Original Research

Dietary Assessment of Arachidonic Acid and Docosahexaenoic Acid Intake in 4 7 Year-Old Children
Vanessa W. Lien, MSc, RD, Michael T. Clandinin, PhD Alberta Institute for Human Nutrition, Edmonton, Alberta, Canada T6G 2H1 Key words: polyunsaturated fat, food records, dietary intake, school age, fat
Objective: To investigate current dietary intakes of arachidonic acid (AA), docosahexaenoic acid (DHA) in healthy children between 4 7 years of age using a 3-day food record. Design: Cross-sectional study investigating dietary intakes using food records. Parents were instructed to document all food and drink consumed by their child for 3 consecutive days. Subjects and Setting: Healthy children (n 91), 4 7 years of age, living in central Alberta, Canada were volunteered by their parents to participate in the study. Seventy-eight children completed the study. Results: AA and DHA intakes were 57 35 mg/day and 37 63 mg/day, ranged between 1.2180 mg/day and 0 350 mg/day and varied day to day at 0 380 mg/day and 0 991 mg/d, respectively. DHA intake was 30 mg/day for 74% of the subjects. Conclusion: Canadian children, 4 7 years of age and not living near a marine environment, have relatively low dietary intakes of AA and DHA. Retinal and neuronal development continues throughout childhood, therefore it is conceivable that low intake of AA and DHA may have a negative impact.

INTRODUCTION
Long chain polyunsaturated fatty acids (LCP), specifically arachidonic acid (20:4 n-6, AA) and docosahexaenoic acid (22:6 n-3, DHA), are important in infant growth and development [1]. These fatty acids are major components of membranes of the brain and retina [2,3]. Dietary sources of AA can be found in liver, poultry, beef, pork, seafood, and eggs, while dietary sources of DHA include fish, shellfish, poultry, and supplemented food items such as eggs [4]. Higher levels of DHA are found in fatty fish such as mackerel, salmon, and herring. During infant development, there is rapid accretion of AA and DHA during the last trimester of gestation [5]. Infants born prematurely are deprived of in utero accretion of AA and DHA unless they are fed AA and DHA [6]. Preterm infants supplemented with DHA showed an improvement in visual acuity [7,8] and speed of information processing [9,10]. In a recent double-blind multi-centre study, feeding preterm infants formula containing AA and DHA for 92 weeks resulted in enhanced growth and higher Bayley mental and psychomotor

development scores than control infants that were provided with unsupplemented formulas during a follow-up assessment at 118 weeks [11]. In term infants, AA and DHA have also been shown to improve vision [1214] and brain development [13,15,16]. The grey matter of the brain, where DHA accumulates in large amounts during growth and development [2], continuously increases from birth through 5 years of age [17]. Although the rate of growth slows, the brain and eye continue to develop throughout childhood [18,19], therefore low levels of AA and DHA availability may have a negative impact on the retinal and neuronal cellular development in children during the early years of life. There is also literature on the beneficial effect of AA and DHA in Attention-deficit/hyperactivity disorder (ADHD), a common childhood behavioural disorder. Children with ADHD are inattentive, impulsive and/or hyperactive [20] and have lower concentrations of AA and DHA in both plasma and red blood cell lipids [21,22]. Children with ADHD that have lower levels of plasma n-3 and n-6 fatty acids compared to subjects with higher levels of n-3 and n-6 fatty acids exhibit signs of

Address reprint requests to: Dr. M. Tom Clandinin, Alberta Institute for Human Nutrition, 823 General Services Building, University of Alberta, Edmonton, AB T6G 2H1, CANADA. E-mail: tom.clandinin@ualberta.ca Abbreviations: AA arachidonic acid, DHA docosahexaenoic acid, LCPUFA long chain polyunsaturated fatty acids, ALA alpha-linolenic acid, LA linoleic acid

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Fat Intakes in 4 7 Year Children

essential fatty acid deficiency including increased thirst, frequent urination, and dry skin [23]. Supplementation with both n-3 and n-6 was effective in reducing ADHD related symptoms in children [24,25]. Other studies have also looked at the role DHA may play in dementia and prevention of macular degeneration [26]. The review paper by Johnson and Schaefer [26] summarizes evidence that supports low dietary DHA intakes and plasma concentrations have been reported to be associated with dementia, cognitive decline and age-related macular degeneration. DHA supplementation has been shown to affect vision and improve visual acuity and slow the progression of the macular dystrophy [27]. A small number of studies exist in the current literature that report dietary AA and DHA intake in healthy children in the past 1520 years. In the late 1980s, there were two national surveys conducted in the United States that reported fatty acid intakes of children to be approximately 100 mg/day for 18: 4AA and 100 mg/day for 20:5DHA [28,29]. In Australia, a national survey in 1995 has estimated that in 4 7 year old children, AA and DHA intakes were 22 mg/d and 47 mg/d respectively [30]. A study done in the Vancouver, Canada (an area where seafood is readily available) has estimated dietary AA and DHA intakes to be 226 mg/d and 96 mg/d respectively [31]. There has not been any new national data published on the dietary intakes of AA and DHA in children. Due to the scarcity of information in the current literature reporting intakes of LCP in healthy children, there is a need to investigate and report current dietary intakes of AA and DHA in these children. Current comparisons of AA and DHA can only be made to what is available in the existing literature. This study helps to add to the current body of knowledge in area of LCP intakes in healthy children. The purpose of our study is to investigate current dietary intakes of AA and DHA in healthy children between 4 7 years of age using a 3-day food record. Geographically children in this study are not living close to a marine environment and would not be expected to have high intakes of fish or seafood high in n-3 fatty acids. It was hypothesized that the diets of children studied provide low intakes of DHA. LCP may play a critical role in brain and eye development throughout childhood, therefore children 4 7 years of age were studied in order to add to our current body of knowledge. approached by the study coordinator and informed about the research study. All Principals and teachers in each of the primary schools agreed to participate in the current research study. Approximately 200 take home information pamphlets were then given to teachers to distribute to children in kindergarten and grade one. No financial incentives were used to encourage participation. Inclusion criteria included male and female children (including siblings) between the ages of 4 7 from both low and higher income families, children with no metabolic abnormalities (e.g. diabetes), written consent from parents on behalf of their child, and at least one parent committed to attending the meetings with the researcher. Children with any chronic illness other than asthma or those with acute illness at the time of the study were not recruited. Children were of normal intellect and age for grade. Intelligence quotient, anthropometric data (e.g. weight and height), if a child was breast or formula fed, and parental dietary habits were not collected for this study as the purpose of this study was solely to investigate dietary habits of these healthy children. From May 2001-June 2002, a total of 91 subjects enrolled in the study. Due to the paucity of data available on LCP intakes in children, there was no basis for estimate of N required to access AA and DHA intake. General information including age, gender, medical information, and parental income was recorded. Low-income cut off for a family of 3 4 persons living in this region was defined as those with total family income of less than $25 000 before taxes [32]. Families with higher incomes were defined as those with a total family income greater than $40 000 before taxes. In 2001, median family income for all families in Stony Plain and Spruce Grove was $62 475 and $64 243 respectively [33]. Thus, it is expected that the income data collected in this study will be skewed towards incomes greater than $40 000 before taxes. Children received a storybook as a token of appreciation after completion of the study. Parents were also given a copy of Canadas Food Guide to Healthy eating and were given the opportunity to ask the dietitian questions regarding the diet of their child.

Dietary Assessment
An initial visit was arranged with the parents where the dietitian outlined the project, reviewed participant responsibilities, obtained informed written consent (parents on behalf of their child), subject information and medical history of the child. Parents were instructed to document all food and drink consumed by their child on a daily basis for a period of 3 consecutive days (two weekdays and one weekend day). The dietitian explained to parents how to record foods consumed by their child on the food record forms and suggested that food items were to be recorded immediately after child had eaten whenever possible. The amount of food/drink consumed by the child was recorded (e.g. cups, milliliters, ounces or pieces), method of preparation of food (e.g. broiled, roasted, fried, etc), brand name of food item, labels and recipes, and use of any

METHODS
Subjects
The Human Ethics Review Committee of the Faculty of Agriculture, Forestry and Home Economics, University of Alberta, Canada approved the study protocol. The Superintendent of the Parkland School County, located in central Alberta was approached, and interested in assisting with the current research study. The superintendent identified 8 primary schools as potential study sites. Principals from each of the schools were

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multivitamins were also collected. Parents were given examples of how to estimate portion sizes if they were unable to weigh food items. In instances where a child brought a lunch to school, the parent would record what was packed for the child, and then have the child bring back any leftovers/garbage and the food record was adjusted as necessary according to the amount actually consumed by the child. Parents would also check with the child if they had consumed any additional snacks/food items while at school. The childs usual food pattern of DHA containing foods was also assessed using a food frequency questionnaire. This questionnaire was developed by study personnel and dietitians and contained detailed questioning on over 41 species of fish and shellfish (including fresh, frozen, canned, smoked and dried). Local foods were included in the questionnaire to develop a locally appropriate food frequency questionnaire. Content and participant instructions were made based on consultation with nutrition experts. Questions regarding the consumption and servings sizes of various fish, shellfish, liver, egg yolks, chicken, turkey, or other poultry on a monthly and/or weekly basis were recorded. Food records were analyzed using the USDA Nutrient Database for Standard Reference, Release 12 [4] in Food Processor 11, version 7.30 (1999, Esha Research, Salem Oregon). AA and DHA values for specific local foods of uncertain fat composition were determined by laboratory analysis (for example arctic char, chicken nuggets, fish sticks, fish rings) and entered into the nutrient database. The quality and quantity of fat (particularly AA and DHA) consumed by the children was determined. To maintain consistent data analysis, all food records and food frequency questionnaires were manually checked and coded by one dietitian who also performed the diet history interview. Macronutrient dietary intakes were compared to the acceptable macronutrient distribution ranges (AMDR) and linoleic acid (LA) and ALA (alpha-linolenic acid) were compared to adequate intake (AI), that is the average daily nutrient intake for a group (or groups) of apparently healthy people based on observed or experimentally determined approximations or estimates outlined by the Food and Nutrition Board and Institutes of Medicine [34]. AI is assumed to be adequate and is used when the RDA cannot be determined [34]. Vitamin A [35], vitamin E [36,37] and iron [38] are also important in normal development and growth of the eye and brain and therefore intakes of these nutrients were also assessed in study. (GLC), .pb10 fatty-acid methyl esters were separated and identified [40]. Fatty acid content was calculated from the internal standard added.

Statistical Analysis
Data was analyzed using the Statistical Analysis System (SAS) for Windows (SAS Institute Version 8.2, Cary NC, USA). Variables are expressed as mean standard deviation, median and range. The Kolmogorov-Smirnov test was used to assess normality for nutrient intake values for all subjects and to assess normality of seasonal intake of DHA using Splus for Windows (Insightful Corporation 6.2, Seattle WA, USA). Oneway analysis of variance (ANOVA) was used to examine differences between gender or between income groups for nutrient intakes (data not displayed). P 0.05 was considered significant. The Kruskal- Wallis test was used to assess the effect of seasons on DHA intake in all subjects. Nutrient values were compared between age groups. There were no significant differences, therefore the dietary intake data was pooled for all subjects. Energy, carbohydrate intakes were rounded to three significant figures, while fat intakes were rounded to two significant figures.

RESULTS
Subjects
A total of 91 subjects enrolled in the study, of which 78 subjects (M 39, F 39) completed. The mean age of subjects was 5.8 0.8 years (range 4.17.9 years). A total of 93.6% of the study population were Caucasian, 3.8% were Aboriginal, and 2.6% were of mixed origin. Ten families (12.8%) were in the low income group, 60 families (76.9%) had incomes $40 000, and 8 families (10.3%) had incomes between $25 000 and $40 000.

Dietary Assessment
Macronutrient, Vitamin, Mineral and Fatty Acid Intakes. The daily nutrient intake was estimated from 3-day food records from all children (Table 1). Protein, carbohydrate, and fat intakes of the children in the current study met the AMDR of 1030%, 4565% and 2535% respectively [34]. For vitamin A, vitamin E, and iron, children met the recommended Estimated Average Requirements, that is the average daily nutrient intake level estimated to meet the requirement of half the healthy individuals between 48 years of age as outlined by the Dietary Reference Intakes [42,43]. LA and ALA did not meet the AMDR specified for these fatty acids, that is 510% of total kcal and 0.61.2% of total kcal respectively. In 64 of 78 subjects, LA intake was lower in study subjects (7.4 3.3 g/day) compared to the AI for LA (10g/day) for those children 48 years of age [34]. In 56 of 78

Fatty Acid Analysis of Specific Foods

Fat from a sample of the specific food items was extracted using the modified Folch procedure [39]. Using thin layer chromatography (TLC), total lipids were separated as described in Clandinin et al [40], samples were methylated [41] using 14% (wt/wt) boron trifluoride in methanol and C17:0 was added as an internal standard. Using gas-lipid chromatography

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Table 1. Average Daily Nutrient Intake for Children Aged 4 7 Assessed by 3-Day Food Records
All subjects (n 78) Nutrients per day Macronutrients Calories (kcal) Protein (g) % of total kcal Carbohydrate (g)* % of total kcal Fat (g) % of total kcal Saturated fat (g)* % of total kcal Monounsaturated fat (g) % of total kcal Polyunsaturated fat (g)* % of total kcal Vitamin and minerals per day Vitamin A (IU) Vitamin A (RE) Vitamin E (mg) Iron (mg) Fatty acids Linoleic acid 18:2 (g)* % of total kcal Linolenic acid 18:3 (g)* % of total kcal Arachidonic acid 20:4 (mg)* Eicosapentaenoic acid 20:5 (mg)* Docosahexaenoic acid 22:6 (mg)* N- 3 (g)* N- 6 (g)* X (SD) 1760 (440) 61 (17) 14 (2.4) 245 (71) 56 (6.0) 63 (17) 33 (4.5) 23 (7.9) 12 (2.4) 22 (6.1) 12 (2.3) 9.8 (3.6) 5.1 (1.6) Median 1730 60 14 230 55 63 33 22 12 22 11 9.4 5 Range 10603370 110 7.020 130510 4270 32110 2243 1153 5.617 9.739 6.117 3.023 2.010

7080 (3990) 1290 (670) 9.5 (5.9) 12 (3.8) 7.4 (3.3) 3.9 (1.6) 0.71 (0.5) 0.36 (0.20) 57 (35) 17 (36) 37 (63) 0.75 (0.5) 7.4 (3.3)

6090 1110 8.1 11 6.8 3.7 0.62 0.35 51 4.6 16.5 0.72 6.7

87619200 2022870 2.034 5.431 2.619 1.08.2 0.173.1 0.0671.32 1.2180 0.0200 0.0350 0.183.1 2.619

* Carbohydrate (g), saturated fat (g), polyunsaturated fat (g), linoleic acid (g), linolenic acid (g), arachidonic acid (mg), eicosapentaenoic acid, docosahexaenoic acid, n-3 (g), and n-6 (g) were considered not normally distributed when assessed using the Kolmogorov-Smirov test. Vitamin A (IU and RE), vitamin E (mg), iron (mg) were considered not normally distributed when assessed using the Kolmogorov-Smirov test

large intra individual (within person) and inter individual (between person) variation. DHA intake was not influenced by seasonal intake of fish and seafood (P NS). DHA Intake Assessed by Food Frequency Questionnaire. The results from the food frequency questionnaire show that average consumption of 3 oz of fish/shellfish, 3 oz of liver, number of egg yolks, or 3 oz of chicken, turkey or other poultry was 3.8 3.7 servings/month, 0.030 0.19 servings/month, 2.6 2.1 servings/week and 2.7 1.5 servings/week respectively. Median intakes of 3 oz of fish/shellfish, 3 oz of liver, number of egg yolks, or 3 oz of chicken, turkey or other poultry was 2.8 servings/month, 0.00 servings/month, 2.0 servings/ week, and 2.5 servings/week respectively. Estimated mean and median DHA intake from food frequency questionnaire was 56 50 mg/day and 43 mg/day respectively. DHA intake ranged from 1.8250 mg/day. Comparison of DHA Intakes Assessed by Food Frequency Questionnaire and Food Record. There is a wide range of DHA intake confirmed by both the food record and the food frequency questionnaire. DHA intakes assessed by the food record is positively correlated with DHA intakes assessed by the food frequency questionnaire (P 0.02, r 0.27). Gender and Income Comparison. Macronutrient and fatty acid intake was not significantly different between males and females. Since income levels of the families were collected during the assessment a comparison of income levels against nutrients was determined. Income data was skewed towards incomes greater than $40 000 before taxes as expected. When macronutrient and fatty acid intakes were determined for the low income group compared with higher income group, all macronutrient intakes met the AMDR (except LA and ALA) and were not significantly different between the two income groups except for AA (P 0.05). AA intake was significantly higher in the lower income group (78.7 33.9 g/day) than in the higher income group (53.3 33.6 g/day).

DISCUSSION
The results reveal that 4 7 year old Canadian children in this study, not living near a marine environment have low intake of LA and ALA, that is 82% and 72% respectively did not meet the AI suggested for LA and ALA [34]. Recent stable isotope studies have suggested that preterm and term infants [44 47] and human adults [48 50] have low ability to convert LA to AA and ALA to DHA. The extent to which de novo synthesis of these LCP are sufficient to support the needs of the growing child has not been investigated. Few studies report AA and DHA dietary intake in children or adults. Of these studies, the data reported varies. In Australian children (aged 4 7 years, n 799), AA and DHA intake was 22 mg/day and 47 mg/day respectively when estimated from the 1995 National Nutritional Survey [30]. The same study also investigated the AA and DHA intake in Australian

subjects, ALA intake was also lower (0.71 0.5 g/day) compared to the AI for ALA (0.9 g/day) for children 48 years of age [34]. The n-6 (g) to n-3 (g) ratio was 10:1. AA and DHA Intake Assessed by Food Record. Mean AA and DHA intake was 57 35 mg/day and 37 63 mg/day respectively. The distribution of AA intake was extensive, and spread out compared to that observed for DHA (Fig. 1). Approximately 74% of the subjects (58 of 78) had DHA intakes 30 mg/day (Fig. 2). Median intake for AA and DHA was 51 mg/day and 17 mg/day respectively. AA and DHA intake calculated as an average of the 3-day intake ranged from 1.2180 mg/day and 0350 mg/day respectively. AA and DHA intake ranged from 0380 mg/day and 0991 mg/day respectively when intakes were calculated per day, which show a

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Fig. 1. Distribution of AA intake (mg/day) assessed by 3-day food records for children aged 4 7. AA intake (mg/day) was expressed in 10 mg/day increments and the number of children in each 10 mg/day increment is displayed in the bar graph.

Fig. 2. Distribution of DHA intake (mg/day) assessed by 3-day food records for children aged 4 7. DHA intake (mg/day) was expressed in 10 mg/day increments and the number of children in each 10 mg/day increment is displayed in the bar graph. Approximately 74% of subjects (58 of 78) had DHA intakes 30 mg/day. Approximately 27 of 78 or 35% had intakes of 10 mg/day.

adults ( 19 years of age, n 10851) and found that AA intakes were similar to the intakes found in the present study, however DHA intakes were 69 mg/day higher than the intakes in the present study. Meyer et al [30] reported the median intakes of AA and DHA in adults to be 24 mg/day and 15 mg/day respectively. Other studies have reported higher AA and DHA nutrient intakes than those estimated in the present study. Jonnalagadda et al [29] estimated fatty acid intake of children (aged 6 11, n 992) determined from data using 1987-1988 US Nationwide Food Consumption Survey and found that intake of 18:4AA was 100 mg/day and for 20: 5DHA was 100 mg/day. Similar results were found by Allison et al [28], who estimated fatty acid intake from the 19891991 US Continuing Survey of Food Intakes of Individuals (n 11 258) in individuals 3 years of age and older. In a study done by Innis et al [31], Canadian children 35 years of age, reported higher AA (226 17 mg/day) and DHA (96 14

mg/day) intakes than in the present study. Raper et al [51] investigated annual per capita food use data in the US food supply and found that the DHA levels have increased from 69 to 78 mg/capita/day between 1935-1939 and 1985 and was the result of increased use of canned tuna, gamefish, and poultry. Using per capital disappearance data of fish, Kris-Etherton [52] estimated DHA intakes in the United States as 0.25 g/day and worldwide as 0.23 g/day. Geographic location where nutrient assessments were completed is one of the reasons why wide variability of AA and DHA intake was observed. Many of the nutrient assessments were national surveys [28 30] which provide the mean intakes of AA and DHA for all subjects in the study. Although it is important to have an idea of the intakes of AA and DHA consumed by a population, it may not indicate areas that may have lower intakes of these fatty acids. In the study by Innis et al [31], DHA intake was expected be to higher than the present

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Fat Intakes in 4 7 Year Children

study, as the preschool children studied were from the Vancouver Coastal Health Authority Region, British Columbia, Canada, an area where seafood is readily available. It is important to compare the median values of AA and DHA, as it represents the middle of a distribution and maybe more informative. In the present study, the median and mean intake of AA are similar, however, the median DHA intake is approximately half of the mean intake for DHA. Our study found and reported that in this sample of children, that the low income group (n 10) had significantly higher intakes of AA than the higher income group (n 60) that is likely attributed to a higher egg consumption. Future studies should investigate the dietary AA and DHA intakes in low and higher income groups using a larger sample size from varying income groups. The wide range of reported AA and DHA nutrient intake (over the 3 days and per day basis) is evidence of inter individual and intra individual variation in these children. Since dietary sources of DHA (e.g. fatty fish and seafood) are not consumed on a daily basis, it was expected that nutrient intakes would vary widely day to day which was confirmed by both the food record and food frequency questionnaire. There is cause for concern that seasonal intakes of fish and seafood (rich sources of DHA) could affect the reported DHA intake. Statistical analysis of food records assessed between May 2001-June 2002 (over the various seasons) showed that DHA intake was not influenced by the seasonal intake of fish and seafood (P NS) between subjects. In this study, the main source of DHA was canned fish, therefore it is not surprising that there is no seasonal variability in intake of this food. Canadas Food Guide recommends that Canadians, including young children eat at least two servings (75 grams each) of fish a week [53]. Certain types of fish (e.g. fresh/frozen tuna, shark, swordfish, marlin, orange roughy and escolar) should be consumed less often as there are concerns as these fish accumulate mercury in their muscles and therefore consumption should be limited to 75 g per month for children 1 4 years of age and 125 g per month and for children 511 years of age [54]. In the current study, the food frequency questionnaire estimated that children consumed on average 324 g of fish per month, and therefore only met 50% of the recommended intake of fish per month suggested by Health Canada. Due to the scarcity of information in the current literature regarding AA and DHA intakes in healthy children, it is uncertain whether these children consume AI of AA and DHA as AI do not currently exist for these fatty acids. Many studies have been conducted in the past on the AA and DHA intakes in infants, therefore the current dietary intakes in these children were compared to intakes observed and reported in the infant literature. The researchers recognize that infants may have a higher requirement of AA and DHA due to their rapid growth and development and that although brain and eye development continues throughout childhood, it is slower in comparison to that of the infant. Even so, a comparison of AA and DHA between infants and children are important as it shows the dietary differences that exist between these two developing groups. In the current study, AA and DHA intake is lower than the intake of AA and DHA in infants consuming mothers milk or formulas containing AA and DHA [40]. From infant to young childhood, results show a dramatic decrease in AA and DHA intakes. To estimate AA and DHA (mg/(kg body weight*day)) intake in infants, the following assumptions can be made: energy requirement of the infant is 100 120 kcal/ (kg body weight*day), human milk provides 55% of calories from fat. Using the AA and DHA levels in human milk reported earlier [40], infant intake level of AA range from 33 40 mg/(kg body weight*day) and DHA intake range from 18.322 mg/(kg body weight*day). Using the AA and DHA levels provided in infant formula (for example: Enfamil A (Canadian brand) or Enfamil Lipil with Iron (US brand) (Mead-Johnson Nutritionals, Evansville, IN)), with assumed energy requirements of an infant is 100 120 kcal/kg, infant intake level of AA ranged from 34 41.2 mg/(kg body weight*day) and DHA level ranged from 17.120.6 mg/(kg body weight*day). In the present study, if the assumed weight of the child, 4 7 years of age is 25 kg, mean AA and DHA intake observed provides approximately 2.3 mg/(kg body weight*day) of AA and 1.5 mg/(kg body weight*day) of DHA respectively. The intakes in the present study are lower than the AA and DHA intake observed in Innis et al [31], where children 35 years of age (assumed weight of a child is 16 kg), mean AA and DHA intake was 14 mg/(kg body weight*day) and 6.0 mg/(kg body weight*day) respectively. The current level of AA and DHA provided by the diet of the children in the present study is approximately 1/15 the amount of AA and 1/12 the amount of DHA that is available to infants fed human milk or formulas containing AA and DHA. An increased intake of AA and DHA may be necessary to support retinal and neuronal cellular development since the brain and the eye continue to develop throughout childhood. The low dietary intake of LA, ALA, AA and DHA may be insufficient to support the needs of the child and requires further investigation. This study may be limited in its methodology, specifically sample size and method of dietary assessment. Due to the paucity of literature on LCP intakes in children that currently exists, there was no basis to estimate the N required to access AA and DHA intake. Although sample size maybe considered small, the information collected from this research study adds to the limited body of knowledge that currently exists in the literature regarding LCP intakes in healthy children. In this study, dietary intakes were assessed using 3-day, unweighed food record. The number of subjects enrolled in the current study is comparable to the number of subjects enrolled in other studies investigating fat intake using a 3-day food record [55,56]. Use of a 7-day weighted food record is considered a more precise method available for estimating usual food and/or nutrient intakes of individuals, so long as respondents do not

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change their usual eating pattern [57]. This method was considered to be too much of a burden to parents participating in the present study and would have resulted in problems with compliance and completion rates. Dierks and Morse [58] estimated nutrient intakes in preschool children using 3-day food records and report a 95% completion rate. The present study had a completion rate of 86%. The food assessment method used in the current study was similar to the method used by Allison et al [28] and Jonnalagadda et al [29] where food intake was collected over a three day period (one 24 hour recall, and 2-day food record). A 24-hour recall method was used by Meyer et al [30] however a single day is not representative of the usual fat consumption of an individual [59,60] and does not estimate AA and DHA intake reliably. Food frequency questionnaires provide an inferior quantitative estimate of intake as this method does not usually define information on a specific food items consumed, exact portion sizes consumed, food preparation methods, brand name and packaging information [28,61,62]. Innis et al [31] used a food frequency questionnaire, and considered this limitation and collected data on specific foods (e.g. frequency food was eaten, portion sizes, brand name or place of purchase, method of preparation, use of fat-reduced foods, types of margarines, shortenings and other fats and oils), therefore, AA and DHA nutrient estimates are more likely to be representative of the preschool children studied. In the current study, a food frequency questionnaire was used to assess intakes of various foods containing DHA. In the current study, the food frequency questionnaire estimated mean DHA intake to be 19 mg/day higher than mean DHA intake estimated using the food record. Results from the food frequency questionnaire showed agreement with food records in that foods which contained higher levels of DHA were consumed in limited quantities over the month. Other studies that use per capita food use data or per capita disappearance data to estimate DHA intake [51,52] overestimates actual consumption as it does not account for spoilage and waste and is not a direct measure of quantity of food actually consumed [63].

ACKNOWLEDGEMENTS
We would like to thank the school administration, teachers, parents and children of the Parkland School Division No. 70, Stony Plain, Alberta, Canada. This work was financially supported by the Natural Sciences and Engineering Research Council of Canada and MTI Meta Tech Inc. Vanessa Lien was supported by a Natural Sciences and Engineering Research Council of Canada Postgraduate Scholarship and the Health Research Studentship from the Alberta Heritage Foundation for Medical Research.

REFERENCES
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CONCLUSION
The diets of these-Canadian children aged 4 7, who do not live near a marine environment have dietary levels of AA and DHA that are low compared to other studies that investigated dietary fat intake in children [28 31]. The current level of AA and DHA provided by the diet of these children is approximately 1/15 the amount of AA and 1/12 the amount of DHA that is available to infants fed human milk or formulas containing AA and DHA. The brain and eye continue to develop throughout childhood, therefore it is conceivable that low dietary intake of AA and DHA may have impact on retinal and neuronal cellular development in children during the early years of life.

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Received October 10, 2006; revision accepted January 12, 2008.

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