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Young vegetarians and omnivores Dietary habits and other health-related aspects Christel Larsson 2001 Department of Food and Nutrition, Umeå University, Sweden Doctoral thesis ISBN 91-7191-983-X Copyright © by Christel Larsson Department of Food and Nutrition Umeå University SE-901 87 Umeå Sweden Printed in Sweden 2001 by Larsson & Co:s Tryckeri AB, Umeå Dedicated to the Department of Food and Nutrition. “The first… … but not the last!” ABSTRACT In the middle of the 1990s many adolescents became vegetarians. There was concern among adults about whether these new young vegetarians got enough energy and nutrients from their dietary intake. The aim of this thesis was to investigate the prevalence of young vegetarians, the food and lifestyle habits, dietary intake and nutritional status of vegetarian and omnivorous adolescents. consumption frequency of fruits/berries, alcoholic beverages, sweets/chocolates and fast foods was seen between vegetarians and omnivores. However, female vegetarians more often than female omnivores consumed dietary supplements. Furthermore, lifestyle characteristics of vegetarians were similar those of omnivores regarding exercise, use of alcohol and smoking habits. The prevalence of adolescents eating a vegetarian school lunch in 124 Swedish secondary schools was investigated by interviewing matrons. Information about prevalence of vegetarians, food and lifestyle habits, of 2041 15-year old students from Umeå, Stockholm and Bergen, was obtained by a questionnaire. The dietary intake and nutritional status of thirty 16-20 year-old vegans were compared with thirty age, sex and height matched omnivores. No significant difference in validity of reported energy expenditure or energy and protein intakes was found between vegans and omnivores. Young vegans (16-20 year-olds) were seen to have a higher calculated intake of vegetables, legumes, and dietary supplements and a lower intake of ice creams, cakes/cookies, and candies/chocolate than omnivores. The dietary intake was below the average requirements of riboflavin for 73% of the vegans, vitamin B12 for all vegans, vitamin D for 43% of the vegans, calcium for 77% of the vegans and selenium for all vegans and 43% of the omnivores. If intake of supplements was included the intake of e.g. calcium and selenium was still lower than the average requirements for 67% and 73% of the vegans respectively. Low iron stores were as prevalent among vegans as among omnivores (20% and 23% with low stores) and three vegans had low vitamin B12 concentrations in blood. Five percent of the adolescents (16-20 years) in Sweden were found to eat vegetarian food at school lunch. In Umeå there was a significantly higher prevalence (15.6%) of 15-year-old vegetarians compared with Stockholm (4.8%) and Bergen (3.8%). It was also found that more females than males (15 years old) chose a vegetarian dietary regime. Even though the female vegetarians consumed vegetables significantly more often than the omnivores, the intake (32 times/month) was not as often as might be expected of a vegetarian population. The male vegetarians reported eating vegetables not even once a day (25 times/month). No difference in the The findings imply that food and lifestyle habits of young vegetarians are different than what previous studies of vegetarians have shown. There is a need for future research of the long-term health effects of being vegetarian. Key words: Adolescent, Biological markers, Dietary intake, Diet history, Doubly labeled water, Energy expenditure, Food habits, Lifestyle, Nutritional status, Questionnaire, Vegan, Vegetarian. CONTENTS LIST OF PUBLICATIONS ............................................................................ 9 ABBREVIATIONS........................................................................................ 10 BACKGROUND ............................................................................................ 11 INTRODUCTION ............................................................................................. 11 DIETARY HABITS OF ADOLESCENTS .............................................................. 11 DEFINITIONS OF VEGETARIANISM ................................................................. 12 PREVALENCE OF VEGETARIANS .................................................................... 13 REASONS FOR VEGETARIANISM..................................................................... 14 HEALTH ASPECTS OF VEGETARIANISM .......................................................... 15 DIETARY ASPECTS OF VEGETARIANISM ......................................................... 16 Nutritional recommendations .................................................................. 16 Energy ...................................................................................................... 18 Protein ...................................................................................................... 18 Vitamins ................................................................................................... 19 Minerals ................................................................................................... 20 DIETARY ASSESSMENT METHODS.................................................................. 22 VALIDITY OF REPORTED ENERGY AND NUTRIENT INTAKE............................. 24 BIOCHEMICAL MARKERS OF IRON STATUS .................................................... 26 AIMS OF THE THESIS................................................................................ 28 SUBJECTS AND METHODS...................................................................... 29 SUBJECTS ...................................................................................................... 29 Definitions................................................................................................ 29 Study samples .......................................................................................... 29 Ethics........................................................................................................ 31 METHODS ...................................................................................................... 31 School Lunch Study................................................................................. 31 Adolescent Questionnaire Study.............................................................. 31 Nutritional Assessment Study.................................................................. 32 Diet history........................................................................................... 32 Validation of protein, sodium and potassium intakes ......................... 34 Validation of energy intake and expenditure ...................................... 34 Physical activity assessment ................................................................ 34 Nutritional assessments in blood......................................................... 35 STATISTICAL METHODS ................................................................................. 36 RESULTS AND DISCUSSION.................................................................... 38 SUBJECTS ...................................................................................................... 38 PREVALENCE OF VEGETARIANS .................................................................... 39 FOOD HABITS ................................................................................................ 43 LIFESTYLE RELATED CHARACTERISTICS ....................................................... 47 VALIDITY OF DIETARY INTAKE AND ENERGY EXPENDITURE ......................... 51 DIETARY INTAKE........................................................................................... 55 BIOCHEMICAL MARKERS OF NUTRITIONAL STATUS ...................................... 63 Iron ........................................................................................................... 63 Vitamin B12 .............................................................................................. 65 Vitamin D................................................................................................. 65 GENERAL DISCUSSION ................................................................................... 66 CONCLUSIONS ............................................................................................ 68 ACKNOWLEDGEMENTS .......................................................................... 70 REFERENCES............................................................................................... 72 PAPER I-V LIST OF PUBLICATIONS This thesis is based on the following papers, which will be referred to by their Roman numerals: I Larsson C, Johansson G. Prevalence of vegetarians in Swedish secondary schools. Scand J Nutr 1997;41;117-20. II Larsson C, Klock K, Nordrehaug Åstrøm A, Haugejorden O, Johansson G. Food habits of young Swedish and Norwegian vegetarians and omnivores Public Health Nutrition 2001;4(5):1005-14. III Larsson C, Klock K, Nordrehaug Åstrøm A, Haugejorden O, Johansson G. Lifestyle related characteristics of adolescent lowmeat-consumers and omnivores in Sweden and Norway (Accepted for publication in J Adolesc Health 2001). IV Larsson C, Westerterp K, Johansson G. Validity of reported energy expenditure and energy and protein intakes of Swedish vegan and omnivore adolescents (Accepted for publication in Am J Clin Nutr 2001). V Larsson C, Johansson G. The dietary intake and nutritional status of young vegans and omnivores in Sweden (Accepted for publication in Am J Clin Nutr 2001). 9 ABBREVIATIONS BMI Body mass index BMR Basal metabolic rate BSE Bovine spongiform encephalopathy DLW Doubly labeled water EEmea Measured energy expenditure EIrep Reported energy intake FIL Food intake level NNR Nordic nutrition recommendation PABA Para-aminobenzoic acid PAL Physical activity level SD Standard deviation TIBC Total iron binding capacity 10 Background BACKGROUND Introduction This thesis focuses on some of the nutritional and health impacts of the dietary regimen and lifestyle of young vegetarians compared with young omnivores. In the middle of the 1990s mass media focused on animal liberation actions, animal transportation, cruelty to animals and young people becoming vegetarians. There was concern among parents, school staff and nutritionally-aware people, that these new young vegetarians did not get enough energy and nutrients from their dietary intake. A public discussion about disadvantages and advantages of becoming a vegetarian at a young age took place and many statements were based on speculation rather than scientific knowledge. To obtain more knowledge about the new and presumably increasing phenomenon of adolescents becoming vegetarians, a study of young omnivores and vegetarians was implemented at the Department of Food and Nutrition, Umeå University. nutrients and irregular meal pattern1, 2, 3, 4, 5. Studies have shown that adolescents have a low and infrequent consumption of vegetables and fruits 2, 6, while milk and bread as well as sweets are consumed daily2, 7. In general, food intake is quite similar between male and female adolescents, but males eat larger amounts of food2, 3. However, the small differences seen point to healthier food intake among females as they eat vegetables more often and consume fewer sweets2. A high-energy intake in combination with low energy expenditure leading to overweight seems to become more prevalent among adolescents in Sweden1. In general, adolescents’ dietary intake of vitamins and minerals are, with some exceptions, adequate for normal health and growth1. Dietary calcium intake has been shown to be high, while the intake of fiber, vitamin D, zinc and selenium and, in females iron, is below the daily recommendations of the Nordic nutrition recommendations 1, 3 (NNR) . Dietary habits of adolescents Snacking and light meals are very common among adolescents and contribute with 25-35% of the daily energy intake1. Questionnaire studies of Swedish 14-15 year-olds Dietary habits of Swedish adolescents are in general characterized by omnivorous food intake, adequate intake of most 11 Background dietary intake, and the most common types of vegetarians are shown in Table 18, 9, 10. found that many adolescents skip breakfast and the free school lunch, while most eat dinner in the evening1, 2, 4. Adolescents who ate no breakfast (8-11%) were found to skip the school lunch more often than those who ate breakfast4, 5. It was also shown that more males than females ate school lunch, otherwise differences of meal patterns with respect to gender were small3, 4, 5. Poor meal habits of adolescents were also found to be associated with other undesirable lifestyle factors, such as smoking and use of alcohol2, 4. Semi-vegetarians are also sometimes called demivegetarians, pesco-vegetarian and near-vegetarians, and some studies also include people in this subgroup who eat poultry or with rare and infrequent meat intake8, 9. Defining a person as a vegetarian, even though the person actually eats some animal products such as fish, poultry or pork, may be controversial, since they are not vegetarians in the strict sense of the word, rather non-red-meat eaters. Other types of vegetarians are fruitarians who are people who only eat those parts of the plant that are cast off or dropped from the plant and that does not involve any destruction of the plant itself8, 9. A macrobiotic diet may be a vegetarian diet, depending on how it is followed8, 9. The followers of Zen macrobiotic diet advance through ten stages of progressive dietary restriction aiming ultimately at a diet composed of cereals only11. Some adolescents choose to eat vegetarian food but few studies have investigated the food intake of vegetarian, especially vegan, adolescents. Definitions of vegetarianism Vegetarianism is the dietary practice of people abstaining from some or all animal products8, 9. The term vegetarian is often used to include people who eat all types of vegetarian diets which, in reality can be highly variable. Today vegetarians are divided into several different subgroups based on their Table 1 Foods eaten by different types of vegetarians. The plus (+) sign indicates that the diet comprises the foods. The minus sign (-) indicates that the diet does not comprise the foods. Type of vegetarians Foods eaten Plant foods Dairy products Eggs Fish Meat Vegans + Lacto-vegetarians + + Lacto-ovo-vegetarians + + + Semi-vegetarians + + + + - 12 Background were vegetarians in 199016. This prevalence is an increase of 23% since the previous Gallup for the Realeat Company Ltd survey in 1988 and a 76% increase since the first survey in 1984. A person may consider him or herself a vegetarian even though the diet has a frequent content of animal products when eating out, although a more appropriate name would be lacto-ovo-fish-steak only when I eat out-vegetarian12. It has been proposed that the criterion for defining a vegetarian diet should more emphasize the presence of vegetables, fruits, grains and other plant foods rather than the absence of animal products13. Further, it has been suggested that people who do not have a daily consumption of fruits, vegetables and whole-grain cereals should not be considered vegetarians, regardless of their consumption of animal products13. In Sweden, as well as in many other countries, few investigations of the prevalence of young vegetarians have been published10, 17. Table 2 shows the results from six different prevalence studies of young vegetarians during 1994-2001. The increase in vegetarianism has been stimulated by media focus on cruelty to animals and more recently bovine spongiform encephalopathy (BSE) also called mad cow disease. It has been reported that women between 1518 years old have the highest tendency to be vegetarian4, 10, 18, and that more adolescent females from high rather than low socioeconomic status areas, have thought about becoming vegetarian10. Prevalence of vegetarians Currently, there is an increasing trend towards vegetarianism in the Western world10. The prevalence of vegetarians among 170 000 British adults was 4.5% in 199514, 15. A British Gallup for the Realeat Company Ltd, based on a sample of 4 162 respondents aged 16 years and over (stratified by region and town) showed that 3.7% 13 Background Table 2 Prevalence of young vegetarians according to different studies during 1994-2001. The prevalence of all vegetarians has been reported by all studies but not always the prevalence of different types of vegetarians. Country Study design Yeara Age nb Sex SVc LVd Ve Allf (years) (%) (%) (%) (%) Australia Questionnaire, 1994 16 536 female 16 21 (south)10 self reported 416 male 6.0 7.0 vegetarianism. England18 Weighed diet 1999 15-18 264 female 10 record. 258 male 1.0 Finland, Questionnaire, (Helsinki)19, g completed by school staff. Sweden4 Questionnaire, self reported vegetarianism. Swedeng, h Telephone interview with school staff. USAi Not presented. a b c d e f g h i 1999 14-16 16-20 8 764 6 731 mixed mixed - - - 4.0 17 1996 15-16 15-16 2 956 3 071 female male 2.0 1.0 2.0 1.0 <1.0 1.0 5.0 3.0 2001 16-20 76 512 mixed - - 0.6 11 2000 17-24 - - - 11 4.0 1 141 female male Year when the study was conducted. Number of people studied. SV = Semi-vegetarians. LV = Lacto-vegetarians. V = Vegans. All types of vegetarians, including: SV, LV, V. Investigated the prevalence of people eating vegetarian food at school lunch. Follow-up study with 104 secondary schools of the same sample as in paper I, plus 19 nearby schools 20. JMA Student Omnibus, January 2000. Survey of students in full time higher education. Reasons for vegetarianism There is most often no single motive for becoming or being a vegetarian. Among the reasons given for being so, are often the following16; Ethical reasons – ethical issues revolving around treatment of animals23. Environmental reasons ecological concern about environment8, 23. Health reasons – to obtain and maintain better health or decrease symptoms or risk of 21, 22, 23 diseases . the Philosophical and religious reasons – Seventh-day Adventists, 14 Background health31, 32. Health studies of groups of vegetarians such as Seventh-day Adventists and Trappist monks show that vegetarianism may not only sustain health, but also improve health33, 34, 35, 36, 37, 38 . These effects may be attributable to diet as well as to other lifestyle characteristics, such as maintaining desirable body weight, regular physical activity and refraining from alcohol, tobacco and drugs23. Hindus, Buddhists, some so called New Age groups, groups of Hare Krishna and Trappist monks emphasize vegetarianism8, 11, 14, 23. Economical reasons – when meat is more expensive than vegetables8, 23 . However, it is rare in Western countries to become vegetarians for economical reasons24. Reasons for eating a vegetarian diet are highly individual and changeable. Health may be the initial motive but ethical issues may become the most important factor for continuing to eat a vegetarian diet or vice versa16. Today, young vegetarians make their choices mainly for ethical reasons, including concern about animal husbandry and animal welfare10, 16, 25, 26. Other reasons why adolescents avoid animal products include fashion, curiosity, rebelliousness, convenience27, liking animals26 and body weight concern28. There may also be the perception that meat is harmful26, 29 and a reaction to food safety ‘scares’ such as the mad cow disease (BSE), Creutzfeldts Jacob’s disease, salmonella, and the use of antibiotics29 and hormones in meat production30. Health aspects of vegetarianism There is a scientific consensus that a higher intake of plant foods and less animal foods promotes 15 Western vegetarians have shown to be leaner than omnivores and have a lower risk of obesity 39, 40, 41. The reduced risk shown among vegetarians of non-insulindependent diabetes may partly be explained by vegetarians being leaner than omnivores23, 24, 39. Blood pressure, hypertensive disorders and other risk factors for coronary heart disease are also reported to be lower among vegetarians, especially vegans, than among the general population42, 43, 44 . Furthermore, it has been shown that a vegetarian regimen reduces mortality from ischaemic heart disease by about 25% 41, 45, 46. Even though it has been concluded that there are no differences in mortality from common cancers (stomach, colorectal, lung, breast and prostate)41, 47, 48 between vegetarians and omnivores, there are studies demonstrating an inverse association between vegetable and fruit intake and many cancers 49, 50. Background Dietary aspects of vegetarianism believe that their diets are somehow guaranteed to be healthy and nutritionally adequate14. Nutrition often comes low on the priority list of adolescents, and poor eating habits sometimes typify this group no matter if they are vegetarians or omnivores. However, previous studies have suggested that adolescents following a vegan diet are at greater risk from nutritional inadequacy than those following a lacto-ovo-vegetarian diet or an The omnivorous diet27, 56. consequence of a more restrictive diet is an increased difficulty in achieving nutritional adequacy, which means that greater care must be taken. Adolescents who consume a wellplanned vegetarian diet can generally meet all their nutritional requirements for growth15, 23. However, those who follow a vegan diet need to make sure that they consume reliable sources of energy, vitamin B12 and vitamin D, if sunlight is limited51. Iron, zinc and calcium intakes may also deserve special attention although intakes are usually adequate when there is a reasonable variety of foods and when energy requirements are met23. Much has been written about adult vegetarianism and nutrition but few studies have examined adolescent vegetarianism10, 51, 52. Nutrient intake during adolescence is important for growth, and longterm health promotion53, 54. Furthermore, since adolescents are becoming more autonomous, behavioral patterns acquired during this period are likely to influence long-term behavior. Nutritional recommendations Well-planned vegetarian diets can effectively meet the Nordic nutrition recommendations (NNR) and can be health-supporting. However, there are distinctions between the various types of vegetarian regimens concerning their impact on nutrient intake, nutritional status, and overall health8. The NNR are, with the exception of zinc, the same for vegetarians as for omnivores. Meat avoidance amongst teenage girls in the UK has, in the mid 1990s, gained in popularity, and there have been concerns about the nutritional adequacy of a meat-free diet in this population group55, 56. Some omnivores are of the opinion that young vegetarians, especially vegans, are generally undernourished whereas some vegans In the NNR a daily recommended nutrient intake (RI) is defined as the average requirements (AR) of a population plus a safety margin of 16 Background is important to obtain enough nutrients to prevent deficiency and thereby adverse symptoms. When evaluating nutritional intake of a group of people, estimated by a dietary assessment method, it is appropriate to use the average requirements (AR) without the safety margin. Having a nutrient intake lower than the AR indicates an increased probability that too little nutrients are being eaten57, 58. Number of people at least two standard deviations (SD) 57, 58 (Figure 1). The lowest recommended intake (LRI) of a given nutrient is defined as the average requirements (AR) of a population minus 2 SD. This is the amount below which there is an increased probability of deficiency symptoms. Even if a person’s daily nutritional intake is lower than the lowest recommended intake (LRI) for a while, deficiency need not follow. However, in the long run it RI = AR + 2 SD AR LRI = AR – 2 SD Requirement of nutrient intake Figure 1 Definitions of the lowest recommended intake (LRI), average requirements (AR) and recommended intake (RI) of a nutrient per day57. 17 Background day60. Vegetarian diets that are not properly planned tend to be characterized by exclusion of animal products without replacement with nutritionally equivalent plant foods. Examples of this are when a person becomes vegan and excludes milk, and water becomes the beverage of choice or when milk based margarines/butter are excluded, and no spread is used. Furthermore, lettuce and cucumber sometimes continue to be the major vegetables instead of, for example, peas, carrots and broccoli. However, in cases where the vegetarian diet has been properly planned it has been concluded that energy intakes are adequate8, 15, 61. It has been suggested that vegetarians have a better understanding of dietary requirements than the general population59. Many vegetarians, especially vegans, are well informed on general nutrition but would like more detailed knowledge, either for their own interest or to answer the questions about their diets that vegans are inevitably asked14. Energy A person’s energy requirements depend on many factors, such as age, sex, body weight and physical activity14. Apart from basal metabolic rate (BMR), physical activity has the greatest impact on energy expenditure, and a correct energy requirement cannot be estimated without taking this into consideration. Vegetarian diets are most often composed of less total fat and protein, but more total complex carbohydrates than omnivorous diets, and consequently vegetarians are likely to consume less total food energy (calories)46. The high carbohydrate content of some vegan diets may also lead to a significant bulk effect with an increased risk of not meeting the energy requirement8. Special attention must be given to meet the energy needs of vegetarian adolescents who consume a lot of low-fat products, and those who limit the number of meals eaten per 18 Protein The traditional concern about vegetarians’ diets has been the adequacy of protein intake, especially of the essential amino acids46. Some authorities suggest a 10% higher protein intake when plant food is the main protein source, since protein from plant sources may be slightly less digestible14. The digestibility of proteins may be affected by the structure of the protein itself, as well as by other factors such as the amount of dietary fiber, tannins and phytates in the food14. The quality of vegetable protein in the diet may be improved by combining food products with different amino acid profiles. For example, cereal grains that are low Background in the essential amino acid lysine but adequate in methionine, can be complemented by legumes having adequate lysine but little methionine. However, it is usually not a problem for vegetarians to obtain an adequate protein intake and it is not necessary, as sometimes believed, to complement amino acids at each meal14, 61. Good plant protein sources for vegetarians are a mixture of grains, legumes, nuts, seeds and soya foods8, 14, 62. Most individuals complement proteins unconsciously e.g. bean-based soups with a slice of bread, combining beans and rice or eating a bean burrito. Vitamins The vitamin intake of vegetarians has been studied by analyzing both food intake and plasma levels and most often vegetarians have an adequate intake of vitamins63, 64, 65. Vitamin B12 is the most likely nutrient to be found lacking in a strict plant-based diet61, since it can only be derived from animal and bacterial sources. Plant foods do not contain the vitamin unless they are contaminated with B12– producing bacteria14. For many years it was thought that fermented soya foods such as soya sauce, tempeh and miso contained some genuine vitamin B12, owing to the presence of certain microorganisms 8, 14 . It was also believed that edible seaweeds e.g. Nori and a blue- 19 green algae called spirulina, which often are contaminated by plankton, contained genuine vitamin B12. However, vitamin B12 analogues (80-94% found in these food products) do not have any bioactivity in the human body, and there is a possibility that their presence may actually block the body’s ability to use genuine vitamin B1214, 23. A better and more reliable dietary source of vitamin B12 for vegans is nutritional yeast8, 14. The yeast produces no genuine vitamin B12 itself (although several inactive analogues), but contains active B12 derived from the B12–enriched molasses medium on which it is grown14. Other reliable sources of B12 are fortified products, such as yeast extracts, oat beverages, some soya beverages, margarines and lately mineral water in Sweden. Intestinal bacteria in humans produce active B12, but mostly in the colon where it cannot be absorbed, rather than in the small intestine where it would have had a chance of being absorbed14. Furthermore, to be absorbed effectively from food, vitamin B12 has to be linked to a so-called intrinsic factor. The most common cause of vitamin B12 deficiency in the general and older population is lack of this intrinsic factor, rather than an inadequate dietary intake14, 23. Vitamin B12 is stored in the liver, which normally contains sufficient amounts (2-5 mg) for a period of three to six years, even in Background the total absence of a food source14. For this reason, although official recommendations are expressed as a daily amount, it is not actually necessary to consume the vitamin every day. However, a long lasting vegan diet ought to be complemented with either vitamin B12 supplement or vitamin B12 fortified foods to ensure adequate intake8, 14. Plant foods, except plankton66 and some mushrooms67, 68, 69 e.g. chantarellus tubaeformis and shiitake, do not contain vitamin D, but some soya drinks, margarines and breakfast cereals acceptable for vegans are fortified with vitamin D2 (ergocalciferol, obtained from yeast and other fungi)14. Vitamin supplements, which contain vitamin D2, are also suitable for vegans. However, food sources of vitamin D may be of minor significance, since vitamin D is produced from the action of ultraviolet B light on sterols in the skin. A cloudy summer day will be enough to stimulate formation of vitamin D in the skin and most people need little or no extra vitamin D from food14. However, the effective light wavelength is probably not present in winter sunlight between October and March in northern Sweden. Thus winter time supplies of vitamin D depend on dietary sources or on the previous summer’s exposure creating adequate stores in the liver. Even though the vitamin D needs may be achieved by exposure to ultra-violet light, people (especially children) living in northern countries are recommended to obtain vitamin D from the diet or supplements during winter time to prevent deficiency58. 20 Minerals Several studies have found mineral intake to be adequate in vegetarians, with the possible exceptions of iron and zinc46. Nonheme-iron is the type of iron found in a strict vegetarian diet while an omnivorous diet contain both nonheme-iron and heme-iron. Only 1-8% of the nonheme-iron in plant foods, while up to 22% of the heme-iron in meat is absorbed14. A low intake of iron in the diet is partly compensated for by increased absorption rate, but in chronic dietary deficiency the body stores run down resulting in iron deficiency. Approximately 20% of the world population have iron deficiency70, 71 and low iron status is also relatively widespread in Sweden, especially among female adolescents. Symptoms of iron deficiency anemia include e.g. tiredness, headache and poor concentration. A study of 179 vegetarians showed that 19% were classified as having iron deficiency72. Another study of 50 vegetarian women (18-45 years) and 24 age-matched omnivores showed that a similar proportion (18%) of vegetarians and Background other organic acids such as malic acid in e.g. plums and apples and citric acid in citrus fruits8, 14. Zinc absorption may be promoted by protein from fish and eggs if those foods are included in the vegetarian diet, or by fermented food76. omnivores (13%) had serum ferritin concentrations <12µg/L and no significant difference in hemoglobin concentrations73. Zinc is important for protein metabolism, cell growth and repair, and immune function74. Because of low bioavailability the NNR gives a 25-30% higher recommendation of zinc for vegans58. Good plant sources include grains, nuts and legumes23. A suboptimal serum zinc level has been shown among 24% of 79 lacto-ovo-vegetarians and 18% of 29 omnivores (14-19 years)75. Vegetarian diets, especially vegan diets, most often contain less calcium than omnivorous diets78. It has been suggested that vegetarians are likely to need less calcium than omnivores since the vegetarian diet is moderate in protein as well as in sodium (salt)79. High protein intake has been shown to lower the acidity (pH) of the urine with an increased urinary excretion of calcium as a consequence80. On the other hand, a recent study showed no association between the intake of protein and calcium 81 absorption . Vegetarians consuming a diet low in calcium have a more efficient absorption of calcium compared with people with adequate intake78. It is not known, however, if this improved absorption compensates fully for a low calcium intake, or to what extent a lower protein, and sodium intake of vegans reduce their calcium requirements. Thus, it is important for those who avoid dairy products to ensure an adequate calcium intake from the diet, by eating enough calcium rich plant foods, using calcium-fortified foods or supplements. Furthermore, it is important not to Although iron and zinc are found in a fairly large number of plant foods, their bioavailability is questionable due to interference of antagonists8. Dietary strategies to enhance the content and bioavailability of iron and zinc in vegetarian diets include decreasing intake of antagonists as well as increasing intake of promoters14, 72, 76, 77 . Examples of an antagonists are tannins from e.g. tea which reduces the absorption of nonheme-iron, as well as phytates found in nuts, cereals, seeds and legumes which reduce the absorption of both iron and zinc. Phytate in plant foods can be decreased by yeast fermentation of whole-wheat flours when baking bread. Examples of promoters, which can increase absorption of nonheme-iron are vitamin C and 21 Background Food records aim to collect data at the time of eating during one or several days. The subject describes every food item eaten by taking notes, and portion sizes are either estimated by piece and household measures, or weighed before being eaten. A most significant problem is the tendency of subjects to eat less the more the days of continuous diet recording go on, indicating a tendency to change dietary habits, more likely to ones that are healthier than usual. This is one of the reasons why the method might not be suitable to use in a comparison study of the nutritional aspects of a vegetarian and omnivorous diet. The method may also be difficult to use on e.g. adolescents who eat a lot of food outside home and lack time and motivation to make a careful food record. It may also be difficult to persuade adolescents to participate in a food record exercise since several days of recording are needed, up to twenty days for some nutrients85. The necessity to repeat the number of measured days is a problem for food records when good estimates of individual nutrient intakes are to be obtained. This is a serious impediment when investigating adolescents. have a too high intake of protein and sodium79, 82. The selenium content of foods varies widely and in plants it is largely dependent on the amount of selenium in the soil78. In Sweden, selenium intakes tend to be low as the soil contains little selenium and locally grown crops therefore are poor sources83. In a Swedish study, a vegan diet contained very low amounts of selenium compared with a lacto-vegetarian and an omnivorous diet, mainly because mostly locally grown rather than imported products were eaten64. In contrast, in another Swedish study, a lacto-vegetarian diet contained rather high values of selenium compared with that of an omnivorous diet. This may be explained by the fact that the vegetarians ate imported cereals, legumes and nuts which seemed to be the major sources of selenium63. Dietary assessment methods The most common dietary assessment methods in nutritional surveys are food records, 24h recalls, diet histories and food frequency questionnaires84. Each method has its own advantages and disadvantages. Which one to use depends upon e.g. the aim of the study, availability of time, money and the number and properties of the subjects. 24h recalls aim to give information about the dietary intake during the preceding 24 22 Background demand and time of participation will be less with diet histories compared with both 24h recalls and food records, since several days of food record or 24h recalls are needed to obtain a valid dietary intake. Therefore diet histories may be more suitable to use on e.g. adolescents. Furthermore, the method is retrospective and will not change the dietary habits of the subjects. The method may also be suitable for vegetarians who eat uncommon food products and dishes since they will have the opportunity to describe the dietary habits in detail. hours. The subject is interviewed about what type and amount of food items have been eaten and portion sizes are estimated in piece, household measures or with pictures of different portion sizes. The major advantage of 24h recalls is that it is quick and simple for both interviewers and subjects. However, to obtain information about day-to-day variation in nutrient intake several 24h recalls and skilled interviewers are needed. The method is dependent on accurate memory, food description and estimates of portion sizes. Unfortunately, this is the method that yields most underreporting86. The method may not be suitable for e.g. young people who have an active life and are difficult to persuade to participate in an interview at short notice, especially when the interviews have to be repeated several times. Diet histories, introduced by Burke et al87, aim to give information about habitual intake during a period of time e.g. one month or six months. Most often it involves a questionnaire, an interview and sometimes a food record. The information collected includes usual meal pattern, frequency of consumption and estimated portion size. The method requires skilled interviewers and is more demanding for the subjects compared with a 24h recall. However, in the end the total 23 Food frequency questionnaires aim to estimate the frequency of consumption of different food items during a period of time e.g. three months or one year84, 88. The questionnaire may be selfadministered or used in an interview and is sometimes focused on specific groups of food and sometimes the total food intake. However, the total food intake is almost impossible to cover. The questionnaire may be semiquantitative by including questions about portion sizes with fixed definitions of, for instance, small, medium, or large portions84. The major advantages of a food frequency method are that it could be low in cost, quick and representative84. The method may be suitable for adolescents since it is simple and quick, which increases the possibility of Background assessment methods there is a need to validate each method used. The validation may be either relative with another dietary method89 or absolute with biological markers such as doubly labeled water (DLW) and nitrogen excretion in 24-h urine collections 90, 91, 92. obtaining a high participation rate. However, the most significant problems with questionnaires are the uncertain validity, the difficulty in developing a relevant and representative questionnaire and problems of obtaining the total food intake. This is especially a problem when the same questionnaire is completed by subjects with different types of diets e.g. omnivores and vegetarians. Validity of reported energy and nutrient intake The doubly labeled water (DLW) method, is used, for instance, to make an accurate assessment of energy expenditure of a free-living person93, 94, 95 and to validate reported energy intake. The principle of the DLW method is that deuterium equilibrates with body water, and oxygen-18 with both water and the bicarbonate pool of the body96. The measured energy expenditure (EEmea) is used to validate reported energy intake (EIrep) and the EIrep/EEmea ratio should equal 1.0 if there is a true estimate of reported energy intake of a person in weight stability. The energy expenditure divided by BMR gives a physical activity level (PAL) and the energy intake divided by BMR gives a food intake level (FIL)93, 97. Both PAL and FIL are multiples of BMR and ought to be >1.2 to be consistent with long-term survival57. The DLW method can be used on anyone and is suitable for studying busy people in a free-living situation, e.g. adolescents who may be difficult to persuade to The dietary methods described are dependent on a food composition database to convert foods into nutrient content. Both the dietary methods and the food composition database are developed to assess an omnivorous diet. When assessing vegetarian diets it is important to know about commonly used vegetarian food products and to include these in the interview and/or questionnaire. Problems often arise when vegetarian food products are not included in the food composition database. If possible the recipes or information about nutrition content in the vegetarian products ought to be entered in the database. Valid information about dietary intake may be difficult to obtain, especially from young people, where the adolescent has irregular meal habits and is not involved in the preparation of the food. Since it is difficult to obtain valid information with dietary 24 Background participate in a study if it is too demanding and time consuming. urine during 24 hours and obtain a complete collection. Estimating the validity of reported nitrogen (protein), sodium and potassium intake involves one or several 24-h urine collections i.e. collection of the total amount of urine excreted during 24 hours98, 99. The amount of nitrogen, sodium and potassium excreted in 24-h urine is analysed and used as biological markers for nitrogen (protein), sodium and potassium intake, respectively. Bingham et al92, showed that 81% of nitrogen intake is excreted in urine, thus excreted nitrogen divided by a factor 0.81 should be equal to nitrogen intake. Sodium excretion in urine and potassium excretion in urine and feces are the biological markers for sodium and potassium intakes, respectively100, 101, 102. Johansson et al100 showed that 77% and 73% of the potassium intake from an omnivorous and a lactovegetarian diet respectively, was excreted in urine. One major problem in validating the intakes of protein, sodium and potassium is the inconvenience for subjects to collect, often several, 24h urine samples. This may lead to the situation where only wellmotivated volunteers take part in validated studies and thus, that people that are not well-motivated will not be studied or take part in studies with low compliance. For instance, it may be difficult to persuade adolescents to collect The completeness of a 24-h urine collection is estimated by analyzing the content of paraaminobenzoic acid (PABA) in urine. The principle of the method is that a subject ingests three 80 mg tablets of PABA, one in the morning, one at midday and one in the evening during the day of urine collection, and a similar amount (240 mg) of PABA ought to be found in the 24-h urine collection if it is complete. Bingham et al92 showed that 93 ± 4% of the PABA was excreted in urine and ±2 SD from the mean excretion resulted in the interval 85-101% of PABA recovery. A 24-h urine collection with a PABA recovery between 85-101% may be classified as a complete 24-h urine collection. Unfortunately, it has been shown that complete 24-h urine collections are very difficult to obtain90, 103. Therefore, to make use of incomplete 24-h collections a linear regression compensation method for collections with PABA recoveries between 50-85% has been developed103. This method is indispensable when studying people, e.g. adolescents, who find it difficult to make complete 24-h urine collections. 25 Background Biochemical markers of iron status The four to five grams of elemental iron present in a normal adult can be divided into three compartments; functional iron compartment (74%) with the essential metabolically active iron, in iron-requiring proteins and enzymes e.g. hemoglobin and myoglobin104. Storage iron compartment (25%) with ferritin as the major storage protein, and the liver and bone marrow as the greatest storage tissues of the human body. The link between the storage and functional iron compartments is the smaller transport iron compartment (<1%), consisting of transferrin. The transferrin iron is delivered to tissues via transferrin receptors expressed on virtually all cells of the body. S-ferritin is a more reliable quantitative laboratory measure of iron status compared with 77 hemoglobin . The main drawback of serum ferritin is the false elevation that occurs with chronic inflammation or infection and therefore the level of different immune cells are also usually estimated in the same blood sample. The function of ferritin in serum is unknown but in most individuals the concentration of Sferritin parallels the total amount of storage iron, and it is the only iron status index that can reflect a deficient, normal or excessive iron status105. S-ferritin levels fall early during iron deficiency, before the characteristic changes in hemoglobin, S-iron and total iron binding capacity (TIBC). Hemoglobin is the oxygen carrying pigment of red blood cells105. Each hemoglobin molecule is a conjugate of a protein (globin) and four molecules of heme, which contains iron. Measurement of the hemoglobin concentration is the most widely used screening test for iron deficiency. However, the hemoglobin concentration in serum is a crude, non-specific index of iron status77 and the hemoglobin concentration falls only in the later stage of iron deficiency (Figure 2)105. S-iron is a measure of the number of atoms of iron bound to the transport protein transferrin105. Each molecule of transferrin can be bound to one or two atoms of iron. Iron deficiency results in a fall in S-iron levels. TIBC is related to the total number of free iron binding sites on the transport protein transferrin105. Iron deficiency is associated with a rise in TIBC from the increase in the number of free iron-binding sites. 26 Background S-iron, TIBC and transferrin saturation are particularly useful for differentiating between nutritional deficiencies of iron and iron deficits arising from chronic infections, inflammation, or chronic neoplastic diseases105. Transferrin saturation is calculated by dividing the S-iron content with TIBC and multiplying by 100 to give a percentage value105. Transferrin saturation is a measure of the iron supply to the erythoid bone marrow. Since Siron decreases and TIBC increases in a situation of iron deficiency, the transferrin saturation decrease. Replete iron stores Depletion of storage iron Depletion of functional iron Transport iron Hemoglobin S-ferritin S-iron Transferrin saturation TIBC Figure 2 Changes in hemoglobin, serum ferritin, serum iron, transferrin saturation and total iron binding capacity (TIBC) at different levels of iron status. Iron status is ranging from replete iron stores, depletion of storage iron, and depletion of functional iron. Revised drawing from Suominen et al, 1998106. 27 Aims of the thesis AIMS OF THE THESIS The general aim of the present thesis was to describe the prevalence of vegetarians, the food habits, the dietary intake and the nutritional status of young vegetarians and omnivores, and health-related lifestyle characteristics. The following objectives were elaborated: To investigate the prevalence of students (16-20 year olds) choosing to eat various types of vegetarian school lunches in Swedish secondary schools (School Lunch Study, paper I). To compare food habits between different types of vegetarians and omnivores among 15-year-old adolescents in Sweden and Norway, and to determine the prevalence of vegetarians (Adolescent Questionnaire Study, paper II). To describe lifestyle related characteristics of 15-year-old low-meatconsumers and omnivores living in Sweden and Norway (Adolescent Questionnaire Study, paper III). To validate a physical activity interview and a diet history interview for estimating energy expenditure, and energy and protein intakes in young Swedish vegans and omnivores, 16-20 years old. The validity was estimated by comparison with measured energy expenditure and biological marker for nitrogen intake (Nutritional Assessment Study, paper IV). To assess the dietary intake and nutritional status of a group of Swedish vegans in the 16 to 20 year old age range living in Umeå, using a validated diet history method and blood samples. Furthermore, and to compare the results with sex-, age- and height-matched omnivores (Nutritional Assessment Study, paper V). 28 Subjects and methods SUBJECTS AND METHODS Subjects Definitions In this thesis adolescents were categorized into vegans, lacto(ovo)-vegetarians (including lactovegetarians and lacto-ovovegetarians), semi-vegetarians (lacto-ovo-fish-vegetarians) and omnivores depending on their dietary intake. The category ‘semivegetarians’, includes adolescents eating sausage, poultry and pork never or seldom during the last 12 months. The term ‘vegetarian’ or ‘low-meat-consumer’ comprises; vegans, lacto-(ovo)-vegetarians and semi-vegetarians. All other subjects were defined as omnivores and most ate meat, fish, eggs and dairy products on a daily basis. particular interest. In addition, 116 schools were randomly selected from the whole country for inclusion in the study. Of the 116 randomly selected schools eight had to be replaced since no information on the students’ dietary intake at lunch was available. The eight new schools were randomly selected using the same procedure as for the other schools. The participants in the Adolescent Questionnaire Study (II, III) were recruited during autumn 1996 from Umeå and Stockholm in Sweden and Bergen in Norway (Figure 4). The Swedish subjects were recruited by a random sample of ninth-grade classes (15-16 year olds) in municipal senior secondary schools. Classes with more than 15 students were selected as the sampling unit, rather than individuals, since the school setting simplified data collection, and improved the possibility of a high response rate. Students from 22 classes in nine out of the ten schools in Umeå, and 22 classes in 17 out of the 50 schools in Stockholm were included. All classes selected in Sweden and all students present on the day of the survey were willing to participate in the study. In Bergen a random Study samples In the School Lunch Study (I) the study sample was taken from the total number of upper secondary school students in Sweden, which was 312 375 students in 19951996. The total sample was 75 210 adolescents in the 16-20 year old age group eating school lunch in 124 Swedish secondary schools (Figure 3). All eight municipal upper secondary schools from Umeå, Skellefteå, Sölvesborg and Vänersborg were selected (Figures 3-4), since these towns were highly influenced by the “Straight Edge” philosophy which, because of its emphasis on vegetarianism, was of 29 Subjects and methods All (8) municipal schools from Umeå, Skellefteå, Sölvesborg and Vänersborg (7 840 students) All Swedish upper secondary schools (312 375 students) Eight schools randomly selected as replacement Eight schools with no information was excluded 116 randomly selected schools (67 370 students) Sample of 124 schools (75 210 students) Figure 3 Sample procedures in the School Lunch Study, 1996. sample of 11 schools was drawn from a total of 24 schools in the municipality. Three schools refused to participate and were replaced by a random selection of three new schools from the remaining 13. In total 2 041 students participated, 578 from Umeå (response rate 95%), 504 from Stockholm (response rate 91%) and 959 from Bergen (response rate 83%). Umeå had 4 212 students in upper secondary schools and the adolescents were recruited by advertisements and visits to schools. The subjects had to be in good health with no chronic diseases and aged between 16 and 20 years. Thirty vegans were recruited and thereafter 30 omnivores were matched by sex, age (± 6 months) and height (± 3.5 cm). Of the 60 subjects participating in the Nutritional Assessment Study (V) 32 subjects, selected at the end of year 1997, conducted an energy expenditure measurement with doubly labeled water (IV). In the Nutritional Assessment Study (IV, V) the study sample was taken from the municipality of Umeå. At the time of the selection in 1997-1998, the municipality of 30 Subjects and methods Skellefteå (I) (I) Umeå (I-V) Bergen (II, III) Stockholm (II, III) Vänersborg (I) Sölvesborg (I) Figure 4 Map of Nordic countries, indicating the study areas of papers I-V. Ethics The study was approved by the Research Ethics Committee of the Medical Faculty, Umeå University. All participants (matrons and not students in paper I) received verbal as well as written information on the purpose and the content of the study and the participation was voluntary. In the Adolescent Questionnaire Study (II, III) permission was obtained from the school authorities and in Sweden the principal of each school and teacher of each class gave their consent to the study. Each participant in the Nutritional Assessment Study (IV, V) gave written informed consent. Methods School Lunch Study In the School Lunch Study (I) a telephone interview with the dining hall matron of each school was performed during March and April in 1996. The matrons were asked to give information about how many students were eating vegan, lacto-ovo-vegetarian or lacto-ovofish-vegetarian food at school lunch. Adolescent Questionnaire Study In the Adolescent Questionnaire Study (II, III), a questionnaire was used to obtain information about food and lifestyle related habits. Each student was asked to complete a questionnaire in a 31 Subjects and methods classroom setting. A pilot-study was performed in 1996 with 44 ninth-grade students from a village close to Umeå and 30 ninth-grade students in Bergen. The Swedish questionnaire had 207 questions and the Norwegian questionnaire 165 questions, and there were 106 identical questions. The questionnaires were divided into eight sections concerning: diet for the 12 months preceding the survey, and the questionnaire asked diet-related questions pertaining to that period of time. Nutritional Assessment Study Diet history In the Nutritional Assessment Study (IV, V) a diet history interview was used to obtain information about typical eating patterns for the preceding three months (Figure 5). The structure of the diet history was elaborated in two pilot interviews before being conducted between August 1997 and June 1998. Twenty-eight of the subjects, 14 vegans and 14 omnivores participated in the diet history during the autumn semester and 32 subjects (IV), 16 vegans and 16 omnivores during the spring semester to obtain seasonal variations in dietary intake. The reported food, beverage and dietary supplement intakes were entered into the dietary analysis program “Stor MATs 4_03e”, which uses the Swedish Food composition data base (version 2_97). For composite foods and dietary supplements that were reported but not listed in the food data base, recipes and information on nutritional content were entered. A nutrient intake below the average requirements (AR), according to the NNR58 (Figure 1), was defined as a nutrient intake indicating an increased risk of deficiency in the present thesis. • Background information and family structure (14 Swedish, 12 Norwegian questions). • Food habits (84 Swedish, 71 Norwegian questions). • Dental health (9 questions). • Smoking and drinking habits (8 Swedish, 11 Norwegian questions). • Physical activity (21 Swedish, 2 Norwegian questions). • Attitudes to school (15 Swedish, 10 Norwegian questions). • Relation to friends and family (19 Swedish, 13 Norwegian questions). • Attitudes to diet and health (37 questions). Answers to the food frequency part of the questionnaire were used to classify subjects as either vegetarians/low-meat-consumers or omnivores, according to the previously mentioned definitions. Subjects were considered vegetarian/low-meat-consumers if they had followed a vegetarian like 32 PHYSICAL ACTIVITY ASSESSMENT (IV) • 32 subjects. • One interview per subject. • Estimated physical activity during 14 days. 33 BLOOD SAMPLE • 60 subjects. • One sample for vitamin D in Nov-Dec 1998. VALIDATION OF ENERGY (IV, V) • 32 subjects got DLW. • Seven urine samples per subject. • Energy expenditure measurement during 14 days. Figure 5 Study design of the Nutritional Assessment Study (IV, V), Umeå 1997-1998. NUTRITIONAL ASSESSMENT (V) • 60 subjects. • Three fasting blood samples per subject. • Assessment of iron, vitamin B12 and folate. DIET HISTORY (IV, V) • 60 subjects (1st in August 1997). • Two 1-2h interviews per subject. • Food intake of the preceding three months. VALIDATION OF PROTEIN, SODIUM AND POTASSIUM (IV, V) • 60 subjects. • Four 24-h urine collection per subject. Subjects and methods Subjects and methods Validation of energy intake and expenditure After the diet history, validation of protein, sodium and potassium, and nutritional assessment were completed, total energy expenditure of 32 subjects (IV) was measured during 14 days by the DLW method (Figure 5)107. Each subject collected three baseline urine samples before ingesting an oral dose of 0.12 g 2H and 0.25 g 18O per kg estimated body water96, 108. Single spot urine samples were taken 24 h after the dose was administrated and thereafter on days 4, 8 and 14. Carbon dioxide production rate was calculated using a multipoint method96 and the respiratory quotient of the diet was taken as 0.85. Validation of protein, sodium and potassium intakes After the subjects had participated in the diet history interview (IV, V) he/she started the first 24-h urine collection (Figure 5). In total each participant completed four 24-h urine collections and the completeness of the collections was verified using PABA92. Each 24-h urine collection was analysed for PABA, nitrogen, sodium and potassium. Sodium and potassium in urine were determined by flame atomic absorption spectrophotometry (Perkin Elmer model 5000), according to routine clinical laboratory procedures. Nitrogen in urine was determined by the Kjeldahl technique (Tecator 1002, Perstorp Analytical, Bristol) and PABA was measured colorimetrically92. The biological marker for sodium intake was taken as sodium excretion in urine and the biological markers for nitrogen (protein) and potassium intakes were taken as excreted amounts divided by 0.81 for nitrogen, 0.77 for potassium of omnivores and 0.73 for potassium of vegans. A linear regression compensation method103 was used for incomplete 24-h collections with PABA recoveries between 5085%. Urine collections containing 85-110% of PABA were classified as complete and those containing less than 50% of PABA and over 110% of PABA were rejected. 34 Physical activity assessment Physical activity interviews with the 32 subjects were conducted (IV) 14 days after ingestion of the oral dose of DLW (Figure 5). Respondents were questioned about their physical activity during the last two weeks, in a 30- to 60minute interview. The activities were categorized according to six activity grades with a given definition and physical activity ratio109; sleep (1.0), very light activity (1.4), light activity (1.7), moderate activity (3.0), heavy activity (4.0) and very heavy activity (6.0). A mean PAL was then calculated and since PAL is claimed to be independent of age, Subjects and methods weight and sex110 total energy expenditure could be calculated by multiplying PAL by the estimated BMR. iron deficiency anemia if P-iron was <7.2 µmol/L (<40 µg/100mL), TIBC was 73.4 µmol/L (410 µg/100mL), transferrin saturation was <15%, and P-ferritin was <10 µg/L. Nutritional assessments in blood Three fasting venous blood samples were taken one week apart from 60 subjects (V). Hemoglobin, S-ferritin, S-iron, TIBC, transferrin saturation and immune cell concentrations were measured in all three blood samples to classify the iron status as “normal”, “early negative iron balance”, “iron depletion”, “iron-deficient erythropoiesis” or “iron deficiency anemia”, according to definitions by Herbert111. A person was classified as having an early negative iron balance if plasma iron (P-iron) was <21.5 µmol/L (<120 µg/100mL), TIBC was 53.7–64.5 µmol/L (300–360 µg/100mL), transferrin saturation was 30%, and plasma ferritin (Pferritin) was <25 µg/L. A person was classified as having iron depletion if P-iron was <20.6 µmol/L (<115 µg/100mL), TIBC was 64.5 µmol/L (360 µg/100mL), transferrin saturation was 30%, and P-ferritin was 20 µg/L. A subject was classified as having irondeficient erythropoiesis if P-iron was <10.7 µmol/L (<60 µg/100mL), TIBC was 69.8 µmol/L (390 µg/100mL), transferrin saturation was <15%, and P-ferritin was 10 µg/L. A person was classified as having Vitamin B12 and folate was measured in the first of the three blood samples taken. A fourth blood sample of the 60 subjects was taken during November and December 1998 for measurement of 1,25-dihydroxy-D-vitamin levels in serum. An adequate blood concentration of vitamin B12, folate, and 1,25-dihydroxy-Dvitamin was defined as a concentration within the reference interval used by the Clinical Chemistry Department, The University Hospital of Northern Sweden, Umeå, Sweden. For Svitamin B12, the reference interval of 95–568 pmol/L was used; for Sfolate, 3.4–47 nmol/L, and for S1,25-dihydroxy-D-vitamin, 26–156 nmol/L. All blood samples were analyzed according to standard methods used at the Clinical Chemistry Department, The University Hospital of Northern Sweden, Umeå, Sweden. Hemoglobin was measured immediately on a specimen collected into ethylenediaminetetracetic acid (EDTA)containing tubes using cyanmethemoglobin and an absorbance measurement. Iron was 35 Subjects and methods measured by a colorimetric assay, ferritin by an immunoturbidimetric assay, and TIBC by an alkaline buffer/reductant solution from Roche Diagnostics, Indianapolis, IN, USA. Vitamin B12 and folate were analyzed by Quantaphase II B12/Folate Radio Assay from BioRad, Diagnostics Group, Alfred Nobel Drive, Hercules, CA, USA. A radio-isotopic assay and chromatographic procedure from Nichols Institute Diagnostics, USA, was used for the determination of 1,25-dihydroxyD-vitamin levels in serum, San Juan Capistrano, CA, USA. Statistical methods Comparisons between dietary groups and statistical analyses were made by using the Mann-Whitney U-test, Pearson’s chi-square, Fisher’s exact test, the Wilcoxon Signed Ranks Test and 2 factor ANOVA (Table 3). A two-tailed P-value of <0.05 was considered statistically significant. All statistical analyses were performed using SPSS, versions 6.1, 9.0.1 or 10.0 (SPSS, Pty Ltd, Chicago, IL, USA). 36 • … background and validation data… • … the validity of the energy and protein intakes… • … background and validation data, food and nutrient intake and nutritional status… • 2 factor ANOVA… • Wilcoxon Signed Ranks Test… • Mann-Whitney U-test… IV V 37 • … physical, health, family, social habit and exercise data… • … physical, health, family, social habit, exercise, alcohol and tobacco data… • Mann-Whitney U-test… • Pearson’s chi-square or Fisher’s exact test for 2x2 tables… III Table 3 Statistical methods used in paper II-V. Paper Statistical method … for comparisons of… II • Mann-Whitney U-test… • … food consumption frequency… • Pearson’s chi-square or Fisher’s • … attitudes, dietary knowledge and meal habits… exact test for 2x2 tables… Subjects and methods … vegans and omnivores of each gender. … vegans and omnivores of each gender. … between… … dietary groups of each gender and for each of the vegetarian groups, as compared with the omnivores. … low-meat-consumers and omnivores of each gender. Results and discussion RESULTS AND DISCUSSION Subjects compared with 251 girls and 342 boys living in Umeå, mean age 16.8 (range 15.9-18.2 years),112 and concerning energy expenditure compared with 374 Swedish 15year-old adolescents113. The subjects in the Adolescent Questionnaire Study may be regarded as representative since ninth grade is compulsory, and since it was a random sample of classes and schools and the mean response rate was high (90%). Also the internal response rate of each question was high with >90% for a majority of the questions. Classes or schools, rather than individuals, were used as sampling units, which probably reduced precision because of clustering, and it is possible that there were differences in ethical considerations between classes or schools. However, the schools selected were distributed throughout the municipalities and may, therefore, be regarded as representative of senior secondary schools. The samples in the Adolescent Questionnaire Study and the Nutritional Assessment Study are different regarding selection method, number of subjects, what was required of the subjects and amount of information obtained. In the Nutritional Assessment Study the sample is less representative than the sample in the Adolescent Questionnaire Study. On the other hand in the Nutritional Assessment Study more was required of the participants and the information collected is, therefore, much more detailed. The subjects participating in the Nutritional Assessment Study were not random and might therefore not be representative for the general population. However, the subjects were regarded as representative concerning height, weight and body mass index (BMI) when The more detailed information collected and the more required of subjects participating in a study, the higher the drop-out rate is most likely to be, which will lead to a less representative sample (Figure 6). 38 Results and discussion • • • Non-random selection. Small number of subjects. A lot required of the subject i.e. high risk of drop-outs. Less representative sample Paper IV and V More representative sample • • • Random selection. Large number of subjects. A little required of the subject i.e. small risk of drop-outs. Paper I, II and III Figure 6 Factors with impact on how representative a sample will be, relevant to the papers in the present thesis. Prevalence of vegetarians The School Lunch Study (I) investigated how many students aged 16-20 years were eating vegetarian food at school lunch in 1996. Assuming that students eating vegetarian food at school lunch really are vegetarians, the prevalence of vegetarians was 5.0% in Sweden (I) (Table 4). Two percent were eating lacto-ovo-fishvegetarian food, 2.9% lacto-ovovegetarian food and 0.1% vegan food. There was a higher occurrence of students eating vegetarian school lunches in the town of Umeå (11.4%) than in Sweden in general (5.0%). old vegetarians in Umeå was 15.6% in 1996, compared with 4.8% in Stockholm and 3.8% in Bergen (Table 4). In Umeå, lacto(ovo)-vegetarians (8.8%) were the most common type of vegetarians but in Stockholm and Bergen semivegetarians were more common (2.6% and 2.3%, respectively). There were a higher percentage of females among vegetarians in Umeå and Stockholm (P<0.01) but no gender difference was found in Bergen (Figure 7). In the Adolescent Questionnaire Study (II), the prevalence of vegetarians was investigated by classifying people as vegetarians according to self-reported dietary intake. The prevalence of 15-year39 1996 Questionnaire completed by subject, categorized 1996 by reported food intake. Telephone interview with school matrons. 1996 Yeara g f e d c b a 40 Questionnaire completed by subject, categorized 1996 by reported food intake. Year when the study was conducted. Number of people investigated. SV = Semi-vegetarians. LOV = Lacto-vegetarians and lacto-ovo-vegetarians. V = Vegans. All types of vegetarians: including SV, LOV, V. Prevalence of students eating vegetarian school lunches. Norway, Bergen (II) Sweden, Stockholm (II) Questionnaire completed by subject, categorized 1996 by reported food intake. Sweden, Umeå (II) Sweden, Umeå (I) Telephone interview with school matrons. Sweden (I)g g Study design Country, city Table 4 Prevalence of young vegetarians in Sweden and Norway, 1996. 451 501 15.4 253 15.5 15.5 244 263 15.7 15.7 293 3 450 67 370 nb 15.6 16-20 16-20 Age (years) Results and discussion male female male female male female mix mix Sex 2.2 2.4 0.8 4.1 0.8 6.1 2.2 2.0 SVc (%) 1.4 1.3 1.2 2.5 4.6 13 7.1 2.9 LOVd (%) 0 0.2 0.4 0.4 1.1 5.5 2.1 0.1 Ve (%) 3.6 4.0 2.4 7.0 6.5 25 11 5.0 Allf (%) Results and discussion Prevalence (%) of vegetarians 30 25 25 20 Females 15 10 Males 6.5 7.0 5 2.4 4.0 3.6 0 Umeå Stockholm Bergen Figure 7 Prevalence (%) of 15-year-old vegetarian females and males in Umeå (Nfemales=293, Nmales=263) and Stockholm (Nfemales=244, Nmales=253) in Sweden and Bergen (Nfemales=451, Nmales=501) in Norway, 1996. The information obtained in the School Lunch Study concerns only what was eaten for school lunch and thus not what was eaten during the rest of the day. There is a possibility that some of the adolescents were eating vegetarian food at school lunch but omnivorous food for other meals. However, since students in most cases (92%) had to notify the matron that they would like vegetarian food every day of the semester, often by having a certificate from a parent, school nurse or doctor, it is likely that the majority of the students were true vegetarians. The subjects in the Adolescent Questionnaire Study were classified into different dietary groups based on the response to the food frequency questions rather than according to their own perception. This provided a measure of prevalence which prevents misunderstanding of how different types of vegetarians are defined. Worsley et al 1998 concluded that the prevalence of vegetarians became higher when vegetarianism was self-reported than if classified according to food intake10. Sources of errors of different methods used for classifying vegetarians need to be considered. The classification in the Adolescent Questionnaire 41 Results and discussion 2001. However, the prevalence of 11% of students eating a vegetarian school lunch is most likely not as well correlated with the prevalence of true vegetarians as the result in the study of 1996. The reason for this is that it has become more common to serve a free-choice vegetarian food alternative for both vegetarians and omnivores at school lunch. In 1996 only 14% of the schools reported serving vegetarian food for everybody, compared with about half of the schools participating in 2001. The increased serving of free-choice vegetarian food made it much more difficult for the matrons interviewed in 2001 to give a valid number of students eating vegetarian food who were also true vegetarians. Therefore, the results of the follow-up study most likely overestimate the prevalence of vegetarians. However, it can be concluded that more vegetarian food is eaten for lunch in Swedish schools, since 98% of the schools in 2001 serve vegetarian food at school lunch, compared with 88% of the schools in 1996 (I). An increase in the number of schools serving vegan food was also seen from 27% in 1996 (I) to 64% in 2001. Study relied on the response on food intake of a limited number of animal products. The major reason for not including specific questions about vegetarianism in the questionnaire was due to the intent to investigate adolescents in general. This made it inappropriate to ask questions specific to vegetarians. Also the questionnaire was very extensive. Instead of calling the subjects vegetarians it may be more correct to call them low-meat-consumers. However, of the 150 subjects that were defined as low-meat-consumers 65 subjects spontaneously noted at the end of the questionnaire that they were either vegan, lacto-vegetarian or semi-vegetarian. Few studies of the prevalence of vegetarian adolescents in Sweden have been made. To be able to conclude that a change in prevalence has occurred, studies with similar design as well as definitions of vegetarians need to be compared. A journalist, A. Asker, made a follow-up study in 2001, with the same methodology as in the School Lunch Study20. Of the 124 schools participating in 1996, 104 participated in the follow-up study. In addition 19 other schools nearby the remaining 20 schools from 1996 were included in the follow-up study. The results of the follow-up indicated that there was an increase from 5% to 11% of students eating vegetarian meals between 1996 and 42 The School Lunch Study found a similar prevalence of vegetarians (5%), as a Swedish questionnaire study of 15-16 year-olds20. The results of that study showed a prevalence of 5% female Results and discussion vegetarians in Umeå (15.6%) compared with Stockholm (4.8%) and Bergen (3.8%) and a majority were females (72.5%). vegetarians and 3% males vegetarians. Thus, both the Swedish questionnaire study and the Adolescent Questionnaire Study (Figure 7) indicate a relationship between gender and vegetarianism, with young females more often choosing vegetarian food than young males. This agrees with other previous studies, which have reported that young as well as adult females are more likely to be vegetarians than young and adult males4, 10, 18. Food habits The Adolescent Questionnaire Study (II) investigated the food habits among 15-year-old vegetarians and omnivores in Umeå, Stockholm and Bergen. Besides meat and other animal products, female vegetarians less often consumed potatoes, soft drinks, chips/peanuts, cakes/cookies, fried and barbecued food while more often consuming rice/pasta, vegetables, coffee, tea, microwave-heated food and dietary supplements than did the female omnivores (Table 5). However, there was no difference between the female groups in consumption frequency of e.g. fruits/berries, ice cream and sweets/chocolates. Male vegetarians showed no major differences in food intake compared with male omnivores, except for eating animal products, margarine, ice cream, cake/cookies, and fried and barbecued food less often (Table 5). One of the reasons for the high prevalence of vegetarians in Umeå might be the active Straight Edge music groups during the 1990s. For example the Straight Edge philosophy emphasizes vegetarianism and spreads its message via music. Another reason may be that the young vegetarians in Umeå got a lot of publicity in the media and there was an intense public debate about vegetarianism, animal welfare etc. In conclusion, the prevalence of adolescents aged 16-20 years eating a vegetarian school lunch in Sweden was 5% in 1996. There was more than three times higher prevalence of 15-year old 43 Results and discussion 1 2 3 4 5 6 7 * ** *** 4.6 4.3 3.7 5.5 0.7 4.2 18 5.2 35 7.5 11 15 3.4 20 24 2.6 6.1 4.4 6.2 2.1 7.6 9.7 3.2 1.6 6.8 5.1 7.4 13 6.1 15 9.5 7.4 2.1 Vegans (n=22) Female omn. (n=880) Female veg. (n=108) 0*** 0*** 0*** 2.7*** 0.4* 3.2** 16** 3.3*** 28** 5.1*** 14*** 12*** 2.7* 32*** 23 3.8 5.0 4.2 6.6 4.2* 15*** 5.9*** 1.7** 2.0 4.1*** 5.4 5.9** 13 2.4 11*** 4.1*** 11* 6.2* Lacto-(ovo)veg. (n=73) 5.7 5.5 4.4 5.9 1.0 5.4 17 5.8 42 8.3 10 16 5.0 17 17 3.8 11 6.5 7.6 5.1 7.6 16 2.3 2.2 9.9 7.6 10 15 12 17 12 9.6 1.9 Semi-veg. (n=55) 0*** 0*** 0*** 2.5*** 0.4* 3.8** 17 6.4 35* 5.1* 9.5 14 5.0 25 19 3.3 9.7 6.5 6.3* 1.7 11 15 0.7 2.1 8.6 4.9** 7.2* 16 8.7 13* 5.1*** 12 5.5 Omn. total (n=1891) Sausages Pork chops/spare ribs/bacon Poultry Fish Seafood (except fish) Eggs Cheese Soft cheese Milk Sour milk and yogurt Rice/pasta Potatoes (boiled, baked, mashed) Fried potatoes Vegetables (except potatoes) Fruits and berries Porridge Cereals Bread (all kinds)2 Margarine3 Coffee4 Tea4 Soft drinks Diet soft drinks Alcoholic beverages Chips, peanuts Ice cream Cakes, cookies Sweets, chocolates Fast foods Fried food Barbecued food5 Microwave-heated food6 Dietary supplements7 Male omn. (n=976) Food item times/month1 Male veg. (n=41) Table 5 Food consumption frequencies of young vegetarians (veg.) and omnivores (omn.) in Umeå, Stockholm and Bergen, 1996. 5.2 4.9 4.1 5.7 0.8 4.9 18 5.5 39 7.9 11 16 4.3 19 21 3.3 8.5 5.5 7.0 3.7 7.7 13 2.9 1.9 8.4 6.4 8.8 14 9.1 16 11 8.5 2.0 0*** 0*** 0*** 7.2* 1.0 4.3 20 6.1 39 7.5 11 18 3.0*** 25** 23 4.6 7.4 4.4* 5.5** 2.3 11 8.6** 1.3 1.7 4.3*** 4.0** 4.5*** 12 4.4** 13 6.4*** 11 4.4* 0*** 0*** 0*** 0*** 0*** 3.7** 18 3.9* 32** 4.7** 15*** 9.3*** 3.1** 31*** 20 2.5** 6.1 5.0 6.5 4.5 13** 8.5*** 1.6 1.9 5.5*** 6.2 8.2 15 4.3 11*** 3.0*** 12 5.3*** 0*** 0*** 0*** 0*** 0** 0*** 0*** 0*** 0*** 0*** 14* 9.6** 4.7 37*** 26 5.1 3.6* 5.1 7.8 3.1 22** 7.9* 0.9 2.8 7.0 4.9 3.6*** 16 3.1 13 3.6*** 11 12*** Unless otherwise indicated, figures show frequency rate (times/month) of consumption of a given food item. Comparisons between dietary groups were made separately for both gender and for each of the vegetarian groups, as compared with the omnivores, by the Mann-Whitney U-test. The data given present slices of bread per day. Grams per slice of bread. Cups per month. During the summer months. Both ready foods that are bought in the supermarket and home-made food. Number of pills per week. Significantly different from omnivores (P≤0.05). Significantly different from omnivores (P≤0.01). Significantly different from omnivores (P≤0.001). 44 Results and discussion legumes, vegetables, nuts/seeds, juice, and dietary supplements and a lower intake of cereals, ice cream, and candy/chocolate than the male omnivores. Also it was shown that vegans (II) consumed potatoes, cereals, soft drinks, cakes/cookies and barbecued food less often than did omnivores (Table 5). They ate rice/pasta and vegetables, drank tea and took dietary supplements more often than their omnivorous peers. A methodological problem of a food frequency questionnaire (II) is that every single food item cannot be included. Therefore, the result that male vegetarians excluded animal products without making significant changes in the consumption frequency of other food items may be explained by a higher consumption of food items not included in the questionnaire. Our questionnaire illustrates the conflict of interest between getting enough detailed information on the one hand, and not making a too lengthy and tiresome questionnaire on the other. Another methodological problem is that it is usually more difficult to remember what was eaten for subjects with irregular food habits. In addition, the format of the question/answer categories may not be suitable for subjects with irregular food habits. Because the students were asked to complete an extensive questionnaire during a short time, there is a risk that towards the end they were becoming less focused114. Young vegetarians (II) generally wanted to find out more about healthy eating than did the omnivores and they preferred to get the information from school and the media. The stricter the vegetarian diet the stronger was the subject’s wish for more information on healthy eating. The 16-20 year old vegan adolescents in the Nutritional Assessment Study (V) had a very heterogeneous dietary intake. Food groups, such as pizza/pies/pastries, nuts/seeds, cereals, porridge, coffee, tea, and dietary supplements, showed a high SD, especially among female vegans. The female vegans had a higher intake of rice, legumes, vegetables, whole-wheat bread, and dietary supplements, and a lower intake of ice cream, cake/cookies, and candy/chocolate than female omnivores (Table 6). The male vegans had a higher intake of 45 Results and discussion Table 6 Reported daily intake (g/day) of 30 vegans and 30 omnivores in Umeå, Sweden 1997–1998a. FOOD GROUP (g/day) FEMALES MALES Median (25th, 75th percentile) Vegans Omnivores Vegans Omnivores (n=15) (n=15) (n=15) (n=15) Meat, and meat dishes Fish, and fish dishes Seafood, and seafood dishes Eggs (not in food dishes) Milk/sour milk/yogurt/cream Cheese Pizza/pies/pastries Potatoes (not in food dishes) Pasta/macaroni (not in food dishes) Rice (not in food dishes) Legumes Vegetables (not potatoes) Fruit/berries (not in food dishes) Nuts/seeds Cereals Porridge Whole-wheat bread White bread Butter/margarine Coffee Tea Juice Soft drinks/fruit drinks Alcoholic beverages Chips/popcorn Ice cream Cake/cookies Candy/chocolate Dietary supplements a * ** *** 0*** 0*** 0** 0*** 0*** 0*** 12 (5, 25) 53 (24, 96) 83 (62, 96) 46* (21, 83) 255*** (136, 400) 292*** (167, 329) 128 (103, 322) 2 (0, 6) 0 (0, 16) 0 (0, 57) 103* (45, 143) 17 (6, 33) 15 (4, 28) 0 (0, 69) 40 (0, 181) 93 (45, 331) 237 (10, 275) 39 (0, 134) 4 (3, 11) 0*** 7* (2, 33) 10* (6, 34) 1* (0, 7) 98 (78, 114) 14 (11, 25) 1.0 (0, 7) 4 (0, 6) 476 (416, 692) 26 (17, 37) 10 (5, 20) 52 (37, 87) 51 (36, 73) 21 (12, 27) 4 (0, 8) 90 (54, 139) 182 (132, 222) 0 (0, 2) 4 (2, 18) 14 (1, 36) 45 (16, 95) 45 (20, 87) 13 (10, 19) 0 (0, 21) 26 (0, 114) 13 (3, 103) 248 (213, 238) 26 (9, 151) 8 (5, 13) 11 (5, 24) 19 (11, 43) 37 (17, 70) 0 (0, 1) 0*** 0*** 0 0** 0*** 0*** 11 (0, 26) 125 (94, 142) 78 (54, 116) 72 (21, 113) 352*** (219, 401) 320*** (184, 404) 127 (49, 216) 5** (1, 11) 0* (0, 10) 32 (0, 71) 150 (93, 225) 87 (27, 199) 28 (27, 45) 0 (0, 0) 26 (0, 107) 148** (86, 411) 297 (0, 172) 29 (0, 172) 23 (10, 43) 0*** (0,0) 11 (7, 31) 21*** (12, 33) 1* (1, 6) 141 (120, 174) 24 (14, 34) 0 (0, 0) 2 (0, 6) 722 (564, 1176) 25 (16, 52) 11 (6, 31) 103 (60, 130) 63 (43, 100) 34 (23, 58) 11 (0, 16) 51 (19, 88) 72 (51, 136) 0 (0, 1) 25 (0, 53) 29 (7, 78) 51 (2, 266) 63 (18, 124) 19 (11, 43) 0 (0, 43) 7 (0, 69) 39 (0, 96) 284 (190, 508) 107 (5, 327) 7 (2, 43) 6 (3, 21) 20 (9, 45) 50 (35, 71) 0 (0, 1) Data are given as medians (25th percentile, 75th percentile). Statistical comparisons between groups with different diets were done using the Mann–Whitney U–test. Significantly different from the data on same sexed omnivores (P<0.05). Significantly different from the data on same sexed omnivores (P<0.01). Significantly different from the data on same sexed omnivores (P<0.001). Even though the female vegetarians (II) consumed vegetables significantly more often than the omnivores, the intake (32 times/month) was not as often as might be expected of a vegetarian population. The male vegetarians (II) did not even eat vegetables once a day (25 times/month). Thirty-three percent of the vegetarians (II), compared with 53% of 106 American vegetarians aged 12-20 years (81% females), reported eating vegetables more than once a day28. Thirty-one percent of the Swedish vegetarians ate vegetables 6-7 times/week while 27% of the American 46 Results and discussion vegetarians reported eating vegetables once a day. The consumption frequency does not give any information about the amount of vegetables eaten, but the vegans (V), both females and males, showed a higher intake (g/day) of vegetables compared with omnivores. This confirms the findings from Donovan and Gibson’s study of Canadian 14-19 year-old adolescents where lactoovo-vegetarians had a higher intake of vegetables than 29 omnivores27. There was no significant difference in fruit and berry consumption among vegetarians (II) nor vegans (V) compared with omnivores. This was also shown by Donovan and Gibson27 but does not verify the results of the above mentioned American study28, which showed that vegetarians were twice as likely to consume fruits. Lower intake of candy and chocolate (g/day) was seen among vegans compared with omnivores (V). The same was found by the previously mentioned Canadian and American study27, 28. However, there were no statistical differences in consumption frequency of candy and chocolate among vegetarians and omnivores (II). The stricter the vegetarian diet, the more often dietary supplements were used (II). This finding is consistent with the results of another questionnaire study comparing health practices of vegetarians and omnivores115. A higher intake of dietary supplements was also shown among vegans compared with omnivores (V). A possible explanation for these findings is that the stricter vegetarians are more concerned about not getting adequate amounts of nutrients from their diet. Lifestyle related characteristics In conclusion, the food habits of vegetarians found in the present studies (II, V) indicate that the vegetarianism of adolescents in Sweden may be a different phenomenon from the vegetarianism shown by previous studies. This is indicated by the fact that in our studies there was a lower vegetable consumption frequency than expected, and a similar fruit consumption frequency as the omnivores’. In the Adolescent Questionnaire Study (III) lifestyle related characteristics of 2 041 low-meatconsumers and omnivores (15year-olds) from Umeå, Stockholm and Bergen were compared. There were no reported differences in weight or calculated BMI between low-meat-consumers and omnivores of either sex. Nor were there any differences in time spent on physical activity or level of activity (high/moderate/low) (Table 7). The consumption 47 Results and discussion common among low-meatconsumers as omnivores (III) (Table 7). This is similar to what was shown in an American questionnaire study of 321 students aged 12-20 years, which showed that physical activity (>3 times/week) was reported by 55% of the vegetarians and 57% of the omnivores, and alcohol and tobacco use (at least weekly) by 23% and 22% respectively28. In contrast to the results in the present study (III), Slattery et al found from interviews that young American adults, eating meat or poultry less than once a week, reported more physical activity and were less likely to drink alcohol than omnivores116. Dwyer et al also found that vegetarians aged 15-35 years were less likely to use alcohol and tobacco117. The health perception data reported by lowmeat-consumers (III) is in contrast to the results of an American study, which found that a larger number of vegetarians than omnivores strongly believed themselves to be healthy115. These findings indicate that the young low-meatconsumers in the present study have a different lifestyle compared to findings from previous studies of vegetarians, and that adherence to a low-meat diet may not correlate with other health promotion practices among adolescents in Sweden and Norway. frequency of alcoholic beverages was similar among vegetarians (II) (Table 5 and 7) and vegans (V) (Table 6) compared with omnivores of both genders. However, the vegan males had a lower intake (g/day) of alcohol compared with male omnivores. There was no difference in frequency of smoking between low-meat-consumers (III) and omnivores but more female lowmeat-consumers used smokeless tobacco (Table 7). Female low-meat-consumers reported (III) more sick-days during the previous year, attached less importance to “being healthy“, and believed they had the ability to affect their own health to a greater extent than female omnivores did (Table 8). Female low-meatconsumers also reported more often that they had been depressed during the last week than did female omnivores. Male low-meatconsumers reported more often than male omnivores that they had been tired without reason, often had headaches, and had been depressed during the last week. There was no difference in frequency of spending time with an organization/club between lowmeat-consumers and omnivores. Physical activity, alcoholic beverage consumption and smoking were reported to be as 48 Results and discussion Table 7 Exercise data and alcohol and tobacco use of 15-year-old low-meat-consumers and omnivores in Umeå, Stockholm and Bergen in 1996. EXERCISE1 Female Female Male Male low-meatOmnivores low-meat- Omnivores consumers consumers (n = 108) (n = 880) (n = 41) (n = 976) Physical activity (hours/week)2 2.9 ± 2.7 3.2 ± 3.0 4.8 ± 3.4 4.3 ± 3.5 Physical activity3 High (≥4h/week) 34 35 62 49 Moderate (2-3h/week) 26 28 18 25 40 37 20 26 Low (≤1h/week) Alcoholic beverage3 Often (≥4/month) 20 15 15 18 Regularly (1-3/month) 28 33 23 26 Seldom (<1/month) 52 52 62 56 Smoking3 Daily 21 17 10 15 Sometimes 18 17 24 14 Never 61 66 66 71 Smokeless tobacco3 Daily 0* 0 12 5 Sometimes 10 2 17 14 Never 90 98 71 81 1 Comparisons between low-meat-consumers and omnivores were made separately for both genders using Pearson’s chi-square and the Mann-Whitney U-test. 2 Mean ± S.D. 3 Response frequencies (%) of the different answer alternatives. * Results significantly different from those for omnivores (P<0.05). 49 Results and discussion Table 8 Health data for 15-year-old low-meat-consumers and omnivores in Umeå, Stockholm and Bergen in 1996. HEALTH DATA1 Female Female Male Male low-meat- omnivores low-meat- omnivores consumers consumers (n = 108) (n = 880) (n = 41) (n = 976) Sick-days during a year2 9.9* ± 12 8.3 ± 12 5.5 ± 6.3 6.6 ± 9.0 Perception of health3, 4 Very good / excellent 25 36 52 62 Good 45 40 44 27 Moderate 23 21 4 10 Poor 7 3 0 1 Importance of being healthy3, 4 Very important 64** 79 65 71 Quite important 27 18 31 22 Neither important nor unimportant 8 3 0 5 Unimportant 1 0 4 2 Ability to affect own health3, 4 Yes 94* 86 83 86 No 6 14 17 14 Tired without reason3-5 Yes 49 52 64* 42 No 51 48 36 58 Often have a headache3-5 Yes 37 34 38* 17 No 63 66 62 83 Sleep badly3-5 Yes 25 23 23 14 No 75 77 77 86 Unable to concentrate3-5 Yes 58 55 59 42 No 42 45 41 58 Stomach disorder3-5 Yes 22 23 32 15 No 78 77 68 85 Depressed3-5 Yes 54** 37 36** 14 No 46 63 64 86 1 Comparisons between low-meat-consumers and omnivores were made separately for both genders using Pearson’s chi-square (or Fisher’s exact test for 2x2 tables) and the MannWhitney U-test. 2 Mean ± S.D. 3 Results pertain to Umeå and Stockholm, with 90 female low-meat-consumers, 447 female omnivores, 23 male low-meat-consumers and 493 male omnivores. 4 Response frequencies (%) of the different answer alternatives. 5 During the last week. * Results significantly different from results for omnivores (P<0.05). ** Results significantly different from results for omnivores (P<0.01). 50 Results and discussion promotion adolescents Norway. The result that low-meatconsumers (III) did not spend time with an organization/club more often than the omnivores, verifies the findings of another study showing vegetarians not joining clubs more often than others118. The reason given in that study was that vegetarians already were part of a subculture that resides within various other subcultures such as the environmental movement. However, Draper et al showed that vegans are more likely to belong to an organization directly connected with their diet compared with other types of vegetarians and 29 omnivores . The explanation given was their numerical minority and that by joining an organization they increased the possibility to link up with like-minded people14. The results of the present study imply that many young low-meatconsumers are part of a subculture and link up with like-minded people in an unorganized way. practices among in Sweden and Validity of dietary intake and energy expenditure In the Nutritional Assessment Study (IV, V) we compared the validity of reported intakes of energy, protein, sodium and potassium, obtained from a diet history method between 30 vegans and 30 omnivores (Table 9). Furthermore, a physical activity interview method was validated (IV) with the DLW method for 16 vegans and 16 omnivores. There was no significant difference in validity of reported energy intake or energy expenditure between the 16 vegans and 16 omnivores (IV), when using a diet history and physical activity interview. The following results are presented as a mean value with a 95% confidence interval. The diet history interview had a bias towards underestimating the energy intake [-1.93 MJ/day (-2.89 to -0.97 MJ/day)], and the physical activity interview had a bias towards underestimating energy expenditure [-1.44 MJ/day (-2.37 to -0.52 MJ/day)]. Furthermore, there was no significant difference in validity of reported protein intakes between the dietary groups. The diet history interview of 32 subjects (IV), showed good agreement for nitrogen (protein) intake and In conclusion, young low-meatconsumers had physical activity, smoked (III) and drank alcoholic beverages (III, V) to the same extent as the omnivores. The study indicates that the lifestyle of young low-meat-consumers differs from the lifestyle found in previous studies of vegetarians. Contrary to findings from other studies, adherence to a low-meat diet may not correlate with other health 51 Results and discussion agreement for reported nitrogen and sodium intake and urinary excretion of nitrogen and sodium. The difference between reported nitrogen intake and excreted nitrogen was +0.31 g/day (-0.23 to 0.85 g/day) and the difference for sodium was -198 mg/day (-538 to 141 mg/day). However, the diet history method underestimated potassium intake [-419 mg/day (-689 to -149 mg/day)] compared with the biological marker. urinary nitrogen excretion, and the difference was -0.40 g/day, (-1.10 to 0.29 g/day). The Nutritional Assessment Study (V) with 59 subjects showed no significant difference in the validity of reported nitrogen or potassium intake between the dietary groups. However, for female vegans the sodium excretion was higher than sodium intake compared with female omnivores (Table 9). The diet history method showed good Table 9 Validation of reported intakes of energy, nitrogen, sodium, and potassium in 30 vegans and 29 omnivores aged 16-20 years using the doubly labeled water method and 24-h urine collections, in Umeå, Sweden 1997–1998a. VALIDATION DATA FEMALES MALES Vegans Omnivores Vegans Omnivores (n=15) (n=15) (n=15) (n=14) FILb 1.41 ± 0.39 1.56 ± 0.37 1.66 ± 0.21 1.71 ± 0.25 PALc 1.41* ± 0.22 1.84 ± 0.44 1.87 ± 0.39 2.05 ± 0.33 d EIrep/EE mea 0.84 ± 0.25 0.92 ± 0.19 0.87 ± 0.19 0.85 ± 0.16 [Nrep x 0.81]/Nmeae 1.00 ± 0.32 0.98 ± 0.13 1.14 ± 0.27 1.04 ± 0.14 Narep/Nameaf 0.78* ± 0.42 1.02 ± 0.21 1.02 ± 0.32 1.15 ± 0.26 [Krep x 0.73 or 0.77] /Kmeag 0.90 ± 0.30 0.88 ± 0.19 1.02 ± 0.27 0.92 ± 0.18 a Data are presented as means ± SD. The 2-factor ANOVA was employed with diet and sex as the two factors for statistical comparisons between groups with different diets and sex. b Food Intake Level = reported energy intake divided by estimated basal metabolic rate. c Physical Activity Level = reported energy expenditure divided by estimated basal metabolic rate of 32 subjects (16 vegans, 16 omnivores and 44% females). d Reported energy intake, of 32 subjects (16 vegans, 16 omnivores and 44% females) divided by energy expenditure, as measured by the doubly labeled water method. 99 e Reported nitrogen intake (Nrep) times the urine excretion factor of nitrogen 0.81 divided by measured nitrogen in urine (Nmea). f Reported sodium intake (Narep) divided by measured sodium in urine (Namea). g Reported potassium intake (Krep) times the urine excretion factor of potassium, 0.73 for 100 vegans and 0.77 for omnivores divided by measured potassium in urine (Kmea). * Significantly different from the data on female omnivores (P<0.05). 52 Results and discussion studies of different dietary methods show that higher validity is obtained with diet history compared with diet records when used on adolescents (Table 10). Thirdly, other dietary assessment methods would be difficult to use on adolescents, e.g. food records or 24h recalls, since these methods require repetition, sometimes up to 20 days, in order to obtain good individual estimates. Therefore these methods may be too troublesome and inconvenient for many adolescents. In general, diet histories have proven to yield the most realistic energy intake compared with other methods86. The 32 subjects (IV) reported a 13% lower energy intake, 10.9 MJ compared with the measured energy expenditure 12.6 MJ (P<0.05) (Table 9). Potassium intake was underestimated by 7% (P<0.05) while the nitrogen and sodium intake showed good agreement with 24-h urine excretion. A diet history method was chosen rather than any other dietary assessment method for three main reasons. Firstly, the aim was to obtain information on typical eating patterns over a long time period. Secondly, validation Table 10 Comparison of different dietary methods food intake level (FIL) and physical activity level (PAL) in studies of adolescents. Data is presented as mean ± standard deviation. Dietary method No. Sex Age FILa PALb EIrep/EEmeac subjects F/M (year) Diet history89 5 F 18.0 1.69d 1.71d 1.01 ± 0.32 d 5 M 18.0 1.98 2.13d 0.94 ± 0.16 7-d food record89 5 F 18.0 1.31d 1.71d 0.77 ± 0.20 5 M 18.0 1.37d 2.13d 0.68 ± 0.31 7-d food record119 25 F 15.0 1.39 ± 3.30 1.79 ± 0.22 0.78 ± 0.16 25 M 15.0 1.56 ± 3.61 1.89 ± 0.16 0.82 ± 0.18 1.39d 1.78d 2w food record120 28 (nonobese) 14F, 14M 14.4 0.80 ± 0.19 d 27 (obese) 14F, 13M 14.7 0.80 1.57d 0.54 ± 0.23 a Food Intake Level = reported energy intake/BMRest. b Physical Activity Level = measured energy expenditure/ BMRest. c Reported energy intake / Measured energy expenditure. d The basal metabolic rate used is based on mean age and weight of the subjects and calculated from standard equations of Schofield 1985121. 53 Results and discussion Bratteby et al reported no significant bias in estimated energy intake in validation studies of the diet history method used on 15and 18-year-old subjects in the UK, USA and Sweden122. However, underestimation of energy intake compared with energy expenditure by the DLW method has been shown repeatedly, ranging from 12% to 31%123, 124. The main advantage with diet histories is that it gives information on habitual intake and thereby decreases the problem with day-to-day variation. Furthermore, it requires a fairly limited effort by the subjects which increases the possibility of achieving a high co-operation rate, compared with repeated recalls or records. A disadvantage with the diet histories is that they have a high subjective impact because the information is based on the competence of the person performing the interview and the subjects’ memory, cognition and motivation in reporting true dietary intake102. Other drawbacks are, that irregular food habits may easily be under- or overreported or forgotten, and portion sizes may be under- or overestimated. collections with >110% PABA recovery were rejected. The reason for exclusion of collections with <50% PABA recovery was that the compensation method is not recommended for cases of PABA recovery levels below 50%. This recommendation is due to the fact that the probability of the subject taking only two of the three PABA tablets is greater below 50% PABA recovery103. Furthermore, the compensated part of the urinary variable should not be larger than the collected part, since it potentially could cause too large an error in the equation. The higher exclusion limit of 110% PABA recovery compared with 101% was chosen since there are food items common in the Swedish diet containing benzoic acids. The Swedish National Food Administration has reported a 5002000 mg/kg of benzoic acid in lingonberry, cloudberry (hjortron) and arctic raspberry (åkerbär), and smaller amounts (10 mg/kg) in several other fruits and berries125. Sour milk products and yogurts containing jam with preserving agent have also been found to contain benzoic acid (100 mg/kg). A large number of the 24-h urine collections were incomplete (168 of 240 collections i.e. 70%) (V). Only 40 collections i.e. 17% (IV, V) were classified as complete and 25 (10%) collections with <50% PABA recovery and 7 (3%) In conclusion, diet history and physical activity interviews showed similar underestimation and validity when used with young vegans as when used with young omnivores. This enables 54 Results and discussion requirements for vegan males even when intake from dietary supplements was included (Table 12, Figure 8). Only 65 % and 98%, respectively, of the selenium and calcium average requirements was obtained from diet and supplements, respectively (Figure 8). The male omnivores had an intake above average requirements of all nutrients except one male who had a lower intake (including supplements) of selenium compared with average requirement (Table 12). comparisons to be made between the vegans and omnivores. Dietary intake The Nutritional Assessment Study (V) investigated the dietary intake by diet history of 30 vegans compared with 30 omnivores. The diet history showed that the vegans had a lower dietary intake (including supplements) than the omnivores of protein, saturated fat, cholesterol, disaccharides, niacin, vitamin D, calcium, phosphorus, and selenium (Tables 11-13). On the other hand, they had a higher intake than the omnivores of energy from carbohydrates, and a higher intake of polyunsaturated fat, monosaccharides, dietary fiber, vitamin C, vitamin E and magnesium. In addition, the male vegans had a lower intake than the male omnivores of alcohol, riboflavin, folate, sodium and zinc (Tables 11 and 12). The female vegans had a lower intake than the female omnivores of total and monounsaturated fat, and a higher intake of vitamin B12 and folate (Tables 11 and 13). A large proportion of male vegans had a lower intake than average requirements for riboflavin and vitamin B12 intake from diet, but this proportion decreased if intake from dietary supplements was included (Table 12, Figure 8). Calcium and selenium intakes from diet were below average 55 A significantly larger number of female vegans had a lower dietary intake (excluding supplements) than average requirements of vitamin A (6 subjects) and riboflavin (10 subjects) compared with female omnivores (Table 13). A significantly larger number of female vegans also had a lower dietary intake of vitamin B12, vitamin D and calcium, both when including and excluding supplements, compared with average requirements (Table 13). At group level the intake of vitamin B12, vitamin D and calcium from the diet of vegan females was below average requirements but a smaller proportion had an intake lower than average requirements when intake from dietary supplements was included (Table 13, Figure 9). The vegan females had too low food intake of selenium and 79% of the average requirement was obtained from diet Results and discussion about half the intake in a typical omnivorous diet. Good sources for vegans are broccoli, soya beans, enriched grains, algae, mushrooms. However, vegans who are not consuming whole or enriched grains and green leafy vegetables may obtain inadequate amounts of riboflavin. and supplements (Figure 9). The selenium intake from diet alone was also below average requirement (91%) for a large proportion of female omnivores, but intakes of all other nutrients were above average requirements at group level. Vegetarians ate more dietary supplements than omnivores (Tables 5 and 6), and the consumption increased the stricter the vegetarian diet was. Several other studies of vegetarians have also shown a higher intake of dietary supplements compared with omnivores52, 115, 126. A large proportion of the young vegans (V) in the present study needed dietary supplements of riboflavin, vitamin B12, vitamin D, calcium and selenium to obtain an intake above the average requirements. The dietary riboflavin intake was below average requirement for 11 male vegans and 10 female vegans (V). A study of 76 vegetarians (mean age 27 years) showed that 73% of the males and 60% of the females had a non-acceptable low urinary riboflavin content40. A tenday diet record of 50 of these 76 vegetarians showed that the mean riboflavin intake was 56% of the recommended dietary allowance40. Riboflavin is widely distributed among the plant and animal kingdom78. Although green leafy vegetables are rich in riboflavin, meat and dairy products supply The vitamin B12 intake from diet was insignificant among vegans (V) since plant foods most often lack the active form of vitamin B12. Vegans have shown to be dependent on supplements to get an adequate intake60, 127. Most vegans (80%) in the study ate, with greater or lesser frequency, dietary supplements containing vitamin B12. The dietary intake of vitamin D among female vegans (V) was below average requirement since their diet did not include enough plankton, certain mushrooms or fortified vegan food items such as soya drinks and margarines to add up to 2.5 µg. There is no average requirement given for vitamin D in the NNR58, but a daily intake above 2.5 µg has been shown to prevent deficiency symptoms of rickets and osteomalacia. 56 The low calcium intake of vegans (V) compared with the average requirement may be explained by the vegans excluding dairy products without replacing them with enough calcium-rich vegan Results and discussion Concern has been raised among parents and other adults about the nutritional adequacy of the diet when young vegetarians exclude animal products without replacing them with equally nutritious vegetarian products132, 133. The vegans in the present study (V) were not “excluders” in a consistent manner, since they were well motivated and interested in finding out about their diet and health. But because of the small non-random sample, we cannot draw the conclusion that excluders do not exist among the young vegetarian population in Sweden. food, such as calcium-fortified soya drinks, legumes and some green vegetables82. However, it is likely that the average requirement of calcium is higher for omnivores than for vegans, since the higher intake of protein and sodium in a omnivorous diet increases the amount of calcium lost in the urine82, 128, 129. A likely explanation for the low intake of selenium among omnivores and vegans (V) is the low content of selenium in the Swedish soil and thus in Swedish crops and animals. Another explanation may be that the data of selenium content in different food items are underestimated. Reports have shown that the selenium content in beef and dairy products is significantly higher than the information given by the food composition data base130, 131. The latter explanation implies that there is not a real low selenium intake among omnivores. In conclusion, the dietary intake of vegans varied considerably and did not comply with the average requirements for riboflavin, vitamin B12, vitamin D, calcium and selenium intakes of a large proportion of vegans. However, this was compensated for by dietary supplements for these nutrients except calcium for males and selenium for males and females. 57 Results and discussion Table 11 Reported intake of macronutrients including supplements. Mean ± SD of 30 vegans and 30 omnivores aged 16-20 years in Umeå, Sweden 1997–1998a. ENERGY AND FEMALES MALES MACRONNRb Vegans Omnivores NNRb Vegans Omnivores NUTRIENTS (n=15) (n=15) (n=15) (n=15) 13.9e 12.2 ± 1.7 13.2 ± 2.0 Energy (MJ) 9.6c 8.9 ± 2.18 9.5 ± 2.0 11.2d 15.7f *** 51 73***± 14 120 ± 22 Protein (g) 47 55 ± 17 80 ± 18 g 10–15 10***± 1.0 15 ± 1.7 E% protein 10–15 10***± 1.5 14 ± 1.3 * 100 88 ± 15 Fat (g) 73 58 ± 23 75 ± 22 100 ± 22 <30 27 ± 2.6 E% fatg <30 24***± 5.5 30 ± 3.8 29 ± 4.2 Carbohydrates (g) 310 340 ± 79 430 460 ± 66 426 ± 75 310 ± 62 55–60 63***± 3.2 55 ± 4.0 E% carbohydratesg 55–60 66***± 5.3 56 ± 3.8 Alcohol (g) 14 3.9 ± 4.4 19 2.1*± 3.1 7.0 ± 7.9 4.9 ± 7.0 g E% alcohol <4.5 1.4 ± 1.6 <4.5 0.5*± 0.7 1.6 ± 1.8 1.6 ± 2.3 *** <33 25***± 5.5 46 ± 9.3 Saturated fat (g) <24 15 ± 6.8 34 ± 10 * 42 32 ± 6.5 Monounsaturated fat (g) 30 20 ± 12 27 ± 8.0 37 ± 8.7 25 21***± 3.8 12 ± 3.7 Polyunsaturated fat (g) 18 15***± 6.5 8.6 ± 2.9 – Cholesterol (mg) – 2.3***± 2.4 230 ± 76 2.1***± 2.9 330 ± 58 * – Monosaccharides (g) – 65 ± 28 48 ± 16 64**± 19 47 ± 17 ** – Disaccharides (g) – 72 ± 29 100 ± 29 77***± 20 140 ± 40 Sucrose (g) 54 69 ± 28 75 73 ± 21 69 ± 22 88 ± 36 *** 38 Dietary fiber (g) 28 34***± 11 21 ± 6.3 44 ± 10 25 ± 8.0 a Data are given as mean ± SD. Statistical comparisons between groups with different diets were done using the Mann–Whitney U–test. b Recommended intake according to the Nordic nutrition recommendations 199658 except for energy. c Energy expenditure (MJ), as measured by the doubly labeled water method, of seven of the female vegans. d Energy expenditure (MJ), as measured by the doubly labeled water method, of seven of the female omnivores. e Energy expenditure (MJ), as measured by the doubly labeled water method, of nine of the male vegans. f Energy expenditure (MJ), as measured by the doubly labeled water method, of nine of the male omnivores. g Percent of total energy intake that is energy derived from protein, fat, carbohydrates or alcohol respectively. * Significantly different from the data on same sexed omnivores (P<0.05). ** Significantly different from the data on same sexed omnivores (P<0.01). *** Significantly different from the data on same sexed omnivores (P<0.001). 58 *** ** * e d c b a 59 Average requirements, according to the Nordic nutrition recommendations. Average requirement of vitamin D was taken as 2.5 µg and lowest recommended intake was used for vitamin E and potassium. Statistical comparisons were employed using Pearson’s chi-square test between groups with different diets, of number of subjects below average requirements. Retinol equivalents. Niacin equivalents. Alpha tocopherol equivalents. Significantly different from the data on same sexed omnivores (P<0.05). Significantly different from the data on same sexed omnivores (P<0.01). Significantly different from the data on same sexed omnivores (P<0.001). Table 12 Dietary intake (mean ± SD) of vitamins and minerals of 15 male vegans and 15 male omnivores, in Umeå, Sweden 1997–1998. Micronutrient ARa No. < ARa, b (excluding supplements) No. < ARa , b(including supplements) Mean ± SD (including supplements) intake of males Vegans Omnivores Vegans Omnivores Vegans Omnivores (n=15) (n=15) (n=15) (n=15) (n=15) (n=15) 750 2 0 2 0 Vitamin A (RE)c 1080 ± 264 1280 ± 365 Thiamine (mg) 1.1 3 0 2 0 2.9 ± 2.0 2.4 ± 0.7 Riboflavin (mg) 1.4 11*** 0 4 0 2.3** ± 1.9 3.0 ± 0.9 15 3 0 0 0 Niacin (NE)d 34*** ± 7.8 48 ± 9.5 Vitamin B6 (mg) 1.3 0 0 0 0 3.6 ± 1.2 3.2 ± 0.9 1.4 15*** 0 4 0 Vitamin B12 (µg) 22 ± 35 6.4 ±1.3 140 0 0 0 0 Folate (µg) 633*** ± 158 944 ± 2460 Vitamin C (mg) 30 0 0 0 0 226*** ± 61 113 ± 44 2.5 2 0 0 0 Vitamin D (µg) 4.9*** ± 1.9 8.6 ± 2.5 4.0 0 0 0 0 Vitamin E (α-TE)e 21*** ± 5.4 11 ± 4.6 Iron (mg) 8.5 0 0 0 0 22 ± 6.7 20 ± 8.1 Calcium (mg) 600 12*** 0 10*** 0 592*** ± 189 1700 ± 443 Phosphorus (mg) 450 0 0 0 0 1370*** ± 288 2180 ± 408 Magnesium (mg) – 572* ± 100 462 ± 112 Sodium (mg) – 3800* ± 856 4660 ± 1110 Potassium (mg) 1600 0 0 0 0 4200 ± 864 4100 ± 696 Zinc (mg) 5 0 0 0 0 14* ± 6.4 18 ± 6.1 30 15*** 2 11*** 1 Selenium (µg) 23*** ± 14 47 ± 17 Results and discussion *** ** * e d c b a 60 Average requirements, according to the Nordic nutrition recommendations. Average requirement of vitamin D was taken as 2.5 µg and lowest recommended intake was used for vitamin E and potassium. Statistical comparisons were employed using Pearson’s chi-square test between groups with different diets, of number of subjects below average requirements. Retinol equivalents. Niacin equivalents. Alpha tocopherol equivalents. Significantly different from the data on same sexed omnivores (P<0.05). Significantly different from the data on same sexed omnivores (P<0.01). Significantly different from the data on same sexed omnivores (P<0.001). Table 13 Dietary intake (mean ± SD) of vitamins and minerals of 15 female vegans and 15 female omnivores, in Umeå, Sweden 1997–1998. a a, b a ,b Micronutrient Mean ± SD (including supplements) AR No. < AR (excluding supplements) No. < AR (including supplements) intake of females Vegans Omnivores Vegans Omnivores Vegans Omnivores (n=15) (n=15) (n=15) (n=15) (n=15) (n=15) 6* 1 5 1 Vitamin A (RE)c 1030 ± 698 1330 ± 713 700 Thiamine (mg) 0.9 3 0 3 0 3.6 ± 6.4 1.8 ± 0.6 Riboflavin (mg) 1.1 10** 1 5 1 3.3 ± 6.4 2.2 ± 0.9 12 4 3 1 0 Niacin (NE)d 26* ± 8.4 33 ± 10 Vitamin B6 (mg) 1.0 0 0 0 0 4.5 ± 6.6 2.3 ± 0.9 1.4 15*** 0 8** 0 Vitamin B12 (µg) 17* ± 41 5.4 ± 2.7 120 0 1 0 1 Folate (µg) 577*** ± 263 251 ± 130 Vitamin C (mg) 30 0 0 0 0 197*** ± 100 127 ± 91 2.5 11*** 0 7** 0 Vitamin D (µg) 3.6* ± 2.8 6.0 ± 2.7 3.0 0 0 0 0 Vitamin E (α-TE)e 17*** ± 7.7 15 ± 3.0 Iron (mg) 10 2 5 2 4 18 ± 9.0 15 ± 9.6 Calcium (mg) 11*** 0 10*** 0 741*** ± 711 1330 ± 372 600 Phosphorus (mg) 0 0 0 0 1040*** ± 306 1540 ± 378 450 Magnesium (mg) – 453** ± 121 332 ± 92 Sodium (mg) – 2590 ± 986 3040 ± 614 Potassium (mg) 0 0 0 0 3460 ± 1240 3160 ± 921 1600 Zinc (mg) 5 1 0 1 0 11 ± 6.5 12 ± 4.3 30 15 11 11 11 Selenium (µg) 24* ± 22 35 ± 36 Results and discussion 50 100 150 200 0 100 150 200 61 Figure 8 Nutritional intakes from diet and supplements, as percentage of average requirements of 16-20 year olds a) 15 male vegans and b) 15 male omnivores. Average requirement of vitamin D was taken as 2.5 µg and lowest recommended intake was used for vitamin E and potassium57, 58. The intakes of the nutrients that reaches 200% of the average requirements indicates that the intakes were above 200% of average requirements. Percent (%) of average requirement 50 b) Male omnivores From supplements. From diet. Thiamine Vitamin A Thiamine Vitamin A Percent (%) of average requirement Niacin Riboflavin Riboflavin Vitamin B6 Vitamin B6 Niacin Folate Vitamin B12 Vitamin C Vitamin C Vitamin B12 Vitamin D Vitamin D Folate Iron Calcium Vitamin E Calcium Phosphorus Vitamin E Phosphorus Potassium Iron Zinc Potassium Zinc 0 Selenium Selenium a) Male vegans Results and discussion 50 100 150 Vitamin D Vitamin C Folate Vitamin B12 Vitamin B6 Vitamin D Vitamin C Folate Vitamin B12 Vitamin B6 Thiamine Vitamin A 0 100 150 Percent (%) of average requirement 50 b) Female omnivores 200 62 Figure 9 Nutritional intakes from diet and supplements, as percentage of average requirements of 16-20 year olds a) 15 female vegans and b) 15 female omnivores. Average requirement of vitamin D was taken as 2.5 µg and lowest recommended intake was used for vitamin E and potassium57, 58. The intakes of the nutrients that reaches 200% of the average requirements indicates that the intake were above 200% of average requirements. From supplements. From diet. Thiamine Vitamin A Riboflavin Percent (%) of average requirement Niacin Riboflavin Niacin 200 Iron Calcium Vitamin E Calcium Phosphorus Vitamin E Phosphorus Potassium Iron Zinc Potassium Zinc 0 Selenium Selenium a) Female vegans Results and discussion Results and discussion Biochemical markers of nutritional status Iron Female vegans (V) had a higher hemoglobin concentration than female omnivores, but there was no difference in the S-ferritin level (Table 14). Among males there were none insufficient iron stores but the prevalence of low iron stores among females was 20% for vegans and 23% for omnivores. These results are in contrast with a study of 79 lacto-ovo-vegetarians (14-19 years) showing a higher number of vegetarians (29%) with low plasma ferritin (<12µg/L) compared with omnivores (17%)75. Similar results are shown in another study of 35 vegetarians and 32 omnivores (mean age 23 years) which showed that 6 vegetarians and 1 omnivore were anemic134. Despite the intake of low absorbed iron and a high intake of dietary fiber and possibly phytate, the vegans in the present study (V) had similar iron status as the omnivores, which confirms the findings of other studies71, 73, 135, 136. Thus, there are conflicting results regarding iron status of vegetarians compared with omnivores. The iron in a vegan diet is nonheme with a lower bioavailability than the heme-iron in an omnivorous diet. However, the female vegans (V) were shown to have a higher nutritional density of iron in their diet compared with omnivores and also a higher density of vitamin C. This may partly explain why there was no difference in iron status between vegan and omnivores. A study in Australia showed that all iron consumed by vegetarians was nonheme-iron and for the omnivores 90% was nonhemeiron137. Research indicates nonheme-iron absorption from a lacto-ovo-vegetarian diet to be 70% lower than from an omnivorous diet, but an associated decrease in fecal ferritin excretion suggests a physiological adaptation to increase the efficiency of iron absorption138. 63 16 ± 6.0 67 ± 8.4 25 ± 11 30 ± 30 452 ± 208 10 ± 4.1 101 ± 39 15 ± 5.4 60* ± 7.7 26 ± 9.1 24 ± 15 287** ± 220 23** ± 10 122 ± 34 7.0–27 35–70 20–50 10–82 96–568 3.4–47 26–156 Serum iron (µmol/L) TIBC (µmol/L)c Transferrin saturation (%) Serum ferritin (µg/L) Serum vitamin B12 (pmol/L) Serum folate (nmol/L) Serum 1,25-dihydroxy- 26–156 3.4–47 96–568 10–200 20–50 35–70 9.0–32 135–168 107 ± 026 18*** ± 6.6 196*** ± 91 56 ± 24 32 ± 7.7 60 ± 5.4 19 ± 4.2 148 ± 7.5 91 ± 23 12 ± 8.1 443 ± 149 61 ± 32 34 ± 7.9 57 ± 3.5 19 ± 4.5 146 ± 6.9 64 D–vitamin (pmol/L) a Data are given as means ± SD. Statistical comparisons between groups with different diets were done using the Mann–Whitney U–test. b According to the Clinical Chemistry Department, The University Hospital of Northern Sweden, Umeå, Sweden. c Total iron binding capacity. * Significantly different from the data on same sexed omnivores (P<0.05). ** Significantly different from the data on same sexed omnivores (P<0.01). *** Significantly different from the data on same sexed omnivores (P<0.001). 120 ± 7.4 127* ± 5.0 119–153 FEMALES MALES Reference rangeb Vegans (n=15) Omnivores (n=15) Reference rangeb Vegans (n=15) Omnivores (n=15) Hemoglobin (g/L) NUTRITIONAL DATA Table 14 Nutritional assessment in blood of 30 vegans and 30 omnivores aged 16-20 years, in Umeå, Sweden 1997–1998a. Results and discussion Results and discussion Vitamin B12 The vegans (V) had an average serum concentration of vitamin B12 within the reference range (Table 14). Three out of 30 vegans (V) had serum vitamin B12 concentration lower than the reference range, which is a lower prevalence than found in other studies52, 65. A study of 93 Australian vegetarians (mean age 29 years) showed that 15 had serum vitamin B12 levels below normal65. None of the vegetarians were vegans and the subjects had practiced their diet for a mean of 7.3 years. Another study of 36 Canadian lacto-ovo-vegetarians (mean age 27 years) showed that almost one third of them had serum vitamin B12 levels below the lower limit of normality52. It was also found that 9 of the 10 vegetarians with low vitamin B12 levels in that study had been vegetarians for more than three years. The vegans in the present study (V) had been vegans for 1.7 years (range 0.5-3.5 years) and vegetarians for 2.8 years and the comparatively low prevalence of low vitamin B12 serum levels could probably partly be explained by the subjects’ few years as vegans and the use of vitamin B12 supplements. Two of the vegans classified as having low vitamin B12 concentrations in the present study (V) did not take any supplements. One did, which indicates an absorption problem caused by e.g. lack of intrinsic factor rather intake14, 23. than inadequate In early B12 depletion, a low level of the vitamin is seen in serum. However, a lower serum level than normal does not necessarily indicate deficiency. The next stage is a decrease in stores of vitamin B12 in the body’s cells, for example the red blood cells. This stage of vitamin B12 depletion may be followed by biochemical changes, such as rise in methylmalonic acid in the blood. Finally, clinical symptoms of vitamin B12 deficiency and megaloblastic anemia occur. However, among vegans neurological symptoms139 are more likely to occur before symptoms of anemia since most often vegans have a high consumption of folate that protects them from anaemia139, 140. In diets, where folate intake is adequate, lowered vitamin B12 intake leads to depleted plasma vitamin B12 concentrations with a concomitant increase in homocystein 141 concentration . The detection of increased homocystein levels is a way to determine vitamin B12 deficiency. 65 Vitamin D The vegans (V) had an average serum concentration of 1,25dihydroxy-D–vitamin within the reference range (Table 14). The dietary intake of vitamin D was lower among vegans compared Results and discussion (20%) as among female omnivores (23%). Three vegans had low serum vitamin B12 levels but further analyses are needed to be able to conclude that vitamin B12 deficiency exists among them. Serum 1,25-dihydroxy-D–vitamin levels were adequate for both vegans and omnivores but this does not exclude a possible inadequate vitamin D status. with omnivores, but they presumably obtained enough vitamin D from supplements and via endogenous production for the conversion of 25-dihydroxyvitamin D to sufficient levels of 1,25dihydroxy-D–vitamin. However, we cannot exclude the possibility that some of the vegans may have inadequate vitamin D status since serum 25-dihydroxyvitamin D levels were not measured. General discussion The concentration of serum 25hydroxyvitamin D is the most accurate measure of vitamin D status, since 1,25-dihydroxy-Dvitamin may be normal in a vitamin D deficient state105. The serum 1,25-dihydroxy-D-vitamin concentration is, however, often used in Swedish clinical work to monitor the vitamin D status of patients with chronic renal failure and distinguish between vitamin Ddependent and vitamin D-resistant rickets142. Before vitamin D supplementation of some food items was started in the 1940s, vitamin D deficiency was fairly common in the Scandinavian countries143. A recent study showed that female omnivores (915 years) living at high latitude, where there is only little sunlight in the winter, had a high prevalence (13.4%) of severe hypovitaminosis D in midwinter (FebruaryMarch)143. In conclusion, low iron stores was as common among vegan females Vegetarianism can be seen as a dietary fad, an ethical choice or a political statement. Therefore eating vegetarian food may not necessarily mean having a healthy diet, as well as eating omnivorous food may not always be adequate for all nutrients. It all depends on what foods and amounts are included in the diet. It is important for adolescents in general and vegetarians in particular to receive more knowledge, both theoretical and practical, about how to combine and prepare a healthy diet. Schools and parents have an important role to play by increasing knowledge and make well-planned food available. The present interest in vegetarianism among adolescents is a great opportunity to fulfil this assignment among both vegetarians and omnivores. The papers in this thesis provide information which indicates that the new generation of vegetarians 66 Results and discussion suggests that healthy vegetarianism improves health. have different dietary and lifestyle habits compared with what previous studies of vegetarians have shown. For vegetarian adolescents, especially vegans, it is essential to be well motivated and take an active interest in the new dietary regime, and to appreciate the fact that it is not simply a matter of excluding food items. Excluded animal products should be replaced with equally nutritious vegetarian products. If there is no interest in nutrition and no nutritious vegetarian food available at home or in school, a poor eating pattern may be the consequence. Some schools choose not to serve a vegan lunch alternative for various reasons, but vegans will most likely not stop eating vegan food just because it is not served for school lunch. Rather, the vegan will probably eat a less nutritious lunch or nothing at all. It is possible for young vegans to have an adequate dietary intake, but vitamin B12 supplements are needed and care needs to be taken to ensure sufficient intake of nutrients, especially selenium and calcium. More over, research This thesis has contributed with new valuable knowledge about young vegetarians and omnivores, but there is still much more to find out. One interesting area for research is studies on long-term effects of, for example, the risk of osteoporosis. The vegans in the present thesis showed a lower intake of calcium, and the energy expenditure indicated less physical activity compared with omnivores, which may result in a lower bone mineral density. To monitor the prevalence of vitamin B12 deficiency in long-term vegetarians is also of high priority. Therefore, biochemical analyses of B12, folate, homocystein and methylmalonic acid should be performed to be able to establish the causes of a possible deficiency. Iodine is also a nutrient interesting to study among vegans since dietary sources are limited, especially if iodinated salt is not used. Another aspect of great interest is to understand the motives of young people for adopting vegetarianism. 67 Conclusions CONCLUSIONS The prevalence of adolescents (16-20 years) eating vegetarian food at school lunch was 5% in Sweden 1996. There was a much higher occurrence of adolescents eating vegan food (2%), lacto-ovo-vegetarian food (7%) and lacto-ovo-fish-vegetarian food (2%) in the town of Umeå compared with Sweden overall (I). There was a significantly higher prevalence of 15-year-old adolescent vegetarians in Umeå (15.6%) compared with Stockholm (4.8%) and Bergen (3.8%) in 1996. A high proportion of the adolescents choosing a vegetarian dietary regime were females (72%) (II). Even though the female vegetarians (15 year olds) consumed vegetables significantly more often than the omnivores, the intake (32 times/month) was not as often as might be expected of a vegetarian population. The male vegetarians reported eating vegetables not even once a day (25 times/month). There was no difference between the intake of fruits/berries, sweets/chocolates and fast foods by vegetarians compared with omnivores (II). However, female vegetarians more often than female omnivores consumed dietary supplements. The lifestyle of young low-meat-consumers differs from the lifestyle shown in previous studies of vegetarians, as they reported being less healthy and having similar exercise, alcohol consumption, smoking frequency and tobacco use as the omnivores. Contrary to findings from other studies, adherence to a low-meat diet may not correlate with other health promotion practices among adolescents in Sweden and Norway (III). There was no significant difference in the validity of reported energy expenditure or energy and protein intakes between vegans and omnivores when using physical activity and diet history interviews. This indicates that it is possible to make a comparison between the vegans and omnivores (IV). 68 Conclusions Young vegans had a higher calculated intake than young omnivores of vegetables, legumes, and dietary supplements, and a lower intake of ice cream, cake/cookies, and candy/chocolate. Vegans had a higher prevalence of dietary intakes below average requirements of riboflavin, vitamin B12, vitamin D, calcium and selenium compared with omnivores. Average intakes of calcium and selenium remained below average requirements even after inclusion of intake from dietary supplements (V). There was no difference in iron status among young vegans and omnivores, 20% of the vegans and 23% of the omnivores had low iron stores. Both vegans and omnivores had adequate levels of 1,25-dihydroxyD–vitamin but three out of 30 vegans had low vitamin B12 concentration in blood (V). 69 Acknowledgements ACKNOWLEDGEMENTS This thesis is not the result of one person’s work. On the contrary a lot of people have contributed in one way or another to bring this work to what it is today. I am grateful to all of you. In particular I wish to express my sincere and warm gratitude to the following: Gunnar Johansson without whom this project would not have been possible. Thank you for introducing me to vegetarian research, for invaluable scientific tutoring and for sharing your knowledge. Brittmarie Sandström for excellent guidance and professional advice. Charlotte Nygren for constructive support and for your trust in my ability. Ola Haugejorden, Kristin Klock, Anne Nordrehaug Åström and Klaas Westerterp for fruitful feedback on the manuscripts. Ulla Rönnlund for being a wonderful mentor. It has been a privilege to work with you. Agneta Hörnell for fruitful feedback and constructive advice during the final part of this thesis. Seppo Salonen for skilful computer help and never ending patient support. Linda Hagfors for valuable help regarding data entry and for creating a warm atmosphere in our room. Teachers and staff at the Department of Food and Nutrition: To each one of you I wish to express my deepest appreciation for your friendship and encouragement. AB PT Datatech Systems Sweden for professional computer support and for undertaking the hard and challenging work with solving our Teleform problem. Elisabet Forsum for generously introducing me to the world of doubly labeled water technique. 70 Acknowledgements Allan Hackett for your wonderful support and reading the manuscripts during my stay in Liverpool. Lars Dahlgren for fruitful discussions about qualitative methods and for showing interest in our work. I look forward to our continuing collaboration. Maria Hedström, Maria Larsson, and Inger Öhlund for valuable help regarding data entry. All the participating subjects, without whom this project would not have been possible, for showing interest and donating your time. Bertil Winther for proofreading my first paper and inspiring me to learn more about the American language. My family Gunnel and Ingvar Larsson, Marita, Helge, Elina and Filip Winther for your never-ending support and encouragement during all these years. Even though you were far away you were always close at heart. Last but not least, my deepest gratitude goes to my partner Peter Tanskanen for standing by my side and providing me with support when times were tough. "May you sail on the sea of success, and land on the shore of happiness". Financial support for this work was given by the Faculty of Social Sciences, Umeå University, the Swedish Foundation for Healthcare Science and Allergy Research, the Swedish Nutrition Foundation, Research Foundation of Bertil Eriksson, Fund of Uppsala Hemsysterskola, JC Kempes Memorial Fund, Patentmedelsfonden för odontologisk profylaxforskning, the Swedish Institute, COST Action-99, Knut and Alice Wallenberg’s fund, NEON and Foundation of Engelbrekts Barnavårds & Husmodersskola. Umeå, 2001 …………………………… 71 References REFERENCES 1 2 3 4 5 6 Samuelson G. Dietary habits and nutritional status in adolescents over Europe. An overview of current studies in the Nordic countries. 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