à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Purpose -Antioxidant vitamins (A, C and E) and antioxidant micronutrients (carotenoids and polyphenols), are assumed to diminish the incidence of several degenerative diseases.

However, absorption efficiency of these compounds is very variable among individuals. We hypothesized that this inter-individual variability is due to genetic variations between individuals. We thus dedicated a clinical study to identify genetic variants associated with fatsoluble antioxidant bioavailability.

Methods -40 healthy male adults consumed, at a miminum 3 wks intervals, meals that contained either a supplement of vitamin E, a supplement of lutein or a tomato purée (as a source of lycopene and β-carotene). Volunteers were genotyped using whole-genome microarrays. Antioxidant concentrations were measured in plasma chylomicrons (CM) isolated at regular time intervals over 8 h postprandially. The association of single nucleotide polymorphisms (SNPs), in or near candidate genes, with the postprandial CM antioxidant responses was assessed by partial least squares (PLS) regression.

Results -

The postprandial CM antioxidant responses to the antioxidant rich test-meals were highly variable among the subjects (CV > 80%). Data obtained allowed us to generate validated significant PLS models, which usually included 20-40 SNPs, and which explained a significant part of the variance of the postprandial CM antioxidant responses.

Conclusions -Minor genetic variants are associated with bioavailability of the studied antioxidants. This partly explains the huge interindividual variability in absorption efficiency of these compounds. Our results allow us to propose genetic scores that may predict the ability of a subject to accurately absorb these antioxidants. This is a first step toward personalize recommendations in antioxidant vitamin and micronutrient intake.

  Obesity and Risk of Thrombosis", UMR 1062 INSERM/1260 INRA/Aix-Marseille University, Marseille, France.


Posi

à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Single nucleotide polymorphisms associated with carotenoid bioavailability.

Patrick Borel 1 1 Nutrition, Obesity and Risk of Thrombosis, UMR 1062 INSERM/1260 INRA/Aix-Marseille University, Marseille, France.

Carotenoids are plant bioactives assumed to participate in the beneficial effects against degenerative diseases attributed to plant products. Their absorption efficiency is very variable among individuals. We hypothesized that this is due, at least in part, to genetic variations between individuals. We thus dedicated a clinical study to identify minor genetic variants associated with carotenoid bioavailability. Healthy male adults (n=40) consumed, at a minimum of 3 weeks intervals, 2 meals that contained either tomato purée (as a source of lycopene and β-carotene) or a lutein supplement. Volunteers were genotyped using whole-genome microarrays. Carotenoid concentrations were measured in plasma chylomicrons isolated at regular time intervals over 8 h postprandial. Partial Least Squares (PLS) regression was used to identify combinations of single nucleotide polymorphisms (SNPs) associated with the variability in carotenoid levels (AUC of the postprandial chylomicron carotenoid concentration).

The carotenoid responses were highly variable among the subjects (CV = 105 % for βcarotene, 75 % for lutein and 70 % for lycopene) and were significantly correlated with fasting plasma carotenoid concentrations. PLS models, which included between 12 and 16 genes and between 25 and 29 SNPs, explained a significant part (69% to 73%) of the variance of the carotenoid responses. Combinations of SNPs are associated with interindividual variation in carotenoid bioavailability which apparently affects the long term carotenoid status. Our results allow us to propose genetic scores that may predict the ability of a subject to accurately absorb these plant bioactives. This is a first step toward personalize recommendations in carotenoid intake.


William Dudefoi

Marie-Hélène Ropers

Suivi des nanoparticules de dioxyde de titane dans les fluides digestifs

teaching and research institutions in France or abroad, or from public or private research centers.

Suivi des nanoparticules de dioxyde de titane dans les fluides digestifs

Dudefoi W. a , Ropers M.H. * a a INRA, Unité Biopolymères Interactions Assemblages, F-44316 Nantes * marie-helene.ropers@nantes.inra.fr

Le dioxyde de titane est un additif alimentaire qui se présente sous la forme d'une poudre blanche et qui est utilisé pour blanchir certaines préparations et notamment les enrobages de confiseries. L'identification d'une fraction de taille nanométrique dans ce type de composé [1] nécessite, au regard des risques de passage de la barrière intestinale, de suivre le devenir de ce composé au cours du tractus gastro-intestinal.

Afin de mieux identifier comment les particules évoluent tout au long du tractus gastro-intestinal, nous avons travaillé sur un modèle de digestion in vitro, qui mime la composition en enzymes et sels, et les conditions de pH des 3 milieux salivaire, gastrique et intestinal. L'application de ce protocole à deux échantillons de dioxyde de titane, différant par leur point isoélectrique a permis d'identifier une agglomération significativement plus importante dans les fluides digestifs que dans l'eau. L'affinité de certains ions et de certaines protéines pour la surface du dioxyde de titane explique l'augmentation de l'agglomération. Les protéines adsorbées en quantité les plus importantes sur le dioxyde de titane ont été identifiées par électrophorèse sur gel. L'amylase est la protéine identifiée systématiquement tandis que la pepsine n'a été identifiée que sur un seul échantillon. Ce comportement a été comparé à celui des particules de référence P25, largement utilisées dans les études de toxicologie mais différant par leurs propriétés physico-chimiques.

[1] Y. Yang, K. Doudrick, X. Bi, K. Hristovski, P. Herckes, P. Westerhoff & R. Kaegi. Environ. Sci. Technol. 48 (2014) 6391-6400


Frédérik Saltré

Joel J Chadoeuf

B W Brook

Damien A Fordham

Marta Rodriguez-Rey

Corey J A Bradshaw

Spatial pattern of Eurasian woolly mammoth extinctions during the Late Pleistocene based on statistical inference from fossil records

à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

  pattern of Eurasian woolly mammoth extinctions during the Late Pleistocene based on statistical inference from fossil records. 9th IALE World Congress, Portland, OR, USA. Title Spatial pattern of Eurasian woolly mammoth extinctions during the Late Pleistocene based on statistical inference from fossil records Authors F. Saltré, B.W. Brook, J. Chadoeuf, D.A. Fordham, M. Rodriguez-Rey, C.J.A. Bradshaw Abstract: Late Pleistocene extinctions affected most large and low-fecundity mammals such as the woolly mammoth (Mammuthus primigenius) in Eurasia. Despite recent advances in both genetic and modelling approaches and increasing fossil data, assessing the relative importance of climate variations and human hunting on the spatial pattern of mammoth extinction remains a challenge. This is because estimating incomplete fossil time series biases the true dates of extinction and appearance, potentially leading to incorrect conclusion of the relative importance of individual and synergistic drivers. We compared outputs of two statistical models that (i) spatially infer mammoth final extinction dates and first human occurrence from sighting records based on the recently developed Gaussian-resampled inverse-weighted McInerny (GRIWM) and (ii) predict the date when mammoths should have gone extinct based on climate change only (climate anomaly and velocity). GRIWM generates a confidence interval of the true extinction time, which suggests a pattern of mammoths extinctions starting in central Europe and spreading toward both western Europe and northeastern Siberia. Climate change had a minor impact on mammoth extinctions, mainly in the northeastern part of Eurasia where the synergistic hypothesis is mainly supported by our results whereas the human hunting hypothesis is more likely to have caused mammoth extinctions elsewhere in Eurasia.


Valérie Guillard Modified

teaching and research institutions in France or abroad, or from public or private research centers.

•

  Pouvoir ajouter un nom de concept directement lors de la saisie // sans intervenir sur les fichiers .csv • Automa>ser l'import de tableau excel • Automa>ser l'import de tableau html?• Comment faire vivre l'ou>l?-Au delà des développements méthodo -Quel communauté? (académique uniquement?) -Quel support?


Paper 1 -Sampling design and sampling unit Sampling unit: the field To

test the effect of landscape metrics on species diversity To investigate the contribution of crop diversity to species diversity Sampling unit: the field To test differences in species diversity response between field border and field centre Sampling unit: the transect

Note that all plant species from all crop types were included in analyses.

Paper 1 -Response variables

Partitioning diversity:

at

Crop_SDHI

ns

Crop_TBL ns ns

Include region as random effect in mixed effect models : TO DO Paper 1 -Perspectives

• Grouping crop types and work at the 'crop type' scale

• Differentiating species diversity between field border and field centre

• Separating flora into functional groups

• Going further in landscape structure characterization Avoid singleton in models; may help to elucidate the strong regional effect Some crops appear only in few regions e.g. almonds -1) arable crops, 2) perennial non woody crops, 3) perennial woody crops … etc -1) winter cereals, 2) summer cereals, 3) grasslands, 4) orchards, … etc Proposals:

Separate data from field border and field centre as they experienced different disturbance level

Deepen the effect of landscape composition/configuration by including % grasslands, % hedges, etc.

Separate annual from perennial plant species Categorize species as rare, harmful, … etc

the FIELD scale at the TRANSECT scale

	Gamma	Total species richness	Total species richness
	Alpha	Mean species richness (=averaging species richness over the two	-
		transects per field)
	Beta	Gamma -Alpha	-

Paper 1 -Response variables Partitioning diversity: at the FIELD scale at the TRANSECT scale

		Paper 1 -Methods and results Paper 1 -Methods and results Paper 1 -Methods and results Paper 1 -Methods and results
		i) To test the effect of landscape metrics on species diversity i) To test the effect of landscape metrics on species diversity ii) To investigate the contribution of crop diversity to species diversity iii) To test differences in species diversity response between field border and field centre
		• Use of simple linear models as: • Use of simple linear models as: Use of mixed effect models (LMMs) as:
	richness Accumulated species	50%	Gamma Alpha Beta Beta_JAC Response variable ~ Crop_SHDI + Crop_TBL + Crop type + Region + Total species richness Total species richness Mean species richness (=averaging species richness over the two transects per field) -Gamma -Alpha -Jaccard index Crop_SHDI:Region + Crop_TBL:Region + Crop_SDHI:Crop + Crop_TBL:Crop Gamma Alpha Beta Beta_JAC Crop_SDHI --ns ns Crop_TBL ns ns --Response variable ~ Crop_SHDI + Crop_TBL + Crop type + Region + Crop_SHDI:Region + Crop_TBL:Region + Crop_SDHI:Crop + Crop_TBL:Crop Gamma Alpha Beta Beta_JAC Crop_SDHI --ns ns Crop_TBL ns ns --Same results Response variable ~ Crop_SHDI + Crop_TBL + Crop type + Region + PVDS Response variable ~ Crop_SHDI + Crop_TBL + Crop type + Region + non crop cover + Crop_SHDI:Region + Crop_TBL:Region + Crop_SDHI:Crop + Crop_TBL:Crop Gamma Alpha Beta Beta_JAC Crop_SDHI --ns ns Crop_TBL ns ns --Camargue Coteaux EastAnglia EastAnglia Ontario East Anglia woody boundary + Crop_SHDI:Region + Crop_TBL:Region + (1| Landscape) + (1| Year) 0 50 100 150 200 250 Armorique Coteaux Lleida Ontario PVDS Accumulated Armorique Lleida Gamma Gamma richness Camargue Goettingen 5 10 15 20 25 Number of crops 7 field border field centre Total species richness per Total species richness per species Goettingen transect « field border » transect « field centre »
			2	4	6	8	10	12
					Number of crops

(=differences in species composition between the two transects per field)

Paper 1 -Methods and results

i) To test the effect of landscape metrics on species diversity

• Use of simple linear models as:

• Testing the robustness of models by adding 'non crop cover ' and 'woody boundary' :

• Testing the robustness of models by adding 'non crop cover ' and 'woody boundary' :

Same results

• Adding random effects in more complex models (LMMs) as:

Response variable ~ Crop_SHDI + Crop_TBL + Crop type + Region + non crop cover + woody boundary + Crop_SHDI:Region + Crop_TBL:Region + (1| Landscape) + (1| Year)

Paper 1 -Methods and results

i) To test the effect of landscape metrics on species diversity


HAL is a

Does wing morphology of the active flyer Calliphora vicina change as it invades islands ruled by flightless insects?

The cosmopolitan blowfly Calliphora vicina established in the sub-Antarctic Kerguelen Islands in the late 1970s, following a warming period that allowed its full development. Although temperature and wind may limit its flight activity, it invaded the archipelago towards sites remote from the introduction point. While most native competitors have converged to flightlessness as a response to local stringent environmental conditions, the flight strategy of C. vicina might be either a handicap or a competitive advantage under ongoing climate change. Using geometric morphometrics, we questioned whether the wing changed over time within the archipelago (1998 vs. 2009), and compared its morphology with a continental population from a temperate area (1983 vs. 2009). Wing shape plasticity to temperature was also experimentally investigated. We found no cues of relaxed selection on flight morphology in the invaded range. However, comparatively rapid changes of wing shape occurred over time in females from the Kerguelen Islands, despite a shorter time-lag between the samples compared to the continental population. We also reported different reaction norms to temperature for wing shape between the populations. These findings are consistent with a fingerprint of local adaptation to the peculiar environmental conditions encountered in the invaded range, but additional studies are necessary to test this hypothesis. From an evolutionary standpoint, sustained flight capability under the novel sub-Antarctic conditions may be critical in the invasive success of C. vicina, given the flightlessness rule observed in native competitors. To address the adaptive significance of the changes recorded, further studies should tackle their aerodynamic consequences and future evolution.

  INRA, UR0633 Zoologie Forestière, 2163 Avenue de la Pomme de Pin, CS 40001, Ardon, 45075 Orléans Cedex 2, France Tel.: +33 2 38 41 78 98 Email: mathieu.laparie@orleans.inra.fr I am a Research Asssociate recently hired at INRA, Forest Zoology Research Unit. My research interests lie in the relationship between the success of invasive and/or expanding species and the way they fit the novel selection regimes they encounter as they expand and disturb newly colonized habitats. I also focus on the effects of climate change as it may facilitate biological invasions or trigger range expansions. The tools I use to study population differentiation as a consequence of the invasion (or range expansion) process range from geometric morphometrics to ecophysiology and metabolomics.

  L E S T U D I U M ® CONFERENCES Insect invasions in a changing world | 17-19 December 2014


Carlos Lopez

Carlos Lopez-Vaamonde

Patterns of invasion of Lepidoptera in Europe

come

Patterns of invasion of Lepidoptera in Europe

Here I provide an overview of Lepidopteran invasions to Europe that result from increasing globalisation and also review expansion of species within Europe. A total of 97 non-native Lepidoptera species (about 1% of the known fauna), in 20 families and 11 superfamilies have established so far in Europe, of which 30 alone are Pyraloidea. In addition, 88 European species in 25 families have expanded their range within Europe and around 23% of these are of Mediterranean or Balkan origin, invading the north and west. Although a number of these alien species have been in Europe for hundreds of years, 74% have established during the 20th century and arrivals are accelerating, with an average of 1.9 alien Lepidoptera newly established per year between 2000-2007. For 78 aliens with a known area of origin, Asia has contributed 28.9%, Africa (including Macaronesian islands, Canaries, Madeira and Azores) 21.6%, North America 16.5%, Australasia 7.2% and the neotropics just 5.2%. The route for almost all aliens to Europe is via importation of plants or plant products. Most alien Lepidoptera established in Europe are also confined to man-made habitats, with 52.5% occurring in parks and gardens.

  INRA, UR0633 Zoologie Forestière, 2163 Avenue de la Pomme de Pin, CS 40001, Ardon, 45075 Orléans Cedex 2, France Tel.: +33 2 38 41 78 61 Email: carlos.lopez-vaamonde@orleans.inra.fr I am an entomologist interested in the patterns and processes of insect evolution, with a particular focus on leaf-mining microlepidoptera and their associated parasitoids. I obtained a PhD on Evolutionary Biology at Silwood Park, Imperial College (2002) and worked as a postdoctoral associate for three years at the Institute of Zoology in London on molecular and behavioural ecology of social insects. I was then hired as an INRA research scientist in July 2005. My work on invasion biology has been focused on using genetic markers to understand the dynamics of invasion. I am also interested in how communities of phytophagous insects and their parasitoids are assembled in their native range and how they are reassembled in the areas where their host plants have been introduced.

  L E S T U D I U M ® CONFERENCES Insect invasions in a changing world | 17-19 December 2014


Julien Haran

Alain Roques

Christelle Robinet

Geraldine Roux-Morabito

Landscape

Landscape Genetics of Monochamus galloprovincialis, vector of the pine wood nematode in Europe

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

I am now working at a different evolutionary scale. My PhD deals with dispersal of the beetle Monochamus galloprovincialis, the vector of the pine wood nematode in Europe. I try to identify the landscape features affecting dispersal behavior of this species to uncover potentials pathways to the spread of the nematode. M. galloprovincialis being a native species, we also explore the micro evolutionary processes involved in its intra-specific diversification.

Landscape Genetics of Monochamus galloprovincialis, vector of the pine wood nematode in Europe.

Julien HARAN, Alain ROQUES, Christelle ROBINET & Géraldine ROUX-MORABITO

The pine wood nematode (PWN), Bursaphelenchus xylophilus (Steiner & Burher) Nickle (Nematoda, Aphelenchoididae) is the causal agent of the pine wilt disease (PWD), a virulent syndrome killing susceptible pines trees within few months. From its native area in North America, it has been introduced in several Asian and European countries causing considerable damages to native pine forests. The PWN was detected for the first time in Europe in Portugal 15 years ago. From its introduction site, it rapidly expanded its range to a large part of the country and entered into Spain. In Europe, the native longhorn beetle Monochamus galloprovincialis (Olivier, Coleoptera, Cerambycidae) is the only known vector for this nematode. This beetle performs its larval development in the wood of declining pine trees and spread the PWN when it emerges from infected wood. Thus, the natural dispersion of the PWN is highly depending on dispersal abilities of the beetle. Given the rapid range expansion of this pest and the threat to forests involved, it is crucial to identify potential barriers to dispersal of M. galloprovincialis to define suitable pest management strategies.

Based on 1043 individuals from Iberian Peninsula, genotyped at 13 microsatellites loci, we conducted a landscape genetics analysis to uncover the landscape features affecting dispersal of M. galloprovincialis. To avoid confounding effect of evolutionary history of this species in the area of study, we used a nested sampling design and assessed population genetic structure in order to select the appropriate populations and the optimal scale for correlation analysis.

Our results show that mountain ranges represent a break to dispersal of M. galloprovincialis, and subsequently potential barriers to the spread of the PWN. We also show that the experimental design (scale and populations selected) strongly influence the results of landscape genetics approach.

  INRA, UR0633 Zoologie Forestière, 2163 Avenue de la Pomme de Pin, CS 40001, Ardon, 45075 Orléans Cedex 2, France Tel.: +33 6 40 90 67 42 Email: julien.haran@gmail.com I am PhD student at INRA, URZF. My research interest focus on macro and micro evolution of phytophagous beetles in relation with their environment. My previous projects at Natural History Museum (UK) aimed at reconstructing the deep phylogeny of weevils based on mitogenome sequences, and to relate diversification of lineages to host plants and larval lifestyle.

  L E S T U D I U M ® CONFERENCES Insect invasions in a changing world | 17-19 December 2014


model counting

• But SAT solvers generally poor at handling large XOR constraints

• Find "input" variables, restrict XORs to these MUS applications -Approximate model counting

• Add a system of large XOR constraints and do complete model counting

• But SAT solvers generally poor at handling large XOR constraints

• Find "input" variables, restrict XORs to these

• MUSes of

F (X) ∧ F (Y) ∧ (x i = y i ) ∧ (x i = y i )
MCS applications

• MaxSAT [Mencía et al, 2015;Bjorner and Narodytska, 2015] • Finding MUSes! [Bacchus and K, 2015] Hitting set duality

(x 1 ) (x 2 ) (x 3 ) (x 1 ∨ x 2 ) (x 1 ∨ x 3 ) (x 2 ∨ x 3 ) (x 1 ) (x 2 ) (x 3 ) (x 1 ∨ x 2 ) (x 1 ∨ x 3 ) (x 2 ∨ x 3 ) (x 1 ) (x 2 ) (x 3 ) (x 1 ∨ x 2 ) (x 1 ∨ x 3 ) (x 2 ∨ x 3 )
• Every (minimal) CS is a hitting set of all (minimal) USes

• Every (minimal) US is a hitting set of all (minimal) CSes [Reiter, 1987] Interesting Queries

• Discover a single MCS/MCS • Enumerate all MCSes/MUSes • Clause set refinement [Nadel et al, 2014] Impact of incremental solver technology • Addition-based [Gregoire et al, 2008;van Maaren and Wieringa, 2008] • Dichotomic [Junker, 2004] • Exploit duality • Addition-based [Gregoire et al, 2008;van Maaren and Wieringa, 2008] • Dichotomic [Junker, 2004] • Exploit duality [Bacchus and K, 2015] ⇒ All applicable to any set minimization problem Deletion based

(x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 1 , x 3 ) (x 2 , x 3 )
Deletion based

(x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 1 , x 3 ) (x 2 , x 3 )
Deletion based

(x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 1 , x 3 ) (x 2 , x 3 ) UNSAT Deletion based (x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 1 , x 3 ) (x 2 , x 3 )
Deletion based

(x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 2 , x 3 ) (x 1 , x 3 ) Deletion based (x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 2 , x 3 ) (x 1 , x 3 ) UNSAT Deletion based (x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 2 , x 3 ) (x 1 , x 3 ) Deletion based (x 1 ) (x 2 ) (x 3 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) Deletion based (x 1 ) (x 2 ) (x 3 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) SAT Deletion based (x 1 ) (x 2 ) (x 3 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) Deletion based (x 1 ) (x 2 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) Deletion based (x 1 ) (x 2 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) UNSAT Deletion based (x 1 ) (x 2 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) Deletion based (x 1 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) (x 2 ) Deletion based (x 1 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) (x 2 ) SAT Deletion based (x 1 ) (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) (x 2 ) Deletion based (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) (x 2 ) (x 1 ) Deletion based (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) (x 2 ) (x 1 ) SAT Deletion based (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) (x 2 ) (x 1 ) Deletion based (x 2 , x 3 ) (x 1 , x 3 ) (x 1 , x 2 ) (x 3 ) (x 2 ) (x 1 ) MUS! (x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 1 , x 3 ) (x 2 , x 3 ) (x 3 ) (x 1 , x 2 ) (x 1 , x 3 ) (x 2 , x 3 ) (x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 1 , x 3 ) (x 2 , x 3 ) (x 1 ) (x 2 ) (x 3 ) (x 1 , x 2 ) (x 2 , x 3 ) (x 1 ) (x 2 ) (x 1 , x 3 )
Exploiting Hitting Set duality

(x 3 ) (x 1 , x 2 ) (x 2 , x 3 ) (x 1 ) (x 2 ) (x 1 , x 3 )
Exploiting Hitting Set duality

(x 1 , x 2 ) (x 2 , x 3 ) (x 1 ) (x 2 ) (x 1 , x 3 ) (x 3 ) (x 1 , x 2 ) (x 2 , x 3 ) (x 1 ) (x 2 ) (x 1 , x 3 ) (x 3 ) (x 1 , x 2 ) (x 2 , x 3 ) (x 1 ) (x 2 ) (x 1 , x 3 ) (x 3 )
No MCS

Exploiting Hitting Set duality

(x 1 ) (x 2 ) (x 1 , x 3 ) (x 3 ) (x 1 , x 2 ) (x 2 , x 3 )

MUS

Is it faster to

• do m SAT checks? -or -

• discover k MCSes?

MCS-MUS Muser-Glucose HaifaMUC

The way forward

• Faster SAT solving

• Better use of existing SAT solvers • Extract more information from the solver

• Improved MCS extraction

• A new algorithm?

More problems

• Enumeration [Liffiton and Sakallah, 2008;Liffiton et al, 2015;Bacchus and K, 2016] • Minimization [Ignatiev et al, 2013;Ignatiev et al, 2015;Liffiton et al, 2009;Marques-Silva and Previti, 2014] • MCS extraction and enumeration [O'Callaghan et al, 2005;Bailey and Stuckey, 2005;Felfernig et al, 2014;N öhrer et al, 2012;Marques-Silva et al, 2013;Bacchus et al, 2014;Bacchus and K, 2015;Mencía et al, 2015] Q?

•

  Deletion-based [Belov and Marques-Silva, 2012]

  [Bacchus and K, 2015] Algorithms• Deletion-based[Belov and Marques-Silva, 2012]

• Help user find the bug in their model

• Help user find the bug in their model • Also for indirectly generated models• Compiler-generated model for type-checking

• Add a system of large XOR constraints and do complete model counting• But SAT solvers generally poor at handling large XOR constraints

• Add a system of large XOR constraints and do complete


Fulgencio Sanchez-Solana

Caridad Ros

Maria Del Mar Guerrero

Victoriano Martínez

Carmen María Lacasa

Ana Hernández

Jeronimo Torres

Dr Alain Palloix

Alfredo Lacasa

Katalin Ertsey-Peregi

Zsuzsanna Füstôs

Gabor Palotas

Gabor Csilléry

Fulgencio Sànchez-Solana'

Caridad Ros'

Maria Del Mar

Victoriano Martinez'

Carmen Maria Lacasa'

Ana Hernândez'

Jer6nimo Torres'

Alfredo Lacasa' J

Could quantitative resistance increase the

Keywords: Capsicum annuum, Meloidogyne incognita, Me genes

à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.


domain: sdv info sde

text:
Nitrogen interactions (Fléchard, 2010)

A

  losses at various scales Animal: CO 2 , H 2 O, CH 4 are released (50% C, H 2 O intake) House: CO 2 , H 2 O, NH 3 , N 2 O, N 2 , CH 4 are released from manure, sometimes after air treatment (20-60% of excreted nitrogen) Farm: gases are also released but fields also collect rainfall and dry deposition => net emissions can be negative (e.g. C storage in forests increased by N gas deposition; CH 4 sink by agricultural soils) Region: farm interacts with other activities; recycling or organic byproducts/biomass production can contribute to emission decrease Introduction 2. C-N are necessary but not sufficient to explain emission variability (to manage emission reductions) Climate influence=> temperature, rainfall Biological activity=> toxicities, synergies explain limiting or accelerating factors Non linearity of biology: feedbacks can be positive then negative around a threshold Scale effects=> analysis of biological & social functions (e.g. food chain) contents are necessary but not sufficient to explain emission variability => quality, dynamics of bioavailability Depending on OM nature, compost will loose more OM or more water => dry matter content can decrease or increase during composting Depending on OM nature, compost volume will be stable or decrease => natural aeration can decrease or increaseIn soils: chemical stability depends on organic species, physical stability depends on particle size and clay content, biological stability depends on T & H 2 O; OM stability affects fertility, C storage, N 2 O emission, N losses as gas or leaching Azote minéral (NH4 + N uréique pour les volailles) Azote organique minéralisable dans l'année Azote organique minéralisable les années suivantes Agricultural reuse of organic by-products Non-food/feed crops for contaminated OM Food/non food needs of urban populations and industries Heat & water availability for crop productions can increase productivity when they are limiting factors Wild animals & functions for non food requirements (recycling, hygienisation, recreation, etc.) hourly time step 5 parameters specific of manure + 25 based on literature and experiments influence of C & N biodegradability, porosity and humidity ., 2011. Ph.D. thesis, Agrocampus Ouest, Rennes. Retrieved from http://tel.archivesouvertes.fr/tel-00662627 , discussing, management Models for data interpolation and data mining Management of model quality, model uses, knowledge integration: long term strategy for model and experimental data development is necessary to integrate various disciplines


Keywords: Numerical Aperture : 1, 4 l: 488 nm Lasers (HeNe), (Arg) ou Diodes

Since molecules are moving driven by diffusion or active transport, bleached molecules exchange their place with un-bleached molecules.

Then the average intensity at the bleached spot recovers.

Kota Miura, EMBL Heidelberg, Germany. Based on the EAMNET Practical Course (20, April, 2004 ) Fluorescence Recovery After Photobleaching (FRAP)

French German summer school 1-07-14 .026

An idealized plot of a FRAP recovery curve. French German summer school 1-07-14

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Recapitulating, FRAP is generally suitable to study and investigate:

• Protein/molecule movement and diffusion (diffusional speed).

• Compartmentalization and connections between intracellular compartments.

• The speed of protein/molecule exchange between compartments (exchange speed).

• Binding characteristics between proteins. Additionally, the effect of mutations that alter individual amino acids on protein association, and the effect of small molecules, such as drugs or inhibitors, on protein pairs can effectively be studies using FRAP.

• Immobilization of proteins that bind to large structures, e.g., DNA, nuclear envelope, membranes, cytoskeletal elements, etc

Fluorescence Recovery After Photobleaching

French German summer school 1-07-14 .029

Colocalization, in a biological manifestation, is defined by the presence of two or more different molecules residing at the same physical location in a specimen. Overlay of both channels (red and green) displays a strong colocalization viewable in yellow.

The ability to determine colocalization in a confocal microscope is limited by the resolution of the optical system and the wavelength of light used to illuminate the specimen. French German summer school 1-07-14 .048

  of kinetic microscopy: FRAP, FLIM/FRET, FCS » Recent developments in advanced microscopy techniques, the so-called F-techniques, including Förster resonance energy transfer, fluorescence correlation spectroscopy and fluorescence lifetime imaging, have led to a wide range of novel applications in biology. The F-techniques provide quantitative information on biomolecules and their interactions and give high spatial and temporal resolution. In particular, their application to receptor protein studies has led to new insights into receptor localization, oligomerization, activation and function in vivo.

French

  I I : initial intensity I 0 : intensity at timepoint t 0 (first postbleach intensity) I 1/2 : half recovered intensity (I 1/2 = (I E -I 0 ) / 2) I E : endvalue of the recovered intensity t half : Halftime of recovery corresponding to I 1/2 (t 1/2 -t 0 ) Mobile fraction F m = (I E -I 0 ) / (I I -I 0 ) Immobile fraction F i = 1 -Fm Fluorescence Recovery After Photobleaching (FRAP)

French

  Widefield fluorescence and confocal microscopes have a theoretical resolution of approximately 200 nanometers olympusconfocal.com Forster Resonant Energy Transfert (FRET) / Fluorescence Lifetime Imaging (FLIM): supplying co-localization Tissular exploration without labeling : • Tissue structural changing (SHG, THG, CARS)German summer school 1-07-14 Biphotonic imaging, what's for in tissular and cellular exploration in vivo? .039 Total Internal reflection fluorescence microscope which applications in gene and cell therapy • Entry of viruses or non viral vector labeled with a fluorescent dye into cells on living or fixed cells and colocalization with cellular compartments, analyze route taken. Effect of chemical inhibitor and small interfering RNA to block specific entry pathway • Endocytic pathway analysis (transferrin, cholera toxin, dextran, clathrin-GFP, caveolin-GFP), discovery of virus entry pathways provides basic knowledge for potential entry portals into cells and contact points for improvements toward more efficient non viral

French German summer school 1-07-14.024

Kota Miura, EMBL Heidelberg, Germany. Based on theEAMNET Practical Course (20, April, 2004 ) Fluorescence Recovery After Photobleaching (FRAP)French German summer school 1-07-14.025

French German summer school 1-07-14.030

We focus a strong laser beam to a spot (red dotted circle).

Let's think of fluorescence molecules dispersed in a field. White circles represent the molecules. Then the strong irradiation BLEACHES the fluorescence at that spot.

Muscle section, Z projection

Detection of Mustems after their injection in the muscle Triple immunolabeling, dystrophine (green), bgal (red), laminine(blue).

Subcellular localisation

K. Rouger et al., Am J Pathol. 2011, 179(5):2501-2518 French German summer school 1-07-14

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G A Oliveira

S Bureau

M-G C Renard

F De Castilhos

DETERMINATION OF INDIVIDUAL SUGARS IN PASSION FRUIT USING NIR AND MIR SPECTROSCOPIC METHODS

In this work, NIR and MIR spectroscopy was investigated and compared for predicting individual sugars (sucrose, glucose and fructose). Spectra of 56 samples of lyophilized passion fruit were collected using an integrating sphere in NIR range and attenuated total reflectance accessory in MIR range. Spectral and reference data were analyzed by principal component analysis. Partial least square regression (PLSR) was used to establish calibration models. MIR technique was better than the NIR technique for glucose (R 2 v = 0.942), fructose (R 2 v = 0.855) and sucrose (R 2 v = 0.818).

INTRODUCTION

Trends in analytical chemistry are towards simple and less time consuming analytical methods, through advances in new technologies, automation, development of qualitative and screening methodologies, and the increasing use of chemometrics [START_REF] Karoui | A comparison and joint use of NIR and MIR spectroscopic methods for the determination of some parameters in European Emmental cheese[END_REF]. Infrared spectroscopy has been proposed as an interesting alternative to traditional methods due to its rapidity, simplicity, cost effectiveness and potential for routine analysis if proper calibration and validation steps were developed. Besides, it does not require special skilled operators [START_REF] Karoui | A comparison and joint use of NIR and MIR spectroscopic methods for the determination of some parameters in European Emmental cheese[END_REF][START_REF] Tavakolian | Differentiation of post harvest date fruit varieties non-destructively using FT-NIR spectroscopy[END_REF].

Compound identification by infrared spectroscopy is based on the molecules property to absorb infrared light and experience a wide variety of vibrational motions characteristic of their composition. Mid-infrared (MIR) spectra (4000 -400 cm -1 range) provide information from frequencies of fundamental molecular vibrations, while near-infrared (NIR) (2500 -700 nm range) spectra contain information from overtones and their combinations [START_REF] Shiroma | Application of NIR and MIR spectroscopy in quality control of potato chips[END_REF]. Compared to MIR, light can better penetrate into the matter and favor applications in bulk or intact samples in NIR range, requiring little or no sample preparation. As light diffusion is much greater in NIR range, spectra are more sensitive to factors that influence light diffusion as physical structure and presence of water in the sample [START_REF] Williams | Near-infrared Technology in the Agricultural and Food Industries[END_REF]. Besides, NIR spectrum is much more encumbered due to abundance of combinations and overtone bands and as a consequence, bands are less specific in the NIR range compared to MIR. Therefore, MIR spectroscopy is highly sensitive to chemical composition of samples compared to NIR [START_REF] Bellon-Maurel | Near-infrared (NIR) and mid-infrared (MIR) spectroscopic techniques for assessing the amount of carbon stock in soils e Critical review and research perspectives[END_REF]. These different features of NIR and MIR spectroscopy suggest that one of them can be more adequate to each application.

Área temática: Engenharia e Tecnologia de Alimentos Assuring fruit quality is gaining attention in recent years. Several reports are available in the literature concerning variables that represent quality in fruits [START_REF] Scibisz | Mid-infrared spectroscopy as a tool for rapid determination of internal quality parameters in tomato[END_REF][START_REF] Tavakolian | Differentiation of post harvest date fruit varieties non-destructively using FT-NIR spectroscopy[END_REF][START_REF] Ribeiro | Prediction of linolenic and linoleic fatty acids content in flax seeds and flax seeds flours through the use of infrared reflectance spectroscopy and multivariate calibration[END_REF][START_REF] Oliveira | Comparison of nirs approach for prediction of internal quality traits in three fruit species[END_REF]. Among the most important variables are the sugar contents, due to their role in fruit quality. Analytical techniques have been developed to evaluate this parameter, however, traditional methods are not well adapted to on-line measurements and require an expensive and time consuming sample preparation. Most of them are based on solvent extraction followed by high performance liquid chromatography (HPLC) separation and/or colorimetric and enzymatic analyses [START_REF] Kelebek | HPLC determination of organic acids, sugars, phenolic compositions and antioxidant capacity of orange juice and orange wine made from a Turkish cv. Kozan[END_REF].

Previous researches have demonstrated the potential of MIR and NIR technology for quantitative characterization of fruits. Pedro and Ferreira (2007) applied the same methodology (NIR) to predict sugar content and acidity in tomato products. [START_REF] Bureau | Application of ATR-FTIR for a rapid and simultaneous determination of sugars and organic acids in apricot fruit[END_REF] used the MIR spectroscopy to determine sugar, organic acid and polyphenols contents in apricot and apples. The Passion fruit species belonging to the Passifloraceae family are mostly native at Latin America and are much used in juices, jams and yogurts manufacturing. In this context, the aim of this work was to assess the performance of NIR and MIR spectroscopy in determining individual sugar, in passion fruits in different ripening stages.

MATERIALS AND METHODS

Selection and preparation of passion fruit samples

Fifty-six yellow passion fruits (Passiflora edulis f. flavicarpa) in different ripening stages were collected in south of Brazil. Pulp was separated from the seed by filtration and then lyophilized. Lyophilized pulp was used in order to eliminate interferences caused by physical features of the fruit, as a recent report of our research group has pointed out [START_REF] Oliveira | Comparison of nirs approach for prediction of internal quality traits in three fruit species[END_REF]. Each sample was characterized in its spectral and biochemical analysis.

Spectra acquisition

A multi-purpose analyser (MPA) spectrometer (Bruker Optics, Wissembourg, France) in 2500 to 800 nm range was used for collecting NIR spectra in reflectance mode. The instrument was equipped with an integrating sphere to provide diffuse reflectance measurements and a TE-InGaAs detector. MPA was completely software-controlled by OPUS software Version 5.0, provided by Bruker Optics. Thirty-two scans were averaged for each sample spectrum. It was acquired with a 2 nm spectral resolution. Lyophilized pulps were transferred into glass vials and placed on automated 30-position sample wheel, each position corresponding to an 18 mm diameter hole. The average spectrum obtained with two measurements of powder from different aliquots was used in calculation.

MIR spectra were collected at room temperature with a Tensor 27 FTIR spectrometer (Bruker Optics, Wissembourg, France) equipped with a single-reflectance horizontal ATR cell with a diamond crystal (Golden Gate, Bruker Optics) and a deuterated triglycine sulfate (DTGS) detector. Lyophilized and homogenized samples were placed at the surface of the diamond crystal and were pressed with a system press tip flap. Samples were scanned at wavenumbers ranging from 4000 to 600 cm -1 and corrected against the background spectrum of air. Thirty-two scans were averaged for each sample spectrum. Two spectra, from two different aliquots of the powder, were performed for each sample. Average spectrum was used in model development.

Determination of soluble solids content

Sugars (glucose, fructose, sucrose) were quantified by enzymatic measurements with kits for food analysis (Boehringer Mannheim Co., Mannhein, Germany). The method was adapted on a 96 well-microplate [START_REF] Vermeir | High-Throughput Microplate Enzymatic Assays for Fast Sugar and Acid Quantification in Apple and Tomato[END_REF] using a spectrophotometer (FLX-Xenius, SAFAS, Monaco) equipped with an automatic injection device. Sugar contents were expressed in mg g -1 DW (dried weight).

Multivariate calibration

MatLab software package (version 6.5, Mathworks USA.) and Origin 6.1® (OriginLab Inc., Northampton, USA) were used for chemometric treatment of data. (principal component analysis) was initially performed for all samples (56 samples) to define calibration (80% samples) and external validation (20% samples) groups In this study, different pretreatments, including mean centering, multiplicative scatter correction (MSC), first-derivative and smoothing, were applied. Quantification models for sucrose, glucose, fructose contents were developed using PLS regression [START_REF] Coury | A method to quantify organic functional groups and inorganic compounds in ambient aerosols using attenuated total reflectance FTIR spectroscopy and multivariate chemometric techniques[END_REF]. Calibrations were performed in order to avoid overfitting data and obtain a reliable model. PLS regression and leaveone-out cross-validation were used in calibration step. Cross-validation estimated the prediction error by splitting all samples into two groups. Leave-one-out cross-validation (on 80% of data) involves using a single observation from the original sample as validation data and the remaining observations as training data. This process was repeated until all the samples had been used once in the validation set (Xie et al., 2007).

To evaluate the results, root mean square error of cross-validation (RMSECV), root mean square error of prediction (RMSEP), correlation coefficient of calibration (R 2 cv ) and of external validation (R 2 v ) were considered.

RESULTS

Chemical and Spectral diversity

Table 1 shows sugars contents determined in lyophilized pulp of passion fruit samples. According to these data, sucrose is the main sugar, presenting the highest content in passion fruit. [START_REF] Haard | Characteristics of edible plant tissues[END_REF] pointed out that main sugars in passion fruits are the three simple sugars: sucrose, glucose and fructose and that sucrose is the main sugar, followed by glucose and fructose. Moreover, these authors affirm that sugars represent about 80% of the soluble solids of passion fruit pulp. 1a andb, respectively. Full spectra presented characteristic peaks. [START_REF] Bureau | Effect of Sample Preparation on the Measurement of Sugars, Organic Acids, and Polyphenols in Apple Fruit by Mid-infrared Spectroscopy[END_REF][START_REF] Bureau | Application of ATR-FTIR for a rapid and simultaneous determination of sugars and organic acids in apricot fruit[END_REF][START_REF] Ruiz | Application of Reflectance Colorimeter Measurements and Infrared Spectroscopy Methods to Rapid and Nondestructive Evaluation of Carotenoids Content in Apricot (Prunus armeniaca L.)[END_REF]. In Figure 1a, MIR spectrum of lyophilized passion fruit pulp showed characteristic absorption zones in 1500-900 cm -1 corresponds to the major components, particularly sugars and organic acids [START_REF] Bureau | Application of ATR-FTIR for a rapid and simultaneous determination of sugars and organic acids in apricot fruit[END_REF][START_REF] Ruiz | Application of Reflectance Colorimeter Measurements and Infrared Spectroscopy Methods to Rapid and Nondestructive Evaluation of Carotenoids Content in Apricot (Prunus armeniaca L.)[END_REF]. Variation around the peak at 1200-900 cm -1 can be associated with C-H group and the peak at 1500-1200 cm -1 can be associated with C-O (Skoog et al., 2002), and may be related to the organic acids and sugars contents [START_REF] Scibisz | Mid-infrared spectroscopy as a tool for rapid determination of internal quality parameters in tomato[END_REF]. Regarding to the NIR spectrum (Figure 1b), four broad absorption peaks can be seen around 1190, 1450, 1790, and 2380 nm wavelengths. The peak at 1190 nm is in agreement with the second and third C-H overtone regions and the 1790 nm peak overlaps with the first C-H overtone region, both associated to sugar solutions (Skoog et al., 2002). The slight peak at 2300 nm falls within the region associated with C-H and O-H-combinations grouping. PCA performed with MID and NIR spectral data using mean center processing is presented in Figure 2. This analysis allowed detecting anomalous spectra and to identify specific sets of samples. In PCA sample map for MID spectra (Figure 2a), the first PC-score (PC1) discriminated the fruits in two sets. The first set, marked with a circle, corresponds to less sweet fruits, and the second set,

market with a square, to the sweetest fruits, with sugar concentration from 300.18 to 762.89 mg g -1 DW.

PCA for NIR spectra (Figure 2b) also identified sample 39, that is a high acid content sample, on the left side and sample 30, a high sugar content sample, on the right side. The specificity of bands is less in NIR range compared to MID [START_REF] Bellon-Maurel | Near-infrared (NIR) and mid-infrared (MIR) spectroscopic techniques for assessing the amount of carbon stock in soils e Critical review and research perspectives[END_REF]. Thus, in Figure 2b, there was no group separation and the difference among samples can be observed through a straight line, which presented an increasing in acid content. More specifically, sample 30 presented a low acid concentration and sample 10, a high acid concentration.

Quantification of individual sugars

A summary of prediction performance of calibration models developed for each chemical parameter in NIR and MID spectral ranges is shown in Table 2. The best results obtained for glucose (R 2 v = 0.941; RMSEP = 8.712%), fructose (R 2 v = 0.855; RMSEP = 8.340%) and sucrose (R 2 v = 0.818; RMSEP = 5.825%) were obtained with MID spectroscopy. Both reasons (spectral specificity and concentration range) contributed to the relative ease and precision of the prediction in the MID range. The number of latent variables for the models were very close. PLS regression models appeared to have an excellent ability to estimate sugars concentration, based on the very high R 2 v and low RMSEP results, which was in agreement with previous works [START_REF] Xie | Quantification of glucose, fructose and sucrose in bayberry juice by NIR and PLS[END_REF][START_REF] Rambla | PLS-NIR determination of total sugar, glucose, fructose and sucrose in aqueous solutions of fruit juices[END_REF].

CONCLUSION

The applicability of spectroscopic techniques in MID and NIR range combined with PLS regression was evaluated for the determination of individual sugars, in lyophilized passion fruit pulp. Calibration models developed in this work covered different maturation stages of passion fruit and thus a wide range of compositions.

In this work, MID spectroscopy was considered better for determination quality parameters in passion fruit. This fact can be attributed primarily because MID spectroscopy contains wavelengths for fundamental rotational molecular vibration, while overtone and combination bands of fundamental transition are associated with near infrared spectrum, making it less reproducible and specific. A second reason that can be assigned is that mid-infrared region is highly sensitive to precise chemical composition of samples compared to near infrared technique.

Figure 1 -

 1 Figure 1 -Spectrum of lyophilized passion fruit pulp: MID (a) and NIR (b)

Figure 2 -

 2 Figure 2 -PCA performed with MID (a) and NIR (b) spectral data.

Table 1 -

 1 Quality traits (mg g -1 DW) of passion-fruit samples.

	Parameter	Range	Median	Standard deviation
	Glucose	23.64 -217.38	96.34	±38.98
	Fructose	28.37 -240.03	97.59	±46.19
	Sucrose	40.43 -396.22	293.92	±65.03
	MID and NIR mean spectra of passion fruit without any treatment are shown in the Figures

Table 2 -

 2 Performance of PLS regression using the MID (cm -1 ) and NIR (nm) spectral ranges for non destructive composition assessment (mg g -1 DW) of freeze-dried passion fruit pulp.

	Parameter	Spectral range	Outliers LV*	Wavelength range	R 2	cv	RMSECV a R 2	v	RMSEP %
	Glucose	MID NIR	2 2	5 6	4000-600 2500-1100	0.945 0.876	13.126 21.102	0.941 0.862	8.712 9.645
	Fructose	MID NIR	2 1	5 5	4000-600 2500-1100	0.934 0.821	15.773 21.343	0.855 0.756	8.340 11.685
	Sucrose	MID NIR	2 1	6 6	4000-600 2500-1170	0.889 0.722	28.531 46.191	0.818 0.691	5.825 7.838

*LV: number of latent variables

Área temática: Engenharia e Tecnologia de Alimentos


Anaïs Le Mauff

Hamza Chouikh

Alison Cartereau

Claude Charvet

Cédric Neveu

Claude Rispe

Olivier Plantard

Emiliane Taillebois

Steeve H Thany

Cedric Neveu

Microtransplantation of tick transmembrane proteins as a

à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Introduction

Ticks are known as harmful parasites and vectors of many microorganisms that could be pathogenic for animal and human. For example, the castor bean tick, Ixodes ricinus, can transmit diseases like Lyme borreliosis and tick-borne encephalitis, becoming a worldwide threat to public health (Boulanger et al., 2019). Due to the lack of effective vaccines or other non-chemical based methods, acaricides remain the main way for tick control. However, there is now increasing evidence of resistance development against currently used acaricides ( Kumar et al., 2019). Thus, new compounds are needed and one strategy could be to target tick neuronal nicotinic acetylcholine receptors (nAChRs). In the present work, we aim to validate the membrane microtransplantation technique as a tool to study the pharmacological profiles of the tick nAChRs expressed in the synganglion. This technique allows to microtransplant the transmembrane receptors maintained in their natural lipidic environment. Thus, the studied native receptors keep their natural properties and regulation (Eusebi et al., 2009).

Conclusion

• We validated and adapted the membrane microtransplantation protocol for the pharmacological characterisation of tick native nAChRs.

• We applied this technique for the screening of four neonicotinoids. Preliminary results highlighted that tick nAChRs are highly sensitive to clothianidin.

• To complete this study, we aim to adapt this technique to other arthropod species and for the screening of other molecules.

This study gives promising results for the screening and selection of new anti-parasitic compounds targeting arthropod transmembrane proteins. We tested the activation of tick nAChRs from synganglion membrane preparation by acetylcholine (ACh) and nicotine (Nic). Both molecules are able to activate the receptors, and nAChRs are more sensitive to nicotine than to acetylcholine. These pharmacological properties are typical of nAChRs. We tested the receptor sensitivity to currently used neonicotinoids: clothianidin (CLT), thiametoxam (TMX), acetamiprid (ACE), imidacloprid (IMI). TMX, ACE, IMI induced similar currents that are lower than nicotine-induced current. On the contrary, CLT is able to induce higher current than those induced by Nic. These results show that tick native nAChRs are differently sensitive to neonicotinoids according to molecules.

  Boulanger N., Boyer P., Talagrand-Reboul E., Hansmann Y., (2019) Ticks and tick-borne diseases. Médecine et maladies infectieuses, 49, pp 87-97. Eusebi F., Palma E., Amici M., Miledi R., (2009) Microtransplantation of ligand-gated receptor-channels from fresh or frozen nervous tissue into Xenopus oocytes: A potent tool for expanding functional information. Progress in Neurobiology, 88, 32-40. Kumar R., (2019) Molecular markers and their application in the monitoring of acaricide resistance in Rhipicephalus microplus. Experimental and Applied Acarology, 78, pp 149-172.Ixodes ricinusElectrophysiological recordings of nAChRs expressed on the oocyte membranePurified membranes solutionTo validate the membrane preparation protocol we used α-bungarotoxin conjugated with Alexa Fluor®488 to label functional nAChRs. The membrane extraction and purification protocol consists in two crucial steps:• The first is the dissociation of the neuronal tissues of I. ricinus (synganglion) and the precipitation of the transmembrane proteins by ultracentrifugation. • The second is the purification of the membrane containing the transmembrane proteins of interest by using a sucrose density gradient.Injected oocytes are placed into an electrophysiology recording system. The two-electrode voltage-clamp (TEVC) technique is used by setting two microelectrodes on the oocyte membrane. One electrode clamp the membrane potential to -60 mV, the second one records the change in voltage after application of molecules. The response resulting from the opening of channels (response of the nAChRs) expressed on the oocyte membrane is recording.

( a )

 a Example of currents obtained after application of ACh and Nic at 1 mM. (b) Effect of the ACh compared with the response induced by 1 mM Nic application; acetylcholine-induced current are smaller than nicotine-induced current (results are the mean of 13 independent experiments; error bars correspond to SEM).

  Ixodes ricinus are sensitive to currently used neonicotinoids (c) Example of currents obtained by application of CLT, TMX, ACE, IMI at 1 mM. (d) Comparison of the maximum currents obtained with different neonicotinoids. Currents are normalised with the nicotine-induced current. Results are the mean of 6 to 9 independant experiments, error bars correspond to SEM.

  An incubation time of 12 to 24h, at 18°C, allows the recycling of tick membrane vesicles. At the end of this step native tick nAChRs are present at the oocyte membrane.Fluorenscent visualisation of the α-bungarotoxin-sensitive nAChRs using αbungarotoxin conjugated with Alexa Fluor®488 in membrane preparation from the tick Ixodes ricinus. (a) membranes labeled with α-bungarotoxin conjugated with Alexa Fluor®488, each fluorescent spot highlighted the presence of functional receptors; (b) same preparation with transmitted light showing the membrane aggregates.Validation of the nAChRs expression on the oocyte membrane Native nAChRs are differently modulated by neonicotinoids according to molecules.Injection of tick membrane solution is made with a glass micropipette using a nanoinjector.


come

Reconstructing colonization routes of invasive species from molecular data: case studies in forest entomology

INRA, UMR CBGP 755 avenue du Campus Agropolis, CS30016 34988 Montferrier-sur-Lez Cedex, France Tel.: +33 4 30 63 04 35 Email: carole.kerdelhue@supagro.inra.fr

The authors belong to a research consortium dealing with insect evolutionary biology and natural genetic structure at different scales. They were involved in various projects studying rangewide phylogeography of several species of forest pests (mainly seed chalcids, scolytids, processionary moths) in Europe and the Mediterranean Basin, and started to collaborate on host-parasitoid systems. In the recent years, they focused on the question of biological invasions, either involving geographic expansions of native species due to climate and landscape changes, or accidental introductions from different regions.

Reconstructing colonization routes of invasive species from molecular data: case studies in forest entomology

Deciphering the colonization processes by which introduced pests invade new areas is essential to limit the risk of further expansion and/or multiple introductions by increasing vigilance against the identified key source populations. It also helps defining the ecological characteristics of introduced populations and predicting the potential extent of their distribution areas. In some cases, it can help choosing strains of potential auxiliary agents to develop biological control strategies. Yet, historical and observational data often provide incomplete, sparse or even misleading information on invasive populations' history. In the last decade, population genetics has been used as an indirect tool to reconstruct routes of introduction, highlighting the complexity and the sometimes counterintuitive nature of the true story. The recent development of new model-based methods, such as approximate Bayesian computation (ABC), has allowed quantitative inferences in case of the complex evolutionary scenarios typically encountered during biological invasions. It specifically allows to compare alternative scenarios regarding the number and genetic composition of sources and to explore the number of successive introduction events from each source, the number of introduced individuals and the dynamics of demographic expansion after each introduction. We describe the principles of the ABC analyses, here applied to microsatellite data and mitochondrial sequences of populations sampled within the native and the invasive range of a species. We will use three main examples in forest entomology, from which we obtained valuable information about the colonization routes and dispersal patterns, namely the cedar seed chalcid Megastigmus schimitscheki, the maritime pine bast scale Matscucoccus feytaudi and the Western conifer seed bug Leptoglossus occidentalis.

  L E S T U D I U M ® CONFERENCES Insect invasions in a changing world | 17-19 December 2014


Maxime Olmos
email: maxime.olmos@agrocampus-ouest.fr

Félix Massiot-Granier

Etienne Prévost

Marie Nevoux

Gerald Chaput

Etienne Rivot

A Bayesian life cycle model to unravel changes in the marine productivity of Atlantic salmon population in the North Atlantic Ocean

Keywords: Atlantic salmon, Bayesian Hierarchical Model, integrated population models, life cycle, population dynamics, life history traits

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


Flavia Pinzari

Loredana Canfora

Alessandro Florio

Melania Migliore

Barbara Felici

Maria Teresa Dell'abate

Rosario Napoli

Fungal translocation of microelements during Fagusand Quercu sleaf litter decomposition in a volcanic soil ecosystem

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


domain: sdv sde

text:
J J Godon

A Galès

E Latrille

J P Steyer

N Wéry

Gradient of bioaerosol impaction/deposition defined by biomimetic passive sensors

Keywords: sensor, deposition, bioaerosol, qPCR

Bioaerosols by their biological nature are potential vectors of pathogens, toxins and thus have a health impact on residents and workers. They also have an ecological and economic impact in the case, for example, of contamination of farm sites and production of fish, crustaceans, shellfish and microalgae.

Currently used bioaerosol sensors are volumetric sensors 'instantaneous' (short periods of time). These sensors cannot take into account the multiple settings that change the dispersion and determine the impacted area. These parameters are variations in the level of emissions, weather conditions (sunshine, temperature, humidity, etc.) and the topography of the site. In the absence of tools, the areas impacted and the level of impact by bioaerosol emission sources are not currently mappable and therefore mapped. To overcome these limitations, we developed the concept of passive sensors for the environmental monitoring of bioaerosols.

On a first trail, we used pine needles as collection surface of bioaerosols released by a composting plant located in the South of France. We quantified the microbial compost indicator Saccharopolyspora rectivirgula (LeGoff, 2010). Samples were collected at distances ranging from 100 m to 4000 m around the composting plant. 16S rDNA abundance from S. rectivirgula and the total bacteria were measured by qPCR to indicate the impact of composting plant bioaerosol. The abundance of 16S rDNA of S. rectivirgula varies from 10 2 to 10 4 copies by g of P. halepensis needles (Fig. 1).

Figure 1: Map corresponding to predicted bioindicator concentration in 16S rRNA gene copies.g -1 of pine needles. White spots correspond to sampled trees. Narrow and thick black lines correspond to altitude and background level contour lines, respectively. The cross corresponds to the location of the composting plant.

The abundance of S. rectivirgula on P. halepensis needles were negatively correlated with distance to the point source. Topography and prevalent wind direction had an influence on the dispersion pattern. The abundance of indicator organism has reached the background level at distances ranging from 2 km to more than 5.4 km from the composting plant (Galès, 2014).

As the pines are not available everywhere, a second trail was performed using bottlebrushes to mimic pine needles. Bottlebrushes located close to a bioaerosol emission source (microalgae pond) collected over time showed an accumulation of bacteria and microalgae (Fig. 2). Accumulation level was similar between leave (indeterminate time) and bottlebrush (up to 20 days). Thus, nowadays we have the tools necessary to draw a map of the areas impacted by any source of bioaerosols. To do that we need (i) the analysis plan (relevant distance between the source and the sensor), (ii) a sensor (bottlebrushes), (iii) a specific microbial indicator (depending of the bioaerosol source), (iv) a qPCR measuring system (depending of the indicator).

Le Goff O, Bru-Adan V, Bacheley H, Godon J-J, Wery N. The microbial signature of aerosols produced during the thermophilic phase of composting. J Appl Microbiol. 2010;108: 325-340. Gales A, Latrille E, Wery N, Steyer J-P, Godon J-J.

Needles of Pinus halepensis as Biomonitors of Bioaerosol Emissions. PLoS One. 2014; 9:

Figure 2 :

 2 Figure 2: accumulation of bacteria over time on bottlebrushes and leaves around a microalgae pond.