Jean-Marc Bardet
email: jean-marc.bardet@univ-paris1.fr

Imen Kammoun
email: imen.kammoun@univ-paris1.fr

Detecting abrupt changes of the long-range dependence or the self-similarity of a Gaussian process

In this paper, an estimator of m instants (m is known) of abrupt changes of the parameter of long-range dependence or self-similarity is proved to satisfy a limit theorem with an explicit convergence rate for a sample of a Gaussian process. In each estimated zone where the parameter is supposed not to change, a central limit theorem is established for the parameter's (of long-range dependence, self-similarity) estimator and a goodness-of-fit test is also built.

technique base sur le priodogramme. Notre approche est fonde sur l'analyse par ondelettes, ce qui prsente plusieurs avantages : c'est une technique non-paramtrique applicable pour des processus trs gnraux, robuste aux tendances polynomiales et, au moins dans le cadre gaussien, s'accompagnant de tests d'adquation de type χ 2 simples et intressants utiliser. Ainsi, un estimateur des m points de ruptures (m ∈ AE * , suppos connu) de la longue mémoire ou d'auto-similarité est conçu pour un échantillon de processus gaussien en se basant sur l'analyse par ondelettes, ce qui permet ensuite de mettre en place des tests d'adquation. Pour ce type de processus, cette méthode a été proposée pour la première fois dans [START_REF] Flandrin | Wavelet analysis and synthesis of fractional Brownian motion[END_REF], puis développée par exemple dans [START_REF] Abry | Long-range dependent: revisiting aggregation with wavelets[END_REF]. La convergence des estimateurs basés sur les ondelettes a été étudiée dans le cas du mouvement brownien fractionnaire (FBM) dans [START_REF] Bardet | Statistical Study of the Wavelet Analysis of Fractional Brownian Motion[END_REF], et dans un cadre semi-paramétrique général de processus gaussiens stationnaires à longue mémoire par [START_REF] Moulines | On the spectral density of the wavelet coefficients of long memory time series with application to the log-regression estimation of the memory parameter[END_REF] et [START_REF] Bardet | Adaptative wavelet based estimator of the memory parameter for stationary Gaussian processes[END_REF]. Ici le principe de l'estimation du paramètre de longue mémoire ou d'auto-similarité est le suivant : dans chaque zone où il n'y a pas de changement, ce paramètre peut être estimé à partir d'une log-log régression de la variance des coefficients d'ondelettes sur plusieurs échelles choisies (voir (1)). Une fonction de contraste définie par la somme des carrés des distances entre ces points et les droites d'ajustement, dans les m + 1 zones possibles détectées, est minimisée (voir (2)), donnant un estimateur des points de ruptures (voir (3)). Sous certaines hypothèses générales, on montre qu'il vérifie un théorème limite avec une vitesse de convergence explicite (voir Theorem 1.1). Dans chacune des zones détectées, les paramètres de longue mémoire, (ou d'auto-similarité) peuvent être estimés, tout d'abord avec la regression des moindres carrés ordinaires (OLS), puis par une regression des moindres carrés pseudo-généralisés (FGLS). Un théorème de la limite centrale est établi pour chacun des deux estimateurs (voir Theorem 1.2 et Proposition 1.1 ci-dessous) et des intervalles de confiance peuvent être calculés. L'estimateur FGLS offre deux avantages : d'une part, sa variance asymptotique est plus petite que celle de l'estimateur OLS, et d'autre part, il permet la construction d'un test d'ajustement très simple basé sur le carré des distances entre les points (d'abscisse, le logarithme d'une échelle choisie et d'ordonnée, le logarithme de la variance empirique des coefficients d'ondelettes pour cette échelle) et les droites de régression pseudo-généralisée correspondantes (voir (8)). La convergence vers une distribution du Chi-deux de ce test est établie dans le Theorem 1.3. Deux cas particuliers de processus gaussiens sont ensuite étudiés dans la section 2. En premier lieu, on s'intéresse aux séries chronologiques stationnaires longue mémoire avec un paramètre de Hurst constant par morceaux. On se place dans un cadre semi-paramétrique contenant par exemple les FGN et les processus FARIMA (voir Figure 1). En second lieu, le cas d'un processus à accroissements stationnaires et autosimilaire par morceaux est traité, ce qui revient à considérer des successions de FBM ayant des exposants de Hurst distincts (voir Figure 1). Pour ces deux exemples, les vitesses de convergence explicites des différents estimateurs et tests sont données et des simulations montrent leurs qualités (voir Table 1). D'autres simulations, preuves des thormes ainsi qu'un exemple plus gnral de dtection de ruptures dans le cadre de processus gaussien localement fractionnaire sont dtaills dans [START_REF] Bardet | Detecting changes in the fluctuations of a Gaussian process and an application to heartbeat time series[END_REF].

Assumptions and main results

Let (X t ) t∈AE be a Gaussian process and assume that X 0 , X 1 , . . . , X N is known. In the sequel, X will be a piecewise stationary long memory time series or a piecewise self-similar time series having stationary increments. Consider ψ : Ê → Ê a function called "the mother wavelet". For (a, b)

∈ Ê * + × Ê, the wavelet coefficient of X for the scale a and the shift b is d X (a, b) := 1 √ a Ê ψ( t-b a )X(t)dt. When only a discretized path of X is available, approximations e X (a, b) are only computable: , e X (a, b) := 1 √ a N p=1 ψ p -b a N X p for (a, b) ∈ Ê * + × AE.
Assume that there exist m ∈ AE (the number of abrupt changes) and

• 0 = τ * 0 < τ * 1 < . . . < τ * m < τ * m+1 = 1 (unknown parameters); • two families (α * j ) 0≤j≤m ∈ Ê m+1 and (β * j ) 0≤j≤m ∈ (0, ∞) m+1 (unknown parameters);
• a sequence of "scales" (a n ) n∈AE ∈ Ê AE (chosen) satisfying a n ≥ a min , with a min > 0, such that for j = 0, 1, . . . , m and k

∈ D * N (j) ⊂ [N τ * j ], [N τ * j+1 ] , e 2 X (a N , k) ∼ β * j a N α * j when N → ∞.
A piecewise sample variance can be the appropriated estimator of such power law. Thus, define

S k ′ k (a N ) := a N k ′ -k [k ′ /aN ]-1 p=[k/aN ] e 2 X (a N , a N p) for 0 ≤ k < k ′ ≤ N.
Now set 0 < r 1 < . . . < r ℓ with ℓ ∈ AE * , and assume that a multidimensional central limit theorem can be established for log

S k ′ k (r i a N ) 1≤i≤ℓ , when [N τ * j ] ≤ k < k ′ ≤ [N τ * j+1 ], i.e. k ′ -k a N log S k ′ k (r i a N ) -log(β * j ) -α * j log r i a N 1≤i≤ℓ L -→ N →∞ |k ′ -k|→∞ N 0, Γ (j) (α * j , r 1 , . . . , r ℓ ) , (1)
with

Γ (j) (α * j , r 1 , . . . , r ℓ ) = γ (j)
pq 1≤p,q≤ℓ a (ℓ×ℓ) matrix not depending on N such that α → Γ (j) (α, r 1 , . . . , r ℓ ) is a continuous function and a positive matrix for all α. Define a contrast function

U N (α j ) 0≤j≤m , (β j ) 0≤j≤m , (k j ) 1≤j≤m = m j=0 ℓ i=1 log S kj+1 kj (r i a N ) -α j log(r i a N ) + log β j 2 (2) with (α j ) 0≤j≤m ∈ A m+1 ⊂ Ê m+1 , (β j ) 0≤j≤m ∈ B m+1 ⊂ (0, ∞) m+1 , 0 = k 0 < k 1 < . . . < k m < k m+1 = N, (k j ) 1≤j≤m ∈ K m (N ) ⊂ AE m .
The vector of estimated parameters α j , β j and k j (and therefore τ j ) is the vector which minimizes this contrast function in A m+1 × B m+1 × K m (N ), i.e., ( α j ) 0≤j≤m , ( β j ) 0≤j≤m , ( k j ) 1≤j≤m := Argmin U N (α j ) 0≤j≤m , (β j ) 0≤j≤m , (k j ) 1≤j≤m

(3)

τ j := k j /N for 1 ≤ j ≤ m. (4)
For a given (k j ) 1≤j≤m , it is obvious that ( α j ) 0≤j≤m and (log β j ) 0≤j≤m are obtained from a log-log regression of

S kj+1 kj (r i a N ) i onto r i a N i , i.e.   α j log β j   = L ′ 1 • L 1 ) -1 L ′ 1 • Y kj+1 kj with Y kj+1 kj := log S kj+1 kj (r i • a N ) 1≤i≤ℓ , L aN :=     log(r 1 a N ) 1 . . . . . . log(r ℓ a N ) 1     . Therefore ( k j ) 1≤j≤m = Argmin U N ( α j ) 0≤j≤m , ( β j ) 0≤j≤m , (k j ) 1≤j≤m , (k j ) 1≤j≤m ∈ K m (N ) .
Remark 1 In this paper, m is supposed to be known. However, if m is unknown, as in [START_REF] Lavielle | Detection of multiple changes in a sequence of dependent variables[END_REF] or [START_REF] Lavielle | The multiple change-points problem for the spectral distribution[END_REF], a penalized contrast Ũm,N = U N + β N × m (with β N an appropriated sequence converging to 0) can be used instead of U N , and by adding a minimization in m, an estimator m of m could be also deduced.

In this paper, parameters (α * j ) are supposed to satisfied abrupt changes: Assumption C : Parameters (α * j ) are such that |α * j+1α * j | = 0 for all j = 0, 1, . . . , m -1. j ) 0≤j≤m such that α * j ∈ [a , a ′ ] and a < a ′ for all j = 0, . . . , m, then if a

1+2(a ′ -a) N N -1 -→ N →∞ 0, for all (v n ) n satisfying v N • a 1+2(a ′ -a) N N -1 -→ N →∞ 0, È v N τ * -τ m ≥ η -→ N →∞
0 for all η > 0.

(

) 5
Remark 2 The proof of this result is provided in [START_REF] Bardet | Detecting changes in the fluctuations of a Gaussian process and an application to heartbeat time series[END_REF]. Unfortunately, the rate of convergence of τ *τ m is only v N = N α with 0 < α < 1 and not N as, for instance, in [START_REF] Lavielle | Detection of multiple changes in a sequence of dependent variables[END_REF] and [START_REF] Lavielle | The multiple change-points problem for the spectral distribution[END_REF]. However the context is not the same: in these papers, the contrast is directly computed from N values of (X) i which do not change following τ . Here, the contrast is computed from only (m + 1)ℓ values of S N which change following τ .

The rate of convergence N can not be reached in such a context (simulations show also this property). This is certainly a drawback of your method, which hopefully does not change the rate of convergence of parameters (α j ) and (β j ).

For j = 0, 1, . . . , m, the log-log regression of S kj+1 kj (r i a N ) 1≤i≤ℓ onto (r i a N ) 1≤i≤ℓ provides estimators of α * j and β * j . However, if τ j converges to τ * j , k j = N • τ j does not converge to k * j , and therefore

È [ k j , k j+1 ] ⊂ [k * j , k * j+1 ]
does not tend to 1. So, define kj and k′ j such that kj = k j + N vN and k′ j = k j+1 -N vN . From ( 5)

with η = 1/2, È [ kj , k′ j ] ⊂ [k * j , k * j+1 ] -→ N →∞ 1. Then, Theorem 1.2 Let Θ * j := α * j log β * j and Θj := (L ′ 1 L 1 ) -1 L ′ 1 Y k′ j kj = αj log βj
. Under the same assumptions as in Theorem 1.1, for j = 0, . . . , m, with Σ (j) (α * j , r 1 , . . . , r ℓ ) := (L

′ 1 L 1 ) -1 L ′ 1 Γ (j) (α * j , r 1 , . . . , r ℓ )L 1 (L ′ 1 L 1 ) -1 , N τ * j+1 -τ * j a N Θj -Θ * j L -→ N →∞ N 0, Σ (j) (α * j , r 1 , . . . , r ℓ ) (6)
A second estimator of Θ * j can be obtained from feasible generalized least squares (FGLS) estimation. Indeed, the asymptotic covariance matrix Γ (j) (α * j , r 1 , . . . , r ℓ ) can be estimated by the matrix Γ(j) := Γ (j) (α j , r 1 , . . . , r ℓ ) and Γ(j) P -→

N →∞ Γ (j) (α * j , r 1 , . . . , r ℓ ). Then, the FGLS estimator Θ j of Θ * j is defined from the minimization among all Θ of the following squared distance,

Y k′ j kj -L aN • Θ 2 Γ(j) = Y k′ j kj -L aN Θ ′ • Γ(j) -1 • Y k′ j kj -L aN Θ .
and therefore define Θ

j := L ′ 1 Γ(j) -1 L 1 -1 L ′ 1 Γ(j) -1 Y k′ j kj .
Proposition 1.1 Under the same assumptions as in Theorem 1.2, for j = 0, . . . , m

N τ * j+1 -τ * j a N Θ j -Θ * j L -→ N →∞ N 0, M (j) (α * j , r 1 , . . . , r ℓ ) (7) with M (j) (α * j , r 1 , . . . , r ℓ ) := L ′ 1 Γ (j) (α * j , r 1 , . . . , r ℓ ) -1 L 1 -1 ≤ Σ (j) (α * j , r 1 , . . . ,

r ℓ ) (for the order's relation between positive symmetric matrix).

Therefore α j is more accurate than αj for estimating α * j when N is large enough. For j = 0, . . . , m, let T (j) be the FGLS distance between points log(r i a N ), log S

T (j) = N τ * j+1 -τ * j a N Y k′ j kj -L aN Θ j 2 Γ(j) L -→ N →∞ χ 2 (ℓ -2). (8)
2. Applications

Piecewise long memory Gaussian processes

Assume that the process X = (X t ) t∈AE is a Gaussian piecewise long-range dependent (LRD) process, i.e. there exists (D * j ) 0≤j≤m ∈ (0, 1) m+1 and for all j = 0, . . . , m and k

∈ [N τ * j ], [N τ * j ] + 1, . . . , [N τ * j+1 ] -1 , X k = X (j) k-[N τ * j ]
, where X (j) = (X (j) t ) t∈AE satisfies the following Assumption LRD(D * j ).

Assumption LRD(D): Y is a centered stationary Gaussian process with spectral density f such that

f (λ) = |λ| -D • f * (λ) for all λ ∈ [-π, π] \ {0} with f * (0) > 0 and with C 2 > 0, |f * (λ) -f * (0)| ≤ C 2 • |λ| 2 for all λ ∈ [-π, π].
Following [START_REF] Bardet | Adaptative wavelet based estimator of the memory parameter for stationary Gaussian processes[END_REF], if the mother wavelet is supposed to be included in a Sobolev ball, then

Corollary 1 Let X be a Gaussian piecewise LRD process defined as above and ψ : Ê → Ê be [0, 1]supported with ψ(0) = ψ(1) = 0 and 1 0 ψ(t) dt = 0 and such that there exists sequence (ψ ℓ ) ℓ∈ satisfying

ψ(λ) = ℓ∈ ψ ℓ e 2πiℓλ ∈ Ä 2 ([0, 1]) and ℓ∈ (1 + |ℓ|) 5/2 |ψ ℓ | < ∞.
Under Assumption C, for all 0 < κ < 2/15, if a N = N κ+1/5 and v N = N 2/5-3κ then ( 5), ( 6), ( 7) and ( 8) hold.

Thus, the rate of convergence of τ to τ * (in probability) is N 2/5-3κ for 0 < κ arbitrary small. Estimators Dj and D j converge to the parameters D * j following a central limit theorem with a rate of convergence N 2/5-κ/2 for 0 < κ. Convincing results of simulations can be observed in Table 1 and Figure 1. 1 Left: Estimation of τ 1 , D 0 and D 1 in the case of piecewise FARIMA(0,d j ,0) (d 1 = 0.1 and d 2 = 0.4) with one change point when N = 20000 (50 realizations). Right: Estimation of τ 1 , τ 2 , H 0 , H 1 and H 2 in the case of piecewise FBM with two change points when N = 5000 and N = 10000 (50 realizations)

Piecewise fractional Brownian motions

Now, X will be called a piecewise fractional Brownian motion if there exist two families of parameters (H * j ) 0≤j≤m ∈ (0, 1) m+1 and (σ * 2 j ) 0≤j≤m ∈ (0, ∞) m+1 such that for all j = 0, . . . , m and t ∈

[N τ * j ], [N τ * j ] + 1, . . . , [N τ * j+1 ] -1 , X t = X (j) t-[N τ * j ]
, where X (j) = (X (j) t ) t∈Ê is a FBM with parameters H * j and σ * 2 j . Following the results of [START_REF] Bardet | Statistical Study of the Wavelet Analysis of Fractional Brownian Motion[END_REF], one obtains, Corollary 2 Let X be a piecewise FBM and ψ : Ê → Ê be a piecewise continuous and left (or right)differentiable, such that |ψ ′ (t -)| is Riemann integrable with ψ ′ (t -) the left-derivative of ψ in t, with support included in [0, 1] and Ê t p ψ(t) dt = 1 0 t p ψ(t) dt = 0 for p = 0, 1.. 5), ( 6), ( 7) and ( 8) hold.

Let A := sup j H * j -inf j H * j . If A < 1/2, under Assumption C, for all 0 < κ < 1 1+4A -1 3 , if a N = N 1/3+κ and v N = N 2/3(1-2A)-κ(2+4A) then (

Remark 3

The dependence of this result on A can be explained by the fact that 2(sup j α * jinf j α * j ) + 1, with α * j = 2H * j + 1, has to be smaller than 3 since a N • N -1/3 -→ N →∞ ∞. However, Corollary 2 is quite surprising: the smaller A, i.e. the smaller the differences between the parameters H j , the faster the convergence rates of estimators τ j to τ * j . If the difference between two successive parameters H j is too large, the estimators τ j do not seem to converge. This is attributable to the influence of the other segments that is even deeper than the involved exponents are different (simulations exhibit this paroxysm in [START_REF] Bardet | Detecting changes in the fluctuations of a Gaussian process and an application to heartbeat time series[END_REF]).

Thus, the rate of convergence of τ to τ * (in probability) can be N 2/3(1-2A)-κ ′ for 0 < κ ′ as small as one wants when a N = N 1/3+κ ′ /(2+4A) . Results of simulations can be observed in Table 1 and Figure 1

Theorem 1 . 1

 11 Define τ * := (τ * 1 , . . . , τ * m ), τ := ( τ 1 , . . . , τ m ) and τ m := max |τ 1 |, . . . , |τ m | . Let ℓ ∈ AE \ {0, 1, 2}. If Assumption C and relation (1) holds with (α *

Theorem 1 . 3

 13 regression line. The following theorem describes the asymptotic behavior of a goodness-of-fit test on each segment [ kj , k′ j [: Under the same assumptions as in Theorem 1.1, for j = 0, . . . , m

Figure 1 .

 1 Figure1. Left: Piecewise FARIMA(0,d j ,0) (with d 0 : 0.1 (D 0 : 0.2), d 1 : 0.4 (D 1 : 0.8) and τ 1 : 0.75). Right: Piecewice FBM(H j ) (τ 1 : 0.3, τ 2 : 0.78, H 0 : 0.6, H 1 : 0.8 and H 2 : 0.5), ( τ 1 : 0.32, τ 2 : 0.77, H0 : 0.5608, H1 : 0.7814 and H2 : 0.4751).

  in a case where A = 0.3 < 1/2.

	6												1.2
													1
	4
													0.8
	2
							Estim. τ 1 : 0.7504					0.6
	0			Estim. D 1 : 0.2083					Estim. D 2 : 0.7510	0.4
	-2												0.2
													0
	-4
													-0.2
	-6	0	0.2	0.4	0.6	0.8	1	1.2	1.4	1.6	1.8	2 4 x 10	-0.4	0	1000	2000	3000	4000	5000	6000	7000	8000	9000	10000


Marion Dohen
email: marion.dohen@icp.inpg.fr

Hélène Loevenbruck
email: helene.loevenbruck@icp.inpg.fr

Auditory-Visual Perception of Acoustically Degraded Prosodic Contrastive Focus in French

Keywords: audiovisual, prosody, perception, whispered speech, contrastive focus

Previous studies have shown that visual only perception of prosodic contrastive focus in French is possible. The aim of this study was to determine whether the visual modality could be combined to the auditory one and lead to a perceptual enhancement of prosodic focus. In order to examine this question, we carried out auditory only, audiovisual and visual only perception tests. In order to avoid a ceiling effect, auditory only perception of prosodic focus being very high, we used whispered speech for which the acoustic prosodic information is degraded. The productions of two different speakers were used. This test showed that adding the visual modality enhances auditory perception of prosodic focus when the acoustic prosodic cues are degraded.

Introduction

Adding the visual modality to the auditory one enhances speech perception. Several studies have shown that audiovisual perception is better than auditory alone perception of speech in noise for example ( [START_REF] Sumby | Visual contribution to Speech Intelligibility in Noise[END_REF][START_REF] Miller | An Analysis of Perceptual Confusions among some English Consonants[END_REF][START_REF] Neely | Effects of visual factors on the intelligibility of speech[END_REF][START_REF] Erber | Auditory-visual perception of speech[END_REF][START_REF] Binnie | Auditory and visual contributions to the perception of consonants[END_REF][START_REF] Summerfield | Use of visual information for phonetic perception[END_REF][START_REF] Macleod | Quantifying the contribution of vision to speech perception in noise[END_REF][START_REF] Grant | Evaluating the Articulation Index for audiovisual input[END_REF][START_REF] Benoît | Effects of Phonetic Context on Audio-Visual Intelligibility of French[END_REF]). An audiovisual advantage was also observed in silent environment for the perception of a non-native language or of semantically complex utterances ( [START_REF] Reisberg | Easy to Hear but Hard to Understand: A Lip-reading Advantage with Intact Auditory Stimuli[END_REF]). These studies dealt with segmental perception of speech i.e. identifying what is being said. It seems reasonable to think that the visual modality could also be useful for the perception of prosody i.e. for the perception of supra-segmental speech features. In previous studies ( [START_REF] Dohen | Audiovisual Production and Perception of Contrastive Focus in French: a multispeaker study[END_REF]), we showed that it is possible to detect prosodic contrastive focus in French from the visual modality alone. This showed that there are potential visual cues to prosodic focus perception. It is however unsure whether the audio and visual modalities can interact to enhance audiovisual perception of prosody.

Swerts & Krahmer examined the possible interactions between the auditory and visual modalities during the perception of focus in Dutch [START_REF] Swerts | Congruent and Incongruent Audiovisual Cues to Prominence[END_REF][START_REF] Swerts | Cognitive processing of audiovisual cues to prominence[END_REF]. Both studies showed that there seemed to be visual information to prosodic focus for Dutch, just as we observed for French ( [START_REF] Dohen | Audiovisual Production and Perception of Contrastive Focus in French: a multispeaker study[END_REF]). The authors observed that the focus perception process appears to result from a combination of the information coming from the auditory and the visual modalities leading to a perceptual decision. When the auditory and visual modalities are conflicting, perception appears to be more difficult and slower. The authors also found that the upper and left parts of the speakers' faces seemed to provide more visual information. We suggest that this may be partly due to the unnaturalness of the task used for recording the material from the speakers (especially the one used to trigger focus) which could have resulted in exaggerated facial gestures or unnatural facial motion. These two studies provide very interesting preliminary information concerning the interactions between the auditory and visual modalities and their relative importance in perception.

The aim of the present study was to test the bimodal perception of French prosodic contrastive focus and to see whether the visual modality, added to the auditory one, could contribute to enhance perception.

Measuring enhancement: the "ceiling effect" problem

It has already been shown that auditory only perception of prosodic contrastive focus in French reaches near to perfect identification scores (e.g., [START_REF] Dahan | Interspeaker Variability in Emphatic Accent Production in French[END_REF]). Therefore, the probability of measuring any significant improvement when adding the visual modality is very low. The challenge was thus for us to design auditorily degraded prosodic stimuli in order to lower the auditory only perception scores and make improvement possible. The classical speech perception paradigm used to measure the audiovisual advantage is the speech in noise perception paradigm. However, even though adding noise to a signal reduces its lexical intelligibility, it does not alter the global fundamental frequency (F0) contour. Actually, [START_REF] Miller | An Analysis of Perceptual Confusions among some English Consonants[END_REF] showed that voicing was the most robust speech feature to noise. This is why we designed a paradigm using whispered speech for which there is no F0 information at all since there is no vocal fold vibration. Moreover, whispering is used when one wants to be understood by the person he/she is speaking to but not overheard by others. The task given to the speakers during the audiovisual recordings described hereafter, was to make themselves understood by someone located at a certain distance from them (and not to whisper in someone's ear). It is therefore possible that the speakers might compensate for the lack of auditory cues by emphasizing visual cues.

Experimental Methods

Materials

Corpus

The corpus used consisted of four sentences with a Subject-Verb-Object (SVO) structure and with CV syllables. An example of a sentence used is given in (1).

(1) Romain ranima la jolie maman. 'Romain revived the good-looking mother.'

Audiovisual recording

We recorded two male native speakers (A and B) of French as they were whispering the sentences from the corpus under four focus conditions: neutral, subject focus (SF), verb focus (VF) and object focus (OF). The two speakers were the same as those recorded for the studies described in [START_REF] Dohen | Audiovisual Production and Perception of Contrastive Focus in French: a multispeaker study[END_REF]. A total of 16 utterances were thus recorded for each speaker. The recordings were done in a sound attenuated room at the Institut de la Communication Parlée (ICP). The speakers were not directly asked to produce focus. A correction task was used instead in order to trigger focus in the most natural way possible. The speakers listened to a prompt in which two speakers (S1 and S2) were talking. S1 first pronounced a sentence from the corpus which S2 then repeated in a question mode because he was not sure to have understood correctly one of the constituents from the sentence (S, V or O). The recorded speaker then had to correct S2 and thus produced contrastive focus on the mispronounced constituent. The recording therefore went as follows (capital letters signal focus): Audio prompt:

S1: Romain ranima la jolie maman. S2: S1 a dit : Denis ranima la jolie maman? 'S1 said: Denis revived the good-looking mother?' Speaker uttered: ROMAIN ranima la jolie maman.

No indication was given to the speakers on how to produce focus (e.g., which syllable(s) was(were) to be focused). When S2 had correctly understood (he produced the correct sentence in a question mode), the recorded speaker was instructed to produce a neutral version (broad focus) of the sentence i.e. without focusing any particular constituent.

The speakers movements were monitored with front and profile cameras. An example of the recorded images is given in Figure 1. The speakers wore blue markers on the lips and chin (see Figure 1) in order to make it possible to use an automatic lip-tracking device designed at ICP [START_REF] Lallouache | Un poste Visage-Parole couleur. Acquisition et traitement automatique des contours de lèvres[END_REF] to extract articulatory features.

Stimuli manipulation

A preliminary informal auditory only perception test was conducted to examine whether the detection of focus using the whispered audio signals was low enough to be possibly improved. This test showed that the perceptual performances were quite high. An acoustic analysis actually showed that the intensity cues seemed to be boosted in whispered speech as if to compensate for the lack of F0 cues. We therefore decided to weigh the intensities of all the utterances recorded in order to bring each constituent of the utterance (S, V and O) to the same level as in the neutral version. A second informal auditory only perception test showed that perception scores were lower using the modified auditory files.

Experimental paradigm

The tests took place in a quiet room at ICP in which the participants were isolated both from outside noise and from the experimenters. The videos were shown on a video monitor placed approximately one meter away from the participants. The speaker's head on the screen was approximately real size. The participants were told that they would be following part of a conversation between two people (S1 and S2). S1 would first utter a SVO sentence. Not having heard the sentence very well, S2 would question S1 by repeating the sentence the way he had understood it, in a question mode. S1 would then repeat the first sentence he had uttered correcting the constituent (S, V or O) that S2 had misunderstood. He would therefore insist on this particular constituent (i.e. focus it).

The participants were told that they would neither hear nor see S1's first utterance as well as S2's. They would either see only (V), hear only (A) or hear and see (AV) S1's correction.

Participants were told that, in some cases, no correction would be performed by S1 because S2 would have correctly understood. In that case, they would just hear or see or see and hear S1 repeating the initial sentence without performing any correction (i.e. neutral version of the sentence). The task was for the participants to identify which constituent (S, V, O or none) had been misunderstood by S2 and thus corrected by S1. They were asked to highlight the constituent they had identified as being corrected on an answer sheet such as the one presented below: Romain ranima la jolie maman.

If they thought that S1 had performed no correction, they were asked to highlight the empty column on the right.

The participants were thus indirectly asked to identify whether a constituent had been focused and which one. They were never told about "focus" or about the experiment's aim.

Three movie clips were elaborated combining the videos recorded and the degraded auditory signals (one clip for each condition: AV, A and V). Each movie consisted of two sequences each corresponding to a random combination of the 32 stimuli. One of the sequences was to be seen with the front view by the participants and the other with the profile view.

Two separate tests (a and b) were designed using the same three movie clips. Each participant went through one of the two tests. For test a, the presentation modality order was: AV, A, V and for test b: A, AV, V. The aim being to analyze the contribution of the visual modality, this paradigm allowed comparisons between the performances corresponding to audio only and audiovisual conditions. For test a, for example, there could indeed be a training effect during the audiovisual session which would affect the performances during the audio only session and vice versa. This is why two tests were necessary. The visual only perception condition represented a control.

A total of 32 stimuli were thus evaluated by the subjects under three conditions (A, V and AV) and two different views (front and profile). This represents a total of 192 stimuli.

A total of 13 native speakers of French (8 men and 5 women) aged 19 to 57 participated in this experiment.

Results

General results

Figure 2 provides the percentages of correct answers for each participant and for each condition (A, V and AV). The means over all the participants for each condition correspond to the thick dark lines. As could have been expected, the perceptual performances are better for the AV condition than for the other conditions. It therefore appears that when the acoustic prosodic cues are lacking, visual information can help recover at least part of the information. For all the conditions, the results are significantly above chance (25%): AV: t=31.478, p<0.001; A: t=13.369, p<0.001; V: t=13.374, p<0.001. The fact that audio only performances turned out to be better than chance was predictable since the durational information was still available for the audio detection. It is also possible that the intensity cues were not entirely "erased".

Figure 2: Percentages of correct answers for each condition (AV, A and V) and each participant (1 bar = 1 participant). The thick dark lines represent the means of these percentages

over all the participants.

Figure 3.a provides the means of the percentages of correct answers over all the participants for each focus condition and for each speaker. It appears that the performances are better for speaker A which reflects what had already been observed in previous visual only perception tests (see [START_REF] Dohen | Audiovisual Production and Perception of Contrastive Focus in French: a multispeaker study[END_REF]). It also appears that the audiovisual advantage is smaller for speaker A than for speaker B (+3.9% for speaker A and +14.4% for speaker B). The performances corresponding to the audio only condition for speaker A are very good (80.8% correct answers) and much better than those for speaker B (63.5%). There must therefore be a ceiling effect for speaker A: the performances are too good in the audio only condition to get improved to a significant extent. This is probably due to the fact that speaker A is a trained speaker. The acoustic cues he produced were very strong and we had great difficulties bringing back the intensity to an average level without distorting the signals. In addition, hyper-articulation has visual as well as acoustic consequences: formant patterns are less reduced for instance, which may be an additional acoustic cue (see [START_REF] Loevenbruck | Pistes pour le contrôle d'un robot parlant capable de réduction vocalique[END_REF] for the difference in auditory perception between hypo and hyperarticulated /iai/ sequences). This may have slightly biased the results and probably explains the small visual supplement measured for speaker A. Since speaker B was a naive speaker, it is possible that the results corresponding to the perception of his productions better reflect what would happen in natural communication. A four intra-subject factor ANOVA was conducted on the results. The four factors were: modality (three levels: AV, A and V), speaker (two levels: speakers A and B), view point (two levels, front and profile) and focus condition (four levels: neutral, SF, VF and OF).

There appears to be a significant modality effect: F(2,24)=7.232, p=.003. The analysis of the ANOVA contrasts shows that the AV performances are significantly higher than those corresponding to the other modalities (p<.001). The results corresponding to the audio and visual only modalities are not significantly different (p=.451). The perceptual performances are thus significantly better when both modalities are available.

There is also a significant effect of the speaker: F(1,12)=121.384, p<.001. As explained before, the general perceptual results are better for speaker A.

Both the view and focus condition factors do not have a significant effect on the perceptual performances (view point: F(1,12)=0.244, p=.63; focus condition: F(1.716,36)=11.231, p<.001).

Further analysis

The mean percentages of correct answers over all the participants were calculated for each stimulus in each condition. The stimuli were then classified into three categories for which explanations were put forward as to what improved or degraded perception:

-"AV≥ ≥ ≥ ≥A,V" category (AV performances are the best):

The majority of stimuli (46.9%) belong to this category. In this case, there were not many acoustic cues to contrastive focus left after degradation except for the durational ones. Auditory perception was thus not very good. Some visual cues were present and contributed to enhancing perception: when both modalities were combined, perceptual performances got better.

-"A≥ ≥ ≥ ≥AV>V" category (auditory only performances are the best and AV perception is better than V perception): 28.1% of the stimuli belong to this category. In this case, it seems quite clear that adding the visual modality impaired perception. Two explanations can be put forward. They correspond to two different cases. The first case (A~AV) corresponds to one for which the AV and A performances are approximately the same. In this case, there must only be few visual correlates and adding the visual modality does not bring anything.

Half of the stimuli from this category correspond to this case. In the second case, the A performances are better than the AV ones (A>AV). The visual cues must not only be absent but they seem to mislead perception. The other half of the stimuli from this category correspond to this case. -"V>AV,A" category (visual only performances are the best): 25% of the stimuli belong to this category. In this case, the visual cues must be present since visual only perception is good but it seems that, when the auditory information is added, performances get lower. If the AV performances are better than the audio only ones (AV>A), it is possible that the auditory signal was poorly manipulated and that it tended to confuse perception and resulted in V>AV. This is the case for half of the stimuli from this category. If AV=A, the auditory signal may have been modified too much resulting for example in an unnatural production. In this case, the visual information was not sufficient to help the participants. This is the case for 25% of the stimuli from this category. Only two stimuli correspond to cases for which AV<A, they are isolated cases for which no explanation could be found.

Conclusions and Discussion

This audiovisual perception test of prosodic contrastive focus in French, conducted with whispered speech for two speakers, showed that visual cues can help detect focus when the auditory information is deteriorated. Perceptual performances are enhanced when the visual modality is added. Performances were better for speaker A than for speaker B which was predictable from the results of the visual only tests performed before ( [START_REF] Dohen | Audiovisual Production and Perception of Contrastive Focus in French: a multispeaker study[END_REF]). The audiovisual advantage is however much stronger for speaker B. It is actually possible that speaker A's auditory performances reached a ceiling that could not be improved.

Figure 1 :

 1 Figure 1: Image from the video signal recorded.

Figure 3 :

 3 Figure 3: Means of the percentages of corrects answers a.(left) for each speaker depending on the condition (AV, A and V) and b.(right) for each view point depending on the condition.

Figure 3 .

 3 Figure 3.b provides the means of the percentages of correct answers across the participants for each view (front and profile) and for each condition (AV, A and V). It shows that there are no differences from one view to another which confirms what had been observed for previous visual only perception tests ([11]).A four intra-subject factor ANOVA was conducted on the results. The four factors were: modality (three levels: AV, A and V), speaker (two levels: speakers A and B), view point (two levels, front and profile) and focus condition (four levels: neutral, SF, VF and OF).There appears to be a significant modality effect: F(2,24)=7.232, p=.003. The analysis of the ANOVA contrasts shows that the AV performances are significantly higher than those corresponding to the other modalities (p<.001). The results corresponding to the audio and visual only modalities are not significantly different (p=.451). The perceptual performances are thus significantly better when both modalities are available.There is also a significant effect of the speaker: F(1,12)=121.384, p<.001. As explained before, the general perceptual results are better for speaker A.Both the view and focus condition factors do not have a significant effect on the perceptual performances (view point: F(1,12)=0.244, p=.63; focus condition: F(1.716,36)=11.231, p<.001).

Acknowledgments

The authors would like to thank Coriandre Vilain, Christophe Savariaux and Alain Arnal for their technical help. We are grateful to our two speakers as well as to all the participants to the test. We also thank Jean-Luc Schwartz and Marie-Agnès Cathiard for their help and comments.


domain: math info sdv

text:
Galf-G Galf-

Test Dataset Test Dataset Test Dataset Test

STAT Motif STAT Motif

Camel, cow and human milk proteins. Camel, cow and human milk proteins.

As described in Kappeler et al. we used the promoters from genes expressed in milk. As described in Kappeler et al. we used the promoters from genes expressed in milk. These genes possess strong TATA signal and typical PolII promoters (Group II in the tree). These genes possess strong TATA signal and typical PolII promoters (Group II in the tree). We extracted the 2000 bp sequences before the annotated TSS for human and cow in We extracted the 2000 bp sequences before the annotated TSS for human and cow in Ensembl, and the 5' sequences available in GenBank for the Camel. Ensembl, and the 5' sequences available in GenBank for the Camel.

TFBS were searched with our TFBS were searched with our pipeline and the best results for pipeline and the best results for each method were clustered and each method were clustered and compared with the TRANSFAC compared with the TRANSFAC database. database.

In the present work we propose the combination of several bioinformatic approaches to identify regulatory motifs in cattle promoters for coregulated genes. We In the present work we propose the combination of several bioinformatic approaches to identify regulatory motifs in cattle promoters for coregulated genes. We integrate results from phylogenetic footprinting (Footprinter) with orthologs from human, mouse and rat genomes and genetic algorithm (GALF-G) and statistical-integrate results from phylogenetic footprinting (Footprinter) with orthologs from human, mouse and rat genomes and genetic algorithm (GALF-G) and statisticalbased motif search (Weeder). based motif search (Weeder).

Deciphering mechanisms of early implantation of the embryo in cattle is of economical and fundamental interest, as these findings could help to reduce the Deciphering mechanisms of early implantation of the embryo in cattle is of economical and fundamental interest, as these findings could help to reduce the implantation failure and also give clues regarding human sterility issues. implantation failure and also give clues regarding human sterility issues. One possible approach is to identify precisely the transcription factors (TF) responsible for gene expression in the uterus-conceptus dialogue. TF are proteins that One possible approach is to identify precisely the transcription factors (TF) responsible for gene expression in the uterus-conceptus dialogue. TF are proteins that interact directly with the DNA molecule by recognizing small and slightly degenerate nucleotide motifs (TFBS) in the promoter region of the genes interact directly with the DNA molecule by recognizing small and slightly degenerate nucleotide motifs (TFBS) in the promoter region of the genes In the present work we propose the combination of several bioinformatic approaches to identify regulatory motifs in cattle promoters for coregulated genes. We In the present work we propose the combination of several bioinformatic approaches to identify regulatory motifs in cattle promoters for coregulated genes. We integrate results from phylogenetic footprinting (Footprinter) with orthologs from human, mouse and rat genomes and genetic algorithm (GALF-G) and statistical-integrate results from phylogenetic footprinting (Footprinter) with orthologs from human, mouse and rat genomes and genetic algorithm (GALF-G) and statisticalbased motif search (Weeder). based motif search (Weeder).

Deciphering mechanisms of early implantation of the embryo in cattle is of economical and fundamental interest, as these findings could help to reduce the Deciphering mechanisms of early implantation of the embryo in cattle is of economical and fundamental interest, as these findings could help to reduce the implantation failure and also give clues regarding human sterility issues. implantation failure and also give clues regarding human sterility issues. One possible approach is to identify precisely the transcription factors (TF) responsible for gene expression in the uterus-conceptus dialogue. TF are proteins that One possible approach is to identify precisely the transcription factors (TF) responsible for gene expression in the uterus-conceptus dialogue. TF are proteins that interact directly with the DNA molecule by recognizing small and slightly degenerate nucleotide motifs (TFBS) in the promoter region of the genes interact directly with the DNA molecule by recognizing small and slightly degenerate nucleotide motifs (TFBS) in the promoter region of the genes

Introduction Introduction Introduction Introduction

Methods

Methods

  mapped onto the corresponding Each EST is mapped onto the corresponding gene in Ensembl. gene in Ensembl. Promoter sequences and their orthologs in Promoter sequences and their orthologs in mouse, human and rat are extracted from the mouse, human and rat are extracted from the Ensembl database through the Compara and Ensembl database through the Compara and Core

MIG -Mathématique, Informatique et Génome MIG -Mathématique, Informatique et Génome Domaine de Vilvert, Domaine de Vilvert, 78352 Jouy-en-Josas 78352 Jouy-en-Josas This work is funded as part of the ANR-08-GENM-037 project MEETAC This work is funded as part of the ANR-08-GENM-037 project MEETAC MIG -Mathématique, Informatique et Génome MIG -Mathématique, Informatique et Génome Domaine de Vilvert, Domaine de Vilvert, 78352 Jouy-en-Josas 78352 Jouy-en-Josas This work is funded as part of the ANR-08-GENM-037 project MEETAC This work is funded as part of the ANR-08-GENM-037 project MEETAC


domain: sdv shs

text:
X Coquil

J L Fiorelli

C Bazard

D Foissy

J M Trommenschlager

A Blouet

Prototyping organic mixed crop-dairy systems at a local scale: a multi-attribute approach

Social demand for Sustainable agriculture and multifunctionality of agriculture since the 2003 CAP reform, leads agricultural systems to be more friendly for the environment. In such a context, research has to work on the conception of environmental friendly agricultural systems. In East of France, a research team is prototyping two complementary mixed crop-dairy systems aiming to respect the environment and to produce milk and crops. Two organic multi-objective prototypes are being built based on (i) respect of land natural properties (ii) use of low external inputs and (iii) limitation of the exchanges between systems at a local scale. Prototyping that kind of systems requires a multi-attribute approach using different methods. Experimentation at farming system level combined to modelling might suit well that purpose. 1

INTRODUCTION

The development of sustainable agriculture is seen as a solution to face environmental injuries caused by intensive farming of the last decades. Sustainable agriculture integrates three main goals: environmental health, economic profitability, and social and economic equity. These notions have been partly integrated in the last reform of the CAP in 2003, with a particular attention given to the environmental respect. The amount of subsidies given to a farm is now partly determined by the ecoconditionality: but do we really know which kind of agricultural system is environmental friendly?

In this context, research on agricultural systems can participate to the design of production systems which are respectful to the environment. That kind of research could (i) give references on environmental friendly agricultural systems and (ii) determine if such sustainable agriculture needs to be subsidized.

Sustainable agriculture has to deal with the land's natural properties and has to assure its conservation and transmission to the future generations. Heterogeneity of agricultural land can be high at a local scale and at a farm scale too. Can that land heterogeneity become an advantage for agricultural activity at a local scale?

To answer these questions, in the INRA Mirecourt station, we are prototyping two organic mixed cropdairy productions systems. Three objectives are assigned to them: (i) respecting biotic and abiotic environment, (ii) producing milk and crops, (iii) and using low external inputs.

In the study, the INRA Mirecourt station, including 240 ha, is seen as a "small scale region" of two edaphic conditions representative of the northern plains of Western Europe (continental climate, two types of soil). This configuration makes this station a good field to work on potential local synergies between farming systems, which could help facing land heterogeneity.

In this text, we will describe the different concepts and methods we are using to prototype these systems.

A CONCEPTION DRIVEN BY LOCAL NATURAL PROPERTIES AND LOW EXTERNAL INPUTS SYSTEMS

The first principle designing the two systems is the local natural properties. This approach is more and more used in developing countries trying to build sustainable systems, suitable to the natural context (Chia et al., 2002;Auricoste et al., 1985). We first referred to the edaphic properties of the experimental station to determine (i) cultivated fields (ii) permanent grassland fields and (iii) cultivated fields able to grow legumes (alfalfa). Secondly, we identified fields accessible to cows by looking at their geographic and spatial disposition relative to the milking parlour. This approach is completely at the opposite of the usual and conventional agronomic approach, developed in the last decades and still used in precision agriculture, consisting in reducing field heterogeneity by technical interventions. Field homogenisation aims at the simplification of the practices at the farm level [START_REF] Soulard | Les agriculteurs et la pollution des eaux[END_REF].

The second principle contributing to the systems design was to have local systems using low external inputs. Lack of access to external inputs implies that only increased use of labour and skills offers a way out (Schiere et al., 2002). Thus, we choose (i) organic mixed crop-dairy farming to valorise the permanent grasslands and to limit external inputs and (ii) complementary systems favouring mutual and equivalent exchanges between them. In those systems the presence of livestock is not contradictory with environmental sustainability. The dairy herd uses and transforms grasslands' energy (Schiere et al., 2002) into human alimentary products. It also contributes to the nutrients cycle by its dejections.

Comment citer ce document : Coquil, X., Fiorelli, J.-L., Bazard, C., Foissy, D., Trommenschlager, J.-M., Blouet, A. (2006). Prototyping organic mixed crop-dairy systems at a local scale: a multi-attribute approach. In:

Joint Organic Congress (p. 34-35). Presented at Joint Organic Congress, Odense, DNK (2006-05-30 -2006-05-31).

Moreover, the two organic local systems that we have built aim at limiting importation and exportation from one to another. These limited local exchanges are costless in terms of energy and, if they are mutual and equivalent, they avoid unbalanced fertility transfers between the systems. That position is opposite to intensive dairy farming which uses high external importations like, for example, massive importations of soybean, leading to high energy expense, soil depletion in soybean exporting countries and causing environmental problems in importing countries.

TWO COMPLEMENTARY PROTOTYPES

Those principles have led to two organic dairy prototypes (Table 1): a Grassland farming system (GFS) and a Mixed-Crop Dairy System (MCDS). The GFS is made-up of permanent grazed by 37 dairy cows and the replacement heifers. Cows are overwintered in cubicles buildings and they are fed by maximising grazing (calving period takes place in late winter) in order to minimize straw, concentrates and energy requirements. The MCDS involves permanent grasslands and cultivated areas. Rotations of six or eight years, including a three year period of mixed grass and legumes cover, are conducted on cultivated areas. Forages are grazed or cut to feed the 60 cows and the replacement heifers. Calving period takes place in late summer in order to valorise all the conserved forages. Cows are overwintered on straw bedding to produce manure that will be spread on cultivated fields. Because housing is really long, straw needs are quite high, so crops are mostly cereals or associations of cereals and legumes. At the small region scale, milk production lasts all year long in spite of a single very short calving period in each system. The GFS prototype needs a minimum of cereals, as energetic concentrates, and straw which are produced on the MCDS. In exchange, the MCDS receive some manure and slurry from the GFS. The exchange is based on nitrogen and carbon contents.

AGRICULTURAL SYSTEM EXPERIMENTATION COMBINED TO MULTI-OBJECTIVE MODELLING

To achieve such multi-objective approach, we have inspired ourselves from Vereijken methodical way of prototyping integrated and ecological arable farming systems (1997). We have declined the objectives and sub-objectives assigned to both systems by confronting the researcher's and experimental farm Technical staff's point of views. We are now defining holistic parameters with threshold values and methods to achieve them. *Montbéliardes (50%) and Holstein Friesian (50%).

These two theoretical prototypes are going to be tested on Mirecourt experimental farm for at least eight years. Decision rules are used to choose and homogenize agricultural operations used from one year to another on the systems. Those rules are created in a participative way with farmers, researchers, advisors and the experimental farm's staff.

Experimental approach will be completed by a multi-objective modelling approach, in order to explore different farming options as described by Ten [START_REF] Ten Berge | [END_REF]. These approaches are complementary (i) we can run protocols to understand and quantify a process in order to model it and (ii) we can test experimentally the prototype proposed by the model at the farm scale.

The model will also be tested in different farming situations in commercial farms from the Lorraine region.

A MULTI-OBJECTIVE APPROACH

To prototype such production systems, a multiobjective approach is always necessary. It is a difficult exercise because even the rules defining the choice of the operating methods have to include the different objectives assigned to the system. For example, in such organic cropping systems, ploughing is an efficient practice for weed management but it enhanced mineralisation and it has a high energetic cost inducing a depressive influence on the environment. An alternative to Vereijken (1997) methodical approach is to use a multi-attribute model mixing empirical and scientific knowledge (Bohanec and Rajkovič, 1999).

Table 1 .

 1 Two complementary organic dairy prototypes setup at the INRA Mirecourt station.

	Organic dairy prototypes	GFS	MCDS
	Cows*	37	60
	Heifers*	55	90
	Natural grasslands (ha)	74,5	45
	Temporal grasslands (ha)	0	48,5
	Crops (ha)	0	72

X.Coquil, INRA-SAD, 662 avenue Louis Buffet, 88500 Mirecourt. France. coquil@mirecourt.inra.fr

A.Blouet. Université H.Poincaré Le Montet, 54600 Villers-les-Nancy. France. andre.blouet@iutnb.uhp-nancy.fr

ACKNOWLEDGEMENT

Thanks to all the staff from Mirecourt INRA experimental research centre and all our stakeholders.


domain: sdv sde

text:
Anne Sophie

Anne-Sophie S Walker
email: walker@versailles.inra.fr

Véronique Decognet

Marc Fermaud

Alexandre Bout

Johann J Confais

Pierre P Leroux

Angelique A Gautier

Fabián Martínez

Jean Roudet

Philippe C Nicot

Angélique Gautier

Fabian Martinez

Marc Bardin

Philippe Robin

Mélody Potron

Elisabeth Fournier

Genetic diversity and structuring factors for Botrytis cinerea french populations

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

Botrytis cinerea has long been considered as a very polymorphic fungus, in terms of morphology, host range, pathogenicity, sensitivity to various xenobiotics, ecology and genetic diversity. This plasticity may explain its large geographic distribution and its ability to colonize various environments and to cause important diseases on a large variety of crops. Despite its agronomic importance, little is known about Botrytis population dynamics and genetics and the available studies mainly analyse one component, often adaptation to host, as a structuring factor for the populations.

First results will be presented from a multifactorial study of Botrytis populations. Samples were collected in 4 distinct French areas (Champagne, Provence, French Riviera and Bordeaux area), on various host plants or decaying substrate (grapevine, tomato, brambles, litter), treated or not with fungicides, in 2 cropping systems (open fields or glasshouses) and during 2 years, in spring and autumn. This sampling plan led to the set up of at least 32 distinct populations per season of collection, and would help to understand the effect of geography and migration at small to large scale, selection by the host-plant and the fungicide programs, effect of the ecological type and of the cropping system on the diversity and genetic structure of Botrytis populations. All the isolates were analysed for their diversity according to 8 neutral microsatellite markers, and part of the collection was screened for its sensitivity to various fungicides. These data are managed through the specifically-developed database POPULABOT.

The first results from autumn 2005 confirmed the occurrence of a sibling species living in sympatry with Botrytis cinerea sensu-stricto. In France, B. cinerea consists of a complex of 2 genetically isolated species: B. cinerea Group I (proposed as B. pseudocinerea) and B. cinerea Group II (or B. cinerea sensu-stricto). In our sampling, strains belonging to Group I were highly differentiated from group II strains and exhibited private alleles for 2 microsatellite loci. They were found in any region but their frequencies never exceeded 10%. Genic diversity was regularly high, indicating an equilibrated distribution of the various microsatellites alleles in the populations, due to migration and/or sexual reproduction. The occurrence of regular recombination was confirmed by the low values of multilocus linkage disequilibrium and by the few numbers of repeated multilocus genotypes (clones) in most of the populations. However, in some Provence tomato glasshouses, genic diversity reached very low levels, whereas the multilocus linkage disequilibrium and the number of clones increased significantly, showing a strong effect of the cropping system on the population dynamics. Pairwise FSTs, Factorial Analyses of Correspondences and Molecular Analysis of Variance were used to characterize the respective influence of the different studied factors on population differentiation.


D Burger¹

F Janett²

M Vidament³

R Stump¹

G Fortier

I Imboden¹

R Thun²

Immunization against GnRH in adult stallions: Effects on semen characteristics, behaviour and shedding of equine arteritis virus

Introduction

There are numerous indications for suppressing reproductive endocrine function in horses, the most frequent being the inhibition of undesirable sexual behaviour. Immunization against GnRH offers a viable alternative to surgical or hormonal castration by circumventing surgical risk, frequent drug administration and residue problems (Stout and Colenbrander, 2004;Stout, 2005). Furthermore, it may prove to be a therapeutic alternative for the treatment of equine arteritis virus (EAV) shedder stallions (Burger et al., 2004), as persistence of EAV in the accessory sex glands has been shown to be androgen dependent. Earlier studies examining GnRH vaccination in stallions have reported varied effects and do not allow a clear conclusion to be drawn. In the present study, 3 experiments were carried out to examine the effect of anti-GnRH vaccination on semen characteristics, libido and shedding of equine arteritis virus in adult stallions.

Materials and methods

Experiment 1:

The experiment was carried out using 9 healthy Franches-Montagnes and Warmblood stallions (9-15 years) of proven fertility. The stallions were randomly allocated to a test group (6 stallions) and a control group (3 stallions). Ejaculates of all 9 stallions were collected and examined weekly for 52 weeks. After the first semen collection, animals in the test group were immunized with 200 µg of a GnRH-conjugate (Equity®; CSL, Australia) intramuscularly in the neck. Control animals received saline solution. The treatment was repeated 4 and 12 weeks later. Side effects to the vaccination were recorded. Simultaneously to weekly semen collection, libido was evaluated (time elapsing between entering the stallion barn and erection and ejaculation, respectively) and the number of mounts before ejaculation recorded. After ejaculation, scrotal width was measured using scrotal calipers. The volume, concentration, total sperm count and motility of the ejaculates were assessed and a morphological examination carried out. Blood samples were collected, always at the same time on the day of semen collection, and plasma testosterone concentration and GnRH antibody titre determined.

Experiment 2:

The effect of GnRH vaccination on libido, sexual behaviour and degree of distraction was evaluated in a retrospective questionnaire based field study, carried out with 22 stallions (7.4 ± 4.3 years old) of various breeds, vaccinated twice at an interval of 28 to 35 days. 18 stallions were vaccinated with 400 µg of a GnRH-conjugate (Improvac®; CSL, Australia) intramuscularly and 4 stallions with Equity®. , 94 (1-4). Presented at 9.

Results

Experiment 1: Scrotal size, total sperm number, percentage of normal sperm and motility of all treated stallions continuously decreased after vaccination. Lowest mean motility values were recorded 6 months after the first immunization (Fig. 1). All the above parameters tended towards prevaccination levels 10 to 12 months after begin of the study. The decrease in libido was also significant but varied individually occuring between 13 and 18 weeks after the first vaccination and lasting between 10 to 32 weeks. While the decrease in one stallion was only slight, the remaining five showed gelding like behaviour. At the end of the experiment, 5 stallions regained their original libido but 1 stallion remained suppressed such that it was not possible to obtain an ejaculate until the end of the study. Mean plasma testosterone concentrations in vaccinated stallions started to decrease significantly after the first injection and remained below 0.7 nmol/L plasma for 10 months (Fig. 2). By the end of the experiment, 3 stallions showed prevaccination levels (> 1 nmol/L plasma) but 3 remained at a basal level. Individual GnRH antibody titers peaked 2 months after the first immunization and remained high until the end of the study. Reactions to injection in the vaccinated group were few: 13 of the 18 vaccinations were without any side effects, the remainder showed mild to painful swellings at the injection site. No increases in body temperature (>38°C) or other complications were observed.

Experiment 2: Among the 19 stallions with a libido problem for the owner, 17 responded with the desired decrease in libido. Among the 15 stallions with a high distraction level, the owners reported less distraction in 13 stallions. Aggressiveness towards humans and/or other horses was markedly reduced in all 7 horses reported to be aggressive.

Experiment 3:

EAV could no longer be isolated from the semen of all four vaccinated stallions 4 -6 months after the first immunization and from all subsequent semen collections. One stallion died for reasons not related to the study 6½ months after the first vaccination. The non-shedding, noncarrier status of the 3 remaining stallions was additionally confirmed by failure of each of the two mares to show seroconversion 28 days after breeding.

Discussion

The efficacy of GnRH vaccination appears to depend to a great extent on the type of vaccine used. Contrary to previous studies (Clement et al., 2005), the use of both Improvac® and Equity® has led to satisfactory results. More than 80% of the stallions in experiment 1 and 2 showed clear loss of libido, the duration of which varied greatly among single individuals. Similar observations regarding libido have also been reported after surgical castration of breeding stallions. This suggests that GnRH vaccination is a useful tool for temporary management of undesirable sexual behaviour in stallions. Evaluation of semen characteristics in experiment 1 showed that although semen quality deteriorated, semen production was never entirely suppressed. At the end of the experiment, all semen parameters were comparable to original values. The number and interval of booster vaccinations required for an optimal effect need further investigation. Failure of 3 stallions to reach plasma testosterone concentrations >1 nmol/L in our study may be due to the third vaccination.

The GnRH antibody titres do not appear to be a useful indicator of immunization effect, either on behaviour or on semen characteristics. 5 stallions showed a full return to reproductive function despite persistent antibody titres. The remaining stallion was surgically castrated 3 months after the study ended and histological examination of the testes revealed normal spermatogenesis.

All four treated EAV-shedding stallions were converted to a non-shedder status. In contrast to findings after short term treatment (35-38 days) with a GnRH antagonist (Fortier et al., 2002), no stallion in our study reverted to a shedding status. After failure of the two test mares to seroconvert, the stallions resumed breeding activity and showed normal fertility. For instance, stallion P (17 years at vaccination in July 2004, EAV shedder since 4 years) had pregnancy rates per cycle of 30% (n=30 cycles) in 2004 and of 37% (n=38 cycles) in 2005, respectively, before and after vaccination. This study demonstrates that vaccination against GnRH could represent a promising method for successfully treating EAV-shedding breeding stallions.

  Four permanent EAV-shedding stallions aged 12 to 17 years were vaccinated twice 4 weeks apart with either Improvac® (1 stallion) or Equity® (3 stallions). Semen was collected and virus content (cell culture and RT-PCR assay) was determined monthly for 6 months and then individually up to 15 months after first immunization. Additionally, 12 to 17 months after the first vaccination, two EAV seronegative mares where bred and serologically tested 4 weeks later. ., Janett, F., Vidament, M., Stump, R., Fortier, D., Imboden, I., Thun, R. (2006). Immunization against GnRH in adult stallions: effects on semen characteristics, behaviour and shedding of equine arteritis virus. In: Proceedings on the Ninth International Symposium on Equine Reproduction (p. 107-111). Animal Reproduction Science

Fig. 1 .

 1 Fig.1. Mean (± SEM) total motility in vaccinated and non vaccinated stallions.

  ., Janett, F., Vidament, M., Stump, R., Fortier, D., Imboden, I., Thun, R. (2006). Immunization against GnRH in adult stallions: effects on semen characteristics, behaviour and shedding of equine arteritis virus. In: Proceedings on the Ninth International Symposium on Equine Reproduction (p. 107-111). Animal Reproduction Science, 94 (1-4). Presented at 9.

Fig. 2 .

 2 Fig. 2. Mean (± SEM) plasma testosterone concentrations in vaccinated and non vaccinated stallions.

  ., Janett, F., Vidament, M., Stump, R., Fortier, D., Imboden, I., Thun, R. (2006). Immunization against GnRH in adult stallions: effects on semen characteristics, behaviour and shedding of equine arteritis virus. In: Proceedings on the Ninth International Symposium on Equine Reproduction (p. 107-111). Animal Reproduction Science, 94 (1-4). Presented at 9.

  1.Burger, D., Janett, F., Imboden, I., Zientara, S., Timoney, P., Thun, R., 2004. Treatment of an equine arteritis virus-shedding stallion by immunization against GnRH. In: 15th International Congress on AnimalReproduction, vol.1, Porto Seguro, Brazil, 281 (Abstract). 2. Clement, F., Vidament, M., Daels, P.,Van der Meer, F., Larry, J.L., Colenbrander, B., Turkstra, J., 2005. Immunocastration in stallions: Effect on spermatogenesis and behaviour. Proceedings of the 4th International Symposium on Stallion Reproduction, Anim. Reprod. Sci. 89, 230-233 (Abstract). 3. Fortier,G., Vidament, M., DeCraene, F., Ferry, B., Daels, P.F., 2002. The effect of GnRH antagonist on testosterone secretion, spermatogenesis and viral excretion in EVAvirus excreting stallions. Theriogenology 58, 425-427. 4. Stout, T.A.E., 2005. Modulating reproductive activity in stallions: A review. Proceedings of the 4th International Symposium on Stallion Reproduction, Anim. Reprod. Sci. 89, 93-103. 5. Stout, T.A.E., Colenbrander, B., 2004. Suppressing reproductive activity in horses using GnRH vaccines, antagonists or agonists. Anim. Reprod. Sci. 82-83, 633-643.

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text:
J.-C Samalens

D Guyon

N Bories

C Moisy

J.-P Wigneron

Spatiotemporal Dynamic of French Forest Phenology from MODIS and GLOBCARBON products

Phenological trends studies based on remote sensing data appear to be a realistic and costeffective way to monitor forest ecosystems over time. Although such vegetation trends have been widely investigated at the global scale, still little is known on pheno-phases anomalies at regional scales. Remote sensing time series at medium spatial resolution (≤ 1km) are now available to fill this gap but the derived phenological products still need to be validated. Our retrospective analysis is based on remotely sensed phenological metrics delivered over 10 years (1998)(1999)(2000)(2001)(2002)(2003)(2004)(2005)(2006)(2007) through the GLOBCARBON project (VGCP product) and from MODIS sensors (MCD12Q2 product). For validation, we took advantage of tree canopy phenological observations of the French Permanent Plot Networks for Forest Monitoring (RENECOFOR) carried out at the ground on the same period. Focusing on two emblematic deciduous broadleaf tree species (beech and oaks) we also investigated phenological responses at the tree species level on a national extent. In-situ comparison of broadleaf forests leaf-on date shows a prediction uncertainty of 8 and 13.8 days and systematics errors of 2.2 and 9.8 days for MODIS and GLOBCARBON. Considering only pure beech forests reduce bias to 1.6 and 5.5 days respectively but no significant relationship could be retrieved for oaks forests. On the France extent, crossproduct comparaison remain stable against tree species. Neither early leaf-on nor senescence stages could accurately be depicted by satellite based metrics. Inter-annual leaf-on trends are consistent with ground observations and spatial patterns of green-up dates are related to latitudinal gradients for oaks forests and altitudinal gradients for beech forests.

INTRODUCTION

Tree phenology reflects their adaptive responses to climate variability and therefore constitutes an efficient bio-indicator of forest condition and productivity. Because warming climate has led to phenological timing shifts over the past half century [START_REF] Chmielewski | Response of tree phenology to climate change across Europe[END_REF][START_REF] Menzel | European phenological response to climate change matches the warming pattern[END_REF], tracking the rhythm of the seasons is still challenging [START_REF] Morisette | Tracking the rhythm of the seasons in the face of global change: phenological research in the 21st century[END_REF] and remote sensing is a key tool for measuring and monitoring forest phenology on a large extent. Vegetation indices derived from satellite data are commonly used for this purpose essentially at global or continental scale. Finer spatial scale studies of vegetation trends can now be retrospectively be performed as medium spatial resolution (≤ 1km) data time series become available over the last decade. Those satellite data are regularly reprocessed to improve information quality (snow, clouds, aerosols correction) or combined via multiple sensors acquisition to get a higher temporal frequency. However, the remote sensing community regularly emphasizes the need for ground validation and multiple sensor comparisons to validate the accuracy of those products [START_REF] Morisette | Tracking the rhythm of the seasons in the face of global change: phenological research in the 21st century[END_REF].

Based on long-term in-situ forest phenological observations, the aim of this study was to evaluate the ability of high-level remote sensing products in monitoring forest foliar dynamics both in time and space. We report here the evaluation of the MODIS and GLOBCARBON "phenological" products (Cf. 2.1). For ground validation, we focus on two major deciduous broadleaf tree species: Beech (Fagus sylvatica) and Oaks (Quercus sp.) because there is significant seasonal trends differences between those two species [START_REF] Davi | Sensitivity of water and carbon fluxes to climate changes from 1960 to 2100 in[END_REF]Vitasse et al., 2009a) but consistent within each one (Vitasse et al. 2009b).

MATERIALS & METHODS

To explore spatio-temporal trends at a national scale we look at long term remote sensing data time series, corrected for atmospheric and bidirectional effects, and for which phenological indicators were already computed.

Remote sensing data

First, we investigate the latest version of the MODIS "land Cover Dynamics" product available on the LPDAAC server, namely MCD12Q2 (V5), which combines the "Terra" and "Aqua" sensors reflectance measurements over the period 2001 to 2006 (see Ganguly et al., 2010 for complete description). The interest of this recent version lies in both a high spatial and temporal resolution (500m and 8 days respectively). Ensuing yearly products gather several land surface phenology indicators derived from Enhanced Vegetation Index (EVI) annuals curves extended to ± 6 months. Those metrics characterize vegetation growth cycle using transition dates namely, Onset Greenness Increase (OGI), Maximum (OGMax), decrease (OGD), minimum (OGMin) and the annual integral of the vegetation index curve (EVI area). We also computed (OGMax-OGI)/2 as an indicator of the date of the growing season inflexion point (hereafter referred as OG).

On another hand, GLOBCARBON v2 "Vegetation Growth Cycle Parameters" product at 1km was used for the years 1998 to 2007. The baseline input combine satellite data from the ESA Earth Observations (ATSR-2, AATSR, MERIS) and SPOT VEGETATION sensors. Complete description of the GLOBCARBON initiative can be found in [START_REF] Plummer | An Update on the GlobCarbon Initiative: Multi-Sensor Estimation of Global Biophysical Products for Global Terrestrial Carbon Studies[END_REF].The generation of annual growth cycle parameters is based on derived LAI profiles smoothed over 54 decades. Authors argue that LAI should be less sensitive to extraneous effects which are often seen on vegetation index time series (e.g. NDVI, EVI). Similarly to the MODIS product, several phenological metrics are provided (n=8) such as the day before the highest first derivative of LAI curve reach 90% of its maximum (hereafter referred as Dbefore). Forest phenological observations are carried out annually since 1997 over the whole French Permanent Plot of the European Network for the Monitoring of Forest Ecosystems (RENECOFOR, see Figure 1). This network encompasses a wide range of bioclimatic conditions of France, from oceanic to Mediterranean and from continental to high elevation mountain context. A subset of this network is composed by 49 stands of 0.5 ha of pure deciduous broadleaved forests. Stands are mostly managed has even-aged high forests (106 years on average, Table 1). Quercus sp. stands are mainly sampled in the northern part of France and homogeneously regarding to longitude. Fagus sylvatica stands are located in north-eastern plains and in southern French mountains. Average elevation is 200 m [20-370 m] for oak plots and 615 m [90-1400 m] for beech plots (Table 1).

In-situ phenological observations

Each year, the same 36 trees/plot are monitored and phenological key stages are recorded weekly in Julian days by local foresters. In spring (March-June), the onset and the end of the green-up phase are defined when leaf-unfolding occurred on 10% and 90% of the trees (hereafter reported as LU10% and LU90%). The same approach is achieved during the autumn season (September-November) for the leaf colouring stages (LC10% and LC90%). For consistent analysis, we selected the 46 stands (17 beech stands and 29 oaks stands, see Table 1) where at least 7 years were effectively observed during the decade 1998-2007 (63% were observed every year of the whole period). Inter-annual average of ground observations dates were plotted against each phenological metrics given by both remote sensing products and simple linear regression was computed. Difference between ground and remotely sensed data was reported as bias and the RMSE was used to evaluate the average satellite records uncertainty relative to in-situ observations.

Forest Cover

Cross-sensor validation was investigated at the national scale for all broadleaf forests and for each studied tree species. To this end, comparative analysis was conducted on image subsets to avoid mixture of landcover classes in a same pixel. As no accurate tree species map was available on this spatial extend, we first used the latest version of Corine Land Cover 2006 to identify pixels covered by a majority of broadleaved forests. Then, we took advantage of the National Forest Inventory database over the period 2005-2008 (9080 NFI plots network based on a systematic sampling design) to locate pure beech or oaks forests. Finally, we combined those data for each tree species to select pixels covered by at least 95% of broadleaved forest (Broadleaf) and for which the NFI observations where pure tree species (Oaks and Beech). Because of different spatial resolution pixels selection was repeated for each remote sensing product.

RESULTS AND DISCUSSION

In-situ comparison

All phenological indicators of both products were compared with ground measurements. For the leaf unfolding phase, significant relationships could only be retrieved using OGa and Dbefore indicators for MODIS and GLOBCARBON respectively (Table 2). The earlier stage of leaf unfolding (LU10%) is overestimated by both indicators with a systematic error greater than 10 days. At the end of this phenological stage (LU90%) bias is reduced by 10 days, ranging from 1.6 to 12.6 days. Bias and RMSE are always lower for OGa as compared to Dbefore. [START_REF] Soudani | Evaluation of the onset of green-up in temperate deciduous broadleaf forests derived from Moderate Resolution Imaging Spectroradiometer (MODIS) data[END_REF] also found that OGa is a good marker of leaf unfolding stage with the earlier version of the product (MOD12Q2, bias = 7.5 days as compared to LU90%). Improved result reported here on the same plots (bias= 2.2 days for LU90%) can be due to the longer time series but also the higher spatial and temporal resolution of the MCD12Q2 product.

Better predictions of the LU90% stage are reached if beech stands are considered separately. Bias is then better for beech (1.6 and 5.4 days) than for oaks (2.6 and 12.6 days) for OGa and Dbefore respectively. Those tree species discrepancies can be related to the potential mixture of different oaks species (and/or understory species) which may affect the spatial phenological variability within a pixel considered her as "monospecific".

Neither MODIS nor GLOBCARBON products can reflect dates of foliar discoloration of tree senescence phase (no correlations with ground data). However, we find that MODIS EVI area can be used as a proxy of the annual length of the growing season (computed as LC90%-LU10%, n=46, R²=0.35, p=1.6 e10 -5 ).

Overall, in-situ comparison shows higher estimations accuracy for MODIS product than GLOBCARBON and for beech forests as compared to oaks ones. Difference in spatial and temporal resolution can be invoked to explain lower accuracy level of the GLOBCARBON estimates as compared to MODIS. Another uncertainty source affecting the GLOBCARBON product might concern the LAI estimation which is based on the GLC2000 land cover map. Nethertheless, both of the satellite-based LU90% estimation accuracy are of the same order of magnitude than in-situ observations which are carried out on a weekly basis.

Spatiotemporal trends

On a wider dataset (NFI plots, Table 3), the correlation between the two satellite-based average leaf unfolding dates (OGa vs Dbefore) are low even if they are highly significative. As previously observed (Cf. Table 2), OGa indicates leaf unfolding dates of about 5 days earlier than Dbefore. Nevertheless, cross-product comparison shows a similar bias and accuracy level irrespective to tree species. In spite of those differences, similar spatial trends can be retrieved. Leaf-unfolding maps reveal spatially consistent results, even for high elevation forests such as beech ones (Figure 2A). This altitudinal trend is quantitatively in accordance with ground observations of Vitasse et al. (2009) and further with satellite-based study of [START_REF] Guyon | Monitoring elevation variations in leaf phenology of deciduous broadleaf forests from SPOT/VEGETATION time-series[END_REF] on the French Pyrénées Mountains. They report a similar trend of 1 day / 100m, just as the slope of Dbefore against elevation (Figure 2A right). This also confirms analysis of Lebourgeois et al. (2010) who found on the RENECOFOR network that elevation is the first spatial component driving unfolding trait. Because of a wider distribution range, a latitudinal variation of spring phenology (1.2 day/°) can also be retrieved within oaks forests (n=621, r²=0.03, p<0.0001), also reported by [START_REF] Ducousso | Latitudinal and altitudinal variation of bud burst in western population of sessile oak (Quercus petrea (Matt) Liebl)[END_REF]. Figure 2B shows that both satellite products correctly depict interannual phenology variability except for the year 2006 for which Dbefore reach peculiarly high values. As compared to in-situ annual observations, the best prediction is achieved for beech forests using the MODIS OGa indicator.

CONCLUSION

Because of high variability of landscape feature but also because of the heterogeneity of climate change throughout France [START_REF] Planton | Expected impacts of climate change on extreme climate events[END_REF], there is a clear expectation for different regional phenological

Modis

GlobCarbon A R²=0.16 , p < 0.0001 y = 0.006 x + 115 R²=0.26 , p < 0.0001 y = 0.01 x + 119

Years

Elevation

Beech

Oaks B responses. Those first results are promising and show the relevance and the spatial consistency of the medium spatial resolution for tracking regional trends in forest phenology. MODIS tends to provide more accurate estimates of leaf unfoldfing dates than GLOBCARBON but these conclusions are closely dependant of ground data observations at the plot level. Discrepancies between phenological responses of tree species emphasize the need for accurate forest type mapping. Further sub-pixels analyses have to be performed to better take into account for tree species composition in forest landscapes.

AKNOWLEDGMENTS

This study was supported by the CNES through a postdoctoral fellowship. We would like to thank the National Forest Inventory and the RENECOFOR network team of the National Forest Office for providing ground observations. Data from the GLOBCARBON project of European Space Agency are distributed by the VITO on the GEOSUCCESS web portal (http://geofront.vgt.vito.be/geosuccess). MODIS data are distributed by the Land Processes Distributed Active Archive Center (LP DAAC), located at the U.S. Geological Survey (USGS) Earth Resources Observation and Science (EROS) Center (lpdaac.usgs.gov).

Figure 1 :

 1 Figure 1: RENECOFOR forest network: location of the deciduous broadleaf forest stands (dots=oaks stands, triangles=beech stands. Grey area represents the Corine Land Cover 2006 broadleaf forest class.

Figure 2 :

 2 Figure 2: Satellite-based average leaf-unfolding dates (OGa, Dbefore) over specific NFI plots. A: regression analysis for beech forests against elevation (m). B: specific interranual variations. RENECOFOR LU90% records by species are also plotted.

Table 1 :

 1 Detailed geographic characteristics of the 46 RENECOFOR deciduous broadleaf forest plots (O=oak, B=beech).

	Plot	Tree Age	Long.	Lat.	Elevation	Slope
		(year)	(°)	(°)	(m)	(%)
	O1	149	4°18'E 48°20'N	115	0
	O2	61	0°50'W 43°44'N	20	5
	O3	85	0°01'W 47°27'N	57	0
	O4	115	5°46'E 49°01'N	220	0
	O5	85	3°45'E 50°10'N	149	3
	O6	69	0°02'W 43°12'N	370	12
	O7	50	6°12'E 47°52'N	240	0
	O8	82	5°14'E 46°58'N	190	0
	O9	103	5°14'E 46°10'N	260	3
	O10	130	2°43'E 46°40'N	260	0
	O11	98	4°27'E 48°17'N	160	0
	O12	93	2°07'E 47°15'N	176	1
	O13	102	5°04'E 47°04'N	220	0
	O14	70	1°30'E 49°21'N	175	0
	O15	116	1°32'W 48°10'N	80	0
	O16	107	1°15'E 47°34'N	127	0
	O17	154	4°57'E 49°02'N	180	2
	O18	100	6°29'E 48°52'N	315	4
	O19	143	7°27'E 49°00'N	320	15
	O20	76	3°39'E 46°58'N	270	7
	O21	75	2°18'E 49°23'N	55	1
	O22	103	0°40'E 48°31'N	220	5
	O23	152	7°28'E 47°41'N	256	0
	O24	79	0°22'E 47°47'N	170	0
	O25	113	1°44'E 44°02'N	300	18
	O26	97	0°29'E 46°37'N	116	4
	O27	144	6°02'E 48°01'N	330	0
	O28	91	7°43'E 48°59'N	350	10
	O29	128	2°43'E 48°27'N	80	0
	B1	68	3°07'E 49°12'N	145	0
	B2	102	2°59'E 46°11'N	590	15
	B3	103	5°48'E 44°07'N	1300	50
	B4	167	1°16'E 42°55'N	1250	32
	B5	98	0°51'W 49°10'N	90	4
	B6	143	4°51'E 47°48'N	400	3
	B7	56	6°16'E 47°11'N	570	2
	B8	173	5°17'E 44°55'N	1320	12
	B9	158	3°32'E 44°06'N	1400	25
	B10	121	5°04'E 47°47'N	440	0
	B11	104	5°00'E 49°10'N	250	0
	B12	77	2°52'E 49°19'N	138	0
	B13	82	0°39'W 43°09'N	400	44
	B14	175	0°26'E 43°01'N	850	25
	B15	102	1°19'E 49°42'N	210	0
	B16	123	2°10'E 43°24'N	700	0
	B17	83	6°14'E 48°06'N	400	3

Table 2 :

 2 Regression analysis in mean Julian days between in-situ observations of leaf unfolding dates (LU10% and LU90%) and satellite-based phenological dates (OGa for Modis and Dbefore for GLOBCARBON). Values shown in bold are Pearson's coefficients of correlation statistically significant at 5% probability level. Average date of Modis Onset Greenness Increase and Onset Greenness Maximum ** Dbefore: Day before the highest first derivative of LAI curve reach 90% of its maximum.

		Oaks (n=29)	Beech (n=17)	Broadleaf (n=46)
		Modis* GlobCarbon** Modis* GlobCarbon** Modis* GlobCarbon**
	In-situ LU10% r²	0.08	0.01	0.58	0.21	0.41	0.03
	Bias	13.8	23.8	10.05	13.8	12.4	20.0
	RMSE	15.2	26.3	12.75	15.0	14.6	22.3
	In-situ LU90% r²	0.01	0.05	0.47	0.27	0.23	0.10
	Bias	2.6	12.6	1.6	5.4	2.2	9.8
	RMSE	8.0	17.0	6.9	8.0	8.0	13.8
	* OGa :

Table 3 :

 3 Cross-comparison on specific NFI plots of the average leaf-unfolding date from MODIS OGa and GLOBCARBON Dbefore (see Table2). Correlations are statistically significant (p<0.0001).

		Oaks	Beech	Broadleaf
	n	816	349	17684
	r²	0.01	0.20	0.12
	bias	5.4	5.8	5.4
	RMSE	8.3	8.9	8.9


In "Making Things Public Atmospheres of Democracy" edited by Bruno Latour and Peter Weibel MIT Press, September 2005, 1072 pp.

Jean-Pierre Le Bourhis, "Water Parliaments : Some Examples", pages 482-485

In the arid places where one could only obtain water from wells, men had to come together to dig them, or at least to reach agreement on their use. This must have been the origin of societies and languages in warm countries. […] There the first ties were formed among families; there were the first rendezvous of the two sexes.[…] There, finally, was the true cradle of nations: from the pure crystal of the fountains flowed the first fires of love.

Jean-Jacques Rousseau

It might seem a little curious to forge links between water, love, the development of societies and language. But what the Rousseauian fable recalls in a lyrical mode is, actually, both obvious and useful: water is so closely related to the development of human groups that it becomes entangled in their social world, for which it constitutes a central link. However, the 'socialization' of water is an extremely complicated process-something the fable does not acknowledge-and is, moreover, one that has hardly been explored.

It also develops more slowly and in a more hazardous way than is supposed by the philosopher, in his rather hasty attempt to reconstitute these socio-hydraulic links. Institutions for water are built up by means of the gradual accumulation of equipment and technical, social, and political tools. This institutional machinery is particularly visible when the survival of the group is dependant upon the water supply; access, allocation, and circulation must be strictly controlled and therefore, corresponding collective (social, cultural and religious) frameworks must be constructed.1 When water is abundant, the intervention of these institutions is more discrete, but they are still present. What is more, if water-management demands a kind of cooperation, equality between users is not immediately essential. Many "water parliaments" do, in fact, operate as sites of collective decision but, for the most part, their functioning still remains nondemocratic. In this domain, as in that of the environment in general, the political debate is still taking shape. 2 The main obstacle to this process of democratization lies in how difficult it is to incorporate elements of democracy into preexisting technical and administrative infrastructures. Institutions for water have existed for a long time now, but they remain in the hands of a few bodies that retain exclusive control over them: states, local authorities or private interests, or various forms of alliance between these. To control water at the level of rivers is a costly undertaking and requires the involvement of organizations that can centralize the resources needed for intervention. Thus, massive hydraulic operations have historically been accompanied by a very heavy bureaucratization, due both to the technicality of the questions that are raised and to how essential it is to be able to mobilize huge resources for such large projects. This hydro-political regime is exemplified by the cases of Ancient Egypt or China, [START_REF] August | Oriental Despotism: A comparative study of total power[END_REF] but it also applies to the contemporary epoch: to the United States where, throughout the course of the 20 th century, the Federal government joined forces with private investors to transform the Californian desert into a fertile plain by means of a massive intervention. [START_REF] Marc | Cadillac Desert: The American West and its disappearing water[END_REF] As well, from 1815 on, as the Rhine in Europe was being navigated, it was continually regulated and subject to changes by means of an accord between riverside-states. [START_REF] Cioc | The Rhine: an eco-biography, 1815-2000[END_REF] Moreover, central and local powers often come together to develop and manage the waters: this was the case after the creation of the Tennessee Valley Authority 6 around the time when the Rhone and the Mediterranean areas were also being outfitted. [START_REF]Cultures, usages, et stratégies de l'eau en Méditerranée occidentale : tensions, conflits et régularisations[END_REF] Finally, on a more local scale, alliances between local powers and private interests make it possible to implement different management activities, such as the water-management associations of the Ruhr that date from the beginning of the 20 th century, 8 the French system of wateringues, or the Dutch Water Boards.

If, in all these cases, group decisions rest in the hands of a very few, we should take note of a more recent development going in the opposite direction. The growth of economic activities in the environmental sector means, in effect, that the environment is subject to greater and greater control. This, alone, is incentive enough to make water into a public good. [START_REF] Burchi | Current Developments and trends in the law and administration of water resources[END_REF] As well, it calls for the greater inclusion of users and citizens within a system of collective control. This development is notably visible in France where, since the 60s and 70s, the governance of water has been opened up in fairly progressive ways to new groups of people. The Water Agencies were created during this period, bringing principle users together with traditional representatives of the public authorities. This assembly (called "the Basin Committee") decides on the sources of financing for the development of policies on large rivers, but has extended its jurisdiction to include the planning of aquatic milieus. This political model is mostly applied to simple rivers, ones that have at their disposal representative assemblies of users who are given the power to intervene.

Other more recent activities were developed in the 90s and are moving in a similar direction. In 1992, an experimental form of public hearing was adopted for one of France's major rivers, the Dordogne. During the course of a "summit" held at a round table, seventeen groups of users met to establish how to manage the river according to a unanimously-voted "Charter" (see illustrations 1 and 2). In another example, a series of public debates was organized in 2004 about a development project that involved the construction of a dam and the management of the river downstream (the Garonne river). During its meetings, a "Special Committee" composed of political representatives, associates, and socio-professionals accompanied by experts, articulated different options and diverse points of view on the issue (illustration 3). The audience could intervene, raise questions and advance propositions (illustration 4). For many, these attempts at democratization remain incomplete; they reveal the difficulties of articulation between democratic debate and public decision. The Water Agency system has been subject to criticism for its lack of democracy (the appointment of its members, its status as a neocorporatist system that favors profit-making sectors of activity). Classic questions about representation must be raised, both in this case and in others: who speaks, in the name of which larger group, with what power and which mandate? Institutional solutions vary according to their plans of action and are not sufficient in their entirety to clarify the exchanges. The participants interrogate each other about their respective statutes: "who do you represent?", "in the name of whom are you speaking?", "who gave you such a mandate?", and so on.

The ways we have of representing the environmental issues are equally contested and uncertain. Data relating to the rivers is provided by studies and reports that align analyses, diagrams, graphs and tables; they are projected as images (see the reverse side of illustration 3) or reproduced on paper (documents covering the table, illustration 4). But these techniques of representation do not suffice as the basis for a collective vision of water (from the rate of flow of a river in m³/second, its power of purification and the species that inhabit the area, its degree of pollution,10 and so on.). There too, a good part of the exchanges are taken up by accusations of manipulation regarding the numbers, or regarding a flawed interpretation or biased selection of the data.

In summary, the institutional framework appears unstable, barely legitimate and easily contestable. Unlike traditional parliaments, the networks of representation here are not wellfounded and legitimate, as they could be between a voter and a representative. As well, the waters and rivers are too poorly equipped with the instruments needed to establish their identity, their development and their requirements in such a way that these could be viewed as incontestable.

Thus, these few burgeoning forms of water parliaments are developments worthy of praisepreviously hidden away in offices, water is a public issue today11 -and, at the same time, they are construction sites where an environmental democracy is being invented. The majority of its parameters are still undefined: communication between established powers and citizens, the representations of political dynamics and of the environment, the organization of on-site structures. 12 Initiating political debates about water seems to be a slow process because it follows the rhythm of historical processes. The stakes, however, are high: the collective governability of water depends on this configuration whose construction is already underway, especially in the face of other forms of (bureaucratic) regulation or non-regulation (laissez-faire economics, or the law of the market).

Translated from the French by Sarah Clift

For an illustration of this point in a work of fiction, cf. Frank Herbert, Dune. Gollancz, London, 1966.

Bruno Latour, Politics of Nature: How to bring the sciences into democracy. Harvard University Press, Cambridge, 2004.

See Christelle Gramaglia's contribution to this volume.

Jean-Pierre Le Bourhis, La publicisation des eaux. Rationalité et politique dans la gestion de l'eau en France, Doctoral Thesis, University of Paris-I Panthéon Sorbonne, Paris, 2004.

All these questions are currently being raised in connection with the application of the Water Framework Directive (adopted in 2000).


Francine De Quelen

Jacques Mourot

Gaëlle Boudry

Supplementation of the maternal diet with alpha-linolenic acid (ALA) modifies the development of the enteric nervous system with functional consequences on intestinal permeability in newborn piglets

Supplementation of the maternal diet with alpha-linolenic acid (ALA) modifies the development of the enteric nervous system with functional consequences on intestinal permeability in newborn piglets

De Quelen F. 1,2 , Mourot J. 1 , Boudry G. The enteric nervous system (ENS) participates in the regulation of various mucosal functions such as intestinal motility and secretion but also permeability. The ENS shares embryologic similarities with the CNS whose perinatal development is known to be modulated by n-3 polyunsaturated fatty acids (PUFA). We hypothesized that n-3PUFA in the maternal diet modify the development of the ENS, by analogy with the CNS, and have consequences on the regulation of jejunal permeability of newborn piglets.

The intestinal permeability was measured on 28 day-old piglets nursed by sows who had been fed either a lard-based diet (LAR, ALA = 3.5% of total fatty acid) or a linseed oil based-diet (LSO, ALA= 28.3%) during gestation and lactation. Basal jejunal permeability to FITCdextran 4000 (FD4) was evaluated in Ussing chambers. Different antagonists of the ENS were used to determine the nervous regulation of intestinal permeability. Additional intestinal tissue were collected to evaluate cholinergic neurons percentages in the ENS by immunohistochemistry and to analyze the expression of tight junction protein and choline acetyl transferase (ChAT) by Western blot and RT-PCR. Inflammatory consequences were also evaluated by analyse of cytokines mRNA in the tissue.

Basal jejunal permeability to FD4 was higher in LSO compared to LAR piglets (621±55 vs 341±45 ng/cm 2 /h, P<0.01). The protein or mRNA expressions of tight junction did not differ between the two groups of piglets. In the LAR group, TTX and VIPra increased jejunal permeability (+ 25%, P<0.05 and +76%, P<0.01, respectively), suggesting a VIPergic tonus.

On the contrary, in the LSO group, VIPra did not modify basal permeability but TTX and atropine/hexamethonium decreased FD4 flux (-20%, P=0.07 and -30%, P<0.05, respectively), suggesting that the high permeability was due to a cholinergic tonus. Moreover, the proportion of cholinergic neurons in the ENS of LSO piglets was greater than in LAR piglets (22.9±1.2% for LAR vs 26.6±0.9% for LSO, P<0.05) and the mRNA relative expression of ChAT was greater in the LSO group compared with the LAR group (1.9±0.6 vs 1.1±0.6, P<0.05). Despite the increase in permeability the cytokine profiles (IL-1β, IL10, TGF-β) did not differ between the two groups.

In conclusion, supplementation of the maternal diet with ALA modified the development of the ENS and the nervous regulation of jejunal permeability, with the induction of a cholinergic tonus. The consequences of such differences on the maturation of the gut immune system of newborn warrant further investigations.