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1 Dietary sources and socio-demographic and economic factors affecting vitamin D and 2 calcium intakes in Flemish preschoolers 3 4 Inge Huybrechts1, Yi Lin1, Willem De Keyzer1,2, Isabelle Sioen1, Theodora Mouratidou3, Luis 5 A. Moreno3, Nadia Slimani4, Mazda Jenab4, Stefanie Vandevijvere5, Guy De Backer1, Stefaan 6 De Henauw1,2 7 8 9 1 Department of Public Health, Faculty of Medicine and Health Sciences, Ghent University, 10 Ghent, Belgium 11 2 12 Gent, Belgium 13 3 14 Zaragoza, C/ Corona de Aragon 42, 50009, Zaragoza, Spain 15 4 16 Lyon Cedex 08, France 17 5 Department of Nutrition and dietetics, University College Ghent, Keramiekstraat 80, 9000 GENUD (Growth, Exercise, NUtrition and Development) Research Group, University of International Agency for Research on Cancer (IARC), 150 Cours Albert Thomas, 69372 Unit of Epidemiology, Scientific Institute of Public Health, Brussels, Belgium 18 19 Running title: vitamin D and Calcium intake in preschoolers 20 21 Funding: The Flanders preschool dietary survey was funded by the Belgian Nutrition 22 Information Center (NICE) 23 1 24 Authors: 25 Inge Huybrechts (PhD), Unit nutrition and food safety, Department of Public Health, Faculty 26 of Medicine and Health Sciences, Ghent University, De Pintelaan 185, B-9000 Ghent, 27 Belgium 28 [email protected] 29 30 Yi Lin (MS, MPH), Unit nutrition and food safety, Department of Public Health, Faculty of 31 Medicine and Health Sciences, Ghent University, De Pintelaan 185, B-9000 Ghent, Belgium 32 [email protected] 33 34 Willem De Keyzer (MS), University College Ghent, department of Nutrition and dietetics, 35 Faculty of Health Care Vesalius, Keramiekstraat 80, B-9000 Ghent, Belgium 36 [email protected] 37 38 Isabelle Sioen (PhD), Unit nutrition and food safety, Department of Public Health, Faculty of 39 Medicine and Health Sciences, Ghent University, De Pintelaan 185, B-9000 Ghent, Belgium 40 [email protected] 41 42 Theodora Mouratidou (PhD), GENUD (Growth, Exercise, NUtrition and Development) 43 Research Group, University of Zaragoza, C/ Corona de Aragon 42, 50009, Zaragoza, Spain 44 [email protected] 45 2 46 Luis Moreno (Prof), GENUD (Growth, Exercise, NUtrition and Development) Research 47 Group, University of Zaragoza, C/ Corona de Aragon 42, 50009, Zaragoza, Spain 48 [email protected] 49 50 Nadia Slimani (PhD), International Agency for Research on Cancer (IARC), 150 Cours 51 Albert Thomas, 69372 Lyon Cedex 08, France 52 [email protected] 53 54 Mazda Jenab (PhD), International Agency for Research on Cancer (IARC), 150 Cours Albert 55 Thomas, 69372 Lyon Cedex 08, France 56 [email protected] 57 58 Stefanie Vandevijvere 59 Unit of Epidemiology, Scientific Institute of Public Health, Brussels, Belgium 60 [email protected] 61 62 Guy De Backer (Prof), Unit nutrition and food safety, Department of Public Health, Faculty 63 of Medicine and Health Sciences, Ghent University, De Pintelaan 185, B-9000 Ghent, 64 Belgium 65 [email protected] 66 3 67 Stefaan De Henauw (Prof), Unit nutrition and food safety, Department of Public Health, 68 Faculty of Medicine and Health Sciences, Ghent University, De Pintelaan 185, B-9000 Ghent, 69 Belgium 70 [email protected] 71 72 73 74 *Corresponding author: Inge Huybrechts, Department of Public Health, Faculty of Medicine 75 and Health Sciences, Ghent University, UZ – 2 Blok A, De Pintelaan 185, B-9000 Ghent, 76 Belgium. Tel: +32 (0)9 332 24 23, Fax: +32 (0)9 332 49 94, email: [email protected] 77 78 4 79 Abstract 80 Background/Objectives: Low calcium and vitamin D intakes have been associated with 81 health risks in childhood and adulthood. This study aims to investigate dietary sources of 82 calcium and vitamin D intake, and its associated socio-demographic, economic and lifestyle 83 factors among preschoolers. 84 Subjects/Methods: Three-day estimated diet records from 696 Flemish preschoolers 2.5-6.5 85 years old (51% boys) were used (66% of 1052 collected diaries). Contribution of 58 food 86 groups to calcium and vitamin D intake were computed. Multiple linear regression was used 87 to examine associations of intakes with socio-demographic, economic and lifestyle factors. 88 Results: Mean calcium intake (844 mg/d) was above, and mean vitamin D intake (2.0 µg/d) 89 largely below the Belgian recommendations. Milk, sweetened milk drinks, and cheese were 90 the main sources of calcium intakes (26%, 25% and 11%, respectively). Butter and margarine 91 were the main vitamin D sources (26%), followed by growth milk (=fortified milk) (20%) and 92 fish (15%). Calcium and vitamin D intake were negatively associated with participants’ age, 93 and calcium positively with parental education and family size. The child’s gender, 94 supplement use and physical activity level, and the employment status and smoking behaviour 95 of the parents were not associated with calcium or vitamin D intake. 96 Conclusions: Flemish preschoolers had too low vitamin D intakes while most had adequate 97 calcium intakes. Milk (including sweetened, fortified/growth milk) was the main food source 98 of calcium intake and the second important source of vitamin D intake after butter and 99 margarine. Calcium intake was positively associated with parental education, while vitamin D 100 intake not. 101 Key words: dietary sources, vitamin D, calcium, preschool, child, socio-demographic, 102 economic 5 103 Introduction 104 Even though clinical consequences of adverse bone health are predominantly seen in older 105 age, accumulating evidence indicates that many predisposing factors arise in childhood 106 (Davies et al, 2005). Bone mineral content at older age is determined by the peak bone mass 107 attained in young adulthood and subsequent adult bone loss (Davies et al, 2005; Kalkwarf et 108 al, 2003). Therefore, interventions designed to maximize bone health should start at an early 109 age. While peak bone mass is mainly genetically controlled, calcium and vitamin D intake 110 during childhood and adolescence are considered to play an important role in bone health 111 (Davies et al, 2005; Johnston et al, 1992; Matkovic, 1992; Rizzoli et al, 2010). 112 Previous findings on calcium and vitamin D intakes of Flemish preschoolers suggest that 113 <70% of the children comply with the recommendations (Hoge Gezondheidsraad, 2003) for 114 calcium intake and <15% with the recommendations for vitamin D intake (Huybrechts & De 115 Henauw, 2007). These borderline calcium and inadequate vitamin D intakes, in comparison 116 to the Belgian Superior Health Council (BSHC) recommendations (Hoge Gezondheidsraad, 117 2003), are cause of concern for the health of Belgian preschool children, and provide evidence 118 of dietary inadequacies in an understudied subgroup of the population. Therefore, it is 119 important to examine the food sources contributing most to daily calcium and vitamin D 120 intakes of Flemish preschoolers, and subpopulations most vulnerable of inadequate intakes in 121 order to enable targeted interventions. 122 The aim of this study is to investigate dietary sources of calcium and vitamin D intakes and its 123 associated socio-demographic, economic and lifestyle factors among Flemish preschoolers. 124 125 6 126 Methods 127 Survey population 128 The study used data from the Flanders preschool dietary survey (data collected from October 129 2002 until February 2003), in which usual dietary intake was estimated from 3-day estimated 130 dietary records (EDR) and a forty-seven item Food Frequency Questionnaire (FFQ), 131 completed by a proxy (parent/caregiver). The sampling design, methods response rate (50%) 132 and representativity of the study sample were described previously (Huybrechts et al, 2008b). 133 In brief, schools were used as primary sampling units and classes as secondary sampling 134 units. A total of 2095 children from forty-three schools were invited to participate. The 135 percentage of under-reporters was low (< 2% of the children when using the individual 136 Goldberg cut-offs adopted for children) and has been described in depth elsewhere 137 (Huybrechts & De Henauw, 2007). 138 139 Socio-demographic, economic and lifestyle information about the child and its family was 140 collected via a parental questionnaire: employment status (employed or unemployed); 141 educational attainment (lower secondary, secondary or higher education); and smoking status 142 (currently smoking: yes/no) of the parents; family size (less or more than two children); 143 children’s physical activity level (light, medium or high) and supplement use (yes/no) of the 144 child. Level of physical activity was assessed via a short parental questionnaire asking about 145 their subjective perception of the physical activity level of their child (very active (e.g. 146 running, swimming, cycling), active (e.g. walking, actively playing) or less active (e.g. mainly 147 sitting or screen activities), and the number of hours their child spends in organized sport 148 activities (e.g. football, ballet, etc.). 149 7 150 The Ethical Committee of the Ghent University Hospital granted ethical approval for the 151 study. Informed written consent was obtained from all parents. 152 153 Assessment of dietary calcium and vitamin D intake 154 For the purposes of the current analysis only food diaries with three completed record days, 155 were included resulting in a final sample of 696 (66% of the 1052 collected) diaries. 156 Using a systematic procedure the following food composition tables (listed in order of 157 priority) were used to perform nutrient calculations: the Belgian food composition tables 158 NUBEL (NUBEL, 2004), Dutch food composition database NEVO (NEVO, 2001), food 159 composition tables of the Belgian Institute Paul Lambin (IPL, 2004), and the McCance and 160 Widdowson’s UK food composition tables (FSA, 2002). Within the context of this study, 161 systematic procedure refers to the use of the Belgian food composition tables as the basic 162 tables and thereafter when insufficient information was available in this Belgian database, the 163 other databases were used. 164 165 In total 936 foods and composite dishes were reported in the food diaries of the 696 children 166 that were included in the study sample. All recipes described in depth as ingredients in the 167 diaries were encoded as ingredients in the database. In order to enable classification of foods 168 into food groups of the Flemish Food Based Dietary Guidelines (FBDG) (Vigez, 2004), eight 169 extra composite dishes had to be disaggregated. After the disaggregating procedures, food 170 items were divided into 58 food groups of similar nutrient content based on the classification 171 of the Flemish FBDG and the expert opinion of the investigators (see table 2). 172 173 The contribution of dietary supplements to total calcium and vitamin D intake was not 174 considered in this study. However, in depth analysis reporting the frequency of consumption 8 175 of dietary supplements among Flemish preschoolers is published elsewhere (Huybrechts et al, 176 2010). 177 178 Statistical analyses 179 The Statistical Package for the Social Sciences for Windows version 14 (SPSS Inc., Chicago, 180 IL, USA) was used to perform statistical analyses. 181 Mean and median usual intakes of the population and the proportion below or above defined 182 cut-offs (Hoge Gezondheidsraad, 2009), were calculated using statistical modeling (NUSSER 183 method, developed at Iowa State University) in order to correct for day-to-day variability in 184 the 3d EDR (Guenther et al, 1997; Nusser et al, 1996). The program used to calculate usual 185 intakes was the Software for Intake Distribution Estimation (C-side) (Iowa State University, 186 USA) (Iowa State University, 2006). Recommendations of the BSHC (revised version 2009) 187 were used to evaluate total calcium and vitamin D intakes (Hoge Gezondheidsraad, 2009). 188 The population proportion formula was used to determine the percentage contribution of each 189 of food group to the intake of calcium and vitamin D (Krebs-Smith et al, 1989) which 190 includes summing the amount of the component provided by the food for all individuals 191 divided by the total intake of that component from all foods for the entire study population 192 (Fox et al, 2006; Krebs-Smith et al, 1989; Royo-Bordonada et al, 2003). 193 194 Multiple linear regression analysis was used to investigate the associations of mean calcium 195 and vitamin D intake (dependent variables) with the socio-demographic, economic and 196 lifestyle variables available from the parental questionnaires (independent variables). 197 Associations were simultaneously controlled for all variables included in the models: total 198 energy intake, sex, age, physical activity level, supplement use, household size, occupational 9 199 status and educational level of the parents and smoking of the parents. Interactions of the 200 independent variables with age and gender were also included. Only significant variables 201 associated with calcium and vitamin D intake were included in the final model. All two-tailed 202 statistical analyses were tested with a significance level set at P < 0.05. 203 204 Results 205 Characteristics of the study population 206 The total study sample of 696 children included 51% boys and 49% girls. 30% of the children 207 were between 2.5-3 years old and 70% between 4-6.5 years old (Table 1). The proportion of 208 children studied in each province (30% in Antwerp, 24% in East-Flanders, 22% in West- 209 Flanders, 15% in Flemish Brabant and 9% in Limburg), compared well with the proportions 210 derived from the target population, namely preschool children in Flanders (28%, 23%, 18%, 211 18% and 13% respectively). Mean calcium intake (95% CI) was 844 (821 – 868) mg per day 212 and mean vitamin D intake was 2.0 (1.8 – 2.2) µg per day. Mean calcium intake was above 213 and mean vitamin D intake largely below the age specific recommendations for children. 214 215 Insert Table 1 216 217 In total, 86% of the children 2-3 years old met the calcium recommendation of 500 mg per 218 day and 64% of the children 4-6 years old met the recommendation of 700 mg per day. Only 219 5% of the children met the vitamin D recommendation of 10 µg per day (4% in the children 2- 220 3 years old and 1% in the children 4-6 years old). 10 221 222 Dietary sources of calcium and vitamin D intake 223 Table 2 lists food and beverage categories and their relative contributions to daily calcium and 224 vitamin D intake in preschool children. Milk, sweetened milk drinks, and cheese were the 225 main sources of calcium intakes (26%, 25% and 11%, respectively), followed by milk 226 desserts, sweet snacks (e.g. child biscuits), cooked vegetables and fruit juice. 227 Butter and margarine (26%), fortified milk drinks (growth milk) (20%) and fish (15%) were 228 the main sources of vitamin D, followed by meat products, sweet snacks, milk, eggs and hard 229 cheese. Very similar results were obtained, for what concerns the main food sources 230 contributing to total calcium and vitamin D intakes, when stratifying for age or gender. 231 232 Insert Table 2 233 234 Associations of calcium and vitamin D intake with socio-demographic and economic 235 factors 236 Calcium and vitamin D intake were both negatively associated with age of the child, and 237 calcium was positively associated with both maternal and paternal educational level and 238 family size (Table 3). Gender, supplement use and physical activity level of the child, and , 239 employment status and smoking behaviour of the parents were not associated with calcium or 240 vitamin D intake. 241 242 Insert Table 3 11 243 244 Discussion 245 Main results 246 Mean calcium intake was above and mean vitamin D intake largely below the 247 recommendation for children (95% of the children had vitamin D intakes below the 248 recommendation) (Hoge Gezondheidsraad, 2009). Milk, sweetened milk drinks (e.g. 249 chocolate milk), and cheese were the main sources of calcium intakes (26%, 25% and 11%, 250 respectively). Butter and margarine were the main sources of vitamin D (26%) probably due 251 to the mandatory fortification of margarine in Belgium, followed by fortified milk drinks 252 (20%) and fish (15%). Meat and poultry also contributed importantly to vitamin D intakes 253 (11% for meat & poultry together). Both nutrients were negatively associated with 254 participants’ age, and calcium was positively associated with parental educational level and 255 family size, but not with the child’s gender, supplement use , the occupational status and 256 smoking behaviour of the parents. The lower calcium and vitamin D intakes in the oldest age 257 group are likely due to the reduced milk intakes in the oldest age group compared to the 258 youngest children (Huybrechts et al, 2008a). 259 It should be taken into consideration that our low estimated vitamin D intakes do not 260 necessarily imply a problem of vitamin D deficiency given that vitamin D is also derived 261 from the action of sunlight on the skin. Unfortunately, sun exposure was not assessed in this 262 study, and also contributions from supplements have not been taken into account. However, 263 studies conducted in similar geographical altitude have reported that the contribution of 264 vitamin D derived from sunlight is insufficient to reach the vitamin D requirement (Stellinga- 265 Boelen et al, 2007; Tylavsky et al, 2006). Previous research showed that the vitamin D intake 266 provided via dietary supplements was about 1.3 µg/d for children < 4 years old and 0.9 µg/d 12 267 for children at least 4 years old (Huybrechts et al, 2010). In that study, it was shown that the 268 group of children taking dietary supplements had still mean vitamin D intakes below 5 µg/d. 269 Therefore, young children in Belgium might risk having an inadequate vitamin D intake 270 during winter months (Davies et al, 1999; Emmett et al, 2002). 271 272 Comparison with the literature 273 Differences in dietary intake assessment methods, study design and food grouping made it 274 difficult to compare our calcium and vitamin D intakes, and food contributions to results 275 derived from other studies. These limitations should be considered when interpreting the 276 comparisons with other studies reported below. 277 Low vitamin D intakes found in our preschool children, were similar to that observed in other 278 European studies (Emmett et al, 2002; Lambert et al, 2004; Serra-Majem et al, 2006). For 279 instance the results concerning vitamin D intake and compliance to the recommendations 280 were comparable to those of Spanish children 2-5 years old (boys: 1.4 µg/d; girls: 1.2 µg/d) 281 and children from the UK aged 1-3 years old (1.8 µg/d) (Emmett et al, 2002; Serra-Majem et 282 al, 2006). In contrast, calcium intakes in the Spanish children (boys: 135 mg/1000 kcal; 283 girls: 116 mg/1000 kcal) were much lower than those observed in our Belgian sample of 284 children (Serra-Majem et al, 2006). Similarly, calcium intakes of the UK (boys: 796; girls: 285 732 mg/d) were lower than those reported for Belgian children (Emmett et al, 2002; Serra- 286 Majem et al, 2006). 287 288 Similarly to our study, milk was the main calcium source in another survey among Spanish 289 children, (26 % in Spain compared with 50 % in Belgium) (Royo-Bordonada et al, 2003). 290 Fish was the main vitamin D source in Spanish children (43 %), while margarine/butter in 13 291 Flemish preschoolers (9% in Spanish children) (Royo-Bordonada et al, 2003). Comparison 292 with main vitamin D contributors among four year old Swedish children showed that in the 293 Swedish preschoolers, the contribution of margarine, butter and oils (20%) and of fish (11%) 294 to total vitamin D intakes was lower in comparison with our Flemish preschoolers (Garemo et 295 al, 2007). 296 297 Both calcium and vitamin D intakes derived from food intake alone was higher for children in 298 the US (calcium: boys: (< 4y) 999 mg/d, (> 4y) 1058 mg/d; girls: (< 4y) 965 mg/d, (>4 y) 951 299 mg/d; vitamin D: boys: (< 4y) 7.2 µg/d, (> 4y) 6.4 µg/d; girls: (< 4y) 6.9 µg/d, (>4 y) 5.5 300 µg/d) than to those of our sample (Bailey et al, 2010). The main calcium food sources in US 301 children 2-5 y old were milk and cheese and the percentage contribution for those two food 302 groups (59% & 11% respectively) was very similar to the percentages reported for Belgian 303 children (Table 2) (Subar et al, 1998). 304 When considering the above comparison with European countries, it appears that the intakes 305 of calcium and vitamin D are lower in European children than in children from the US. This 306 could be attributed to differences in fortification policies or food composition. In addition, 307 within Europe, differences in fortification policies might be causing differences in vitamin D 308 and calcium intakes and in the main food sources contributing to these intakes (e.g. due to 309 margarine fortification, the main vitamin D sources in Belgian children were margarine/butter 310 compared to fish in Spain). 311 Although no information was found of the influence of parental education on the intake of 312 children’s calcium and vitamin D intakes, Galobardes et al. showed that calcium and vitamin 313 D intakes were negatively associated with the educational level of the study subjects 314 (Galobardes et al, 2001). A recent study in Polish children and adolescents (7-16 y old) also 14 315 showed more frequent fish, fruit and vegetable intakes among children & adolescents of 316 higher socio-economic status (SES) families (Suliga, 2010). Given the gap in data 317 investigating the influence of socio-demographic and economic factors influencing calcium 318 and vitamin D intakes among preschool aged children, future studies investigating 319 preschoolers dietary intakes should look at possible socio-demographic and economic factors 320 influencing these intakes to evaluate the results obtained in our study. 321 322 Strengths & limitations 323 Although, willingness to participate leads to some selection bias, these data represent a more 324 general population of preschool children in Flanders compared to other food consumption 325 surveys mostly restricted to local areas. Nonetheless, as shown previously (Huybrechts et al, 326 2008b), the study sample was subject to some selection bias, with lower socio-economic 327 classes being slightly underrepresented. Although this slight selection bias might have 328 influenced our results concerning the association between calcium and vitamin D intakes and 329 socio-economic factors, it is not possible to prevent such selection bias in large-scale surveys. 330 331 Similarly, to other dietary assessment methodologies, diet records are prone to a degree of 332 misreporting which possibly had an influence in the classification of compliance and non- 333 compliance with the recommendations. However, the percentage of under-reporters in the 334 final sample for analysis was less than 2%. In addition, a 3d diet record does not necessarily 335 reflect individual’s usual intake. For this reason,, statistical modelling (NUSSER method) that 336 accounts for within-individual variability (Iowa State University, 2006) was used in order to 337 calculate usual calcium and vitamin D intake. Unfortunately, it was impossible to correct for 338 seasonal variations, because the fieldwork was conducted only during autumn and wintertime. 15 339 No data were found about potential seasonal influences on nutrient intake in this population 340 group in Belgium. However, from the results of the National food consumption survey 341 (2004), it could be concluded that seasonal variations have a small effect on nutrient and food 342 intakes (De Vriese et al, 2006), likely due to the widespread availability of most foods all year 343 round. In addition it should be noted that food composition data, used for calculating nutrient 344 intake might also introduce some bias in dietary surveys reporting nutrient intake. Therefore, 345 the authors would like to emphasize the growing requirement for good quality food composition 346 data. 347 Finally, it should also be mentioned that the SES and lifestyle variables used in the analyses 348 were obtained via a parental questionnaire that included some basic questions about the 349 family situation and some lifestyle habits of the child. These subjective SES and lifestyle 350 measure could also introduce some bias (e.g. due to social desirable answers). 351 352 Recommendations 353 Our study findings and published literature indicate unfavourably low vitamin D intakes 354 among childhood populations. Enhancing the daily amount of spreadable margarine/butter on 355 preschool children’s bread and increasing fatty fish intake could contribute importantly to 356 higher vitamin D intakes. While spreadable margarine for preschool children’s bread was 357 under-consumed in comparison with the FBDG (Huybrechts et al, 2008a), it was still the main 358 source of vitamin D intakes. Although spreadable fat is an item that might be more prone to 359 underreporting in diaries, the results from the FFQ also revealed that more than one-third of 360 the children never consume spreadable fat on their bread (Huybrechts et al, 2008a). However, 361 in the battle against obesity one should keep in mind that the total energy intake should not be 362 influenced by the enhancement of these fatty food groups what could necessitate a 363 concomitant decrease in other (energy dense) foods containing invisible fat or sugar (e.g. soft 16 364 drink and candy bars). In the Flemish FBDG children are recommended to use about 5g 365 margarine per slice of bread (3-5 slices of bread per day are recommended in preschool aged 366 children and margarine is recommended above butter because of its more advantage fatty acid 367 contents). 368 Also the consumption of fortified (growth) milk in children could be recommended to 369 increase children’s vitamin D intake. However, most of these growth milks contain added 370 sugar which makes them energy dense and less appropriate in the fight against obesity. 371 If public health policies for the increase in vitamin D intake are to be effective, policy 372 development and implementation needs to target the main sources of dietary vitamin D in 373 various populations (including higher as well as lower educated populations). In addition to 374 the mandatory fortification of margarines, manufacturers could also contribute by 375 reformulating existing products to incorporate more calcium and/or vitamin D. 376 377 Conclusion 378 Flemish preschoolers surveyed between 2002 and 2003, had mean vitamin D intakes largely 379 below the recommendation, whereas mean calcium intake was above the minimum 380 recommended intake level. This situation could be improved by promoting the consumption 381 of food categories with high vitamin D contents such as margarine, fish and fortified dairy 382 products. Both calcium and vitamin D intakes were negatively associated with age, and 383 calcium intake was positively associated with the educational level of the mother and the 384 father and family size. This finding suggests that prevention strategies should especially focus 385 on children from lower educated parents. 386 387 17 388 Acknowledgements 389 We thank all the parents and teachers who participated into this project and generously 390 volunteered their time and knowledge. The Flanders preschool dietary survey was funded by 391 the Belgian Nutrition Information Center (NICE). 392 Contributors: Inge Huybrechts performed and interpreted statistical analyses and drafted the 393 article. Yi Lin contributed to the statistical analyses and the interpretation of the results. All 394 other authors helped in the evaluation of the results and the writing of the manuscript. 395 Moreover, Inge Huybrechts and Stefaan De Henauw were responsible for the study protocol 396 and the fieldwork. All authors have read and have approved the manuscript as submitted. 397 398 Conflict of interest 399 The authors declare no conflict of interest. 400 401 402 18 403 Table 1. Characteristics of the children participating in the Flanders preschool dietary survey Children with 3d diet record n£ (%) Boys 339 (51.2) Girls 323 (48.8) 2.5-3 years old 197 (29.8) 4-6.5 years old 465 (70.2) Antwerp 211 (30) Limburg 62 (9) East-Flanders 167 (24) Flemish-Brabant 101 (15) West-Flanders 155 (22) Lower secondary education 26 (4.0) Secondary education 250 (38.2) Higher education 379 (57.9) Gender: Age: Province: Maternal education 19 Paternal education Lower secondary education 49 (7.7) Secondary education 279 (43.8) Higher education 309 (48.5) Two children or less in the family 480 (72.7) More than 2 children in family 180 (27.3) Smoker 98 (14.9) Non-smoker 561 (85.1) Smoker 160 (24.9) Non-smoker 483 (75.1) Employed 488 (74.5) Unemployed 167 (25.5) Employed 566 (88.9) Unemployed 71 (11.1) Brothers and sisters in the family Smoking habits mother Smoking habits father Maternal employment Paternal employment Physical activity level of child 20 Less physically active 246 (37.7) Moderate physically active 303 (46.5) Highly physically active 103 (15.8) Special diet for diabetes 0 - Gluten-free diet (gluten-intolerance) 1 (0.1) Lactose-free diet (lactose-intolerance) 3 (0.4) Special diet for food-allergies 8 (1.1) Energy-restricted diet 2 (0.3) 3 (0.4) 17 (2.4) Vegetarian or semi-vegetarian (no red meat) 3 (0.4) Lacto-ovo-vegeterian 1 (0.1) Ovo-vegeterian 0 - Lacto-vegeterian 0 - Biological diet 6 (0.9) Religious or other eating pattern 1 (0.1) Total 11 (1.6) 9 (1.3) Taking special diet: Other diet (e.g. laxative diet, high energy diet, fibre restricted diet) Total Following special eating pattern: Used dietary supplements during previous month: Iron 21 Calcium 9 (1.3) Fluor 0 - Multiminerals 3 (0.4) Vitamin C 34 (4.9) Vitamin B-complex 0 (0.0) Fat-soluble vitamins 3 (0.4) Multi-vitamins 96 (13.8) Multi-minerals & -vitamins 80 (11.5) Echinacea 15 (2.2) Total‡ 226 (32.5) Mean 95% CI 844.3 (820.6-868.0) 2.0 (1.8-2.2) 584.1 (569.5-598.6) 1.4 (1.3-1.5) Nutrient intake* Calcium (mg/d) Vitamin D (μg/d) Calcium(mg/1000kcal) Vitamin D (µg/1000kcal) £ n 696 (number of children who completed diet record) 404 *Distribution was corrected for within person variability and for day of the week using Cside 405 (Iowa State University, 2006) 406 †Belgian Superior Health Council (BSHC) (Adequate intake = AI) 407 ‡ The total number of children using dietary supplements is not equal to the sum of the 408 children using the different types of dietary supplements as some of the children were using 409 more than one type of dietary supplement 22 Table 2 - Dietary sources of calcium and vitamin D intakes of Flemish preschoolers. Food intake (g) Food group Subgroup Beverages (incl. juices but no drinks from restgroup) Water Mean Median Total Vitamin D (SD) 486.2 % order 0.7 6.0 2.2 0.0 (90.1) 0.0 0.2 8.2 0.0 (43.5) 0.0 0.0 172.8 150.0 (209.3) 0.0 2.6 Vegetable juice 0.2 0.0 (6.0) 0.0 0.0 Soup / bouillon 57.7 0.0 (101.7) 0.7 1.0 Fruit juice Bread and cereals Bread / rolls / crackers / rice cakes 23.1 order 0.0 Tea and coffee without sugar 150.0 % (226.4) Light beverages 224.2 Total Calcium 86.7 0.0 62.5 (46.8) 0.0 2.0 Sweetened bread 7.5 0.0 (22.5) 0.0 0.3 Breakfast cereals (ready-to-eat / hot) 8.9 0.0 (20.0) 0.0 2.1 Pasta / noodles 86.7 15.4 1.4 0.0 (41.0) 7 4.3 70.3 Potatoes and grains 9 10 1.2 0.0 0.2 23 Food intake (g) Food group Subgroup Rice Potatoes Vegetables Mean Median Total Vitamin D (SD) % order 6.3 0.0 (25.5) 0.0 65.0 50.0 (69.3) 1.4 66.5 Total Calcium % order 0.1 10 0.0 1.0 3.4 Cooked vegetables 53.7 40.0 (60.1) 0.0 3.1 Raw vegetables 12.8 0.0 (38.3) 0.0 0.3 Fruit (sweetened / unsweetened) 109.9 0.0 1.3 Fresh fruit 94.0 68.8 (102.7) 0.0 1.2 Canned fruit 15.4 0.0 (45.4) 0.0 0.1 Dried fruit 0.4 0.0 (3.7) 0.0 0.0 Olives 0.1 0.0 (1.5) 0.0 0.0 Milk, milk products and calcium enriched soy milk 433.9 24.9 Milka 170.3 112.5 (193.5) Sweetened milk drinks (e.g. Fristi, chocolate 171.3 125.0 (195.7) milk, …) 6 60.3 2.3 0.03 6 25.1 1 22.8 2 24 Food intake (g) Food group Subgroup Growth milk (fortified with vitamins & Mean Median Total Vitamin D (SD) % Total Calcium order % 2 order 19.7 0.0 (88.2) 19.7 2.4 4.5 0.0 (25.3) 0.0 0.8 Sweetened or aromatised yoghurt 14.2 0.0 (46.9) 0.0 2.3 Soy drinks 15.7 0.0 (82.5) 0.8 1.7 Milk desserts 19.9 0.0 (56.2) 1.4 Milk desserts based on soy 2.3 0.0 (19.1) 0.0 0.2 Probiotics (e.g. Actimel, Yakult, …) 0.7 0.0 (7.4) 0.0 0.0 15.3 0.0 (43.3) 0.7 1.7 minerals) Yoghurt White (fresh) cheese Cheese 14.5 4.0 11.8 0.0 (22.6) 3.5 Cheese spread 2.7 0.0 (8.8) 0.6 8.6 Butter / margarine 8.3 7 (9.5) 4 25.9 11.1 3 1.1 25.9 6.0 3.3 12.2 Hard cheeseb Fat & oilc 9 8 0.3 1 0.3 25 Food intake (g) Food group Subgroup Mean Median Total Vitamin D (SD) % Total Calcium order % order Oil 0.3 0.0 (1.4) 0.0 0.0 Frying oil 0.0 0.0 (0.6) 0.0 0.0 Meat / poultry / fish / egg / meat alternates 90.3 Meat, game and meat products 37.2 20.0 (46.1) 7.7 4 0.9 Chicken / turkey 15.9 0.0 (34.7) 3.2 9 0.2 8.5 0.0 (28.7) 14.6 3 0.3 Cold cuts (from meat products) 20.7 6.8 (30.2) 3.3 8 0.3 Cold cuts (from fish products) 0.9 0.0 (6.8) 1.3 Eggsd 5.1 0.0 (18.2) 4.1 Meat substitutes (e.g. tofu, tempe, …) 1.7 0.0 (11.6) 0.0 0.2 Nuts and seeds 0.3 0.0 (3.4) 0.0 0.0 Fish / shellfish Restgroup (snacks & desserts) Brioches Sweet snacks 34.2 201.8 2.4 0.0 6 9.2 0.3 8.3 3.5 0.0 (17.0) 0.0 43.6 32.0 (43.5) 7.5 0.2 5 3.3 5 26 Food intake (g) Food group Subgroup Mean Median Total Vitamin D (SD) % Total Calcium order % order Salty snacks 2.1 0.0 (9.8) 0.0 0.1 Tea and coffee with sugar 3.2 0.0 (26.6) 0.0 0.0 Soft drinks 97.7 0.0 (169.4) 0.0 0.6 Salty sauces 12.5 0.0 (24.9) 0.6 0.9 Cream 0.3 0.0 (2.6) 0.1 0.0 Sweet sauces 0.1 0.0 (2.5) 0.0 0.0 Chocolate 3.1 0.0 (9.5) 0.0 0.7 Chocolate spread 9.4 0.0 (13.9) 0.0 1.3 Other sweet spread (e.g. jam, honey, …) 5.3 0.0 (11.6) 0.0 0.1 Sugar 0.1 0.0 (0.9) 0.0 0.0 Fried snacks 0.1 0.0 (2.6) 0.0 0.0 14.6 0.0 (37.7) 0.0 0.4 6.2 0.0 (23.2) 1.0 0.6 French fries / croquettes Sweet desserts (e.g. ice cream, tiramisu, …) Miscellaneous 4.2 0.5 0.4 27 Food intake (g) Food group Subgroup Mean Median (SD) Total Vitamin D % order Total Calcium % order Pizza & quiches 2.2 0.0 (17.8) 0.3 0.3 Other miscellaneouse 2.0 0.0 (21.3) 0.2 0.1 The contributions of each food group are expressed in percentage of daily calcium and vitamin D intake. a Includes cow's milk and goat's milk b Excludes cream cheese c Includes lard / animal fats and regular / low-fat / fat-free versions of cream cheese / sour cream / half-and-half d includes only eggs reported separately and eggs included in disaggregated food mixtures e includes foods or components with negligible contributions to total nutrient intakes that could not be categorized in the above food groups (e.g. herbs and spices / monosodium glutamate / starch / plain gelatin / artificial sweeteners / pectin / cocoa powder / etc.) 28 Table 3. Associations of calcium and vitamin D intake with socio-demographic and economic characteristics of participants. β Type I Wald Dependent variable: Coefficients 95% Confidence Interval Lower Upper Bound Bound Chi-Square P-value β SE Intercept 0.503 0.435 -0.349 1.4 1.4 0.247 Energy 0.002 0.000 0.001 0.002 45 <0.001 -0.191 0.0071 -0.330 -0.051 7.2 0.007 Vitamin D (n=594)† Age Calcium (n=594) ‡ Intercept Energy Age 17.7 103 -184 220 0.030 0.863 0.495 0.040 0.417 0.572 156 <0.001 -28 12.0 -51 -4.0 5.3 0.022 29 β Type I Wald Dependent variable: Coefficients β 95% Confidence Interval SE Lower Upper Bound Bound Chi-Square P-value Secondary educated mother* 111 58 -3.5 225 3.6 0.057 Higher educated mother* 123 59 8.0 239 4.4 0.036 67 42 -15.3 149 2.6 0.110 Higher educated father* 139 44 53 225 10.0 0.002 Family size (>2children)§ 61 25 12.0 5.9 0.015 Secondary educated father* 110 *Maternal and paternal educational level, using lower education as a reference. § Family size, using two children or less as a reference. †Non-significant variables with standardized coefficients β for vitamin D intake. Gender (girl) : β=-0.077, P=0.636; 30 Maternal education : secondary: β=-0.777, P=0.051; higher: : β=-0.702, P=0.073; Paternal education : secondary: β=0.436, P=0.150; higher: : β=0.462, P=0.143; Physical activity: moderate : β=0.045, P=0.794; higher; β=-0.023, P=0.923; Family size (>2 children): β=0.005, P=0.977; Maternal smoking status (no): β=-0.420, P=0.058; Paternal smoking status (no): β=0.172, P=0.378; Maternal employment (unemployed): β=-0.239, P=0.225; Paternal employment (unemployed): β=0.196, P=0.449; Supplement (no): β=-0.321, P=0.052 ‡Non-significant variables with standardized coefficients β for calcium intake. 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