The role of bacteria in pine wilt disease: insights from microbiome analysis.

Pine Wilt Disease (PWD) has a significant impact on Eurasia pine forests. The microbiome of the nematode (the primary cause of the disease), its insect vector, and the host tree may be relevant for the disease mechanism. The aim of this study was to characterize these microbiomes, from three PWD-affected areas in Portugal, using Denaturing Gradient Gel Electrophoresis, 16S rRNA gene pyrosequencing, and a functional inference-based approach (PICRUSt). The bacterial community structure of the nematode was significantly different from the infected trees but closely related to the insect vector, supporting the hypothesis that the nematode microbiome might be in part inherited from the insect. Sampling location influenced mostly the tree microbiome (P < 0.05). Genes related both with plant growth promotion and phytopathogenicity were predicted for the tree microbiome. Xenobiotic degradation functions were predicted in the nematode and insect microbiomes. Phytotoxin biosynthesis was also predicted for the nematode microbiome, supporting the theory of a direct contribution of the microbiome to tree-wilting. This is the first study that simultaneously characterized the nematode, tree and insect-vector microbiomes from the same affected areas, and overall the results support the hypothesis that the PWD microbiome plays an important role in the disease's development

Review of the genus Bursaphelenchus Fuchs, 1937 (Nematoda, Aphelenchida) with some conclusions on the host-parasite and vector-parasite evolution.

INTRODUCTION AND GOALS: Genus Bursaphelenchus includes several pests of the world importance for the rural economy, the most dangerous are the Bursaphelenchus xylophilus (the pinewood nematode caused decline of the pine trees in south Asia and in one spot area in Europe, Portugal, Peninsula de Setubal) and the Bursaphelenchus cocophilus, causing the decline of coco-palm plantations in Carribean and Latin American regions. The peculiarity of the host-parasite association of the genus that the nematode life cycle includes three trophic components: plant (mostly a tree), insect vector and a fungus. Goals of the presentation is to list all species of the world fauna and all efficient diagnostic characters, then create the identification tool and analyze the similarity of species and possible ways and causes of the host-parasite evolution of the group. RESULTS: Complete list of species with synonymy and a catalogue of all efficient diagnostic characters with their states, selected from papers of the most experienced taxonomists of the genus, are given for the genus Bursaphelenchus. List of known records of Bursaphelenchus species with names of natural vectors and plants and their families is given (for world pests the most important groups of trees and insects are listed). The tabular, traditional and computer-aided keys are presented. Dendrograms of species relationships (UPGMA, standard distance: mean character difference) based on all efficient taxonomic characters and separately on the spicule characters only, are given. Discussion whether the species groups are natural or purely diagnostic ones is based on the relationships dendrograms and the vector and associated plant ranges of Bursaphelenchus species; the xylophilus species group (B. xylophilus, B. abruptus, B. baujardi, B. conicaudatus, B. eroshenkii, B. fraudulentus, B. kolymensis, B. luxuriosae; B. mucronatus), the hunti group (B. hunti, B. seani, B. kevini and B. fungivorus) are probably the natural ones. CONCLUSIONS: The parasitic nematode association includes three trophic components: plant, insect vector and fungus. The initial insect-plant complex Scolytidae-Pinaceae is changeable and only in rare occasions the change of the preferred vector to Cerambycidae (the xylophilus group), Hymenoptera (the hunti group) led to formation of the natural species-groups. From the analysis it is clear that although the vector range is changeable it is comparatively more important for the evolution of the genus Bursaphelenchus than associations with plants at the family level. Data on the fungi species (3rd component in natural Bursaphelenchus associations) are insufficient for the detailed comparative analysis

NO BUG: biobased mosquitoes repellent textiles

This research work is part of the FP7 No-Bug project (Novel release system and biobased utilities for insect repellent textiles). The main interest of the project is personal protective textiles against insects (mosquitoes) for application not only in tropical areas where vector borne diseases are a major threat to the public health but also in European countries where the presence of mosquitoes can be nuisance. Malaria and dengue fever are well known diseases that cause a lot of deaths in the world today

Biological vector control with the sterile insect technique for the chilungunya disease

Chikungunya is a vector-borne Disease, usually localized in Asia and East-Africa, with Aedes albopictus mosquito as the principal vector for the Chikungunya virus. In 2005 and 2006, Réunion Island faced two epidemics of Chikungunya: the 2006's epidemic was particularly dramatic. This was the _rst time that a developed country, like Réunion Island, was a_ected by this virus. In July 2007, a small outbreak occurred in Italy, indicating that the South of Europe is potentialy threatened. In recent works [1,2], we proposed and studied a mathematical model to explain the outbreak of 2005 and possible links with the explosive epidemic of 2006. These studies speci_cally focus on the comparison of di_erent mosquito control tools (adulticide, larvicide, and mechanical control) in order to know if it would have been possible to contain or to completely avert the 2006 epidemic. We showed that the combination of the three control tools (with a suitable period of release and a su_cient duration of the treatment) would have been useful to control the explosive epidemic of 2006 [2]. As far as we know, Aedes albopictus in Réunion Island is yet sensitive to Deltamethrin, the only authorized adulticide, but can become resistant, like in Martinique, a West Indies French Island. Moreover, Réunion Island is a hot spot of endemicity and, thus, the use of chemical control tools can be limited. It is also necessary to study and to check the feasibility of other vector control tools such as the Sterile Insect Technique (SIT). To this e_ect, a project called TIS (Technique d'Insecte Stérile), funded by the French Ministry of Health, the European Union and the Regional Council is ongoing in Réunion Island. The aim of this talk is to give a short introduction to the TIS project and to present some recent mathematical results related to the SIT-LSIR model considered for the Chikungunya disease. Moreover, because mechanical control (destruction of breeding sites) is a very cheap and sustainable alternative, we combine mechanical control and SIT control. We present several numerical simulations to assess the e_cacy of the SIT vector-control in comparison with the Chemical vector control, studied in [2]. We show that SIT (impulse) control could be useful to control the wild mosquito population and thus lower the risk of an epidemic. (Résumé d'auteur

Biological Control of the Chagas Disease Vector Triatoma infestans with the Entomopathogenic Fungus Beauveria bassiana Combined with an Aggregation Cue: Field, Laboratory and Mathematical Modeling Assessment

Background: Current Chagas disease vector control strategies, based on chemical insecticide spraying, are growingly threatened by the emergence of pyrethroid-resistant Triatoma infestans populations in the Gran Chaco region of South America. Methodology and findings: We have already shown that the entomopathogenic fungus Beauveria bassiana has the ability to breach the insect cuticle and is effective both against pyrethroid-susceptible and pyrethroid-resistant T. infestans, in laboratory as well as field assays. It is also known that T. infestans cuticle lipids play a major role as contact aggregation pheromones. We estimated the effectiveness of pheromonebased infection boxes containing B. bassiana spores to kill indoor bugs, and its effect on the vector population dynamics. Laboratory assays were performed to estimate the effect of fungal infection on female reproductive parameters. The effect of insect exuviae as an aggregation signal in the performance of the infection boxes was estimated both in the laboratory and in the field. We developed a stage-specific matrix model of T. infestans to describe the fungal infection effects on insect population dynamics, and to analyze the performance of the biopesticide device in vector biological control. Conclusions: The pheromone-containing infective box is a promising new tool against indoor populations of this Chagas disease vector, with the number of boxes per house being the main driver of the reduction of the total domestic bug population. This ecologically safe approach is the first proven alternative to chemical insecticides in the control of T. infestans. The advantageous reduction in vector population by delayedaction fungal biopesticides in a contained environment is here shown supported by mathematical modeling.Fil: Forlani, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Girotti, Juan Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Mijailovsky, Sergio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Cardozo, Rubén Marino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina. Provincia de Salta. Ministerio de Salud Pública. Coordinación de Gestión Epidemiológica; ArgentinaFil: Gentile, Alberto G.. Provincia de Salta. Ministerio de Salud Pública. Coordinación de Gestión Epidemiológica; ArgentinaFil: Hernández Suárez, Carlos. Universidad de Colima. Facultad de Ciencias; MéxicoFil: Rabinovich, Jorge Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Juarez, Marta Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentin

Bursaphelenchus pinophilus Brzeski & Baujard, 1997 (Nematoda: Parasitaphelenchinae) associated with nematangia on Pityogenes bidentatus (Herbst, 1783) (Coleoptera: Scolytinae), from the Czech Republic

The occurrence of Bursaphelenchus species in the Czech Republic is poorly known, the first report of the genus being made by Kubátová et al. (2000) who reported the association of B. eremus with the hyphomycetous microfungus, Esteya vermicola, and the bark beetle, Scolytus intricatus, collected from Quercus robur, in central Bohemia. To date, four other species have been reported from the country, namely B. fungivorus (Braasch et al., 2002), B. hofmanni (see Braasch, 2001), B. mucronatus (see Braasch, 2001) and B. vallesianus (Gaar et al., 2006). More recently, a survey for Bursaphelenchus species associated with bark- and wood-boring insects in the Czech Republic identified B. pinophilus Brzeski & Baujard, 1997 from the Moravia region. Although this represents a new country record, it was also associated with nematangia on the hind wings of a new insect vector. A total of 404 bark- and wood-boring insects were collected from declining or symptomatic trees and screened for the presence of Bursaphelenchus. Bark and longhorn beetles were captured manually after debarking parts of the trunk displaying symptoms of insect attacks. Longhorn beetle larvae were also collected together with logs cut from the trunk. Logs were kept at room temperature in the laboratory until insect emergence. Each adult insect was individually dissected in water and examined for nematodes. All nematodes resembling dauer juveniles of Bursaphelenchus were collected and identified by molecular characterisation using a region of ribosomal DNA (rDNA) containing the internal transcribed spacer regions ITS1 and ITS2. ITS-RFLP analyses using five restriction enzymes (AluI, HaeIII, HinfI, MspI, RsaI) were performed to generate the species-specific profile according to Burgermeister et al. (2009). Species identification was also confirmed by morphological data after culture of the dauers on Botrytis cinerea Pers. ex Ft., growing in 5% malt extract agar. During this survey, only species belonging to the Curculionidae, subfamily Scolytinae, revealed the presence of nematodes belonging to Bursaphelenchus. Dauers of this genus were found aggregated under the elytra in nematangia formed at the root of the hind wings (Fig. 1). The dauers were identified from 12 individuals of Pityogenes bidentatus (Herbst, 1783) (Coleoptera: Scolytinae) collected under the bark of Pinus sylvestris trunks. Each insect carried ca 10-100 dauers. The ITS-RFLP patterns of the dauers so obtained confirmed the identification of B. pinophilus associated with this insect species. Bursaphelenchus pinophilus has been found mainly in Europe and has been reported from various countries such as Poland (Brzeski & Baujard, 1997), Germany (Braasch, 2001), and Portugal (Penas et al., 2007). The recent detection of this species associated with dead P. koraiensis in Korea (Han et al., 2009) expands its geographical distribution and potential importance. It has been found associated only with Pinus species, but very little is known about the insect vector. The bark beetle, Hylurgus ligniperda, was initially suggested as the insect vector by Pe-nas et al. (2006), although the nematode associated with this insect was later reclassified as B. sexdentati by morphological and molecular analysis (Penas et al., 2007). According to the literature, P. bidentatus has been cited as a vector of Ektaphelenchus sp. (Kakuliya, 1966) in Georgia, and an unidentified nematode species in Spain (Roberston et al., 2008). Interestingly, B. pinophilus was found in the nematangia formed at the root of the hind wings of P. bidentatus. Although this phenomenon is not so common in other Bursaphelenchus species, B. rufipennis has been found recently in such a structure on the hind wings of the insect Dendroctonus rufipennis (Kanzaki et al., 2008). Although other nematode species (e.g., Ektaphelenchus spp.) are frequently found associated within the same nematangia (see Kanzaki et al., 2008), in this particular case, only dauers of B. pinophilus were identified. The association between B. pinophilus and P. bidentatus represents the first report of this biological association and the association with the Scolytinae strengthens the tight and specific links between this group of Bursaphelenchus species and members of the Scolytinae (Ryss et al., 2005)

Sterile insect technology for control of Anopheles mosquito: a mathematical feasibility study

Anopheles mosquito is a vector responsible for the transmission of diseases like Malaria which a_ect many people. Hence its control is a major prevention strategy. Sterile Insect Technology (SIT) is a nonpolluting method of insect control that relies on the release of sterile males. Mating of the released sterile males with wild females leads to non hatching eggs. Thus, if sterile males are released in su_cient numbers or over a su_cient period of time, it can leads to the local reduction or elimination of the wild population. We study the e_ectiveness of the application of SIT for control of Anopheles mosquito via mathematical modeling. Our main result is that there exists a threshold release rate ^_ depending only on the basic o_spring number R and the wild mosquito equilibrium for males such that a release rate higher than ^_ results in elimination of the mosquito population irrespective of its initial size. A release rate _ which is lower than ^_ eliminates the mosquito populations only if it is su_ciently small. If the population is at the wild equilibrium it is reduced by a percentage depending on _ and R only. (Résumé d'auteur

Postembryonic RNAi in Heterorhabditis bacteriophora: a nematode insect parasite and host for insect pathogenic symbionts

Background: Heterorhabditis bacteriophora is applied throughout the world for the biological control of insects and is an animal model to study interspecies interactions, e.g. mutualism, parasitism and vector-borne disease. H. bacteriophora nematodes are mutually associated with the insect pathogen, Photorhabdus luminescens. The developmentally arrested infective juvenile (IJ) stage nematode (vector) specifically transmits Photorhabdus luminescens bacteria (pathogen) in its gut mucosa to the haemocoel of insects (host). The nematode vector and pathogen alone are not known to cause insect disease. RNA interference is an excellent reverse genetic tool to study gene function in C. elegans, and it would be useful in H. bacteriophora to exploit the H. bacteriophora genome project, currently in progress. Results: Soaking L1 stage H. bacteriophora with seven dsRNAs of genes whose C. elegans orthologs had severe RNAi phenotypes resulted in highly penetrant and obvious developmental and reproductive abnormalities. The efficacy of postembryonic double strand RNA interference (RNAi) was evident by abnormal gonad morphology and sterility of adult H. bacteriophora and C. elegans presumable due to defects in germ cell proliferation and gonad development. The penetrance of RNAi phenotypes in H. bacteriophora was high for five genes (87-100%; Hba-cct-2, Hba-daf-21, Hba-icd-1; Hba-nol-5, and Hba-W01G7.3) and moderate for two genes (usually 30-50%; Hba-rack-1 and Hba-arf-1). RNAi of three additional C. elegans orthologs for which RNAi phenotypes were not previously detected in C. elegans, also did not result in any apparent phenotypes in H. bacteriophora. Specific and severe reduction in transcript levels in RNAi treated L1s was determined by quantitative real-time RT-PCR. These results suggest that postembryonic RNAi by soaking is potent and specific. Conclusion: Although RNAi is conserved in animals and plants, RNAi using long dsRNA is not. These results demonstrate that RNAi can be used effectively in H. bacteriophora and can be applied for analyses of nematode genes involved in symbiosis and parasitism. It is likely that RNAi will be an important tool for functional genomics utilizing the high quality draft H. bacteriophora genome sequence

Status Penelitian Serangga Vektor Penyakit Kerdil Pada Tanaman Lada

Research Status on Insect Vector of Stunted Disease on Black PepperStunted disease is one of the important diseases of black pepper. Two viruses, i.e. Piper Yellow Mottle Virus (PYMV) and Cucumo Mottle Virus (CMV) are associated with this disease. The disease is spread through seed as well as insect vectors. Two mealybugs, Planococcus minor and Ferrisia virgata; are known as insect vectors of PYMV in Indonesia and Aphis gossypii is an insect vector of CMV. The two mealybugs are polyphagous insects and efficient vectors.of stunted disease. Preliminary control of insect vectors has been conducted at the green house and field. Neem and tobacco extracts have showed effective control against Planococcus as also shown on monocrotophos and carbofuran treatments. Vector management is needed to reduce disease spread, through controlling insect vectors based on understanding their ecobiology. In addition, examining other potential insect vectors and screening existing hybrid lines to the disease and insect vectors need further examination

Vector control for the chikungunya disease: chemical control versus biological control: a mathematical point of view.

The aim of this talk is to present recent investigations on the Chikungunya Disease that hitted Réunion Island, a French territory in Indian Ocean, in 2005 and 2006. Chikungunya is a vectorborne Disease, usually localized in Asia and East-Africa. In 2005, it was the first time that a developed country was affected by this virus. In July 2007, a small outbreak raised in Italy, indicating that the South of Europe is potentialy threatened. In recent works, we have proposed and studied a mathematical model to explain the outbreak of 2005 and possible links with the explosive epidemic of 2006. We also have focused our study in the comparison on different mosquito control tools (adulticide, larvicide, and mechanical control), in order to know if it would have been possible to contain or to stop the epidemic of 2006. Recently, a new project has began to check the feasability of the Sterile Insect Technique (SIT) as a tool for vector-control in Réunion Island. After a short review on the Chikungunya virus, its principal Vector, Aedes albopictus, and recent biological results, we will present the mathematical models developed to assess the efficacy of the vector-control tools used in Réunion Island. We will introduce the SIT project, provide some recent results, and compare them to the previous ones. Finally, we will end the presentation with some prospective works. (Texte intégral