Molecular and morphological characterization of heterorhabditid entomopathogenic nematodes from the tropical rainforest in Brazil

Despite massive losses of primary forest, the Amazonian rainforest remains an extremely rich source of biodiversity. In recent years, entomopathogenic nematodes (EPNs) have been isolated from soil in various parts of the world and used successfully as biological control agents against numerous insect pests. Therefore, a sampling in the rainforest of Monte Negro, Rondônia, Brazil was conducted with the aim of discovering new strains and/or species of EPNs for future development as biological control agents. From 156 soil samples taken at nine collecting sites, 19 isolates were obtained, all of them belonging to the genus Heterorhabditis. Four strains were subjected to detailed morphological and molecular evaluation. Based on morphometrics and internal transcribed spacer (ITS) sequence data, the strains LPP1, LPP2 and LPP4 were identified as Heterorhabditis indica, whereas LPP7 was considered Heterorhabditis baujardi. Comparative analysis of the ITS1 sequence of H. indica and H. baujardi isolates showed a polymorphic site for the restriction enzyme Tth 111 that could be used to distinguish the two species. Consequently, strains LPP1, LPP2, LPP3, LPP4, and LPP9 were identified as H. indica, whereas LPP5, LPP7, LPP8 and LPP10 were identified as H. baujardi.

Resultats de dos anys de Lluita Obligatòria pel control de la mosca de la fruita (Ceratitis capitata) en presseguers de les comarques de l’Alt i Baix Empordà

Estudi sobre el treball realitzat durant els anys 2005 i 2006 en més de 300 ha de presseguers de l'Alt i el Baix Empordà que contempla aspectes biològics de C. capitata, la metodologia de seguiment i control utilitzada durant els dos anys, avaluacions realitzades a camp així com l’anàlisi de les dades obtingudes

Determinació de la dosi potencial de pseudotecis de l'ascomicet Pleospora allii per millorar els programes de control de l'estemfiliosi de la perera

L’objectiu principal d’aquest treball és relacionar el nivell d’estemfiliosi en plantacions comercials de perera, a caiguda de fulla, amb la DPP o dosi potencial de pseudotecis de Pleospora allii (fase sexual del patogen Stemphylium vesicarium). També es pretén determinar aspectes relacionats amb la biologia del fong P. allii, com la dinàmica de producció de pseudotecis en el temps i l’efecte de la temperatura en aquesta formació. El coneixement del DPP en plantacions de pera i la predicció del desenvolupament inicial de la malaltia poden permetre perfeccionar els programes de control de l’estemfiliosi, reduint-ne el cost i l’impacte ambiental

Análise da riqueza da fauna de Braconidae (Hymenoptera, Ichneumonoidea) em remanescentes naturais da Área de Proteção Ambiental (APA) de Descalvado, SP

Analysis of the Braconidae (Hymenoptera: Ichneumonoidea) fauna richness in natural remnants of the Área de Proteção Ambiental (APA) of Descalvado, SP. A survey of the Braconidae fauna on fragmentary natural environments at northeast São Paulo State was conducted using Malaise traps installed in five sites of Área de Proteção Ambiental of Descalvado. A total of 2,262 specimens, representing 22 subfamilies and 94 genera, was sampled throughout a period of sixteen months (from May, 1999 to August, 2000). Biological data were obtained from current literature and behavior patterns of host utilization for the studied fauna, mainly Braconidae which have been successful in control of agricultural insect pests. Statistical analyses indicated that the asymptote of the genera richness has been approached using this sampling method. This is the most complete survey yet available from natural fragmentary areas of the Descalvado Braconidae fauna. Patterns of distribution and richness of the Braconidae genera were established. Cluster Analysis was adopted taking as attribute the number of Braconidae genera.

Nature, evolution and characterisation of rhizospheric chemical exudates affecting root herbivores

Similar to aboveground herbivores, root-feeding insects must locate and identify suitable resources. In the darkness of soil, they mainly rely on root chemical exudations and, therefore, have evolved specific behaviours. Because of their impact on crop yield, most of our knowledge in belowground chemical ecology is biased towards soil-dwelling insect pests. Yet the increasing literature on volatile-mediated interactions in the ground underpins the great importance of chemical signalling in this ecosystem and its potential in pest control. Here, we explore the ecology and physiology of these chemically based interactions. An evolutionary approach reveals interesting patterns in the response of insects to particular classes of volatile or water-soluble organic compounds commonly emitted by roots. Food web analyses reasonably support that volatiles are used as long-range cues whereas water-soluble molecules serve in host acceptance/rejection by the insect; however, data are still scarce. As a case study, the chemical ecology of Diabrotica virgifera virgifera is discussed and applications of belowground signalling in pest management are examined. Soil chemical ecology is an expanding field of research and will certainly be a hub of our understanding of soil communities and subsequently of the management of belowground ecosystem services.

Oral insecticidal activity of plant-associated pseudomonads.

Biocontrol pseudomonads are most known to protect plants from fungal diseases and to increase plant yield, while intriguing aspects on insecticidal activity have been discovered only recently. Here, we demonstrate that Fit toxin producing pseudomonads, in contrast to a naturally Fit-deficient strain, exhibit potent oral activity against larvae of Spodoptera littoralis, Heliothis virescens and Plutella xylostella, all major insect pests of agricultural crops. Spraying plant leaves with suspensions containing only 1000 Pseudomonas cells per ml was sufficient to kill 70-80% of Spodoptera and Heliothis larvae. Monitoring survival kinetics and bacterial titres in parallel, we demonstrate that Pseudomonas fluorescens CHA0 and Pseudomonas chlororaphis PCL1391, two bacteria harbouring the Fit gene cluster colonize and kill insects via oral infection. Using Fit mutants of CHA0 and PCL1391, we show that production of the Fit toxin contributes substantially to oral insecticidal activity. Furthermore, the global regulator GacA is required for full insecticidal activity. Our findings demonstrate the lethal oral activity of two root-colonizing pseudomonads so far known as potent antagonists of fungal plant pathogens. This adds insecticidal activity to the existing biocontrol repertoire of these bacteria and opens new perspectives for applications in crop pest control and in research on their ecological behaviour.

Uncovering Cryptic Parasitoid Diversity in Horismenus (Chalcidoidea, Eulophidae).

Horismenus parasitoids are an abundant and understudied group of eulophid wasps found mainly in the New World. Recent surveys based on morphological analyses in Costa Rica have quadrupled the number of named taxa, with more than 400 species described so far. This recent revision suggests that there is still a vast number of unknown species to be identified. As Horismenus wasps have been widely described as parasitoids of insect pests associated with crop plants, it is of high importance to properly establish the extant diversity of the genus, in order to provide biological control practitioners with an exhaustive catalog of putative control agents. In this study, we first collected Horismenus wasps from wild Phaseolus bean seeds in Central Mexico and Arizona to assess the genetic relatedness of three morphologically distinct species with overlapping host and geographical ranges. Sequence data from two nuclear and two mitochondrial gene regions uncovered three cryptic species within each of the three focal species (i.e., H. missouriensis, H. depressus and H. butcheri). The monophyly of each cryptic group is statistically supported (except in two of them represented by one single tip in which monophyly cannot be tested). The phylogenetic reconstruction is discussed with respect to differences between gene regions as well as likely reasons for the differences in variability between species.

Avaluació del sistema de confusió sexual pel control de carpocapsa (Cydia pomonella L.) en plantacions de pomeres de Fructícola Empordà S.L. durant el període 2003-2011

La carpocapsa o corc de la poma (Cydia pomonella L.) és la plaga clau de lesplantacions comercials de pomera de la zona de Girona. Per controlar-la amb eficàcia,anualment són necessaris diferents tractaments insecticides que a més del cost econòmicque representen, afavoreixen fenòmens de resistències, aparició de plagues induïdes iproblemes de toxicitat, residus i contaminació ambiental.La combinació de diferents metodologies per combatre aquesta plaga, algunes menysagressives amb la fauna auxiliar i el medi ambient, s’ha convertit en una pràcticaimprescindible per aconseguir-ne un control efectiu i sostenible. La confusió sexual ésuna tècnica de lluita biotecnològica que per les seves característiques d’especificitat,toxicologia i respecte al medi ambient, representa una alternativa a la problemàticagenerada pels productes químics tradicionals.Fructícola Empordà S.L., empresa agroalimentària situada al municipi de Sant PerePescador (Alt Empordà, Girona) dedicada a la producció i comercialització de fruitafresca, com a entitat participant del projecte Àrea Pilot de Reducció d’Insecticides enplantacions comercials de pomera de Girona (APRI) emmarcat en el programad’actuacions específiques de les Associacions de Defensa dels Vegetals (ADV’s) de lazona, introdueix per primera vegada la tècnica de la confusió sexual a l’estratègia decontrol de carpocapsa en determinades parcel•les comercials on la lluita químicaexclusiva no és suficient (2003). Durant el període 2003-2011, l’ús de la confusió sexual per al control de carpocapsa esgeneralitza progressivament a les finques dels agricultors de Fructícola Empordà S.L.amb l’objectiu principal de reduir el percentatge de dany per atac de carpocapsa en elsfruits a collita i, conseqüentment, disminuir la pressió del corc de la poma a la zona. Atal efecte es va plantejar un estudi amb l’objectiu d’avaluar l’eficàcia de l’estratègia decontrol de la carpocapsa basada en la combinació del mètode de la confusió sexual ambaplicacions fitosanitàries de reforç durant el període d’expansió de la tècnica (2003-2011) en les plantacions comercials de pomera de Fructícola Empordà S.L. així com,fer una valoració d’aquest respecte al sistema de control paral•lelament utilitzat a lazona, la lluita química assessorada

Feromônios associados aos coleópteros-praga de produtos armazenados

One strategy to overcome risks of insecticide-based control in agriculture is to use semiochemicals. In the case of pheromones, these specific compounds can be applied in traps to detect and monitor the occurrence, abundance and distribution of insect pests. Reliable detection helps to time insecticide sprays, to decide the quantity of insecticide that will be used and the place where it will be applied. This manuscript aims to give an overview of the pheromones associated to coleopteran pests in stored products, and their utilization in integrated pest management.

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.

Gypsy moths (Lymantria dispar) in the Niagara Region : population density variation, introduction of an entomopathogenic fungus (Entomophaga maimaiga) and occurrence of nuclear polyhedrosis virus

The gypsy moth, Lymantria dispar, a major defoliator of broad leaf trees, was accidentally introduced into North America in 1869. Much interest has been generated regarding the potential of using natural pathogens for biological control of this insect. One of these pathogens, a highly specific fungus, Entomophaga maimaiga, was accredited with causing major epizootics in populations of gypsy moth across the north-eastern United States in 1989 and 1990 and is thought to be spreading northwards into Canada. This study examined gypsy moth population densities in the Niagara Region. The fungus, .E.. maimaiga, was artificially introduced into one site and the resulting mortality in host populations was noted over two years. The relationship between fungal mortality, host population density and occurrence of another pathogen, the nuclear polyhedrosis virus (NPV), was assessed. Gypsy moth population density was assessed by counting egg masses in 0.01 hectare (ha) study plots in six areas, namely Louth, Queenston, Niagara-on-the-Lake, Shorthills Provincial Park, Chippawa Creek and Willoughby Marsh. High variability in density was seen among sites. Willoughby Marsh and Chippawa Creek, the sites with the greatest variability, were selected for more intensive study. The pathogenicity of E. maimaiga was established in laboratory trials. Fungal-infected gypsy moth larvae were then released into experimental plots of varying host density in Willoughby Marsh in 1992. These larvae served as the inoculum to infect field larvae. Other larvae were injected with culture medium only and released into control plots also of varying host density. Later, field larvae were collected and assessed for the presence of .E.. maimaiga and NPV. A greater proportion of larvae were infected from experimental plots than from control plots indicating that the experimental augmentation had been successful. There was no relationship between host density and the proportion of infected larvae in either experimental or control plots. In 1992, 86% of larvae were positive for NPV. Presence and intensity of NPV infection was independent of fungal presence, plot type or interaction of these two factors. Sampling was carried out in the summer of 1993, the year after the introduction, to evaluate the persistence of the pathogen in the environment. Almost 50% of all larvae were infected with the fungus. There was no difference between control and experimental plots. Data collected from Willoughby Marsh indicated that there was no correlation between the proportion of larvae infected with the fungus and host population density in either experimental or control plots. About 10% of larvae collected from a nearby site, Chippawa Creek, were also positive for .E.. maimaiga suggesting that low levels of .E.. maimaiga probably occurred naturally in the area. In 1993, 9.6% of larvae were positive for NPV. Again, presence or absence of NPV infection was independent of fungal presence plot type or interaction of these two factors. In conclusion, gypsy moth population densities were highly variable between and within sites in the Niagara Region. The introduction of the pathogenic fungus, .E.. maimaiga, into Willoughby Marsh in 1992 was successful and the fungus was again evident in 1993. There was no evidence for existence of a relationship between fungal mortality and gypsy moth density or occurrence of NPV. The results from this study are discussed with respect to the use of .E.. maimaiga in gypsy moth management programs.

The influence of organophosphorus pesticide-methylparathion on protein, lipid metabolism and detoxifying enzymes in rohu (Labeo rohita)

Methylparathion (MP) is an organophosphorus insecticide used world wide in agriculture due to its high activity against a broad spectrum of insect pests. The aim of the study is to understand the effect of methylparathion on the lipid peroxidation, detoxifying and antioxidant enzymes namely catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione Stransferase (GST), total reduced glutathione (GSH), lipid peroxidation (LPO), acetylcholinesterase (AChE) and disease diagnostic marker enzymes in liver, sarcoplasmic (SP) and myofirbirllar (MF) proteins in muscles, lipids and histopathlogical changes in various organs of Labeo rohita of size 75 i 6g at lethal and sublethal level of exposure. The probit analysis showed that the lethal concentration (LC 50%) for 24, 48, 72 and 96h were 15.5mg/L, 12.3mg/L, 11.4mg/L and 10.2mg/L respectively which is much higher compared to the LC50 for juvenile fish. The LPO level and GST activity increased five folds and two folds respectively on exposure to methylparathion at 10.2 mg/L and the level of the enzymes increased, on sub lethal exposure beyond 0.25mg/L. AChE activity was inhibited by 74% at a concentration of 1.8mg/L and 90% at 5.4mg/L. The disease diagnostic marker enzymes AST, ALT, ALP and LDH increased by about 2, 3 ,3 and 2 folds respectively at pesticide concentration of 10.2mg/L when compared to control. On sub lethal exposure, however the enzymes did not show any significant changes up to 0.5mg/L. At a concentration of 10.2 mg/L, there was a three fold increase in myofibrillar proteins while the increase in sarcoplasmic protein was above 1.5 fold. On sub lethal exposure, significant alteration was noticed up to 30 days up to 1mg/L of methylparathion concentration. Further exposure up to 45 days increased sarcoplasmic proteins (upto 0.5mg/L). ln the case of myofibrillar proteins, noticeable changes were observed at 1mg/L concentration right from 15th day. The cholesterol content in brain tissues increased by about 27% at methylparathion concentration of 5.4 mglL. However at 0.25mg/L sub lethal concentration, no significant alteration was observed in enzyme activity, muscle proteins, lipids and histopathology of the tissues. The results suggest that methylparathion has the potential to induce oxidative stress in fish, and that liver, muscle and brains are more sensitive organs of Labeo rohita, with poor antioxidant potentials at higher concentrations of the pesticide. The various parameters studied in this investigation can also be used as biomarkers of methylparathion exposure.

Resultats de dos anys de Lluita Obligatòria pel control de la mosca de la fruita (Ceratitis capitata) en presseguers de les comarques de l’Alt i Baix Empordà

Estudi sobre el treball realitzat durant els anys 2005 i 2006 en més de 300 ha de presseguers de l'Alt i el Baix Empordà que contempla aspectes biològics de C. capitata, la metodologia de seguiment i control utilitzada durant els dos anys, avaluacions realitzades a camp així com l’anàlisi de les dades obtingudes

Determinació de la dosi potencial de pseudotecis de l'ascomicet Pleospora allii per millorar els programes de control de l'estemfiliosi de la perera

L’objectiu principal d’aquest treball és relacionar el nivell d’estemfiliosi en plantacions comercials de perera, a caiguda de fulla, amb la DPP o dosi potencial de pseudotecis de Pleospora allii (fase sexual del patogen Stemphylium vesicarium). També es pretén determinar aspectes relacionats amb la biologia del fong P. allii, com la dinàmica de producció de pseudotecis en el temps i l’efecte de la temperatura en aquesta formació. El coneixement del DPP en plantacions de pera i la predicció del desenvolupament inicial de la malaltia poden permetre perfeccionar els programes de control de l’estemfiliosi, reduint-ne el cost i l’impacte ambiental

Use of entomopathogenic bacterium Pseudomonas putida (Enterobacteriaceae) and it secretions against the Greater Wax Moth Galleria mellonella pupae

The bacterium from Pseudomonas putida from Steinernema abbasi and its metabolic secretions caused the mortality of the Galleria mellonella pupae. Experiments were conducted in sand and filter paper on time exposure, temperature, moisture, dose and time of penetration of bacterium in pupae and tested stored or dried toxic metabolites using G. mellonella pupae as a test target organism. Death of pupae was probably due to the toxic metabolites. Pseudomonas putida cells were recovered from the haemocoele when bacterial cells were applied to the G. mellonella pupae indicating that bacterial cells can enter the haemocoele in the absence of nematode vector. Penetration of bacterium was found rapidly after application on G. mellonella pupae. Pseudomonas putida or its toxic secretions can be used as a microbial control for insect control. The experimental results indicate that there is possibility of using P. putida and its toxic secretions as a biopesticide and can contribute in the development of new microbial and biological control against insect pests.