F Phocas

Genetic analysis of breeding traits in a Charolais cattle population segregating an inactive myostatin allele

Keywords: beef cattle, calving difficulty, double muscling, fertility, genetic parameter, myostatin

ABSTRACT:

The objective of this study was to quantify the effects of the Charolais-specific inactive myostatin allele on phenotypic means and genetic parameters of heifer breeding traits. Records were registered from 1996 to 2006 in 282 herds dedicated to the on-farm French Charolais purebred progeny test. Data consisted of 36,867 female calf records, including 17,518 inseminated heifers that were bred by 186 genotyped sires, of which 43 were heterozygous and 6 were double muscled bulls. Six traits were analyzed under a univariate animal model accounting for maternal effects and myostatin sire genotype: calving difficulty, birth and weaning weights, muscle and skeleton scores at weaning, and fertility of artificially inseminated heifers. The inactive myostatin allele had a large positive effect on weaned heifer muscle score, unfavorable effects on calving difficulty and skeleton score, and small effects on birth and weaning weight. This allele did not induce an adverse effect on heifer fertility. Univariate estimates of polygenic variance parameters were almost unaffected by the estimation of the myostatin sire genotype, except for heifer morphology traits. Direct and maternal genetic variances and covariances were significantly reduced under a model accounting for the myostatin sire genotype effect on muscle score. The myostatin genotype explained 45% of the direct genetic variance and had a pleiotropic action across both direct and maternal effects on muscle score. Because the myostatin sire genotype had no significant effect on birth weight, the multitrait sire analysis concerned only the 5 other traits. Accounting for the myostatin sire genotype, estimates of polygenic correlation between skeleton score and muscle score, on the one hand, and calving difficulty on the other hand, were largely modified: from a negative estimate of -0.3 to 0.0 and from a positive estimate of 0.4 to 0.7, respectively.

INTRODUCTION

The so-called double muscling phenotype is a muscular hypertrophy due to an increase in the volume of muscle cells, which is explained by disruptive mutations in the myostatin gene [START_REF] Grobet | A deletion in the bovine myostatin gene causes the double-muscled phenotype in cattle[END_REF][START_REF] Grobet | Molecular definition of an allelic series of mutations disrupting the myostatin function and causing doublemuscling in cattle[END_REF]. It is associated with some breeding problems such as reduced fertility, dystocia, reduced calf survival, and milking cow ability [START_REF] Ménissier | Muscle hypertrophy of genetic origin and its use to improve beef production[END_REF][START_REF] Hanset | Genetic analysis of some maternal reproductive traits in the Belgian Blue cattle breed[END_REF]. However, cattle showing the syndrome have greater meat yield and meat quality (lean and tender meat) as reviewed by [START_REF] Arthur | Double muscling in cattle: A review[END_REF]. Literature information on the effects of double muscling is mainly available for the Belgian Blue breed [START_REF] Ménissier | Muscle hypertrophy of genetic origin and its use to improve beef production[END_REF][START_REF] Hanset | Genetic analysis of some maternal reproductive traits in the Belgian Blue cattle breed[END_REF], crossbred populations [START_REF] Arthur | Heterosis, maternal and direct effects in double-muscled and normal cattle: I. Reproduction and growth traits[END_REF][START_REF] Casas | Quantitative analysis of birth, weaning, and yearling weights and calving difficulty in Piedmontese crossbreds segregating an inactive myostatin allele[END_REF][START_REF] Casas | Association of myostatin on early calf mortality, growth, and carcass composition traits in crossbred cattle[END_REF][START_REF] Short | Pleiotropic effects in Hereford, Limousin, and Piemontese F 2 crossbred calves of genes controlling muscularity including the Piedmontese myosatin allele[END_REF], and South Devon cattle [START_REF] Wiener | Muscle-related traits in cattle: The role of the myostatin gene in the South Devon breed[END_REF]. However, the double-muscled syndrome is found in many breeds of cattle and is often due to different alleles of the myostatin gene. In the Charolais breed, a specific inactivated myostatin mutation, named Q204X, segregates and corresponds to a cytosine to thymine substitution in the second exon of the myostatin gene [START_REF] Grobet | Molecular definition of an allelic series of mutations disrupting the myostatin function and causing doublemuscling in cattle[END_REF][START_REF] Dunner | Haplotype diversity of the myostatin gene among beef cattle breeds[END_REF]. There is no evidence that different alleles of the myostatin gene playing a similar role in muscular hypertrophy should also have similar pleiotropic effects on other traits. Furthermore, the literature is mainly focused on dystocia, growth, conformation, and carcass traits. Few references focus on female breeding traits, and only in the Belgian Blue breed [START_REF] Hanset | Genetic analysis of some maternal reproductive traits in the Belgian Blue cattle breed[END_REF] or crossbreds [START_REF] Arthur | Heterosis, maternal and direct effects in double-muscled and normal cattle: I. Reproduction and growth traits[END_REF][START_REF] Casas | Quantitative analysis of birth, weaning, and yearling weights and calving difficulty in Piedmontese crossbreds segregating an inactive myostatin allele[END_REF][START_REF] Short | Pleiotropic effects in Hereford, Limousin, and Piemontese F 2 crossbred calves of genes controlling muscularity including the Piedmontese myosatin allele[END_REF]. Consequently, the first aim of the study was the estimation of myostatin pleiotropic effects on Charolais breeding traits, such as calving difficulty (CD), birth weight, weaning weight (WW), morphology, and fertility of heifers. A second objective was to assess the impact of the myostatin genotype on the estimation of genetic pa-rameters, in particular on genetic correlations between breeding traits.

MATERIALS AND METHODS

Animal Care and Use Committee approval was not obtained for this study because the data were obtained from the existing French national database of cattle herds named SIG.

Data

Records were extracted from the French national database of cattle. Records concerned 282 Charolais herds located in Burgundy and Vendée that are dedicated to on-farm purebred progeny testing. Those herds had a large number of progeny born in 1996 to 2006 bred by 186 sires genotyped for the myostatin gene. Data editing consisted of selecting records from those sires and from contemporary groups with over 4 heifer records per herd-year. The final research data set included data of 36,867 female calves, from 24,213 dams, for CD, birth weight (WO) and WW, muscle (MS) and skeleton (SS) scores at weaning, and fertility. Fertility was measured by calving success following the first insemination on only 17,518 heifers selected for reproduction after weaning. Birth traits (WO and CD) were directly recorded by breeders. Calving difficulty was scored 1 for unassisted calving and 4 for a cesarean section. Weaning weight was adjusted to 210 d. Muscle and skeleton scores were scored over 100 points from visual appraisal. Muscle score was a sum of scores of 5 different subjective measurements to assess the calf muscular frame: width at withers, width behind withers, hindquarter width, curve of rump, and loin thickness. Skeleton score was a sum of scores of 5 other subjective measurements to assess the calf skeletal frame: stature, canon bone circumference, withers hooks length, hooks spins length, and spins width. Technicians belonging to agricultural institutes, which are independent of the breed societies, measured the WW and performed the scoring for MS and SS. Table 1 presents the numbers of records and raw means of traits according to myostatin sire genotypes and the female populations that are concerned by the measurements: either heifers alive at birth (birth traits) and at weaning (weaning traits) or inseminated heifers (fertility). The small number (6) of double muscled sires is explained by the French Charolais breeding goal: the Q204X allele frequency should not increase in the purebred Charolais cow population, where this frequency is estimated at about 12% [START_REF] Allais | Qualités des carcasses et des viandes de jeunes bovins hétérozygotes pour les mutations Q204x et nt821 du gène de la myostatine dans trois races allaitantes françaises[END_REF]. Consequently, only 2 double muscled sires had over 30 daughters recorded at birth, corresponding to 10 and 6 heifer reproductive records, respectively.

Statistical Analyses

Data were analyzed by animal linear models using ASREML software [START_REF] Gilmour | ASREML Reference Manual[END_REF]. To easily derive genetic correlations across traits, discrete traits (like CD and fertility) were analyzed under linear models, knowing that estimates of genetic correlations are not affected by the statistical treatment (linear or threshold model) of the categorical trait [START_REF] Kadarmideen | Genetic parameters and evaluations from single and multiple trait analysis of dairy cow fertility and milk production[END_REF].

The relationship matrix included 3 generations of ancestors that accounted for 69,463 animals in total. Apart from the animal effects, other random effects were maternal genetic and dam permanent environmental effects for all traits except fertility. Those effects had near zero variances for fertility, and, consequently, were ignored. Random mating bull effects were fitted for fertility, although they explained less than 0.3% of the phenotypic variance.

Fixed effects were the contemporary group herdbirth year for CD and WO; herd-year-weaning group for WW, MS, and SS; and herd-year-reproduction group for fertility. Because no reproduction group was indicated, following the strategy of [START_REF] Robinson | Days to calving in artificially inseminated cattle. 55-58[END_REF], the reproduction group was defined within herd-year by 60-d intervals between last AI of a first group and first AI of a second group. The age of the dam was also fitted for all traits into 6 classes: 3 classes for primiparous dams (<25 mo, 25 to 30 mo, >30 mo), 2 classes for the 2nd calving dams (below and over 42 mo) and the last class for multiparous cows over 42 mo.

For birth and weaning traits, the last fixed effect was the birth period (5 levels: January to February, March to June, July to August, September to October, and November to December). For fertility, the heat synchronization treatment (no/yes), AI agent, and week day of the AI were also fitted, as well as the female age class at first AI (12 to 18 mo, 19 to 27 mo, and 28 to 30 mo) and the breeding period (5 levels: September to October, November to January, February to March, April to May, and June to August). Univariate analyses were performed to determine the impact of myostatin sire genotype on each of the 6 traits, separately. A likelihood ratio test was performed to test the significance level of the sire genotype on each heifer trait. The P-value was derived from a chi-square distribution with 2 degrees of freedom (3 possible sire genotypes). Genetic parameters were estimated with both models accounting or not for the sire genotype as a fixed effect. Because 88% of the heifer dams are expected to be normal homozygous (and the last 12% only heterozygous) at the myostatin gene, the allelic substitution effect can be approximated by the effect estimated for the double muscled sire genotype because 94% of their daughters are expected to be heterozygous and only 6% double muscled. Whatever the mutant allele frequency, the contrast of progeny from double muscled sires vs. progeny of normal sires would cleanly estimate the allelic substitution effect under 3 conditions: additive gene action, no assortative mating, and a large sample of sires from each genotype. When considering that double muscled sires and heterozygous sires are drawn from the same genetic population, the heterozygous sire genotype effect should be one-half the effect observed for double muscled sires. Hence, another way to approximate the allelic substitution effect is to double the heterozygous sire genotype effect. Due to the small number of double muscled sires accounted for in our analysis, this last alternative should be preferred. The dominance, additive, or recessive effects of the Q204X allele on heifer traits could not be assessed because heifer genotypes were not available.

A multivariate analysis was performed for CD, WW, MS, SS, and fertility because they were the 5 traits exhibiting significant differences (P-value up to 10%) according to sire genotypes. For multivariate analysis, the maternal genetic effect was not accounted for, but a global maternal effect (including both genetics and permanent environment) was fitted for CD, WW, MS, and SS. Indeed, data information was not sufficient to estimate maternal genetic correlations [START_REF] Heydarpour | Influence of population structure on estimates of direct and maternal parameters[END_REF]. Ignoring maternal genetic effects under an animal model may induce large overestimation of direct genetic variances [START_REF] Meyer | Variance components due to direct and maternal effects for growth traits of Australian beef cattle[END_REF], and in consequence, may provoke bias in the estimation of direct genetic correlations. Because the focus was on getting unbiased estimates of direct genetic correlations across heifer breeding traits, a sire model was considered for the multitrait analysis. The sire relationship matrix included 3 generations of ancestors that accounted for 784 sires in total. For each trait, the same fixed effects as in the univariate analysis were fitted in the multitrait model.

RESULTS AND DISCUSSION

Myostatin Sire Genotype Effects on Heifer Birth Traits

The goodness-of-fit of the model accounting for the effect of myostatin sire genotype on CD was not clearly better than the fit of the simple polygenic model: the P-value of the likelihood ratio (LR) test was only 10% (Table 2). However, this low significance level is probably due to the joint facts of a small effect of heterozygous sire on CD and a low number of records from double muscled sires. The effect of double muscled sire genotype on the birth condition of daughters was as large as the effect of the parity of the dams (i.e., primiparous versus second-calving dams). [START_REF] Short | Pleiotropic effects in Hereford, Limousin, and Piemontese F 2 crossbred calves of genes controlling muscularity including the Piedmontese myosatin allele[END_REF] found an allelic substitution effect equal to about 0.6 phenotypic SD on dystocia score of Piedmontese crossbred calves. This effect seemed to be additive. Here, the allelic substitution is only estimated to be 0.2 phenotypic SD on CD. [START_REF] Casas | Quantitative analysis of birth, weaning, and yearling weights and calving difficulty in Piedmontese crossbreds segregating an inactive myostatin allele[END_REF] did not find any significant contrast between CD of heterozygous and normal Piedmontese crossbred calves, but a significant increase of 19% in the proportion of CD for double muscled calves compared with heterozygous ones.

The LR test did not reveal any significant effect of the myostatin genotype on WO. Because of the nonsignificant LR test, it was decided to drop the WO records for running the multitrait analysis.

The allelic substitution effect was also estimated to about 20% of phenotypic SD on heifer birth weight (i.e., +1 kg is expected for a heterozygous compared with a normal female calf). [START_REF] Casas | Quantitative analysis of birth, weaning, and yearling weights and calving difficulty in Piedmontese crossbreds segregating an inactive myostatin allele[END_REF] estimated this contrast to +3 kg in Piedmontese crossbreds accounting for male and female calves. They did not find any significant contrast of birth weights between double muscled and heterozygous calves. However, most of the results in the literature found rather an additive gene action on WO with the magnitude of the allelic substitution effect ranging from 0.2 to 0.4 phenotypic SD. [START_REF] Short | Pleiotropic effects in Hereford, Limousin, and Piemontese F 2 crossbred calves of genes controlling muscularity including the Piedmontese myosatin allele[END_REF] found a larger and additive allelic substitution effect equal to about 0.4 phenotypic SD on birth weight. In a crossbred experimental population, [START_REF] Casas | Association of myostatin on early calf mortality, growth, and carcass composition traits in crossbred cattle[END_REF] looked at the effect of the inactive myostatin allele on WO within the Charolais sire line (Q204X allele) or Belgian Blue × Bristish Breed sire line (nt821 allele). They found an allelic substitution effect of +2 kg at birth for both alleles. [START_REF] Vinet | Bilan génétique de 15 ans de sélection du troupeau souche INRA95[END_REF] compared the effects of the myostatin gene polymorphism on calf WO and conformation at 4 mo in a double muscled synthetic sire line, INRA95. In this population, 6 different inactive alleles of the myostatin gene were segregating. The authors found a significant contrast between 134 normal and 228 Q204X/Q204X calves of 0.5 phenotypic SD for WO, and 0.2 phenotypic SD between WO of 337 heterozygous and the 134 normal calves, corresponding to +1.5 kg at birth. This last result was consistent with our finding on Charolais purebred female calves.

Myostatin Sire Genotype Effects on Heifer Weaning Traits

The maximum significant LR test was for MS (Table 2). The allelic substitution effect was estimated to be 0.6 phenotypic SD of MS from the contrast between heterozygous and normal sire genotypes and 0.9 phenotypic SD when considering the contrast between double muscled and normal sire genotypes. [START_REF] Vinet | Bilan génétique de 15 ans de sélection du troupeau souche INRA95[END_REF] estimated the contrast between Q204X/Q204X and normal calves to be equal to 0.85 phenotypic SD on calf MS at 4 mo, whereas the contrast between heterozygous and normal calves was only 0.2 phenotypic SD. The Q204X allele effect seemed to be partially recessive on MS.

The LR test for skeletal score (Table 2) was also highly significant. The allelic substitution effect was estimated to be -0.15 phenotypic SD of SS from the contrast between heterozygous and normal sire genotypes and -0.60 phenotypic SD when considering the contrast between double muscled and normal sire genotypes. No information was found in the literature to discuss that result, apart from the well-known reduction in the weight of the skeleton of double muscled animals [START_REF] Arthur | Double muscling in cattle: A review[END_REF]. Given that SS was highly correlated with WW, the results were consistent with those presented below for WW.

The LR test indicated a significance level of 2% for the myostatin sire genotype on WW, although the contrast between heterozygous and normal sire genotypes was null. The contrast was -8 kg (i.e., about -0.3 phenotypic SD of WW) between double muscle and normal sire calves. The discrepancy between the 2 estimates of the allelic substitution effect may be due to the sampling genetic background of the 6 double muscled sires. The data seemed rather to indicate a null effect of the Q204X allele on WW. [START_REF] Wiener | Muscle-related traits in cattle: The role of the myostatin gene in the South Devon breed[END_REF] in South Devon cattle and [START_REF] Vinet | Bilan génétique de 15 ans de sélection du troupeau souche INRA95[END_REF] in the INRA95 sire line also found a nonsignificant contrast for WW between genotypes. On the contrary, [START_REF] Casas | Quantitative analysis of birth, weaning, and yearling weights and calving difficulty in Piedmontese crossbreds segregating an inactive myostatin allele[END_REF] found that heterozygous Piedmontese crossbred calves were significantly heavier than the 2 homozygous types, which had similar WO at weaning. However, their results were not confirmed by [START_REF] Short | Pleiotropic effects in Hereford, Limousin, and Piemontese F 2 crossbred calves of genes controlling muscularity including the Piedmontese myosatin allele[END_REF] in Piedmontese crossbreds, where no significant effect of the mutant allele was detected for WW. Once again, [START_REF] Casas | Association of myostatin on early calf mortality, growth, and carcass composition traits in crossbred cattle[END_REF] for the 11-pair deletion mutation of the Belgian Blue Breed and the Q204X mutation of the Charolais breed found a significant increase in the WW of the heterogeneous calves (+10 kg), 2 homozygous types having similar WW in the Belgian Blue crossbreds.

Myostatin Sire Genotype Effects on Heifer Fertility

The LR test (Table 2) did not clearly select the model accounting for the effect of myostatin sire genotype on fertility: the significance threshold was only 7%. The Q204X allele seemed to have a null effect, or even a slightly favorable effect (1 to 10% of phenotypic SD), on heifer fertility. Because information came from breeders and not from the experiment station, it was checked that these results were not biased by MS phenotypic selection of the normal heifers. Average MS of the inseminated heifers (Table 3) were very close to the average MS of all animals at weaning (Table 1) recorded before selection occurred for breeding. Heifer selection was clearly performed on greater SS and WW, and no counter-selection of double muscled or heterozygous heifers seemed to occur. On raw data (Table 1), fertility of heifers bred by heterozygous sires is less (-1.5%) than fertility of heifers bred by normal sires. However, the corresponding BLUP estimate is +0.3% with SE of 1.53%, clearly indicating no effect of sire genotype on fertility. The large, but nonsignificant, estimate of +4.7% for fertility of heifers sired by double muscled bulls was consistent with raw data, but is probably due to sampling error of the 2 sires or their 16 daughters. Indeed, this result seemed to be inconsistent with the literature (e. g., [START_REF] Ménissier | Muscle hypertrophy of genetic origin and its use to improve beef production[END_REF][START_REF] Arthur | Heterosis, maternal and direct effects in double-muscled and normal cattle: I. Reproduction and growth traits[END_REF]; [START_REF] Arthur | Double muscling in cattle: A review[END_REF], where double muscled Belgian Blue heifers had a 4% less nonreturn rate [START_REF] Hanset | Genetic analysis of some maternal reproductive traits in the Belgian Blue cattle breed[END_REF] than normal heifers. It also can be thought from the experiment of [START_REF] Arthur | Heterosis, maternal and direct effects in double-muscled and normal cattle: I. Reproduction and growth traits[END_REF] that the inactivated myostatin gene has rather a recessive effect on female fertility. However, no information based on genotypes rather than visual classification of animals was found in the literature of the last 10 yr. Fertility is a difficult trait to analyze due to its huge phenotypic variability (Table 4). In most analyses of on-farm data, the environmental effects considered for modeling heifer fertility are only the age of the heifer, her mating group, and her bull mate. In the present study, other large effects were also fitted such as the weekday of the AI (an AI on Monday had -2.7% less success than an AI on Wednesday) or the synchronization treatment (-4.2% on heifer fertility).

Impact of Myostatin Sire Genotype on Direct and Maternal Genetic (Co)variances Within Trait

Tables 4 and5 present genetic and phenotypic parameters estimated by a univariate animal model for birth traits, WW, and fertility, and for morphology traits, respectively. Table 4 presents the results for a model not accounting for the myostatin genotype because almost the same variance components were estimated for CD, WO, WW, and fertility whether or not the myostatin sire genotype was accounted for. Parameters for weaning traits and fertility are quite similar to those previously estimated on the French Charolais population [START_REF] Phocas | Genetic parameters for birth and weaning traits in French specialized beef cattle breeds[END_REF][START_REF] Phocas | Fertility in beef cows[END_REF]. On the con-trary, the direct-maternal genetic correlations for birth traits are clearly not statistically different from zero in this study, whereas previous estimates were negative (about -0.4). This was probably due to the fact that only female traits were recorded here, whereas male and female calves were both accounted for in the previous analysis.

For morphology traits (Table 5), and in particular for MS, genetic parameters were significantly different when accounting for the myostatin sire genotype in the model. As expected, the phenotypic and direct genetic variances were reduced under such a model. The variance reductions would have probably been larger if heifer genotypes, rather than sire genotypes, could have been fitted [START_REF] Hoeschele | Genetic evaluation with data presenting evidence of mixed major gene and polygenic inheritance[END_REF]. Maternal heritabilities of MS and SS, and direct heritability of SS were kept almost constant at about 0.04, 0.06, and 0.25, respectively. The direct heritability of MS dropped from 0.38 to 0.25. The direct-maternal genetic correlation for SS was kept almost constant at about -0.45, whereas the corresponding correlation for MS increased from -0.45 to -0.11 as the maternal genetic variance was reduced from 4.2 to 3.4. This last phenomenon revealed some pleiotropic and negative action of the myostatin gene across maternal and direct effects of MS. However, only a small amount (4%) of the phenotypic variability is explained by the maternal genetics.

Impact of Myostatin Sire Genotype on Direct Genetic Correlations Across Traits

For the multivariate sire analysis, maternal genetic effects were ignored, but they were still accounted for Calving difficulty score ranges from 1 for unassisted calving to 4 for cesarean section. as a part of the permanent maternal random effect for CD, WW, MS, and SS of heifers. Table 6 presents the results both for the polygenic sire model and for the sire model accounting for the myostatin genotype as a fixed effect. Heritabilities were kept constant across models for CD, WW, SS, and fertility and were very similar to previous univariate estimates under the animal model. On the contrary, the drop in MS heritability when accounting for myostatin sire genotype was larger than in the univariate animal model analysis.

Under the animal model, the myostatin sire genotype explained about one-third of the additive genetic variance, whereas it explained 45% of the genetic variance under a sire model. This last figure is a better estimate of the contribution of the myostatin gene to the genetic variance of MS due to the lack of knowledge of heifer genotypes under the animal model. Under a polygenic model, the genetic correlations statistically different from zero (absolute value over twice the SE) were mainly the correlations between SS and the 4 other traits. The maximum correlation was estimated between SS and WW (over 0.8). Intermediate positive correlations were estimated between SS and fertility (0.5) or CD (0.4), and between CD and WW (0.4). Moderate negative correlations were estimated between MS and SS (-0.3) or fertility (-0.4).

When considering the myostatin sire genotype, the correlation estimate between WW and SS stayed re-markably constant (over 0.8) due to the absence of a sufficient effect of the myostatin sire genotype on heifer WW. On the contrary, some large differences were observed in other correlation estimates. In particular, the correlation between MS and SS tended toward zero, indicating that the previous negative estimate was only due to the pleiotropic effect of the myostatin gene on MS and SS. The correlation between SS and CD was also greatly increased with a new estimate of about 0.7; the previous estimate was less due to the opposite sign of the myostatin genotype effect on both traits. Such a result questioned the general tendency to consider that estimates of genetic correlation close to zero correspond to traits that are genetically unlinked. In fact, the mathematical rule applying to independent variables should always be kept in mind: the correlation between 2 independent variables is zero, but a null correlation does not mean that 2 variables are independent.

The only literature information available on differences in genetic correlations when accounting for the myostatin genotype concerned the South Devon cattle where the same allele (nt821) as in the Belgian Blue Breed is segregating [START_REF] Wiener | Muscle-related traits in cattle: The role of the myostatin gene in the South Devon breed[END_REF]. Three traits of their paper were almost comparable with CD, MS, and WW. Contrary to the present study, the authors found no significant correlation between CD and WW, whatever the genetic model. As in the present study, they found almost no correlation between MS and WW whatever the genetic model and also between MS and CD under a polygenic model. This last correlation became moderately positive (0.2) when the myostatin genotype was accounted for in their study, whereas the correlation remained null in the present analysis.

Implications for Beef Cattle Selection Schemes

Estimates of genetic correlations across breeding traits validated the traditional behavior of Charolais breeders, which consists of selecting replacement heifers among calves with high SS and moderate MS, to maximize the weaned calf crop per cow. For decades, breeders put emphasis on female SS and selected against double muscling genotypes in the Charolais cow population. Hence, breeders indirectly reduced CD risk, but also improved WW and heifer fertility. Identification of the myostatin gene and its variation in cattle has been an important finding of the end of the 1990s because it enables management of its allele frequency in the beef cattle population, depending on the economic breeding goals. A deliberate effort was made in the last century either to increase the frequency of double muscling (Belgian Blue, Piedmontese cattle) when considering that the favorable effects on beef traits are overwhelming the unfavorable ones on breeding traits, or to eradicate it when the breeding goal favored breeding traits as in hardy breeds such as Gascon cattle. Another system proposed was a terminal sire breeding system whereby normal females are mated to double muscled sires and all progeny slaughtered, the breeding female herd being kept free from double muscling. Further investigation is still needed to propose alternative and efficient methods of selection within beef cattle breeds for high female breeding performance and high quality and quantity of beef production. For instance, accounting for the myostatin genotype separately, Charolais breeders should be aware that selection on a high polygenic component of the SS would provoke a strong increase in CD due to the high genetic correlation between the polygenic components of the 2 traits.

2

  Muscle score is the sum of 5 subjective measurements assessing the calf muscular frame. It ranges from 0 to 100 points.3 Skeleton score is the sum of 5 subjective measurements assessing the calf skeletal frame. It ranges from 0 to 100 points.

Table 1 .

 1 Number of records according to sire myostatin genotype and corresponding phenotypic means for breeding traits recorded on Charolais heifers

	Item	Total data set	Normal sires	Heterozygous sires	Double muscled sires
	Number of sires	186	137	43	6
	Average number of daughters/sire	198	216	166	17
	Total birth records	36,867	29,629	7,136	102
	Total weaning records	31,442	25,349	6,010	83
	Total reproductive records	17,518	14,229	3,273	16
	Mean calving difficulty 1	1.34	1.38	1.30	1.40
	Mean birth weight, kg	45.2	44.6	45.9	45.6
	Mean weaning weight, kg	267.1	265.7	271.5	250.4
	Mean muscle score 2	59.8	59.0	62.7	68.4
	Mean skeleton score 3	65.5	65.4	65.9	58.9
	Mean fertility, %	56.6	56.9	55.3	68.7

Table 2 .

 2 Estimates of sire myostatin genotype effects on heifer traits (in contrast to normal sire genotype effects)

	Trait	Heterozygous genotype (SE)	Double muscled genotype (SE)	Likelihood ratio test	P-value
	Calving difficulty 1	+0.06 (0.017)	+0.19 (0.065)	5.6	0.10
	Birth weight, kg	+0.5 (0.26)	+0.6 (0.82)	1.0	0.84
	Weaning weight, kg	+0.1 (1.43)	-8.2 (4.40)	8.0	0.02
	Muscle score 2	+2.8 (0.51)	+9.0 (1.71)	45.4	<0.001
	Skeleton score 3	-0.6 (0.46)	-5.1 (1.42)	12.4	<0.001
	Fertility, %	+0.3 (1.53)	+4.75 (13.6)	6.2	0.07

Table 3 .

 3 Phenotypic means for breeding traits recorded on artificially inseminated Charolais heifers according to their sire myostatin genotype

	Mean trait	Total data set	Normal sires	Heterozygous sires	Double muscled sires
	Calving difficulty 1	1.33	1.36	1.24	1.12
	Birth weight	45.7	45.3	47.1	44.1
	Weaning weight, kg	271.1	269.4	278.5	258.9
	Muscle score 2	60.8	59.9	64.2	68.3
	Skeleton score 3	67.0	66.9	67.7	64.3
	Fertility, %	56.6	56.9	55.3	68.7
	1

Table 4 .

 4 Genetic

	parameters for calving difficulty (CD), birth weight (WO), weaning weight (WW), and fertility
	under polygenic animal univariate models (SE in parentheses)
	Parameter	CD	WO	WW	Fertility
	Phenotypic variance	0.310	28.72	790.0	2,410
	Direct genetic variance	0.012	5.70	133.6	26.0
	Maternal genetic variance	0.023	1.57	61.83	0
	Permanent environment/s p 2	0.11 (0.012)	0.12 (0.010)	0.13 (0.012)	0
	Direct heritability	0.04 (0.010)	0.20 (0.031)	0.17 (0.030)	0.01 (0.005)
	Maternal heritability	0.07 (0.014)	0.05 (0.013)	0.08 (0.017)	0
	Direct-maternal genetic correlation	-0.07 (0.165)	-0.27 (0.118)	-0.39 (0.104)	-

Table 5 .

 5 Genetic parameters of morphology traits at weaning (SE in parentheses) for univariate animal models

			Muscle score		Skeleton score
	Parameter 1	Polygenic model	Model with myostatin gene	Polygenic model	Model with myostatin gene
	s p 2	101.1	97.70	67.93	67.54
	s d 2	38.72	24.89	17.25	15.70
	s m 2	4.24	3.43	3.84	4.03
	C 2	0.04 (0.011)	0.04 (0.011)	0.07 (0.011)	0.07 (0.011)
	h 2 d	0.38 (0.051)	0.25 (0.038)	0.25 (0.040)	0.23 (0.038)
	h 2 m	0.04 (0.013)	0.03 (0.011)	0.06 (0.014)	0.06 (0.015)
	Rdm	-0.45 (0.119)	-0.11 (0.153)	-0.46 (0.104)	-0.44 (0.104)
	1 s p 2 = phenotypic variance; s d 2 = direct genetic variance; s m 2 = maternal genetic variance; C 2 = permanent environment/s p 2 ; h 2 d = direct heri-tability; h 2 m = maternal heritability; Rdm = direct-maternal genetic correlation.

Table 6 .

 6 Heritabilities (h 2 ), maternal contribution to phenotypic variance (C 2 ), and direct additive genetic correlations across heifer traits: figures with asterisks (*) for a sire model accounting for myostatin genotype, polygenic sire model otherwise (SE in parentheses)

	Trait	h 2	C 2	CD	WW	MS	SS	Fertility
	CD 1	0.03 (0.008)	0.18	0.03 (0.008)*	0.51 (0.128)*	-0.10 (0.158)*	0.68 (0.105)*	0.15 (0.255)*
	WW 2	0.19 (0.033)	0.07	0.38 (0.147)	0.17 (0.032)*	0.23 (0.126)*	0.82 (0.046)*	0.44 (0.226)*
	MS 3	0.31 (0.049)	0.09	0.23 (0.154)	0.03 (0.125)	0.17 (0.032)*	-0.02 (0.131)*	-0.53 (0.208)*
	SS 4	0.28 (0.046)	0.02	0.40 (0.149)	0.83 (0.044)	-0.28 (0.112)	0.23 (0.039)*	0.44 (0.226)*
	Fertility	0.01 (0.006)	-	0.07 (0.267)	0.24 (0.248)	-0.44 (0.240)	0.47 (0.242)	0.01 (0.006)*

1 CD = calving difficulty score. 2 WW = weaning weight. 3 MS = muscle score.

Corresponding author: florence.phocas@jouy.inra.fr Received August

22, 2008. Accepted February 6, 2009.

at INRA Institut National de la Recherche Agronomique on September 3, 2013 www.journalofanimalscience.org Downloaded from

Calving difficulty score ranges from 1 for unassisted calving to 4 for cesarean section.

Muscle score is the sum of 5 subjective measurements assessing the calf muscular frame. It ranges from 0 to 100 points.

Skeleton score is the sum of 5 subjective measurements assessing the calf skeletal frame. It ranges from 0 to 100 points.

SS = skeleton score.Phocas

This article has been cited by 1 HighWire-hosted articles: at INRA Institut National de la Recherche Agronomique on September 3, 2013 www.journalofanimalscience.org Downloaded from


PhD Carla Estaquio
email: c.estaquio@uren.smbh.univ-paris13.fr

PhD Nathalie Druesne-Pecollo

PhD; LUC DAUCHET Paule Latino-Martel

MD, PhD Serge Hercberg

S Bertrais

P Latino- Martel

Socioeconomic Differences in Fruit and Vegetable Consumption among Middle-Aged French Adults: Adherence to the 5 A Day Recommendation

Background Numerous studies support the protective effect of high fruit and vegetable consumption on chronic disease risk, mainly against cancer and cardiovascular diseases. The increase of fruit and vegetable intake has become a public health priority in many countries. Objective The aim of the study was to investigate the relationships of socioeconomic, demographic, and behavioral factors with both quantity and variety of fruit and vegetable consumption. Design/subjects Fruit and vegetable intake was assessed using repeated 24-hour dietary records collected during a 2-year period from 4,282 French subjects (2,373 men and 1,909 women), aged 45 to 62 years, who participated in a large prospective study. Statistical analysis Both education level and occupation categories were used as socioeconomic indicators. Logistic regression models were applied to assess factors related to meeting the 5 A Day fruit and vegetable recommendation. Covariance analyses were performed to compare the fruit and vegetable variety scores and the contributions of fruit and vegetables to the total daily diet cost across socioeconomic indicators within each sex.

Results

Meeting the 5 A Day recommendation was more likely in subjects aged 50 years and older, higher education levels, nonsmokers, moderate alcohol drinkers and in women engaging in regular physical activity. The odds ratio (95% confidence interval) for the lower vs higher education level was 0.70 (0.54 to 0.92) in men and 0.65 (0.48 to 0.85) in women. No significant difference was observed between occupation categories. A positive relationship between vegetable variety and education level was found in both sexes. Fruit variety was positively associated with both education and occupation categories, but only in men. The contribution of fruits to the total daily diet cost increased with occupation (PϽ0.02) and education (PϽ0.0001) in men, but decreased with occupation in women (PϽ0.05). Conclusions Although cost constraints may explain the lower fruit and vegetable intake in lower socioeconomic groups, the relative influence of budgetary resources, nutrition knowledge, and social and environmental barriers in socioeconomic disparities need further investigation.

H

igh consumption of fruits and vegetables has been suggested to be associated with a lower mortality risk [START_REF] Agudo | Fruit and vegetable intakes, dietary antioxidant nutrients, and total mortality in Spanish adults: Findings from the Spanish cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC-Spain)[END_REF] and a reduced incidence of many chronic diseases such as cardiovascular disease [START_REF] Dauchet | Fruit and vegetable consumption and risk of coronary heart disease: A meta-analysis of cohort studies[END_REF], stroke [START_REF] He | Fruit and vegetable consumption and stroke: Meta-analysis of cohort studies[END_REF], and certain cancers (4). A recent report of the World Health Organization stated that there is convincing evidence that fruit and vegetable consumption also decreases risk of diabetes and obesity [START_REF]Diet, Nutrition, and the Prevention of Chronic Diseases[END_REF]. The report recommended a minimum intake of 400 g fruits and vegetables per day.

Increasing fruit and vegetable intake by the general population has become a public health priority in many countries (4-6), with government agencies around the world asserting that everyone should eat at least five servings of fruit and vegetables daily (ie, Ն400 g/day) [START_REF]Diet, Nutrition, and the Prevention of Chronic Diseases[END_REF]. The Healthy People 2010 health objectives in the United States include increasing to 75% the number of people who eat at least two daily servings of fruit and increasing to 50% the number of those who eat at least three daily servings of vegetables. In France, fruit and vegetables are targeted by one of the nine nutrition priority goals specified in the Programme National Nutrition Santé for the 2001-2010 period. More precisely, the objective is to reduce the frequency of low consumers (Ͻ3.5 servings per day [START_REF] Beer-Borst | Dietary patterns in six european populations: Results from EURALIM, a collaborative European data harmonization and information campaign[END_REF]) by 25%. As in several other countries, a national campaign has been launched to promote the consumption of at least five servings of fruits and vegetables per day.

Identifying the correlates of fruit and vegetable consumption is a first step that may help health policy makers target at-risk populations and guide the development of specific actions to improve fruit and vegetable intake by low consumers. According to previous studies, socioeconomic factors like occupation categories and education level are important determinants of low fruit and vegetable intake [START_REF] Irala-Estevez | A systematic review of socio-economic differences in food habits in Europe: Consumption of fruit and vegetables[END_REF][START_REF] Kamphuis | Environmental determinants of fruit and vegetable consumption among adults: A systematic review[END_REF]. Compared to the United States and other European countries [START_REF] Irala-Estevez | A systematic review of socio-economic differences in food habits in Europe: Consumption of fruit and vegetables[END_REF][START_REF] Kamphuis | Environmental determinants of fruit and vegetable consumption among adults: A systematic review[END_REF][START_REF] Roos | Disparities in vegetable and fruit consumption: European cases from the north to the south[END_REF], there are few data regarding the consumption of fruit and vegetables and its relation with socioeconomic status in France.

The aim of this study was to investigate relationships of socioeconomic, demographic, and behavior factors with both quantity and variety of fruit and vegetable consumption in a sample of middle-aged French subjects. This study also quantified the contribution of fruit and vegetables to total diet cost according to socioeconomic status. The hypothesis was that due to budgetary constraints people with low education and from low occupation categories have lower fruit and vegetables intake in terms of both consumed quantity and variety.

MATERIALS AND METHODS Study Population

This study was conducted using data from the SUpplémentation en VItamines Minéraux et AntioXydants (SU.VI.MAX) cohort, a large sample of middle-aged adults living all over French territory. The design, methods, and rationale of the SU.VI.MAX study have been described elsewhere [START_REF] Hercberg | A primary prevention trial using nutritional doses of antioxidant vitamins and minerals in cardiovascular diseases and cancers in a general population: The SU.VI.MAX study-Design, methods, and participant characteristics. SUpplementation en VItamines et Mineraux AntioXydants[END_REF]. The SU.VI.MAX study was initially designed as a randomized, double blind, placebocontrolled, primary prevention trial to test the efficacy of daily supplementation with antioxidant vitamins and minerals at nutritional doses in reducing the incidence of ischemic heart disease and cancers. A total of 13,017 subjects (5,141 men aged 45 to 62 years; 7,876 women aged 35 to 62 years) were included in 1994-1995 for a planned follow-up of 8 years. The SU.VI.MAX study was approved by the Ethical Committee for Studies with Human Subjects at the Paris-Cochin Hospital (CCPPRB No. 706) and the Commission Nationale Informatique et Liberté (CNIL No. 334641).

For our analyses, the only subjects included were those aged 45 to 62 years who completed at least six 24-hour dietary records during the first 2 years of follow-up and with available data for all sociodemographic and behavioral characteristics. Furthermore, to take into account seasonal variations in fruit and vegetable consumption [START_REF] Capita | Differences in reported winter and summer dietary intakes in young adults in Spain[END_REF], the study sample was restricted to subjects having at least two 24-hour dietary records in the autumn-winter period (November to April) and at least two 24-hour dietary records in the spring-summer period (May to October).

Dietary Assessment

Dietary data were collected using the Minitel Telematic Network. The Minitel is a small terminal that was widely used as an adjunct to the telephone in France at the beginning of the SU.VI.MAX study. Participants were invited to transmit dietary data every 2 months. At enrollment, they received a scheduled calendar of 24-hour dietary data recording and a tiny central processing unit specifically developed for the study containing specialized software that allowed subjects to fill out the computerized dietary record off-line and to transmit data during brief telephone connections. The 24-hour records were distributed randomly for 2 weekend days and 4 weekdays per year, so that each day of the week was covered in all seasons for the mean intake of each participant. Diet has been shown to be very stable across the years in this cohort so diet measured over the first 2 years was considered to be representative of usual diet. Subjects were assisted by the conversational features of the software and an instruction guide for codification of foods, including photographs to facilitate estimation of portion size. This manual includes photographs of more than 250 foods (corresponding to 1,000 generic foods) represented in three different portion sizes. Along with the two intermediate and two extreme quantities, there are seven choices of amounts. Edible portions corresponding to each food were previously validated in a pilot study [START_REF] Moullec | Validation du manuel-photos utilisé pour l'enquête alimentaire de l'étude SU.VI.MAX {Validation of photographic document used to estimate the amounts of foods eaten by subjects in the SU.VI.MAX study[END_REF].

The nutritional values of the diet were estimated using a recently published French food composition table [START_REF] Hercberg | Table de composition des aliments SU.VI.MAX [French food composition databank from the SU.VI.MAX study[END_REF]. This database was completed by the unit cost of each food, based on the mean retail prices in France in 1997, obtained from marketing research, from the French National Institute of Statistics and Economic Studies, and from supermarket Web sites. To account for potential seasonal and weekly variations, all values reported here are based on the average dietary intakes of each subject including beverages.

The fruit and vegetable group includes fruits, vegetables, 100% fruit or vegetable juices, and mixed foods whose ingredients contain fruits or vegetables. To comply with French guidelines, dried fruits, potatoes, and legumes were not included in the fruit and vegetable group [START_REF] Hercberg | The French National Nutrition and Health Program: 2001-2006-2010[END_REF]. French recipes validated by food and nutrition professionals were used to assess the amounts consumed from mixed foods. To assess the compliance with fruit and vegetable recommendations (at least five servings per day), one serving was defined as 80 g fruit and vegetable (ie, a common portion size described in the literature) [START_REF] Williams | Healthy eating: Clarifying advice about fruit and vegetables[END_REF][START_REF]for Cancer Research. Food, Nutrition, Physical Activity, and the Prevention of Cancer: A Global Perspective[END_REF]. Furthermore, fruit and vegetable were classified using the International Agency for Research on Cancer classification (4) adapted to French dietary habits (see the Figure ). Methodology used to calculate fruit or vegetable variety scores was similar to those described by Giskes and colleagues [START_REF] Giskes | Socio-economic differences in fruit and vegetable consumption among Australian adolescents and adults[END_REF]. Separate scores were calculated for fruits and vegetables. Variety was measured as the number of different types of fruits/vegetables consumed. Each type of fruits/vegetables consumed was given a score of 1. Mixed dishes were given a score of 2 when the recipe contained at least two types of fruits/ vegetables.

Assessment of Sociodemographic and Behavior Characteristics

At enrollment subjects were asked to fill in a questionnaire to provide sociodemographic and lifestyle information. Level of education was coded in three categories: elementary school, secondary school (junior and high school), university, or equivalent. Occupation categories were used as proxy for income ranges because direct measure of material resources was not available. In France, subjects are generally unwilling to provide their income. The official French classification [START_REF]Les nomenclatures des professions et catégories socioprofessionnelles[END_REF] was used to classify subjects into four occupation categories according to their self-reported occupation or last occupation if they were retired or unemployed. The incomes of these different occupation categories are generally quite close for intermediate professions, employees, farmers, and manual workers and higher for managerial staff.

The location of residence was classified in two categories (urban/rural) according to the definition of the French National Institute of Statistics and Economics Studies, which is based on both population density and economic activity of the area corresponding to each subject's zip code [START_REF] Jeannic | Pôles urbains et périurbanisation, le zonage en aires urbaines [Urban and peri-urban zoning[END_REF]. Smoking status was categorized as current, former, or never. Overall level of habitual physical activity was assessed using the following questions: "Do you get regular physical activity (transportation, work, and leisure combined): yes/no? If yes, is it equivalent to at least 1 hour of walking per day?" Responses were coded in three categories (irregular, Ͻ1 hour of walking per day, Ն1 hour of walking per day) [START_REF] Czernichow | Indicators of abdominal adiposity in middle-aged participants of the SU.VI. MAX study: Relationships with educational level, smoking status and physical inactivity[END_REF]. Alcohol intake was estimated in grams of alcohol per day, from a short validated semiquantitative dietary questionnaire [START_REF] Lasfargues | Validity of a short self-administered dietary questionnaire compared with a dietetic interview[END_REF].

Statistical Analyses

Data were analyzed by gender to account for differences in dietary behaviors between men and women. All statistical analyses were performed using SAS software (version 8.2, 2001, SAS Institute Inc, Cary, NC). Descriptive results are reported as percentages or meanϮstandard deviation. The Student t test and 2 test were used to compare sexes, as appropriate. Logistic regression models were applied to assess factors related to meeting the recommendation to consume five servings of fruits and vegetables per day. Multivariate models included age, education level, occupation categories, marital status, household location, smoking status, physical activity categories, alcohol consumption, and total daily energy intakes. Adjusted odds ratios and their 95% confidence intervals are reported. Covariance analyses were performed to compare average fruit and vegetable variety scores, expenses to eat fruit and vegetables, and contributions of fruit and vegetables to the total cost of the daily diet, across socioeconomic status (ie, education level and occupation categories) within each sex after adjustment for total energy intake. Global and linear trend tests were performed. Because of skewed distribution of variety score values, comparisons were based on squareroot transformed data. For all analyses, the significance level was set at PϽ0.05.

RESULTS

Table 1 summarizes the general characteristics of the subjects. Mean age was 52.1Ϯ4.7 years in men and 51.1Ϯ4.6 years in women (PϽ0.0001). All sociodemographic and behavior factors significantly differed between sexes. More than 83% of men and women lived in an urban area (PϽ0.35 for sex difference).

Consumption of Fruit and Vegetables

Mean daily energy intakes were 2,431Ϯ537 kcal/day in men and 1,775Ϯ421 kcal/day in women (PϽ0.0001). As shown in Table 1, more than 50% of men and women consumed at least five servings of fruit and vegetables per day (ie. Ն400 g per day). After controlling for total energy intakes, women had higher consumption than men for both fruits and vegetables. However, the contributions of fruit and vegetable groups to total fruit and vegetable intake and the fruit and vegetable variety scores were very similar in both sexes (Table 1). Apple, pear, and other pome fruits; fruit juices; and citrus fruits (eg, orange and grapefruit) were the most consumed. Fruits used as vegetables (eg, tomato and cucumber) (Ͼ25%), root vegetables (Ͼ19%), green salads (Ͼ14%), and green Separate variety scores were calculated for fruits and vegetables. Methodology used was similar to those described by Giskes and colleagues [START_REF] Giskes | Socio-economic differences in fruit and vegetable consumption among Australian adolescents and adults[END_REF]. Variety was measured as the number of different types of fruits/vegetables consumed. Each variety of fruits/vegetables consumed was given a score of 1. Mixed dishes were given a score of 2 when the recipe contained at least two types of fruits/vegetables.

beans and peas (Ͼ13%) were the major contributors to total vegetable intake.

Intake of Composite Foods in Daily Fruit and Vegetable Consumption

The mean intake of vegetables from composite foods was around 50 g/day, which represents 25% of total vegetable consumption. For fruit, composite foods were a less important contributor, at 20 g/day (9% to 10% of total fruit intake). Fruit juices represented 40 g/day (20% of total fruit intake).

Meeting the 5 A Day Recommendation

Results from univariate associations showed that, in both sexes, the percentages of subjects who consumed at least five servings per day increased with age and education level (Table 2). Borderline significant differences between occupation categories were found in men (PϽ0.06). In both sexes, achieving the fruit and vegetable recommendation was negatively related to smoking status and positively to physical activity. A negative association was also observed with alcohol consumption, although only significant in men. No relationship was found with household location and marital status. In multivariate analyses, occupation categories in both sexes, and physical activity in men, were no longer related to fruit and vegetable intakes. Marital status was weakly associated with achieving recommended levels in men (Pϭ0.07).

Variety Score

Older subjects had higher vegetable variety scores in both sexes (Table 2). In contrast, no difference in fruit variety was observed between age groups. Fruit variety scores significantly increased with both education level and occupation categories in men, whereas vegetable variety score was positively associated with education in both sexes. Moreover, being married/living with a partner was associated with a higher fruit variety in women, and a higher vegetable variety in both men and women. Fruit variety was also associated with a more healthful lifestyle, including nonsmoking in both sexes, and low alcohol consumption and regular physical activity in men. Vegetable variety was inversely related to smoking status in men.

Contribution of Fruit and Vegetables to Total Cost of Daily Diet

Mean total cost of daily diet was €6.92Ϯ1.8 in men and €5.36Ϯ1.5 in women (Table 3). Managerial staff had more expensive diets than farmers/self-employed or manual workers in men, whereas both occupation and education were positively associated with total daily diet cost in women. The contribution of fruit and vegetable to the total diet cost was 16% of total diet cost (€1.09) and 20% (€1.05), respectively, in men and women. Significant differences were found in men for education (mainly due to the contribution of fruits), whereas in women an association was found for occupation categories. In men, managerial staff and intermediate professions had the highest fruit cost contribution compared to manual workers (Table 3). In contrast, in women, managerial staff had the lowest fruit and vegetable contribution to total daily diet cost.

Contribution of Fruit and Vegetable Categories to Total Cost of Daily Diet

Analyses of the relationship between the two indicators of socioeconomic status (ie, occupation and education) and the various fruit and vegetable groups showed that the contribution of apple, pear, and other pome fruits to fruit expenses decreased with increasing education in women (Table 4). In both sexes, less-educated subjects spent more money to eat tropical fruits (other than banana).

The contributions of fruit juices to daily fruit cost increased with education in both sexes and with occupation categories in men. A positive association with education was also observed for the contribution of fruits used as vegetables and green salads to vegetable cost, in women. Conversely, the contributions of root vegetables, green beans and peas, bulb, and stem vegetables to daily vegetable cost decreased with education. Occupation and education categories were positively related to green salad cost contribution only in women. No vegetable categories differed significantly according to education and occupation in men.

DISCUSSION

In this study, achieving the 5 A Day fruit and vegetable recommendation was positively related to education level in both sexes. Older subjects, nonsmokers, and moderate alcohol drinkers were more likely to meet fruit and vegetable recommendations. In women, a positive relationship between fruit and vegetable consumption and physical activity level was also observed. No association was found with occupation categories, after adjustment for education, sociodemographic, and behavior characteristics. Variety scores increased with education level for fruits and vegetables in men and only for vegetables in women. Occupation categories were positively associated with fruit variety in men. Contribution of fruit and vegetable to total daily diet cost also differed between socioeconomic groups with some sex-specific relationships.

Intake of Composite Foods in Daily Fruit and Vegetable Consumption

Most studies do not take into account composite foods and fruit juices, although composite foods are an important sources of fruit and vegetable intake. In our study, the results demonstrate that composite foods and fruit juices should be included when estimating fruit and vegetable intakes to avoid bias in estimates of intake, as mentioned elsewhere [START_REF] O'brien | The importance of compos-December 2008 • Journal of the AMERICAN DIETETIC ASSOCIATION 2029 ite foods for estimates of vegetable and fruit intakes[END_REF]. It is interesting to note that the higher consumption of fruit and vegetable found for managerial and intermediate professions vs manual workers was due to the higher consumption of fruit juices. This result was also found among British adults [START_REF] Billson | Variation in fruit and vegetable consumption among adults in Britain. An analysis from the dietary and nutritional survey of British adults[END_REF].

Socioeconomic Status and Adherence to the 5 A Day Recommendation

The observed relationship between adherence to the 5 A Day recommendation and education level is consistent with previous literature [START_REF] Irala-Estevez | A systematic review of socio-economic differences in food habits in Europe: Consumption of fruit and vegetables[END_REF][START_REF] Roos | Disparities in vegetable and fruit consumption: European cases from the north to the south[END_REF][START_REF] Perrin | Ten-year trends of dietary intake in a middle-aged French population: Relationship with educational level[END_REF]. However, it has been suggested that relationships with several indicators of socioeconomic status should be addressed simultaneously to provide a better understanding of the social inequalities in dietary habits [START_REF] Galobardes | Diet and socioeconomic position: does the use of different indicators matter?[END_REF][START_REF] Lallukka | Multiple socio-economic circumstances and healthy food habits[END_REF][START_REF] Turrell | Measuring socioeconomic position in dietary research: Is choice of socio-economic indicator important?[END_REF]. Occupation category is another widely used indicator of socioeconomic status. In Separate variety scores were calculated for fruits and vegetables. Methodology used was similar to those described by Giskes and colleagues [START_REF] Giskes | Socio-economic differences in fruit and vegetable consumption among Australian adolescents and adults[END_REF]. Variety was measured as the number of different types of fruits/vegetables consumed. Each variety of fruits/vegetables consumed was given a score of 1. Mixed dishes were given a score of 2 when the recipe contained at least two types of fruits/vegetables. c ORϭodds ratio. OR determined by logistic regression analyses. Variables included in the model were age, education level, occupation categories, marital status, household location, smoking status, physical activity categories, alcohol consumption, and total daily energy intakes. d CI-confidence interval. CI determined by logistic regression analyses. Variables included in the model were age, education level, occupation categories, marital status, household location, smoking status, physical activity categories, alcohol consumption, and total daily energy intakes. e The official French classification [START_REF]Les nomenclatures des professions et catégories socioprofessionnelles[END_REF] was used to classify subjects into four occupation categories according to their self-reported occupation or last occupation if they were retired or unemployed. The incomes of these different occupation categories are generally quite close for intermediate professions, employees, farmers, and manual workers and higher for the managerial staff. The overall level of habitual physical activity was assessed using the following questions: "Do you get regular physical activity (transportation, work, and leisure combined): yes/no? If yes, is it equivalent to at least 1 hour of walking per day?" [START_REF] Czernichow | Indicators of abdominal adiposity in middle-aged participants of the SU.VI. MAX study: Relationships with educational level, smoking status and physical inactivity[END_REF]. Responses were coded in three categories. h Alcohol intake was estimated in grams of alcohol per day, from a short validated semiquantitative dietary questionnaire [START_REF] Lasfargues | Validity of a short self-administered dietary questionnaire compared with a dietetic interview[END_REF].

our study, occupation was not an independent correlate of fruit and vegetable consumption. Education level reflects other aspects than socioeconomic status, such as dietary knowledge and ability to interpret health education messages [START_REF] Galobardes | Diet and socioeconomic position: does the use of different indicators matter?[END_REF][START_REF] Turrell | Measuring socioeconomic position in dietary research: Is choice of socio-economic indicator important?[END_REF]. That may be another explanation of the differential relationships of adherence to the recommendation to consume five servings of fruits and vegetables per day with education level and occupation category.

Sociodemographic and Lifestyle Characteristics and the 5 A Day Recommandation

Numerous studies have reported inverse associations between fruit and vegetable consumption and other lesshealthful behaviors, including smoking status [START_REF] Billson | Variation in fruit and vegetable consumption among adults in Britain. An analysis from the dietary and nutritional survey of British adults[END_REF][START_REF] Birkett | Intake of fruits and vegetables in smokers[END_REF][START_REF] Pollard | Motivations for fruit and vegetable consumption in the UK Women's Cohort Study[END_REF], alcohol consumption [START_REF] Agudo | Vegetable and fruit consumption associated with anthropometric, dietary, and lifestyle factors in Spain. EPIC Group of Spain. European Prospective Investigation into Cancer[END_REF][START_REF] Padrao | Smoking, alcohol, and dietary choices: Evidence from the Portuguese National Health Survey[END_REF], and low physical activity level [START_REF] Agudo | Vegetable and fruit consumption associated with anthropometric, dietary, and lifestyle factors in Spain. EPIC Group of Spain. European Prospective Investigation into Cancer[END_REF][START_REF] Oppert | Leisure-time and occupational physical activity in relation to cardiovascular risk factors and eating habits in French adults[END_REF]. The results from our study are in line with previous data, even though in men physical activity level was not related to the adherence of the 5 A Day recommendation. Previous studies have shown that married people have higher fruit and vegetable intakes [START_REF] Kamphuis | Environmental determinants of fruit and vegetable consumption among adults: A systematic review[END_REF]. In our study, being married/living with a partner was weakly inversely associated with meeting fruit and vegetable recommendation in men (borderline significant) and no association was observed in women. No difference in fruit and vegetable consumption was found according to urban/rural location, whereas Johansson and colleagues [START_REF] Johansson | Healthy dietary habits in relation to social determinants and lifestyle factors[END_REF] reported that subjects living in rural areas had lower fruit and vegetable intakes than their counterparts living in cities, even after adjustment for age, sex, and education. However, good local access to food stores and availability of fruit and vegetable may be a more important environmental determinant of fruit and vegetable consumption as suggested by a growing body of literature [START_REF] Kamphuis | Environmental determinants of fruit and vegetable consumption among adults: A systematic review[END_REF]. Moreover, Giskes and colleages [START_REF] Giskes | Socioeconomic inequalities in food purchasing: The contribution of respondent-perceived and actual (objectively measured) price and availability of foods[END_REF] recently showed that perceived availability of healthful foods may be a major contributor of food purchasing.

Socioeconomic Status and Fruit and Vegetable Variety

In addition to encouraging fruit and vegetable consumption, promotion of their variety may be important to ensure sufficient intakes for certain nutrients [START_REF] Giskes | Socioeconomic differences among Australian adults in consumption of fruit and vegetables and intakes of vitamins A, C and folate[END_REF]. Only one study has previously examined the differences in fruit or The official French classification [START_REF]Les nomenclatures des professions et catégories socioprofessionnelles[END_REF] was used to classify subjects into four occupation categories according to their self-reported occupation or last occupation if they were retired or unemployed. The incomes of these different occupation categories are generally quite close for intermediate professions, employees, farmers, and manual workers and higher for the managerial staff.

vegetable varieties according to socioeconomic status.

Giskes and colleagues reported a lower variety in adults from low-income socioeconomic groups [START_REF] Giskes | Socio-economic differences in fruit and vegetable consumption among Australian adolescents and adults[END_REF]. The observed fruit variety score is approximately the same as those found in Australian adults, whereas the variety in vegetables consumed was one point lower in this sample [START_REF] Giskes | Socio-economic differences in fruit and vegetable consumption among Australian adolescents and adults[END_REF]. This discrepancy could be related to differences in dietary assessment. In fact, the choice to select at least six 24hour dietary records over a 2-year period allows a more reliable diet estimation and limits misclassification by decreasing intraindividual variation [START_REF] Sempos | Effects of intraindi-vidual and interindividual variation in repeated dietary records[END_REF]. Precision of data from other studies is limited because they used only food frequency questionnaires or one 24-hour dietary recall. Another possibility to this discrepancy is eventually the fruit and vegetable classification used, which was not specified in the article by Giskes and colleagues. However, in accordance with this previous study, a negative relationship of vegetable variety with education level was found in both sexes. In addition, fruit variety was posi-tively associated with both education and occupation categories, but only in men. These results support the hypothesis that economic constraints may be one of the barriers to fruit and vegetable consumption, especially in low-income populations [START_REF] Drewnowski | Food choices and diet costs: An economic analysis[END_REF]. Men from the highest occupation categories had higher total daily diet cost and spent more money to eat fruit and vegetable. In women, diet cost and fruit and vegetable expenses were positively related to education and negatively to occupation. Differences in fruit and vegetable product prices (38) may also explain some differential fruit and vegetable consumption patterns because subjects with a primary school level or having a manual occupation consumed more lowpriced fruit and vegetables, such as pome fruits, fruits used as vegetables and beans/peas. Several studies also suggest that access to food stores and availability and variety of fruit and vegetable may be lower in poorer neighbourhoods [START_REF] Morland | The contextual effect of the local food environment on residents' diets: The atherosclerosis risk in communities study[END_REF]. In addition to influence of local environment, results from a recently published study showed that several dimensions of the social context might also influence fruit and vegetable consumption [START_REF] Sorensen | The influence of social context on changes in fruit and vegetable consumption: Results of the healthy directions studies[END_REF]. Several limitations should be addressed. Because our study was performed in a French context, these results should be compared to data obtained in other populations with caution. More than half of subjects achieved the five servings a day of fruit and vegetables recommendation. Fruit and vegetable consumption seems particularly high compared to other populations [START_REF] Guenther | Most Americans eat much less than recommended amounts of fruits and vegetables[END_REF][START_REF] Naska | Fruit and vegetable availability among 10 European countries: How does it compare with the '5-A-Day' recommendation? DAFNE I and II projects of the European Commission[END_REF]: In a study based on the 1989-1991 Continuing Surveys of Food Intakes by Individuals, 32% of US adults older than age 20 years old met the 5 A Day objective [START_REF] Krebs-Smith | US adults' fruit and vegetable intakes, 1989 to 1991: A revised baseline for the Healthy People 2000 objective[END_REF]. In 1999-2000, it has been estimated that only 40% of Americans ate an average of five or more servings of fruits and vegetables per day [START_REF] Guenther | Most Americans eat much less than recommended amounts of fruits and vegetables[END_REF]. However, our study was limited to the age range 45 to 62 years and previous studies have shown that the percentages of subjects meeting the 5 A Day fruit and vegetable recommendation were lower in younger age groups [START_REF] Guenther | Most Americans eat much less than recommended amounts of fruits and vegetables[END_REF][START_REF] Agudo | Dietary intake of vegetables and fruits among adults in five regions of Spain. EPIC Group of Spain. European Prospective Investigation into Cancer and Nutrition[END_REF]. Another limitation is that subjects were participants of a nutrition intervention study [START_REF] Hercberg | .VI.MAX Study: A randomized, placebo-controlled trial of the health effects of antioxidant vitamins and minerals[END_REF], thus they may have had more healthful lifestyle and dietary patterns. Another potential limitation is the bias that can be derived from over-reporting consumption among those with higher levels of education [START_REF] Irala-Estevez | A systematic review of socio-economic differences in food habits in Europe: Consumption of fruit and vegetables[END_REF].

CONCLUSIONS

In this study of middle-aged French subjects, socioeconomic disparities in both quantity and variety of fruit and vegetable consumed were found. Higher socioeconomic groups also spent more to eat fruit and vegetables. This suggests that price intervention may be a valuable strategy for increasing the proportion of subjects who attain the recommended levels, especially in low-income socioeconomic groups. A reduction in the price of fruit and vegetables, which can be achieved by means of public policies, could lead to an increase in fruit and vegetable intake [START_REF] Claro | Income, food prices, and participation of fruit and vegetables in the diet[END_REF]. In addition, for underprivileged people with particularly low income, distribution of fruit and vegetable vouchers (or checks), has been demonstrated as an useful measure to encourage low-income consumers to increase the consumption of such products [START_REF] Anderson | 5 A Day fruit and vegetable intervention improves consumption in a low income population[END_REF][START_REF] Herman | Choices made by low-income women provided with an economic supplement for fresh fruit and vegetable purchase[END_REF]. Achieving the recommendation to consume five servings per day of fruit and vegetables was related to education but not to occupation categories, suggesting that factors other than budgetary resources may explain the socioeconomic disparities in fruit and vegetable consumption.

To develop effective interventions and policies, there is a need to better understand the relative influence of budgetary constraints, nutrition knowledge and skills, social context, and environmental barriers. Whatever the country, nutrition education is a practical intervention that can be applied by many health professionals until global policy changes can be accomplished.

Figure .

 . Figure.Classification of fruits and vegetables according to French dietary habits. Methodology adapted from reference (4).

f

  The location of residence was classified in two categories (urban/rural) according to the definition of the French National Institute of Statistics and Economics Studies, which is based on both population density and economic activity.

  g

Table 1 .

 1 Basic characteristics of selected participants in the Supplémentation en Vitamines et Minéraux Antioxydants study

	Characteristic	Men		Women
	Sociodemographic	(n)373,2؍		(n)909,1؍
		4 ™™™™™™™™™™ n % ™™™™™™™™ 3
	Age group (y) a
	45-49	866 36.5	860 45.1
	50-54	727 30.6	560 29.3
	55ϩ	780 61.5	489 38.5
	Education level a
	University or equivalent	959 40.4	676 35.4
	Secondary school	843 35.5	834 43.7
	Elementary school	571 24.1	399 20.9
	Occupation categories ab
	Managerial staff	1,007 42.4	355 18.6
	Intermediate professions,
	employees	1,074 45.3 1,420 74.4
	Farmers, self-employed	127	5.4	87	4.6
	Manual workers	165	7.0	47	2.5
	Marital status a
	Living alone	224	9.4	360 18.9
	Cohabiting	2,149 90.6 1,549 81.7
	Household location c
	Urban	1,989 83.8 1,620 84.9
	Rural	384 16.2	289 15.1
	Smoking status a
	Nonsmokers/former smokers	2,056 86.7 1,696 88.8
	Current Smokers	317 13.4	213 11.2
	Physical activity ad
	Irregular	550 23.2	473 24.8
	Ͻ1 h/d of walking	586 24.7	671 35.2
	Ն1 h/d of walking	1,237 52.1	765 40.0
	Dietary intake
	Alcohol consumption
	(g/d) ae
	Ͻ20 g/d	821 34.6 1,763 92.4
	Ն20 g/d	1,552 65.4	146	7.6
	>5 fruits and vegetables
	per day	54%		53%
		4™ meanϮstandard deviation ™ 3
	Energy intake (g/d) af	2,431.0Ϯ537	1,775.0Ϯ421
	Fruits and vegetables
	(g/d) ag	424.4Ϯ194.5	492.4Ϯ163.5
	Fruits (g/d) ag	228.3Ϯ152.0	266.4Ϯ119.0
	Apple, pear, and other pome
	fruits (% of fruit intakes) a	33.8Ϯ21.4	31.2Ϯ18.3
	Citrus fruits (% of fruit
	intakes) a	10.9Ϯ12.6	12.7Ϯ11.0
	Grapes (% of fruit intakes)	4.5Ϯ8.6	4.1Ϯ6.8
	Berries (% of fruit intakes) a	8.5Ϯ10.2	9.5Ϯ10.2
	Stone fruits (% of fruit
	intakes) a	9.1Ϯ11.1	9.9Ϯ10.3
	Melon (% of fruit intakes)	5.3Ϯ9.3	5.5Ϯ8.8
	Banana (% of fruit intakes) a	5.6Ϯ9.6	4.1Ϯ6.7
	Other tropical fruits (% of fruit
	intakes)	9.3Ϯ11.9	9.1Ϯ10.2
	Fruit juices (% of fruit intakes)	15.4Ϯ20.3	15.0Ϯ18.5
				(continued)

Table 1 .

 1 Continued

	Characteristic	Men	Women
	Sociodemographic	(n)373,2؍	(n)909,1؍
	Variety score (% of fruit
	intakes) ah	2.1Ϯ0.5	2.4Ϯ0.5
	Vegetables (g/day) ag	196.1Ϯ89.8	226.0Ϯ83.6
	Green salads (% of
	vegetable intakes)	14.2Ϯ9.8	14.3Ϯ9.1
	Leafy vegetables (% of
	vegetable intakes)	7.8Ϯ8.4	8.3Ϯ8.9
	Fruits used as vegetables
	(% of vegetable intakes)	26.3Ϯ17.8	25.7Ϯ13.1
	Root vegetables (% of
	vegetable intakes)	19.5Ϯ11.0	19.4Ϯ10.4
	Green beans and peas (% of
	vegetable intakes)	13.4Ϯ10.6	13.1Ϯ10.0
	Bulb and stem vegetables
	(% of vegetable intakes)	9.6Ϯ6.6	9.2Ϯ6.2
	Flowering vegetables (% of
	vegetable intakes)	5.9Ϯ7.6	7.1Ϯ8.5
	Mushrooms (% of vegetable
	intakes) a	4.6Ϯ5.8	3.8Ϯ4.5
	Sprouts (% of vegetable
	intakes)	0.4Ϯ1.8	0.4Ϯ1.5
	Vegetable juices (% of
	vegetable intakes)	0.3Ϯ2.2	0.4Ϯ3.2
	Variety score (% of
	vegetable intakes) ah	1.9Ϯ0.3	2.1Ϯ0.3

a Test of difference between sexes. P value from

2

test for categorical variables and from Student t test for continuous variables Ͻ0.05.

b

The official French classification

[START_REF]Les nomenclatures des professions et catégories socioprofessionnelles[END_REF]

was used to classify subjects into four occupation categories according to self-reported occupation or last occupation if subject was retired or unemployed. The incomes of these different occupation categories are generally quite close for intermediate professions, employees, farmers, and manual workers and higher for the managerial staff. c The location of residence was classified in two categories (urban or rural) according to the definition of the French National Institute of Statistics and Economics Studies, which is based on both population density and economic activity of the area corresponding to each subject's zip code

[START_REF] Jeannic | Pôles urbains et périurbanisation, le zonage en aires urbaines [Urban and peri-urban zoning[END_REF]

. d

The overall level of habitual physical activity was assessed using the following questions: "Do you get regular physical activity (transportation, work, and leisure combined): yes/no? If yes, is it equivalent to at least 1 hour of walking per day?"

[START_REF] Czernichow | Indicators of abdominal adiposity in middle-aged participants of the SU.VI. MAX study: Relationships with educational level, smoking status and physical inactivity[END_REF]

. Responses were coded in three categories. e Alcohol intake was estimated in grams of alcohol per day, from a short validated semiquantitative dietary questionnaire

[START_REF] Lasfargues | Validity of a short self-administered dietary questionnaire compared with a dietetic interview[END_REF]

. f Mean energy intakes estimated from repeated 24-hour dietary records collected during the 1995-1997 period. g Adjusted for total daily energy intakes.

h

Table 2 .

 2 [START_REF]Diet, Nutrition, and the Prevention of Chronic Diseases[END_REF] A Day consumption and variety score, in both sexes, according to sociodemographic and behavior characteristics a

		Men (n)373,2؍			Women (n)909,1؍
		Fruits and Vegetables			Fruits and Vegetables
		>5 Servings per Day	Variety Score b	>5 Servings per Day	Variety Score b
	Characteristic	%	O R c (95% CI d )	Fruits	Vegetables	%	OR (95% CI)	Fruits	Vegetables
	Age (y)
	45-50	49.2	1.00 (Referent)	2.13	1.88	48.8	1.00 (Referent)	2.43	1.99
	50-55	53.7	1.26 (1.02-1.56)	2.10	1.93	52.7	1.19 (0.95-1.50)	2.43	2.1
	55ϩ	60.5	1.75 (1.41-2.16)	2.16	2.02	60.7	1.75 (1.37-2.23)	2.43	2.13
	P for trend	Ͻ0.0001	Ͻ0.0001	0.77	Ͻ0.0001	0.0001	Ͻ0.0001	0.94	0.001
	Education level
	University or equivalent	58.1	1.00 (Referent)	2.28	2.02	56.7	1.00 (Referent)	2.34	2.1
	Secondary school	52.6	0.74 (0.59-0.92)	2.13	1.88	53.2	0.88 (0.69-1.11)	2.53	2.07
	Elementary school	50.4	0.70 (0.54-0.92)	1.96	1.88	46.4	0.65 (0.48-0.88)	2.37	1.96
	P for trend	0.007	0.0008	0.0002	0.002	0.005	0.009	0.78	0.002
	Occupation categories e
	Managerial staff	55.6	1.00 (Referent)	2.16	1.93	54.9	1.00 (Referent)	2.43	2.1
	Intermediate professions, employees	55	1.13 (0.91-1.40)	2.19	1.96	52.2	0.96 (0.73-1.27)	2.43	2.07
	Farmers, self-employed	46.5	0.77 (0.51-1.18)	1.74	1.82	58.6	1.34 (0.78-2.30)	2.43	1.99
	Manual workers	47.3	0.86 (0.58-1.28)	1.96	1.88	53.2	1.36 (0.68-2.72)	2.19	1.99
	P for trend	0.06	0.60	0.005	0.19	0.57	0.66	0.71	0.63
	Marital status
	Cohabiting	53.8	1.00 (Referent)	2.13	1.96	53.7	1.00 (Referent)	2.5	2.1
	Living alone	59.4	1.31 (0.97-1.77)	2.16	1.82	50.0	0.94 (0.73-1.20)	2.16	1.93
	P	0.11	0.07	0.78	0.02	0.20	0.61	0.0003	0.0009
	Household location f
	Urban	50.3	1.00 (Referent)	2.16	1.93	51.6	1.00 (Referent)	2.37	2.07
	Rural	55.1	1.19 (0.94-1.52)	2.13	1.93	53.3	1.17 (0.89-1.55)	2.43	2.07
	P	0.08	0.12	0.62	0.66	0.59	0.36	0.53	0.98
	Smoking status
	Nonsmokers/former smokers	57.0	1.00 (Referent)	2.19	1.96	55.4	1.00 (Referent)	2.46	2.07
	Current smokers	36.6	0.44 (0.34-0.57)	1.74	1.74	33.8	0.46 (0.33-0.63)	2.10	1.99
	P	Ͻ0.0001	Ͻ0.0001	Ͻ0.0001	Ͻ0.0001	Ͻ0.0001	Ͻ0.0001	0.001	0.09
	Physical activity g
	Irregular	49.8	1.00 (Referent)	2.02	1.93	45.2	1.00 (Referent)	2.34	2.02
	Ͻ1 h/d of walking	53.6	1.10 (0.86-1.41)	2.16	1.93	55.4	1.33 (1.04-1.71)	2.46	2.07
	Ն1 h/d of walking	56.6	1.13 (0.91-1.41)	2.19	1.93	55.7	1.44 (1.13-1.85)	2.46	2.07
	P for trend	0.03	0.28	0.03	0.77	0.0005	0.004	0.17	0.16
	Alcohol consumption h
	Ͻ20 g/d	57.5	1.00 (Referent)	2.31	1.96	53.4	1.00 (Referent)	2.43	2.07
	Ն20 g/d	52.6	0.67 (0.56-0.81)	2.04	1.93	48.0	0.66 (0.45-0.95)	2.25	1.99
	P	0.02	Ͻ0.0001	0.0002	0.65	0.20	0.03	0.15	0.18

a Data from selected participants of Supplémentation en Vitamines et Minéraux Antioxydants prospective study. Mean dietary intakes estimated from repeated 24-hour dietary records collected during 1995-1997.

b

Table 3 .

 3 Contribution of fruits and vegetables to total daily diet cost in both sexes of participants in the Supplémentation en Vitamines et Minéraux Antioxydants prospective study, according to education and occupation characteristics a Costs were based on the mean retail prices in France in 1997, obtained from marketing research, from the French National Institute of Statistics and Economic Studies, and from supermarket Web sites. 1 €ϭ1.49 US$ (as of January 2, 2008).

		Total Daily Diet	Fruits	Vegetables	Fruits and Vegetables
	Characteristic	€/d b	€/d	%	€/d	%	€/d	%
	Men (n,)373,2؍ overall	6.92	0.55	8.2	0.54	8.0	1.09	16.0
	Education level c
	University or equivalent	6.9	0.59	8.8	0.55	8.2	1.14	16.9
	Secondary school	6.97	0.54	7.9	0.53	7.8	1.07	15.7
	Elementary school	6.84	0.51	7.6	0.53	7.9	1.04	15.5
	P for trend	0.4	Ͻ0.0001	Ͻ0.0001	0.21	0.2	Ͻ0.0001	Ͻ0.0001
	Occupation categories de
	Managerial staff	7.09	0.57	8.2	0.54	7.9	1.1	16.0
	Intermediate professions, employees	6.8	0.56	8.4	0.53	8.0	1.09	16.4
	Farmers, self-employed	6.66	0.49	7.5	0.53	8.1	1.01	15.6
	Manual workers	6.87	0.5	7.5	0.54	8.3	1.04	15.8
	P for trend	0.0001	0.006	0.02	0.88	0.48	0.06	0.21
	Women (n,)909,1؍ overall	5.36	0.54	10.2	0.5	9.7	1.05	19.9
	Education level c
	University or equivalent	5.48	0.56	10.5	0.51	9.6	1.07	20.0
	Secondary school	5.35	0.55	10.4	0.51	9.7	1.05	20.2
	Elementary school	5.19	0.5	9.7	0.49	9.7	0.99	19.4
	P for trend	0.0002	0.0001	0.02	0.07	0.72	0.0001	0.13
	Occupation categories de
	Managerial staff	5.56	0.51	9.6	0.5	9.3	1.01	18.9
	Intermediate professions, employees	5.31	0.54	10.4	0.5	9.7	1.05	20.1
	Farmers, self-employed	5.52	0.57	10.6	0.56	10.4	1.14	21.0
	Manual workers	5.14	0.55	10.6	0.51	10.0	1.06	20.6
	P for trend	0.003	0.14	0.05	0.04	0.08	0.03	0.008
	a Mean dietary intakes estimated from repeated 24-hour dietary records collected during 1995-1997.

b Including beverages. c Adjusted for age, total daily energy intake, and occupation categories.

d

Table 4 .

 4 Contribution of the major fruit and vegetable categories to total daily fruit and vegetable costs, based on data from selected participants in the Supplémentation en Vitamines et Minéraux Antioxydants prospective studyab The official French classification[START_REF]Les nomenclatures des professions et catégories socioprofessionnelles[END_REF] was used to classify subjects into four occupation categories according to their self-reported occupation or last occupation if they were retired or unemployed. The incomes of these different occupation categories are generally quite close for intermediate professions, employees, farmers, and manual workers and higher for the managerial staff.

			Contribution to Total Fruit Cost (%)		Contribution to Total Vegetable Cost (%)
		Apple, pear,						Fruits			Green	Bulb and
		and other	Fruit	Citrus	Tropical	Stone		used as	Root	Green	beans	stem
	Characteristic	pome fruits	juices	fruits	fruits c	fruits Berries	vegetables	vegetables	salads	and peas	vegetables
	Men (n,)373,2؍ overall 33.8	15.3	11.3	9.3	9.1	8.5	27.5	19.3	14.3	13.7	9.6
	Education level d
	University or equivalent	33.2	16.4	11.3	8.7	9.0	8.5	28.7	19.3	14.2	12.8	9.3
	Secondary school	33.7	15.3	10.9	9.4	9.3	8.6	26.8	19.8	13.8	14.0	9.8
	Elementary school	35.0	14.2	10.3	10.1	9.0	8.4	27.7	19.7	14.5	13.7	9.8
	P for trend	0.14	0.06	0.14	0.04	0.94	0.91	0.47	0.52	0.61	0.13	0.18
	Occupation categories ef
	Managerial staff	32.6	16.3	11.0	9.4	9.1	8.7	28.8	19.1	14.4	12.8	9.4
	Intermediate professions,
	employees	34.3	15.7	10.7	9.1	9.2	8.4	26.8	19.6	14.2	13.8	9.8
	Farmers, self-employed	37.0	9.9	12.3	10.2	8.7	9.0	25.6	20.7	14.7	12.9	10.0
	Manual workers	35.1	12.9	10.7	8.7	9.3	7.9	28.3	21.3	12.1	14.7	9.3
	P for trend	0.13	0.006	0.6	0.72	0.96	0.77	0.07	0.13	0.07	0.14	0.47
	Women (n,)909,1؍
	overall	31.2	15.2	12.2	9.1	9.9	9.6	27.3	19.6	14.1	12.8	9.5
	Education level d
	University or equivalent	29.7	16.1	12.2	8.8	10.2	10.3	28.9	18.2	14.8	11.9	9.0
	Secondary school	31.8	14.7	13.1	8.6	9.9	9.2	26.6	19.6	14.3	13.3	9.6
	Elementary school	32.5	13.6	12.7	10.7	9.5	8.9	24.7	20.8	13.5	14.6	10.0
	P for trend	0.02	0.04	0.47	0.007	0.35	0.05	Ͻ0.0001	0.0001	0.03	Ͻ0.0001	0.01
	Occupation categories ef
	Managerial staff	31.2	13.9	12.8	10.2	8.7	9.9	28.2	18.3	15.8	12.1	9.3
	Intermediate professions,
	employees	30.9	15.5	12.6	9.1	10.1	9.5	26.9	19.6	14.1	13.2	9.5
	Farmers, self-employed	33.3	13.3	12.7	7.5	12	9.2	24.5	19.8	13.0	13.9	10.8
	Manual workers	36.2	11.9	14.5	6.9	10.1	9.2	26.4	19.5	12.7	15.9	8.8
	P for trend	0.18	0.24	0.68	0.08	0.05	0.9	0.15	0.24	0.01	0.09	0.16

a Mean dietary intakes estimated from repeated 24-hour dietary records collected during 1995-1997. b Costs were based on mean retail prices in France in 1997, obtained from marketing research, from the French National Institute of Statistics and Economic Studies, and from supermarket Web sites. 1 €ϭ1.49 US$ (as of January 2, 2008). c Excluding banana. d Adjusted for age, total daily energy intake, and occupation categories. e Adjusted for age, total daily energy intake, and education level. f 2028 December 2008 Volume 108 Number 12

December 2008 • Journal of the AMERICAN DIETETIC ASSOCIATION 2023

December 2008 • Journal of the AMERICAN DIETETIC ASSOCIATION 2025

e Adjusted for age, total daily energy intake, and education level. December 2008 • Journal of the AMERICAN DIETETIC ASSOCIATION 2027

The first author (C.E.) was supported in part by the Fondation Louis Bonduelle, France.

The authors thank Stacie Chat-Yung for reviewing and editing the manuscript.


J.-F Hocquette

Didier Boichard

Isabelle Cassar-Malek

Elisabeth Laville

Gilles G Renand

Hubert H Levéziel

Brigitte B Picard

Functional and positional genomics in beef cattle : current programs and potential progress

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