtubules were concentrated around the nuclei of the nurse and follicular cells. In contrast, the oocytes and the external tunica showed faint staining for tubulin.

Comportamento de três protetores auriculares tipo concha, em ambientes com ruídos em baixa freqüência

Esta dissertação consiste no estudo comparativo entre três tipos básicos de protetores auriculares do tipo concha (modelo simples de baixo preço e baixo NRR, modelo sofisticado, de maior custo e com NRR elevado, e modelo ativo com circuito eletrônico de cancelamento de ruído), tanto quanto ao conforto quanto às reais atenuações oferecidas pelos mesmos, num ambiente industrial com elevados níveis de ruído e de baixa freqüência (com predominância na faixa de 500 Hz). Para a análise dos protetores foram utilizadas a técnica MIRE (Miniature Microphone in Real-Ear, ou Microfone Miniatura no Ouvido Real) de avaliação do nível de atenuação, o uso de questionários de conforto, e medições de cortisol urinário, para a avaliação do estresse no trabalho. Os resultados indicaram que o protetor ativo não apresentou os resultados esperados em termos de atenuação, o que não justifica o seu elevado custo, e apresentou a pior condição de conforto entre os três tipos estudados. As medições do cortisol urinário, por sua vez, não apresentaram resultados que pudessem ser úteis na escolha do melhor modelo de protetor tipo concha.

Escola da família agrícola: uma alternativa para o ensino rural

Este trabalho descreve a experiência educativa de três Es colas da Família Agrícola do Movimento de Educação Promocional do Espírito Santo, existentes no meio rural do Estado do Espírito Santo. Duas escolas são masculinas e uma feminina. Uma escora masculina localiza-se no Município de Rio Novo do Sul, na região sul e outra no povoado de Bley, Município de São Gabriel da Palha, região norte. A escola feminina localiza-se no distrito de Nestor Gomes, Município de são Mateus, ao norte. A Escola da Família Agrícola adota a metodologia da Maison Familiale. Seus princípios básicos norteadores, a alternância e a educação geral orientada para o meio rural. fazem com que ela seja uma alternativa da escola pós-primária viável para o meio rural dos países emvia de desenvolvimento. No Espírito Santo, essa experiência educativa foi implantada em regiões agrícolas tradicionais em transição. Nos seus primeiros anos de funcionamento ela sofreu influências de várias forças para adaptar-se ao meio onde atua e ã legislação de ensino. A descrição e análise do seu processo de adaptação ã real idade espiritossantense oferecem indicadores valiosos ao planejamento e administração do ensino rural para regiões agrícolas exploradas por mão-de-obra familiar. Com esse propósito são descritos e analisados os instrumentos metodológicos, o plano de formação, a composição da equipe de monitores, o prédio escolar, a propriedade agrícola e as características principais ,de sua clientela: ida de, procedência geográfica. percurso da residência até a escola e a forma de trabalho das famílias . os agricultores e os 11ão-agricultores. Em razão da original idade de sua metodologia, analisam-se ainda os motivos de sua aceitação entre os alunos ~~ suas famílias. A estrutura da alternância, seu baixo custo operacional , a correspondência do seu currículo e processo de avaliação às escola!. de ensino regular urbanas (ginásio e escola polivalente), o aprofundamento de estudos voltados para o meio rural. entre outros, são os moth'os mais significativos para que ela seja aceita como uma escola adequada para o meio rural. As aspirações profissionais dos alunos e a concretização profissional dos egressos em ocupações vinculadas ao meio rural e urbano demonstram que só a ação isolada da Escola da Família Agrícola torna-se incapaz para fixar o jovem no meio rural.

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

Biological aspects of the immature stages of Ceraeochrysa everes (Banks) (Neuroptera: Chrysopidae)

Para o estabelecimento de um programa de controle biológico, o conhecimento de alguns aspectos biológicos e de comportamento dos inimigos naturais é de extrema importância. Os aspectos biológicos do desenvolvimento larval e pupal do predador Ceraeochrysa everes (Banks) foram estudados. Larvas oriundas de adultos da geração F1 foram mantidas em laboratório a 25 ± 21°C, 70 ± 10 % UR e fotofase 14 horas, sendo alimentadas com ovos de Sitotroga cerealella (Olivier). A duração e viabilidade do período embrionário, estágios imaturos de desenvolvimento e o período de ovo a adulto foram avaliados. O período embrionário foi em média de 5,0 dias, enquanto que as durações médias para o primeiro, segundo e terceiro instares foram de 5,1 ± 0,03; 4,3 ± 0,05 e 4,5 ± 0,05 dias, respectivamente, com viabilidade superior a 90 %. Os estágios larval, pré-pupal e pupal apresentaram duração média de 13,9 ± 0,07; 5,7 ± 0,07 e 9,6 ± 0,12 dias, respectivamente. A duração do ciclo biológico foi de 34 ± 0,11 dias em média. Ovos de S. cerelella não foram adequados para a manutenção de C. everes em laboratório, por interferir no desenvolvimento do predador.

Parasitismo natural de ovos do curuquerê-do-algodoeiro por Trichogramma pretiosum em diferentes estágios fenológicos de variedades de algodoeiros em Ipameri, GO

The cotton can be damaged by many pests, standing out among them cotton leafworm, responsible for major losses in production, if not controlled properly. A group of natural enemies is the most studied of the family Trichogrammatidae, mainly species of the genus Trichogramma, because of its importance in biological control at the initial stage of development of their hosts. In order to study the natural egg parasitism of Alabama argillacea (Hubner, 1818) (Lepidoptera: Noctuidae) by Trichogramma pretiosum Riley, 1879 (Hymenoptera: Trichogrammatidae) in different phenological stages of transgenic and conventional varieties of cotton (Gossypium hirsutum L.) in the region of Ipameri, GO, it was carried out this experiment from december 2007 to april 2008, in the UEG, State of Goias. The experimental design was in randomized blocks with five treatments consisting of the conventional varieties DeltaOPAL, FMX 966, FMX 993, FMX 910 and NuOPAL transgenic variety, in four replications. In the beginning of oviposition of the A. argillacea it was started the 27 old days plant and the parasitism increased as the availability of eggs of the host, regardless of variety. It was found that the varieties over the vegetative cycle did not show statistical differences on the preference for oviposition by the moth neither on the parasitism. The level of parasitism of eggs of A. argillacea by T. pretiosum varied depending on the density of eggs on plants, not in terms of varieties and phenological stages.

Produtividade agrícola de variedades de cana-de-açúcar e incidência de broca-comum e cigarrinha-da-raiz em canavial colhido sem queima

Com o uso de variedades adaptadas e o manejo do solo pós-colheita da soqueira de cana-de-açúcar em sistema de colheita mecanizada, poderá ser afetada a incidência de pragas e a produtividade do canavial. Assim, desenvolveu-se o presente trabalho com o objetivo de se avaliar a influência da palhada de dezoito variedades de cana-de-açúcar submetidas a diferentes sistemas de manejo de colheita mecânica, sem queima, na produtividade agrícola e na incidência de broca-comum e cigarrinha-das-raízes na cultura de cana-de-açúcar. Os tratamentos foram constituídos pela presença da palha sem trituração/sem cultivo (PST/SC); palha sem trituração/com cultivo com escarificador (PST/CC); palha triturada/com cultivo com escarificador (PT/CC). O rendimento de colmos na cana-soca é favorecido pelo manejo, com cultivo, da palhada da planta crua, colhida mecanicamente, sem alteração na incidência de broca-comum e da cigarrinha das raízes. A variedade SP87-365 de cana-de-açúcar é opção viável e rentável, em qualquer sistema de manejo da cana crua, com adaptação à colheita mecânica e às condições edafoclimáticas da região de Ribeirão Preto, SP. A incidência de broca-comum e da cigarrinha das raízes pode ser bastante reduzida com a utilização das variedades SP88-817 e SP76-112 de cana-de-açúcar, em qualquer sistema de manejo da palhada da cana crua colhida mecanicamente e minimizada na colheita mecânica da cana crua, seguida de trituração da palhada e cultivo com escarificador na entrelinha.

Controle de pragas do pessegueiro através do ensacamento dos frutos

Objetivou-se, no trabalho, buscar opções ao ensacamento de pêssegos, visando o controle eficiente das principais pragas do pessegueiro. Para isso, instalou-se um experimento em um pomar comercial de pêssego do cultivar Aurora 2, conduzido em sistema de vaso e espaçamento de 6 x 4 m. Os tratamentos foram os seguintes: T1 - Sacolas de TNT branco (45 g/m²) fechado, T2 - Sacolas de TNT branco (45 g/m²) aberto, T3 - Sacolas de TNT branco (20 g/m²) fechado; T4 - Sacolas de TNT branco (20 g/m²) aberto; T5 - Sacolas de polipropileno microperfurado transparente (furos de 1mm) fechado; T6 - Sacolas de polipropileno microperfurado transparente (furos de 1mm) aberto; T7 - Sacolas de polipropileno microperfurado transparente (furos de 2mm) fechado; T8 - Sacolas de polipropileno microperfurado transparente (furos de 2mm) aberto; T9 - Sacolas de polietileno microperfurado leitoso (furos de 1mm) fechado; T10 - Sacolas de polietileno microperfurado leitoso (furos de 1mm) aberto; T11 - Sacolas de polietileno microperfurado leitoso (furos de 2mm) fechado; T12 - Sacolas de polietileno microperfurado leitoso (furos de 2mm) aberto; T13 - Sacolas de papel impermeável fechado; T14 - Sacolas de papel impermeável aberto; T15 - Testemunha (sem ensacamento). de acordo com os resultados concluiu-se que todas as embalagens foram eficientes no controle de moscas-das-frutas e pássaros, porém não para mariposa oriental. As sacolas com abertura no fundo podem ser utilizadas como opção ao ensacamento de pêssegos.