Photodynamic and Antibiotic Therapy Impair the Pathogenesis of Enterococcus faecium in a Whole Animal Insect Model

Enterococcus faecium has emerged as one of the most important pathogens in healthcare-associated infections worldwide due to its intrinsic and acquired resistance to many antibiotics, including vancomycin. Antimicrobial photodynamic therapy (aPDT) is an alternative therapeutic platform that is currently under investigation for the control and treatment of infections. PDT is based on the use of photoactive dye molecules, widely known as photosensitizer (PS). PS, upon irradiation with visible light, produces reactive oxygen species that can destroy lipids and proteins causing cell death. We employed Galleria mellonella (the greater wax moth) caterpillar fatally infected with E. faecium to develop an invertebrate host model system that can be used to study the antimicrobial PDT (alone or combined with antibiotics). In the establishment of infection by E. faecium in G. mellonella, we found that the G. mellonella death rate was dependent on the number of bacterial cells injected into the insect hemocoel and all E. faecium strains tested were capable of infecting and killing G. mellonella. Antibiotic treatment with ampicillin, gentamicin or the combination of ampicillin and gentamicin prolonged caterpillar survival infected by E. faecium (P = 0.0003, P = 0.0001 and P = 0.0001, respectively). In the study of antimicrobial PDT, we verified that methylene blue (MB) injected into the insect followed by whole body illumination prolonged the caterpillar survival (P = 0.0192). Interestingly, combination therapy of larvae infected with vancomycin-resistant E. faecium, with antimicrobial PDT followed by vancomycin, significantly prolonged the survival of the caterpillars when compared to either antimicrobial PDT (P = 0.0095) or vancomycin treatment alone (P = 0.0025), suggesting that the aPDT made the vancomycin resistant E. faecium strain more susceptible to vancomycin action. In summary, G. mellonella provides an invertebrate model host to study the antimicrobial PDT and to explore combinatorial aPDT-based treatments.

Modelling fungus dispersal scenarios using cellular automata

This study focused on representing spatio-temporal patterns of fungal dispersal using cellular automata. Square lattices were used, with each site representing a host for a hypothetical fungus population. Four possible host states were allowed: resistant, permissive, latent or infectious. In this model, the probability of infection for each of the healthy states (permissive or resistant) in a time step was determined as a function of the host's susceptibility, seasonality, and the number of infectious sites and the distance between them. It was also assumed that infected sites become infectious after a pre-specified latency period, and that recovery is not possible. Several scenarios were simulated to understand the contribution of the model's parameters and the spatial structure on the dynamic behaviour of the modelling system. The model showed good capability for representing the spatio-temporal pattern of fungus dispersal over planar surfaces. With a specific problem in mind, the model can be easily modified and used to describe field behaviour, which can contribute to the conservation and development of management strategies for both natural and agricultural systems. © 2012 Elsevier B.V.

Insect/Bt-cotton interactions: Are immunological variables and hemocyte ultrastructure in Podisus nigrispinus (Hemiptera: Pentatomidae) affected?

Recent studies have shown that ingestion by the army worm Spodoptera frugiperda of Cry1Ac toxin from Bt cotton promotes histochemical and ultrastructural changes in the digestive cells of the predatory pentatomid bug Podisus nigrispinus. Therefore, mindful of the changes in the midgut of the predator, which represents the first line of defence in this insect, our aim was to test the hypothesis that the consumption of Bt cotton-fed S. frugiperda by P. nigrispinus might lead to alterations in components of the immune system of P. nigrispinus. The Cry1Ac toxin level in the leaves of Bt cotton, nitric oxide, phenoloxidase activity, and total proteins were quantified by ELISA. Total and differential hemocyte counts were evaluated, and hemocyte ultrastructure analysis was undertaken. We found that ingestion of the prey fed daily with approximately 23 ± 0.70 ng g-1 Cry1Ac by wet weight of leaves, and expressed by the Bt cotton, induces small ultrastructural changes in the predator's granulocytes and plasmatocytes. However, these changes did not affect the total number and differential and humoral variables analyzed for the bug's hemocytes. © 2013 Copyright Taylor and Francis Group, LLC.

Responses of aquatic insect functional diversity to landscape changes in Atlantic forest

The Atlantic Forest domain, one of the 25 world's hotspots for biodiversity, has experienced dramatic changes in its landscape. While the loss of species diversity is well documented, functional diversity has not received the same amount of attention. In this study, we evaluated functional diversity of insects in streams utilizing three indices: functional diversity (FD), functional dispersion (FDis), and functional divergence (FDiv), seeking to understand the roles of three predictor sets in explaining functional patterns: (1) bioclimatic and landscape variables; (2) spatial variables; and (3) local environmental variables. We determined the amount of variation in different measures of functional diversity that was explained by each predictor set and their interplays using variation partitioning. Our study showed that variation in functional diversity is better explained by a set of variables linked to different scales dependent on spatial structures, indicating the importance of landscape and mainly environmental variables in the functional organization of aquatic insect communities, and that the relative importance of predictor sets depends on the indices considered. Variation in FD was better explained by the interplay among the three predictor sets and by local environmental variables, whereas variation in FDis was better explained by spatial variables and by the interplay between environmental and spatial variables. Variation in FDiv was not significantly explained by any predictors. Our study adds more evidence on the harmful effects caused by landscape changes on biodiversity in the Atlantic Forest, suggesting that these effects also influence the functional organization of stream insect communities. © 2013 The Author(s) Journal compilation © 2013 by The Association for Tropical Biology and Conservation.

Specialized Seed Dispersal in Epiphytic Cacti and Convergence with Mistletoes

Mistletoes represent the best example of specialization in seed dispersal, with a reduced assemblage of dispersal agents. Specific dispersal requirements mediated by the specificity of seed deposition site have apparently led to the evolution of such close relationships between mistletoes and certain frugivores. Here, we provide evidences for another case of specialization involving epiphytic cacti in the genus Rhipsalis, and small Neotropical passerines Euphonia spp., which also act as the main seed dispersers of mistletoes in the family Viscaceae. With field observations, literature search, and observations on captive birds, we demonstrated that Rhipsalis have specific establishment requirements, and euphonias are the most effective dispersers of Rhipsalis seeds in both quantitative and qualitative aspects, potentially depositing seeds onto branches of host plants. We interpret the similar dispersal systems of Rhipsalis and Viscaceae mistletoes, which involve the same dispersal agents, similar fruit morphologies, and fruit chemistry as convergent adaptive strategies that enable seeds of both groups to reach adequate microsites for establishment in host branches. © 2013 by The Association for Tropical Biology and Conservation.

A survey of DNA methylation across social insect species, life stages, and castes reveals abundant and caste-associated methylation in a primitively social wasp

DNA methylation plays an important role in the epigenetic control of developmental and behavioral plasticity, with connections to the generation of striking phenotypic differences between castes (larger, reproductive queens and smaller, non-reproductive workers) in honeybees and ants. Here, we provide the first comparative investigation of caste- and life stage-associated DNA methylation in several species of bees and vespid wasps displaying different levels of social organization. Our results reveal moderate levels of DNA methylation in most bees and wasps, with no clear relationship to the level of sociality. Strikingly, primitively social Polistes dominula paper wasps show unusually high overall DNA methylation and caste-related differences in site-specific methylation. These results suggest DNA methylation may play a role in the regulation of behavioral and physiological differences in primitively social species with more flexible caste differences. © 2013 Springer-Verlag Berlin Heidelberg.

Seed dispersal by the lek-forming white-bearded manakin (Manacus manacus, Pipridae) in the Brazilian Atlantic forest

The movement patterns of males, females and juveniles of lekking species often differ due to differences in the commitment to lek activities, which may lead to differences in the spatial distribution and dispersal distances of seeds they eat. By sampling seeds in three lek and non-lek areas of the white-bearded manakin (Manacus manacus), we tested whether this lekking species increased the abundance and species richness of seeds in lek areas and, at a finer scale, in 21 displaying courts within lek areas. Combining data on seed defecation or regurgitation rates by free-ranging individuals, the number of seeds in droppings or regurgitations of mist-netted birds, and the distances travelled by birds equipped with radio-transmitters, we estimated the potential spatial distribution of seeds generated by six resident males and six females or juveniles during the morning peak of lek activity and when lek activity decreased in the afternoon. There was no difference in the species richness (46 and 44 morphospecies, respectively) and abundance of seeds (15.4 ± 7.3 seeds and 14.0 ± 1.1 seeds, respectively) between lek and non-lek areas. Within leks both parameters increased in courts (45 spp., 17.6 ± 14 seeds) compared with non-court sites (22 spp., 1.9 ± 1.8 seeds), likely as a consequence of the longer time spent by resident males in perches in or near display courts. Distances moved by juveniles and females per 60-min period (183 ± 272 m) were greater than resident males (42.6 ± 22.0 m) in the mornings, while the opposite happened in the afternoons (55.2 ± 40.7 m and 157 ± 105 m, respectively). We conclude that the spatial aggregation of seeds in lek areas of M. manacus occurs at the court level, and the spatial distribution of deposited seeds varies with manakin lekking status and the daily period of foraging. © Cambridge University Press 2013.

Action of the insect growth regulator fluazuron, the active ingredient of the acaricide Acatak®, in Rhipicephalus sanguineus nymphs (Latreille, 1806) (Acari: Ixodidae)

The present study evaluated the efficacy of fluazuron (active ingredient of the acaricide Acatak®) and its effects on Rhipicephalus sanguineus nymphs fed on rabbits exposed to different doses of this insect growth regulator. Three different doses of fluazuron (20 mg/kg, 40 mg/kg, and 80 mg/kg) were applied on the back of hosts (via pour on), while distilled water was applied to the Control group. On the first day of treatment with fluazuron (24 h), hosts were artificially infested with R. sanguineus nymphs. Once fully engorged, nymphs were removed and placed in identified Petri dishes in Biochemical Oxygen Demand (BOD) incubator for 7 days. After this period, engorged nymphs were processed for ultramorphological analysis. The results revealed alterations in the ultramorphology of many chitinous structures (smaller hypostome and chelicerae, less sclerotized scutum, fewer sensilla, fewer pores, absence of grooves, marginal and cervical strips and festoons in the body, even the anal plaque was damaged) that play essential roles for the survivor of ticks and that can compromise the total or partial development of nymphs and emergence of adults after periodic molting. Our findings confirm the efficacy of fluazuron, a more specific and less aggressive chemical to the environment and human health, and that does not induce resistance, in nymphs of the tick R. sanguineus in artificially infested rabbits treated with this arthropod growth regulator (AGR), indicating that it could be used in the control of this stage of the biological cycle of the tick R. sanguineus. © 2013 Wiley Periodicals, Inc.

Dispersão de machos de Diatraea saccharalis (Fabricius) (Leipdoptera: Crambidae) em cana-de-açúcar

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Isolamento e caracterização de leveduras de Polybia ignobilis (Hymenoptera: Vespidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Distância e padrões de dispersão contemporânea de pólen e sistema de reprodução em pequeno fragmento isolado de Copaifera Langsdorfii Desf. (Leguminosae – Caesalpinoideae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Insetos predadores de sementes e suas relações com a qualidade e a morfologia de frutos e sementes

Pós-graduação em Ciências Biológicas (Botânica) - IBB

Processos auto-organizados em agregados de larvas de moscas-varejeiras em dispersão pós-alimentar: experimentação, modelagem e validação

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

Dispersal of Cotesia flavipes in sugarcane field and implications for parasitoid releases

Diatraea saccharalis Fabr. (Lepidoptera: Crambidae) is a major sugarcane pest in Brazil. The management of infested areas is based on the release of Cotesia flavipes (Cameron) (Hymenoptera: Braconidae), a parasitoid of D. saccharalis larvae, but there are doubts about the effectiveness of C. flavipes, primarily regarding its rate of dispersal in sugarcane fields. Thus, the objective of this study was to evaluate the dispersal of C. flavipes in a sugarcane field and suggest a release method that provides higher parasitoid efficiency. The study was carried out in four areas of approximately 1 ha, in which stalk pieces containing 20 D. saccharalis larvae were distributed in a rectangular grid, and 12,000 C. flavipes adults were released at four points, that were 50 m apart and 25 m from the field border. Three days later, the D. saccharalis larvae were recovered and kept in the laboratory until they reached pupal stage or C. flavipes emergence. Parasitism varied from 13.2% to 42.8%. The random distribution of parasitized larvae was found in one assay. In three areas, the parasitized larvae showed an aggregated distribution, with a range of 15 to 25 m. Since the parasite's success is directly linked to parasitoid dispersion, it would be interesting to move the release points to 30 m from each other because the dispersal may happen in a 15 m radius.