The temperate Burkholderia phage AP3 of the Peduovirinae shows efficient antimicrobial activity against B. cenocepacia of the IIIA lineage

Burkholderia phage AP3 (vB_BceM_AP3) is a temperate virus of the Myoviridae and the Peduovirinae subfamily (P2likevirus genus). This phage specifically infects multidrug-resistant clinical Burkholderia cenocepacia lineage IIIA strains commonly isolated from cystic fibrosis patients. AP3 exhibits high pairwise nucleotide identity (61.7%) to Burkholderia phage KS5, specific to the same B. cenocepacia host, and has 46.7% - 49.5% identity to phages infecting other species of Burkholderia. The lysis cassette of these related phages has a similar organization (putative antiholin, putative holin, endolysin and spanins) and shows 29-98% homology between specific lysis genes, in contrast to Enterobacteria phage P2, the hallmark phage of this genus. The AP3 and KS5 lysis genes have conserved locations and high amino acid sequence similarity. The AP3 bacteriophage particles remain infective up to 5 h at pH 4-10 and are stable at 60°C for 30 min, but are sensitive to chloroform, with no remaining infective particles after 24 h of treatment. AP3 lysogeny can occur by stable genomic integration and by pseudo-lysogeny. The lysogenic bacterial mutants did not exhibit any significant changes in virulence compared to wild-type host strain when tested in the Galleria mellonella moth wax model. Moreover, AP3 treatment of larvae infected with B. cenocepacia revealed a significant increase (P < 0.0001) in larvae survival in comparison to AP3-untreated infected larvae. AP3 showed robust lytic activity, as evidenced by its broad host range, the absence of increased virulence in lysogenic isolates, the lack of bacterial gene disruption conditioned by bacterial tRNA downstream integration site, and the absence of detected toxin sequences. These data suggest the AP3 phage is a promising potent agent against bacteria belonging to most common B. cenocepacia IIIA lineage strains.

Selection and Floral Evolution in Platanthera bifolia and P. chlorantha (Orchidaceae)

Natural selection mediated by pollinators has influenced the evolution of floral diversity of the flowering plants (angiosperms). The scope of this thesis was to study: 1) phenotypic selection, 2) mating systems, and 3) floral shifts involved in plant speciation. Model plant species were Platanthera bifolia and P. chlorantha (Orchidaceae). These orchids are moth-pollinated, strictly co-sexual (bisexual flowers), and produce a spike that displays 10-20 white flowers. I explored the influence of characters on plant fitness by using multiple linear regressions. Pollen removal (male fitness) and fruit set (female fitness) increased with more flowers per plant in three P. bifolia populations. There was selection towards longer spurs in a dry year when average spur length was shorter than in normal-wet years. Female function was sensitive to drought, which enabled an application of the male function hypothesis of floral evolution (Bateman's principle). The results show that selection may vary between populations, years, and sex-functions. I examined inbreeding by estimating levels of geitonogamy (self-pollination between flowers of an individual) with an emasculation method in two P. bifolia populations. Geitonogamy did not vary with inflorescence size. Levels of geitonogamy was 20-40% in the smaller, but non-significant in the larger population. This may relate to lower number of possible mates and pollinator activity in the smaller population. Platanthera bifolia exhibits the ancestral character state of tongue-attachment of pollinia on the pollinator. Its close relative P. chlorantha attaches its pollinia onto the pollinator's eyes. To explore the mechanism of a floral shift, pollination efficiency and speed was compared between the two species. The results showed no differences in pollination efficiency, but P. chlorantha had faster pollen export and import. Efficiency of pollination in terms of speed may cause floral shifts, and thus speciation.

Low doses of a neonicotinoid insecticide modify pheromone response thresholds of central but not peripheral olfactory neurons in a pest insect

International audience

Microlepidoptera of Illinois hill prairies

Among insects, which are the most diverse eukaryotic group on earth, Lepidoptera is one of four enormously diverse orders, with approximately 10,000 described species in North America. Within the order, Nearctic “microlepidoptera,” which represent an overwhelmingly large percentage of diversity within the order, remain poorly known despite their ecological importance in many plant communities. In this thesis, I undertook several studies of microlepidoptera diversity in a natural community type (hill prairie) and a managed community type (biofuel feedstock). In two Illinois hill prairies differing in size, latitude, and plant composition, alpha diversity of Pyraloidea and Tortricidae was similar, but the prairies were found to support different sets of species of these moth groups. It is concluded that the similarity in alpha diversity occurs because the larger prairie supports primarily a complement of moth species that feed as larvae on prairie plants (especially species of Asteraceae and Fabaceae), whereas the moths collected in the small prairie represent relatively few prairie-associated species, plus a large component of species that feed as larvae on deciduous trees that surround the prairie. This agrees with the finding of high beta diversity of moths between the sites, which reflects a high level of larval hostplant specificity in most species of Pyraloidea and Tortricidae. Based on published information plus observations made on microlepidoptera collected during the course of this study, 31 families of basal microlepidoptera were reviewed with an aim toward evaluating the likelihood of their including species that are dependent on tallgrass prairie. Of these families, 12 were evaluated as possible, and two as likely or certain, to include prairie-dependent species. In a comparison of moth diversity in light-trap samples from corn, miscanthus, switchgrass, and native prairie, alpha diversity was highest in prairie and was higher in switchgrass than in the other two biofuel crops. Moth species complements generally were similar among the biofuel crops, and prairie shared higher species complementarity with switchgrass than with corn or miscanthus. These findings suggest that large-scale conversion of land to biofuel crops may, to a substantial degree, detrimentally affect arthropod biodiversity, with a resulting loss of valuable arthropod-derived ecosystem services both within the cropping systems and in the surrounding landscape. During the course of this study, rearing efforts yielded two species of moths of the family Gelechiidae, both of which are monophagous leaf feeders on leadplant, Amorpha canescens (Fabaceae). Because these moths are restricted to tallgrass prairie, they are likely to be of interest to conservation biologists. In the interest of naming the moths to facilitate communication regarding them, and to augment our taxonomic knowledge of their respective genera, the moths are described, and diagnoses are provided to differentiate them from similar, related species.

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

Ecological infrastructures in sustainable olive growing: studies about Prays oleae (Bernard) and its natural enemies

Doutoramento em Engenharia Agronómica - Instituto Superior de Agronomia - UL

Effects de l'application d'acide gibbérellique (GA3) sur la qualité de raisins et de vins produits en climat tropical au Nord-Est du Brésil.

The honeydew moth Cryptoblabes gnidiella is the main problem for the wineries in the Northeast of the Brazil, because it attacks the bunch and reduces the quality of the grapes and the wines. In order to stretch out the bunch to facilitate the penetration of the insecticides, it was used gibberellic acid. Six treatments with different concentrations and different dates of application, and the control were compared. The bunches are compact, characteristic of the "Syrah" grapes in the region. The grape berries were analysed at harvest and wines were made by microvinifications. The grape berries showed different qualitative characteristics, as berry weight, number of berries. °Brix, total acidity and heterogeneity of the maturation. The microvinifications were carried out with 50 kg of grapeberries into glass bottles of 20 L at 22°C, for the alcoholic and malolactic fermentations, then stabilized and bottled. The wines were tasted by a panel of ten people and compared on smell and taste plans. The tasting results showed that the control treatment was the best graded wine. The application of gibberellic acid allowed to control the honeydew moth attack, but it caused a heterogeneity on grape maturation, with a lower quality of the grapes and wines compared to the control.

Pollination, herbivory, and habitat fragmentation: Their effects on the reproductive fitness of Angadenia berteroi, a native perennial plant of the south Florida pine rocklands

Angadenia berteroi is a tropical perennial subshrub of the pine rocklands with large yellow flowers that set very few fruits. My dissertation seeks to elucidate the factors that affect the reproductive fitness of Angadenia berteroi a native species of the south Florida pine rocklands. I provide novel information on the pollination biology of this native species. I also assess the effects of herbivory on growth and the reproductive success of A. berteroi. Finally, I elucidate how habitat fragmentation and quality are correlated with reproductive fitness of this native perennial plant.^ Using a novel experimental approach, I determined the most effective pollinator group. I used nylon fishing line of widths corresponding to proboscis diameter of the major groups of visitors to examine pollen removal and deposition. In the field, I estimated visitation frequency and efficacy of each pollinator type. Using potted plants, I exposed flowers to single visit from different types of pollinators to measure fruit set. I performed artificial defoliation with scissors on plants growing in the greenhouse to assess the effects of defoliation before flowering as well as during flowering. Additionally, I used structural equation modelling (SEM) to elucidate how A. berteroi reproductive fitness was affected by habitat fragmentation and quality. ^ My experiments provide evidence that Angadenia berteroi is specialized for bee pollination; though butterflies, skippers and others also visit its flowers, A. berteroi is exclusively pollinated by two native bees of the South Florida pine rocklands . This research also demonstrated that herbivory by the oleander moth may have direct and indirect effects on Angadenia berteroi growth and reproductive success. The SEM results suggested that habitat quality (litter depth and subcanopy cover) may favor reproduction in native species of the South Florida pine rocklands that are properly maintained by periodic fires and exotic control. Insights from this threatened and charismatic species may provide impetus to properly manage remaining pine rocklands in South Florida for this and other endemic understory species.^

Review of the Psychodinae from Mallorca, Spain, with description of Pericoma unipennata, sp. n. (Diptera, Psychodidae)

We review the Psychodinae of Mallorca, recognising fifteen species based on recent collections and available literature. Previously unpublished data is presented for eleven species, of which Neoarisemus ibericus Wagner, 1978, Mormia tenebricosa (Vaillant, 1954), Clogmia albipunctata (Williston, 1893), Lepiseodina rothschildi (Eaton, 1913), Paramormia ustulata (Walker, 1856), Philosepedon pyrenaicus Vaillant, 1974 and Psychoda (Psycha) grisescens Tonnoir, 1922 are first records for Mallorca. An old record of Pericoma trifasciata (Meigen, 1804) is considered doubtful. Pericoma unipennata sp. n is described and illustrated based on a male collected at Deía. Distributional data are reviewed for all newly recorded species. Based on the Psychodinae fauna, the zoogeographical affinities of Mallorca are briefly discussed.