Molecular analysis of a novel gene cluster encoding an insect toxin in plant-associated strains of Pseudomonas fluorescens

Pseudomonas fluorescens CHA0 and the related strain Pf-5 are well-characterized representatives of rhizosphere bacteria that have the capacity to protect crop plants from fungal root diseases, mainly by releasing a variety of exoproducts that are toxic to plant pathogenic fungi. Here, we report that the two plant-beneficial pseudomonads also exhibit potent insecticidal activity. Anti-insect activity is linked to a novel genomic locus encoding a large protein toxin termed Fit (for P. fluorescensinsecticidal toxin) that is related to the insect toxin Mcf (Makes caterpillars floppy) of the entomopathogen Photorhabdus luminescens, a mutualist of insect-invading nematodes. When injected into the haemocoel, even low doses of P. fluorescens CHA0 or Pf-5 killed larvae of the tobacco hornworm Manduca sexta and the greater wax moth Galleria mellonella. In contrast, mutants of CHA0 or Pf-5 with deletions in the Fit toxin gene were significantly less virulent to the larvae. When expressed from an inducible promoter in a non-toxic Escherichia coli host, the Fit toxin gene was sufficient to render the bacterium toxic to both insect hosts. Our findings establish the Fit gene products of P. fluorescens CHA0 and Pf-5 as potent insect toxins that define previously unappreciated anti-insect properties of these plant-colonizing bacteria

Management of slips and its effect on growth and production of 'Pérola' pineapple plants

The lack of good quality planting material has limited the expansion and contributed to yield reduction of the Brazilian pineapple culture. Alternatives of 'Pérola' pineapple slips management were studied aiming at obtaining superior planting material within a shorter time period and making good use of healthy slips of low vigor, that are commonly discarded by growers. Two experiments were carried out at the Experimental Field of Embrapa Cassava & Fruits, Cruz das Almas, Bahia, Brazil, and another one in a commercial plantation in the region of Itaberaba, BA, using blocks or entirely randomized designs with at least four replications. In the first one, the development of slips of different initial sizes (6 to 20 cm long), when grown on mother plants after fruit harvest, was compared with that of slips grown in a nursery after their removal from the mother plants. It became clear that larger slips grow more vigorously and that the removal from the mother plant delays their growth. However, results from the second study showed that those slips grown in nursery had vegetative and agronomic performance similar to that of conventional slips and close to that of plantlets produced from plant stem sections. Independently from the type of planting material used, the larger ones presented a more vigorous growth and produced higher yields. In the third study, it was observed the influence of mineral fertilization, pest control and growth regulator application after fruit harvest on slip development. The treatments applied did not significantly accelerate slip growth. Slips reached 50 cm length and at least 300 g fresh weight within 90 days after fruit harvest, indicating that vigorous plants have enough nutritional storage material for slips development.

Toxicity of allyl esters in insect cell lines and in Spodoptera littoralis larvae

We investigated the effects of five allyl esters, two aromatic (allyl cinnamate and allyl 2-furoate) and three aliphatic (allyl hexanoate, allyl heptanoate, and allyl octanoate) in established insect cell lines derived from different species and tissues. We studied embryonic cells of the fruit fly Drosophila melanogaster (S2) (Diptera) and the beet armyworm Spodoptera exigua (Se4) (Lepidoptera), fat body cells of the Colorado potato beetle Leptinotarsa decemlineata (CPB) (Coleoptera), ovarian cells of the silkmoth Bombyx mori (Bm5), and midgut cells of the spruce budworm Choristoneura fumiferana (CF203) (Lepidoptera). Cytotoxicity was determined with use of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and trypan blue. In addition, we tested the entomotoxic action of allyl cinnamate against the cotton leafworm Spodoptera littoralis .The median (50%) cytotoxic concentrations (EC50s) of the five allyl esters in the MTT bioassays ranged between 0.25 and 27 mM with significant differences among allyl esters (P = 0.0012), cell lines (P < 0.0001), and the allyl estercell line interaction (P < 0.0001). Allyl cinnamate was the most active product, and CF203 the most sensitive cell line. In the trypan blue bioassays, cytotoxicity was produced rapidly and followed the same trend observed in the MTT bioassay. In first instars of S. littoralis, allyl cinnamate killed all larvae at 0.25% in the diet after 1 day, while this happened in third instars after 5 days. The LC50 in first instars was 0.08%. In addition, larval weight gain was reduced (P < 0.05) after 1 day of feeding on diet with 0.05%. In conclusion, the data provide evidence of the significant but differential cytotoxicity among allyl esters in insect cells of different species and tissues. Midgut cells show high sensitivity, indicating the insect midgut as a primary target tissue. Allyl cinnamate caused rapid toxic effects in S. littoralis larvae at low concentrations, suggesting further potential for use in pest control.

Phenotipic comparison of clinical and plant beneficial strains of Pantoea agglomerans

Certain strains of Pantoea are used as biocontrol agents for the suppression of plant diseases. However, their commercial registration is hampered in some countries because of biosafety concerns. This study compares clinical and plant-beneficial strains of P. agglomerans and related species using a phenotypic analysis approach in which plant-beneficial effects, adverse effects in nematode models, and toxicity were evaluated. Plant-beneficial effects were determined as the inhibition of apple fruit infection by Penicillium expansum and apple flower infection by Erwinia amylovora. Clinical strains had no general inhibitory activity against infection by the fungal or bacterial plant pathogens, as only one clinical strain inhibited P. expansum and three inhibited E. amylovora. By contrast, all biocontrol strains showed activity against at least one of the phytopathogens, and three strains were active against both. The adverse effects in animals were evaluated in the plant-parasitic nematode Meloidogyne javanica and the bacterial-feeding nematode Caenorhabditis elegans. Both models indicated adverse effects of the two clinical strains but not of any of the plant-beneficial strains. Toxicity was evaluated by means of hemolytic activity in blood, and genotoxicity with the Ames test. None of the strains, whether clinical or plant-beneficial, showed any evidence of toxicity

Feromônios de insetos: tecnologia e desafios para uma agricultura competitiva no Brasil

Pheromones are chemical compounds used by species to communicate intra and inter specifically. As strategy of integrated pest management (IPM), the use of these compounds allows the monitoring of target insects that results in more reliable interventions and consequently avoiding unnecessary use of pesticides. In certain crops these compounds are used as a control measure, not only monitoring. The Brazilian agriculture has a portfolio of 49 major crops that are attacked by 447 species of insects. Of this total, 103 species have already been the subject of study in the research with pheromones. Currently, in the Brazilian market, 28 pheromone products are indicated for the control of 19 insect pests. However, these products are not used regularly in major crops of the country. This stems from the lack of implementation of IPM of these crops. While the research focused on the main species of agribusiness pests, marked of pheromones products is serving to more marginal crops like apples and peaches at the expense of major crops like soybean and corn.

Compostos orgânicos voláteis na defesa induzida das plantas contra insetos herbívoros

The identification and manipulation of chemical compounds involved in vital activities of arthropods have the potential for developing less aggressive pest control strategies. Herbivory induces the emission of volatile organic compounds involved in the recruitment of natural enemies, plant-plant interactions and repellency of other herbivores. In this report, we review the main chemical groups of volatile organic compounds and their ecological functions, provide an overview of the signal transduction pathways activated upon herbivory, and review the current state of knowledge for practical applications in pest management. We conclude by proposing perspectives for future research.

Noctuidae-induced plant volatiles: current situation and prospects

Noctuids are phytophagous lepidopterans with some species causing significant damage to agriculture. The host plants, in turn, have developed defense mechanisms to cope with them, for instance chemical defenses. In this study we review the literature on plant volatiles induced by noctuids, and discuss the methodologies used to induce the production of volatiles that are usually employed in plant defense mechanisms. Future prospects involving this line of research in pest control are also discussed.

Développement, application et validation d’une nouvelle stratégie de mesure des indicateurs biologiques de l’exposition aux pyréthrinoïdes et aux pyréthrines chez l’humain

Les pyréthrinoïdes et les pyréthrines sont des insecticides neurotoxiques auxquels on attribue également des effets néfastes sur les systèmes immunitaire, hormonal et reproducteur. Ils sont abondamment utilisés en agriculture, mais aussi en horticulture, en extermination et dans le traitement d’infestations parasitaires humaines et animales (gale, poux). Il y a donc un intérêt en santé environnementale à connaître l’ampleur de l’exposition humaine à ces pesticides. La mesure des métabolites urinaires des pyréthrinoïdes et des pyréthrines apparaît une approche de choix pour arriver à cette fin puisqu’elle permet, en théorie, d’obtenir un portrait global de l’exposition. Or,traditionnellement et par soucis de simplicité les concentrations volumiques ou ajustées pour la créatinine) de ces biomarqueurs dans des urines ponctuelles sont déterminées, mais l’effet de l’utilisation de ces unités sur la validité des estimations de dose quotidienne absorbée n’a jamais été vérifié. L’objectif général de cette thèse était donc de développer, appliquer et valider une nouvelle stratégie de mesure et d’analyse de biomarqueurs pour améliorer la précision et la fiabilité des évaluations de l’exposition individuelles et populationnelles aux pyréthrinoïdes et pyréthrines. Les objectifs spécifiques étaient : i) de caractériser l’exposition humaine à ces contaminants en région urbaine et rurale au Québec et ii) de comparer la validité de la nouvelle stratégie de mesure et d’analyse de biomarqueurs urinaires proposée avec les autres méthodes plus usuelles utilisées pour estimer l’exposition individuelle et populationnelle et les doses absorbées de pyréthrinoïdes. Des adultes et des enfants, recrutés dans la population de l’Île de Montréal et de la Montérégie ont recueilli leurs urines pendant une période d’au moins douze heures et complété un questionnaire documentant les sources potentielles d’exposition. Les quantités de métabolites de pyréthrinoïdes et pyréthrines (pmol) mesurées dans les urines ont été ajustées pour une période de douze heures exactement et pour le poids corporel. Les quantités excrétées en région urbaine ont été comparées à celles excrétées en région rurale et les données individuelles et populationnelles ont été comparées à celles qui auraient été obtenues si des concentrations volumiques ou ajustées pour la créatinine avaient été mesurées. Les résultats montrent que l’exposition à ces pesticides est ubiquiste puisque plus de 90% des participants excrétaient les principaux métabolites de pyréthrinoïdes et pyréthrines à un niveau supérieur au seuil de détection analytique. Les résultats suggèrent que l’alimentation pourrait être à l’origine de ce niveau de base puisque les autres sources d’exposition connues n’ont été que rarement rapportées. Au Québec, l’exposition en milieu rural apparaissait légèrement plus importante qu’en milieu urbain et certains facteurs d’exposition, comme l’utilisation de pesticides domestiques, ont été rapportés plus fréquemment en milieu rural. Enfin, il a été démontré que la mesure des concentrations volumiques ou ajustées pour la créatinine de biomarqueurs est une approche qui peut entraîner des biais importants (jusqu’à 500% d’erreur et plus) en particulier lors de l’évaluation de l’exposition individuelle. Il est évident que les autorités de santé publique et de santé environnementale employant des biomarqueurs urinaires afin d’évaluer l’exposition aux pyréthrinoïdes et aux pyréthrines (ainsi qu’à d’autres molécules ayant des demi-vies d’élimination similaire)devraient diriger leurs efforts vers la mesure des quantités excrétées pendant une période d’au moins douze heures pour obtenir un portrait le plus valide possible de l’exposition. Il serait aussi souhaitable de mieux documenter la toxicocinétique de ces molécules chez l’humain afin d’établir avec une plus grande confiance la relation entre les quantités excrétées de biomarqueurs et les doses absorbées de ces contaminants.

Processing, procurement and marketing of pepper with particular reference to co-operative sector

The present Study is designed to gather, record and analyse data on history of pepper, pepper production, procurement and marketing with particular reference to Kerala. The main emphasis is given to study the'role of cooperative sector with regard to procurement and export efforts and also the services rendered by cooperative sector agencies under MARKETFED and NAFED to pepper trade. The scope of the Study covers the botany, methods of cultivation, fertilizer application, pest control management and other related aspects of pepper. Taking into consideration Kerala's supremacy in pepper cultivation and production, detailed study of its production, procurement, internal and export marketing with reference to Kerala has been given importance. As Kerala accounts for 96 per cent1 of the pepper cultivation and 94 per cent of the pepper production, the present study is entirely confined to Kerala

Optimización ambiental y sanitaria en los casinos del Club Campestre El Rancho en la ciudad de Bogotá

El plan de saneamiento básico empleado en el Club El Racho de la ciudad de Bogotá, busca primordialmente mejorar las operaciones dentro de los espacios de preparación de alimentos del club, brindando en si una serie de lineamientos y parámetros básicos para lograr un óptimo desarrollo del mismo. Dentro de este plan se estudian y se exponen una serie de fases que requieren no sólo una implementación adecuada, sino también un proceso de seguimiento posterior a dicha implementación, con el único objetivo de controlar y mitigar reacciones y posibles alteraciones del programa. Llevando a cabo programas de limpieza y desinfección, de manejo de residuos sólidos y de control de plagas en los casinos del club, se busca también, lograr una concientización general tanto en empleados como socios y demás usuarios de los establecimientos y así darle al plan de saneamiento básico una ruta exitosa que garantice, así mismo, un desarrollo sostenible y una responsabilidad social.

Biosecurity in agriculture: an economic analysis of coexistence of professional and hobby production

One component of biosecurity is protection against invasive alien species, which are one of the most important threats worldwide to native biodiversity and economic profitability in various sectors, including agriculture. However, agricultural producers are not homogeneous. They may have different objectives and priorities, use different technologies, and occupy heterogeneous parcels of land. If the producers differ in terms of their attitude towards invasive pests and the damages they cause, there are probably external effects in the form of pest spread impacts and subsequent damages caused. We study such impacts in the case of two producer types: profit-seeking professional producers and utility-seeking hobby producers. We show that the hobby producer, having first set a breeding ground for the pest, under-invests in pest control. We also discuss potential policy instruments to correct this market failure and highlight the importance of considering different stakeholders and their heterogeneous incentives when designing policies to control invasive alien species.

Pathogenicity of the bacterium Pseudomonas putida from the entomopathogenic nematode Steinernema abbasi and its secretion against Galleria mellonella larvae

The Entomopathogenic bacterium Pseudomonas putida from Steinernema abbasi and its metabolic secretions were lethal to the Galleria mellonella larvae. Different laboratory experiments on time interval, substrate, moisture, temperature, dose, penetration of cells, stored and dried metabolites were conducted in sand and filter paper bioassays. It was concluded that death was probably due to the toxic metabolites. This bacterium and its metabolites were found very effective at 30 degree C. Penetration of bacterium was rapid after application on G. mellonella larvae. P. putida cells were recovered from the haemocoele when suspensions containing bacterial cells were applied to the G. mellonella indicating that bacterial symbionts do have a free-living existence and can enter the haemocoele in the absence of nematode vector. Stored metabolite and dried metabolites were found persistent for long time. This bacterium or its toxic secretions can be used for insect control that can be important component of integrated pest management against different insect pests. P. putida and its secretions are suggested as the most appropriate suspension to apply against insect pest control program in tropical ecological regions.

The value of sown grass margins for enhancing soil macrofaunal biodiversity in arable systems

The presence of a grass strip was found to be beneficial to soil macrofauna, increasing the species densities and abundances of earthworms, woodlice and staphylinid beetles. The biodiversity of the three main feeding groups - predators, soil ingesters and litter consumers - was also significantly higher in the grass strips than in the field edges without strips, indicating that establishment of grassy margins in arable fields may enhance ecosystem services such as soil fertility and pest control. The grass strip habitat contained a large number of species of soil macrofauna, being second only to hedgerow habitat, with 10% of the total species list for the farm found only within the margins. Of the rare species recorded on the farm, five of the nine were from the grass strips, four of which were found only there. This study shows that establishing grassy strips in the margins of arable fields increases the biodiversity of the soil macrofauna, both within fields (alpha diversity) and across the farm (beta diversity). (C) 2008 Elsevier B.V. All rights reserved.

The value of arable field margins for enhancing soil macrofaunal biodiversity in agroecosystems

The presence of a grass strip was found to be beneficial to soil macrofauna, increasing the species densities and abundances of earthworms, woodlice and staphylinid beetles. The biodiversity of the three main feeding groups – predators, soil ingesters and litter consumers – was also significantly higher in the grass strips than in the field edges without strips, indicating that establishment of grassy margins in arable fields may enhance ecosystem services such as soil fertility and pest control. The grass strip habitat contained a large number of species of soil macrofauna, being second only to hedgerow habitat, with 10% of the total species list for the farm found only within the margins. Of the rare species recorded on the farm, five of the nine were from the grass strips, four of which were found only there. This study shows that establishing grassy strips in the margins of arable fields increases the biodiversity of the soil macrofauna, both within fields (alpha diversity) and across the farm (beta diversity).