Marion Tiradon

Adrien Bonnet

Hong Do
email: hong.do@sophia.inra.fr

Thi Khanh

Etty Colombel

Maurane Buradino

Elisabeth Tabone
email: elisabeth.tabone@sophia.inra.fr

Evaluation of a new biological pest control method against the palm borer, Paysandisia archon using oophagous parasitoids

Keywords: Trichogramma, Paysandisia archon, biological pest control, oophagous parasitoids, palm tree Trichogramma, Paysandisia archon, lutte biologique, parasitoïdes oviphages, palmier

des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

INTRODUCTION

Since 2009, the palm borer Paysandisia archon (Burmeister, 1880) is classified as a quarantine pest in France (Huguet and Fouilleux, 2010). Native from South America (Argentina, Brazil, Paraguay, Uruguay), it was first reported in France in the regions of Hyères and Toulon and would have been imported via palm trees (Trithrinax campestris) from Argentina in the 1990s (Montagud Alario, 2004). P. archon larvae cause significant damages on palm trees by feeding on the stipe. Moreover, pest is able to attack different palm species as Chamaerops humilis, Livistona chinensis, L. decipiens, Sabal spp., Phoenix canariensis, P. dactylifera, Trachycarpus fortunei and Washingtonia filifera [START_REF] Sarto I Monteys | The Castniid Palm Borer, Paysandisia archon (Burmeister, 1880), in Europe: Comparative biology, pest status and possible control methods (Lepidoptera: Castniidae)[END_REF].

Human activities have greatly contributed to the expanding of the pest. In France, the moth has been observed for the first time in the Var region. Now, it can be seen throughout South of France from the "Alpes-Maritimes" to the "Pyrénées-Orientales". It is also found in many European countries as Italy, Spain, Greece, Slovenia, Switzerland and Cyprus (EPPO, 2011). The lack of controls and vigilance during trade allowed the moth to gain ground. Furthermore, it appears that P. archon can travel large distances by flying. Its biology makes detection and control difficult: larvae remain endophagous throughout their life, the adults do not feed and are active during the day, females do not secrete long range pheromones [START_REF] Sarto I Monteys | Moths Behaving like Butterflies. Evolutionary Loss of Long Range Attractant Pheromones in Castniid Moths: A Paysandisia archon Model[END_REF], making attractant-based trapping presently impossible. A new research is in progress (CNRS) concerning pheromones secretion by the male butterflies. Mechanical control methods such as bagging or the use of birdlime have been advised as well as the use of chemicals. Regarding biological pest control, tests were performed and means established. The use of nematodes Steinernema carpocapsae as biopesticide, or products based on the entomopathogenic fungi Beauveria bassiana, are known to control the larvae. To date, no attempt of control using parasitoids or predators have been introduced. In Argentina, natural enemies such as birds or Ichneumonidae parasites are likely controlling the pest (Sarto i Monteys and Aguilar, 2005).

The INRA PACA MFEU aims to find new natural enemies, particularly oophagous parasitoids. Tests were set up and collects carried out with two distinct objectives:  In the laboratory, a screening of different Trichogramma strains was performed. Strains with physiological and morphological benefits were previously selected. The strains were then raised for several generations on hosts laying bigger eggs than the host Ephestia kuehniella usually used. Trichogramma produced on big eggs were then tested on P. archon eggs.

 In the field, collects of P. archon eggs have started in order to look for native oophagous parasitoids which would naturally parasitized eggs of this butterfly. The oldest sites attacked will be preferred. The aim is to raise those parasitoids on large lepidopteran eggs and then, to study their potential parasitism on P. archon eggs This article informs the first results obtained by INRA PACA during the first year of Palm Protect program.

MATERIAL AND METHODS

REARING OF TRICHOGRAMMA

Selected strains

Nineteen strains of trichogramma belonging to 9 species and taken from the INRA PACA collection (Figure 1) were selected on biological criteria that could facilitate their potential on big eggs of P. archon: -Strains fertility, or their ability to parasitize a large number of eggs.

-Originated host from which the strains have been collected before rearing on E. kuehniella eggs. Strains from biggest host eggs were selected.

-Others benefits such as large ovipositor (which could help to penetrate chorion of big eggs more easily), or the ability to parasitize on high vegetation layers, have also been taken into account. The different selected strains are shown in Table 1. Those strains, initially reared on Ephestia kuehniella eggs, were reared during several generations on Philosamia ricini (Drury) (Lep., Saturniidae), and Bombyx mori (L. ) (Lep., Bombycidae) eggs, in order to accustom the parasitoids on larger eggs and stronger chorion.

Table 1: Species / strains of trichogramma initially selected. Espèces / souches de trichogrammes pré-sélectionnées

Specie / strain Author

Trichogramma chilonis SB Ishii

Trichogramma chilonis Th Ishii

Trichogramma chilonis Ca Ishii

Trichogramma chilonis P Ishii

Trichogramma chilonis C Ishii

Trichogramma cordubensis Pt

Vargas & Cabello

Trichogramma cordubensis Et

Vargas & Cabello

Trichogramma cordubensis Ct

Vargas & Cabello

Trichogramma pretiosum U Riley

Trichogramma pretiosum HA Riley

Trichogramma pretiosum HB Riley

Trichogramma dendrolimi I Matsumura

Trichogramma dendrolimi L Matsumura

Trichogramma buesi C Voegelé

Trichogramma exiguans U Pinto & Platner

Trichogramma embryophagum It Hartig

Trichogramma embryophagum I Hartig

Trichogramma euproctidis A Girault

Trichogramma daumaleae C Dugast & Voegelé

Picture : Tiradon Marion The rearing of Philosamia ricini (Figure 2) was undertaken, in order to provide big eggs necessary for trichogramma rearing. The first larvae were ordered at the OPIE association. The breeding was then maintained on privet leaves over several generations, in order to have available adults and eggs for maintaining trichogramma rearing. Tubes containing 60 eggs of P. ricini for about 30 trichogramma were used. The eggs renewal was performed by migration of the emerging trichogramma in a new tube (7.5 × 1.2 cm) with honey, for each generation of each strain. In the same way, a B. mori rearing was undertaken and the selected trichogramma were bred on those eggs.

Tests on the pest P. archon

Strains with the best parasitism on big eggs were tested on Paysandisia archon eggs in tubes and fed with honey. The pest eggs were regularly provided by our partner CIRAD of Montpellier. Several tests have been done following different modalities showed in Figure 3.

In a first time, emergence of trichogramma from P. archon eggs proposed to the parasitoids for oviposition was then examined.

COLLECTS OF OOPHAGUS PARASITOIDS

First collects on the field were done. The aim was to let incubate the collected eggs until emergence of any oophagous parasitoids. Parasitoids will be then grouped by type and then reared on large eggs.

FIRST RESULTS

TESTS ON BIG EGGS

Rearing results on Philosamia ricini eggs Concerning trichogramma rearing on P. ricini eggs, the different strains initially selected all have been raised on this host over several generations. The Trichogramma size has significantly increased in the first generation.

Rearing results on Bombyx mori eggs Breeding of the different trichogramma strains on B. mori eggs was also a success. However, although the size of trichogramma from these eggs is greater than those reared on E. kuehniella, they remain significantly smaller than those produced on P. ricini eggs.

Screening on Paysandisia archon eggs

To date, no emergence of trichogramma could have been observed on eggs of P. archon. The eggs have hatched, giving a larva, or aborted. Whatever the host, trichogramma females showed no real interest in the moth eggs.

COLLECTS

First results of collects for P. archon eggs evidenced difficulty of finding eggs in situ. Other collects are nevertheless considered in areas where P. archon attacks are denser.

DISCUSSION

Trichogramma are widely used in biological pest control around the world. They are rather generalist parasites capable of parasiting many host eggs with variable shapes and sizes. Indeed, trichogramma are observable on a large number of moth family, they can be found on Sphingidae, Noctuidae, Tortricidae, Saturniidae, etc. depending on the species of trichogramma (Consoli, 2010). The first step of our protocol, which covers trichogramma rearing on P. ricini and B. mori eggs showed positive results. These parasitoids were able to develop over several generations on these hosts, with a larger size. Females were found interested in these eggs from their first contact with them. In addition, laboratory production has supply fresh eggs that are more attractive for trichogramma.

Unfortunately, in this study, we were unable to demonstrate their ability to parasitize P. archon eggs. Several factors come into play in the choice of oviposition and can cause a rejection from the female:

-egg size, -egg resistance to the ovipositor, -chemical factors, -age of the egg: a too old egg reduces the attractiveness of trichogramma and decreases their success of parasitism.

-secretion of toxins (Consoli, 2010).

P. archon eggs have a very characteristic shape (rice grain) which differs greatly from the original host of rearing. Moreover, they are very large (about 1.5 mm wide and 4.5 mm long).

Also, the selected alternative hosts for rearing was perhaps not the better choice. Additional assays have to be realized on others lepidopteran host families. Indeed, trichogramma gain experience at the larval stage regarding their host preference. This preference goes toward a host from which they originate (Consoli, 2010) and as there is no native Castniidae in France, phylogenetically or physiologically close families will be used for our next experiments. These assumptions could be the cause of lack of interest of tested trichogramma toward P. archon eggs.

CONCLUSION AND PERSPECTIVES

To date, parasitism tests on P. archon eggs using different trichogramma strains have not yielded positive results. However, the interest to continue the breeding of these parasitoids on other major lepidopteran hosts before testing on P. archon eggs remains. Families potentially close of Castniidae will be preferred. An European trichogramma specie (Trichogramma gicai), naturally more robust than those we usually harvest will be sought from the spring of 2013.

A new campaign for collecting P. archon eggs in heavy infested areas is already planned. Similarly, P. archon eggs will be placed in different climate and environmental zones, chosen in order to trap any native oophagous parasitoids potentially adapted to this pest eggs. Thus, many new avenues are still developing in the field of oophagous parasitoids.

Figure 1 :

 1 Figure 1 : Trichogramma collection of INRA PACA Collection de trichogrammes à l'INRA PACA

Figure 2 :Figure 3 :

 23 Figure 2 : Philosamia ricini

ACKNOWLEDGEMENTS

The present work was supported by the EU under project Number FP7-KBBE-2011-5-289566. We thank the FREDON PACA and LR, the CIRAD of Montpellier for their help and collaboration.


Romain Barillot

Didier Combes

Pierre Huynh

Abraham J Escobar-Gutiérrez

• Need for growing legume species in Europe (vegetable proteins, sustainable

Keywords: Ascochyta Blight (Mycosphaerella pinodes), stem and pod rot (Sclerotinia sclerotiorum), powdery mildew

•••

  Variability of architectural parameters of pea: plant height, spatial distribution of leaf area, lateral branching… (Béasse et al., 2000; Le May et al., 2009) • Virtual plant models: suitable tools for studying plant-pathogen interactions and diseases dispersal (Robert et al., 2008) Arumingtyas et al. (1992) Kusnadi et al. (1992) Gourlay et al. Model developed on the platform L-Py: L-System coupled to a dynamic language Python (Boudon et al., 2012) Parameters: random selection on data (mean, SD) • Take into account the variability of the population L-Pea -Vegetative module • PAR intercepted by apex used to estimate the R/FR ratio (Escobar-Gutiérrez et al., 2009)• Tropism of stems depending on PAR and R

REPRODUCTIVE


The concept of dietary bioactive peptides brings a new dimension that should be considered in the description of dietary protein quality. Bioactive peptides have been defined as protein fragments that have, in addition to their nutritional value, a positive impact on body functions (vascular system, immunity, metabolism …) and may ultimately influence health. Most dietary proteins contain bioactive sequences, but the corresponding peptides need to be released by proteolysis during digestion, and absorbed from the gut.

The purpose of this study was to develop the analytical tools required for peptide isolation, characterization and identification by mass spectrometry, in plasma and serum samples.

A rapid and robust sample preparation method, based on a protein precipitation using acetonitrile, was optimized. Peptides profiles were then determined using Ultra Performance Liquid Chromatography coupled on-line to a LTQ-Orbitrap Velos mass spectrometer. The separation was performed on an Aeris peptideXB-C18 column and using a 17 min gradient. Data were collected in positive ion mode in scan and data dependent MS/MS modes. Two approaches depending on peptide masses were developed to perform the data processing and the identification. Peptides of masses between 400-800 m/z were identified with proteomic tools: ions were extracted using Progenesis LC-MS software, and Mascot (http://www.matrixscience.com/) was used to identify peptides in the merged peaklist generated by Progenesis. For peptides of lower masses between 100-500 m/z, not listed in proteomic databases, metabolomic tools were used: the raw data were transformed to centroid mode and mass corrected before being analyzed using XCMS The data were peak detected and noisereduced. Identification was performed using an in-house database built from peptide data extraction from the Human Metabolome DataBase ((http://redpoll.pharmacy.ualberta.ca/hmdb/HMDB/) and from the BIOPEP website (http://www.uwm.edu.pl/biochemia). The validation of the putative identification was performed by analysis of their fragmentation with the combination of CID and HCD.

The developed method allowed us characterizing the peptide fraction in plasma or serum samples, to study the bioavailability of peptides and their kinetics of appearance in blood. following the intake of a complete meal. ((http://redpoll.pharmacy.ualberta.ca/hmdb/HMDB/) and from the BIOPEP website (http://www.uwm.edu.pl/biochemia).

The validation of the putative identification was performed by analysis of their fragmentation with the combination of CID (collision induced dissociation) and HCD (Higher-energy C-trap dissociation) a good compromise will be possible to obtain better fragmentation informations.

The developed method allowed us characterizing peptide fraction from plasma or serum samples, to study the bioavailability of peptides and their kinetics of appearance in blood following the intake of a complete meal.

  Auvergne, Unité de Nutrition Humaine, BP10448, F-63000Clermont-Ferrand, France


Impact of age and intestinal microbiota on the expression of avian defensins in the chicken gut

  Anne-Christine A.-C. Lalmanach, Djihad Bencherit, Hélène Marty, Angélina Trotereau, Annie Brée, Catherine C. Schouler To cite this version: Anne-Christine A.-C. Lalmanach, Djihad Bencherit, Hélène Marty, Angélina Trotereau, Annie Brée, et al.. Impact of age and intestinal microbiota on the expression of avian defensins in the chicken gut. International Symposium on Alternatives to Antibiotics (ATA) : Challenges and Solution in Animal Production, World Organisation for Animal Health (OIE). Paris, INT., Sep 2012, Paris, France. hal-


est destinée au dépôt

Assessing potential expansion of the Pine Wood Nematode (Bursaphelenchus xylophilus) from the spatial genetic structure of the vector (Monochamus galloprovincialis)

Julien Haran, Alain Roques, Geraldine Roux-Morabito

To cite this version: Since its introduction in Portugal, the PWN has rapidly expanded its range to a large part of the country and will probably keep expanding to the rest of Europe. Estimation of dispersal abilities of M. galloprovincialis over various landscapes and across mountains is a key point to predict the invasion of the PWN in Europe, and will help to set up management for this pest.

Microsatellites are highly variable genetic markers. Their polymorphism provides information on genetic structure of organisms at a broad scale (phylogeography), but also at local scale (migration of individuals). We developed a set of 12 microsatellites loci specific to M. galloprovincialis. First assessments conducted on six populations along a European North-South transect reveal a reduction of the genetic diversity northward, with a maximum of allelic richness in Spain. This seems consistent with post glacial recolonization of Mediterranean species associated with Pines trees. We have also observed a significant differentiation between some Iberian populations.

Our perspectives are to use the microsatellites markers (i) to construct the European phylogeography of M. galloprovincialis, (ii) to estimate the effect of the Pyrenees and the landscape structure on dispersal abilities of this species, and finally, (iii) to look at the effect of PWN invasion on its genetic structure in Portugal.

Assessing potential expansion of the Pine Wood Nematode (Bursaphelenchus xylophilus) from the spatial genetic structure of the vector (Monochamus galloprovincialis)

  Haran J, Roques A, Roux G, Assessing potential expansion of the Pine Wood nematode (Bursaphelenchus xylophilus)from the spatial genetic structure of the vector (Monochamus galloprovincialis). In: Schröder, T. (ed.), Pine Wilt Disease Conference 2013, pp. 13, Braunschweig, ISSN: 1866-590X

	Haran J,	Roques A, Roux G
	INRA Orléans, URZF-2163 Avenue de la Pomme de Pin 45160 ARDON, France

Julien Haran, Alain Roques, Geraldine Roux-Morabito. Assessing potential expansion of the Pine Wood Nematode (Bursaphelenchus xylophilus) from the spatial genetic structure of the vector (Monochamus galloprovincialis). Pine Wilt Disease Conference 2013, Julius Kühn-Institut. DEU., Oct 2013, Braunschweig, Germany. 142 p., 10.5073/berjki.2013.169.000. hal-02746336

Email: julien.haran@orleans.inra.fr Monochamus galloprovincialis (Coleoptera, Cerambycidae) is the main factor involved in the natural spread of the Pine Wood Nematode (PWN), a serious pest for pine forests.