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28. Protein requirements of Filipino children 22-29 months old consuming local diets


Carmen Intengan

Food and Nutrition Research Center, National Science Development Board, Manila, Philippines

Objective

To determine the protein requirements of young children on a diet based mainly on rice gruel and mung bean when dietary energy is not a limiting factor.

Experimental Design

Environment

Tropical climate; temperature 25 to 29 C; relative humidity, 50 to 58 per cent.

Subjects

Five apparently healthy male Filipino children 22 to 29 months old participated in the study. They were recruited from a welfare institution. All children had just been subjects in another metabolic study. Their characteristics are shown in table 1. Using body weight as a parameter. three of the children were classified as normal by Philippine standards 11); the other two subjects were in the upper limit of firstdegree malnutrition. Using the same standards, all subjects were below their ideal height for age by 5 to 9.7 cm. Blood analyses indicated good-to-borderline levels.

Diets

The diets were essentially based on rice and mung bean with added fruits and vegetables low in protein content (table 2). The diets were calculated to provide 100 kcal/kg body weight/day using proximate composition and Atwater factors. A blend of coconut-corn oil (50:50) and the small amounts of fat from other foods provided about 30 per cent of the energy intake. One-third of the dietary protein was provided by mung bean.

TABLE 1. Characteristics of Subjects

Subject Age
(months)
Body
Weight
(kg)
Height
(cm)
Hb
(g/100 ml)
Hct
(Vol. %)
Serum
Albumin
CH Ia Classifi
cation by weight
R.D. 23 13.1 84.0 11.95 37.0 4.62 0.95 Nb
P.D. 22 12.4 83.6 12.50 35.0 3.04 0.86 N
S.L. 29 11.4 83.1 13.38 37.2 3.99 0.98 1c
P.J. 29 11.2 85.9 12.50 39.0 4.11 0.86 1
M.P. 26 12.3 85.0 12.55 38.0 - 0.94 N

a. Creatinine height index.
b. Normal.
c First-degree undernourished or - 10 % underweight.

TABLE 2. Diet Composition Per 1,000 kcala

Food Items Weight Calories Protein Fat
Rice 57.0 209 4.80 0.29
Mango 18.3 65 4.17 0.18
Sweet potato 100.0 136 1.10 0.40
Mung-bean starch 25.0 88 0.05 0.05
Gourd 50.0 8 0.25 0.05
Potato 25.0 5 0.15 0.05
Squash 25.0 8 0.48 0.10
Banana 100.0 92 1.20 0.30
Papaya 50.0 24 0.30 0.01
Coconut-corn oil (1:1) 31.8 280 - 31.80
Sugar 21.2 85 - -
Total 503.3 1,000 12.50 33.23

a Sample diet for a 10 kg child at 100 kcal/kg and at a level of 200 mg N/kg body weight (1 25 9 protein)

Four levels of dietary protein were given to each subject. Adjustment in protein level was done by substituting protein calories with starchy roots, fruits, or mung-bean starch. Two children (R.D. and P.D.) followed the descending design, that is, starting from the highest level followed by the other levels in 0.25 9 decrements; the three other children followed the ascending design.

Vitamin and mineral supplements given daily provided the following: vitamin A (3,000 IU), 0,9 mg; vitamin D2 (400 IU), 10 mg; vitamin C, 50 mg; thiamine, 1.5 mg; riboflavin, 1.2 mg; pyridoxine, 1 mg; vitamin B12, 3 mg; niacinaminde, 10 mg; iron, 3 mg; iodine, 75 mg; calcium, 40 mg; phosphorus, 43 mg; magnesium, 3 mg; manganese, 0.5 mg; zinc, 3.071 mg; choline, 5 mg; dexpanthenol, 5 mg and inositol, 5 mg. In addition, 2 teaspoons of Cetrin were given daily. Each 30 ml contains 250 mg vitamin C.

Duration

Each dietary protein level lasted for seven days with a three-day break period between two experimental levels. During this period the subjects were fed the regular diet (250 mg N/kg/day).

Indicators and Measurements

Nitrogen Balance

Apparent N balance was calculated during the last four days of each seven-day feeding period at each N level for each child. Aliquots of daily urine excretions, two-day pooled faecal specimens, and diets were analysed by the macro-Kjeldahl method.

Apparent N absorption was calculated from data on dietary and faecal nitrogen. Apparent N retention was estimated by subtracting urinary and faecal nitrogen from total nitrogen intake and dividing this by the total intake. Biological value was estimated by dividing the percentage of N retained by the corresponding apparent absorption.

Protein Requirements

This was calculated as the zero balance intercept from the regression equation of true N balance (y) on N intake (x), pooling all data points (2). The safe level of protein intake was calculated from the upper 95 per cent confidence band.

An allowance of 10 mg N/kg/day was made for integumental and miscellaneous N losses to estimate the true N balance.

Urinary Creatinine and Urea Nitrogen

Determined by Folin's method and a modified Van Slyke and Cullen's method, respectively.

Anthropometry

Body weight was taken daily every morning before breakfast, post-voiding, with minimal clothing. The following measurements were obtained on day four and day 10 of each dietary period; height, arm, waist, chest and tricipetal and subscapular skinfold thicknesses.

Main Results

N balance and Absorption

The summary of individual mean nitrogen and energy intakes, apparent N balance, N absorption, and biological values are presented in table 3. At the lowest level of nitrogen intake (1 9 protein/kg), two subjects were in negative nitrogen balance, but when these were corrected for sweat nitrogen (10 mg N/kg), all subjects except one remained positive. At higher levels all the subjects were in positive balance.

N absorption, N retention, and biological values decreased as levels of N in the diet were lowered. The low percentage of N absorption may be accounted for by the large amounts of N lost in faeces.

Protein Requirements

By regression analysis of true N balance y = - 120. + 0.751 x (r = 0.8604). The mean N requirement (PRm) was calculated as 160.9 mg N, or 1.0 9 protein/kg/day. The safe level of intake for 97.5 per cent of the population was calculated as 221.9 mg N or 1.39 9 protein/kg/day.

Anthropometric Measurements

Changes in body weights of the subjects for each experimental period are also shown in table 3. Although energy intake was kept constant, the subjects lost body weight on N intake levels of 200 mg to 280 mg but gained at the 160 mg N/kg body-weight intake level. There was, however, one subject (P.D.) who lost weight at this level, which could not be explained from his daily record during this period.

The relationship of increased weight gain with a higher percentage of PE does not seem to apply to the diets given in this study. Other body measurements did not show a consistent pattern when correlated with changes in body weight.

Conclusions and Comments

The mean protein requirement of young children obtained when given rice and mung bean as the main sources of protein was 1 g/kg/day, and the safe level of intake was 1.39 g/kg/day. This requirement is much higher than a comparative study on a rice-fish diet where PRm was 0.7 g/kg/day and the safe level was 1 g/kg/day.

TABLE 3. Nitrogen and Energy Intakes, Nitrogen Balance, Digestibility, Biological Value and Changes in Body Weights at Each Level of Nitrogen Intake

Subject N
Intake
Mean
Daily
N Intake)
(mg/kg)
Mean Daily Energy Intake Mean Daily N Excretion Balance
(mg/kg)
Absorption
(%)
Retention
(%)
BV Changes
Body
Weight
(g/day)
(kcal/kg) (% PE) Urine
(mg/kg)
Faeces
(mg/kg)
R.D. 258 100 6.5 93 42 123 83.7 47.7 56.9 - 16
241 101 6.0 81 67 93 72.2 38.6 53.4 - 22
196 99 4.9 62 63 71 67.9 36.2 53.4 + 26
150 98 3.8 56 93 1 38.0 0.7 1.8 + 34
P.D. 290 100 7.3 96 81 113 72.1 39.0 54.1 + 12
252 100 6.3 85 100 67 60.3 26.6 44.1 + 06
191 100 4.8 69 59 63 69.1 33.0 47.7 + 19
162 98 4.1 78 76 8 53.1 4.9 9.3 - 40
S.L. 247 102 6.3 87 85 75 65.6 30.4 46.3 - 31
232 100 5.7 80 91 61 60.8 26.3 43.3 - 35
197 98 4.9 81 91 25 53.4 12.7 23.6 - 41
151 99 3.8 81 91 - 28 53.8 12.7 0.0 + 12
P.J. 276 102 7.0 102 113 61 59.1 22.1 37.4 - 36
240 99 5.9 87 113 40 52.9 16.7 31.5 - 35
204 100 5.2 70 107 27 47.6 13.2 27.8 - 18
146 98 3.7 61 87 - 2 40.4 0.0 0.0 - 2
M.P. 288 98 7.3 87 100 101 65.3 35.1 53.7 - 9
238 98 6.1 70 92 76 61.3 31.9 52.0 - 6
201 99 5.1 62 107 32 46.8 15.9 34.0 + 13
163 100 4.1 60 80 23 50.9 14.1 27.7 + 24

TABLE 4 Comparison of Results of this Study with the Study Carried Out by Roxas et al. (3)

  Roxas Study This Study
Age of children, months
Energy intake, kcal/kg body weight
18-24
100
22-29
100
Number of children 4 5 5 5
N intake (mg/kg body weight) 197 235 200 243
Faecal N (mg/kg body weight) 65 56 85 92
Urinary N (mg/kg body weight) 90 124 67 81
N retention (mg/kg body weight) 42 55 44 67
Apparent N digestibility (%) 67 76 57 62
Apparent N retention (%) 22 23 22 28
Changes in body weight (g/day) 3 10 5 - 5 - 24

Losses in body weight of the subjects when N levels in the diet were 200 mg and above could have been caused by one or several of the following factors.

1. The use of whole bean in the diet. In a comparable study reported by Roxas et al. (3), the mung beans used were dehulled and pulverized. A comparison of the results of the Roxas study with this study is shown in table 4.

At comparable levels of N intake, faecal losses in this study are higher, N digestibilities are lower, and subjects lost more weight at 241 mg N intake than at 198 mg N. On the other hand, in Roxas' study the weight gains observed were adequate at a level of 197 mg N/kg/day and increased at the higher N intake.

2. The dietary fibre of the rice/mung-bean diet used in this study is most likely higher than the rice/dehulled mung-bean diet used by Roxas. In a study on the Guatemalan diet reported by Calloway and Kretsch (4), the high dietary fibre of the black-bean and lime-treated tortillas reduced the digestibility of energy and protein by 3 to 4 per cent. Similar losses in energy could have occurred with the rice-mung bean diet in this study.

3. The chemistry department of IRRI (International Rice Research Institute) has reported the presence of poorly digested protein from cooked milled rice, with the major fraction having MW 16,000 (5).

4. The presence of trypsin inhibitor in mung bean cannot be discounted. This may explain the poor utilization of nitrogen as levels are increased. In the diet used by Roxas, the dehulling of the bean made use of dry heat that could have destroyed any trypsin inhibitor.

The results of this study could be an important consideration in the planning of ricebased diets or weaning foods for young children.

References

1. Boundaries of Classification by Weight of Filipino Children /Males and Females Combined) Ages 18 Months to 72 Months (FNRI Publication No. 137A, 1971).

2. W.M. Rand, N.S. Scrimshaw. and V.R. Young. "Determination of Protein Allowances in Human Adults from Nitrogen Balance Data," Am. J. Clin. Nutr, 30:1129-1134 (1977).

3 B.V. Roxas, C.L. Intengan, and B.O. Juliano, "Protein Content of Milled Rice and Nitrogen Retention in Preschool Children Fed Rice-Mung-bean Diets." Nutr. Rep. Int., 11: 393 (1975)

4. D.H Calloway and M.J. Kretsch, "Protein and Energy Utilization in Men Given a Rural Guatemalan Diet and Egg Formulas With and Without Added Oat Bran, Am. J. Clin. Nutr, 31: 1118-1126 (1918).

5. B.V. Roxas, C L. Intengan, and B.O. Juliano, Annual Report (Grant No. AID/D SAN-G-0157. Agricultural/Nutrition Branch, AID, Washington D.C.).


29. Long-term study on the adequacy of usual Thai weaning food for young children


Kraisid Tontisirin, Nissawan Ajmanwra, and Aree Valyasevi

Institute of Nutrition, c/d Ramathibodi Hospital. Mahidol University, Bangkok. Thailand

Summary of Short-term Study Phase 1 as Background Information

Nine children, aged 9 to 36 months, weighing 8.1 to 11.1 kg, and living in a metabolic unit, were given usual Thai weaning diets at three levels of energy intake varying from 87 to 118 kcal/kg/day. These diets consisted of rice, fish, and bananas. The protein intake was fixed at the "safe level," 1.7 g/kg/day, derived from rice and fish in a ratio of 70:30, and having an amino-acid score of 94.8 with assumed digestibility of 93 per cent.

Each level of energy intake was fed for seven days: the first four days for adaptation to a new intake level and the last three for balance studies. There was a four-day resting period between each treatment. Fat intake was kept constant throughout the study at about 10 per cent of energy intake. Vitamin and mineral supplements were given daily.

The adequacy of protein intake was evaluated by measuring N retention and weight gain. Other measurements included fat and energy absorption; blood constituents were also measured. A summary of the data in study phase 1 is shown in tables 1 and 2.

At the lowest level of energy intake, 87 kcal/kg/day, apparent N retention and weight gain were quite low, being 43.7 mg N/kg/day and 3.5 g/day respectively. At the two higher levels of energy intake (100 and 118 kcal/kg/day), apparent N retention was greater than 60 mg/kg/day and weight gain was about 20 g/day or more.

Biological value (BV) and net protein utilization (NPU) were clearly affected by the changes in energy intake. They decreased significantly with decreased energy intake. N absorption and digestibility, however, were not affected by changes in energy intake.

The results from this study suggest that at the "safe level" of protein intake, as recommended by FAO/WHO in 1973, the usual Thai weaning food provides adequate protein for the needs of young children if energy intake is supplied at 100 kcal/kg/day or higher.

TABLE 1 Summary of Study Phase 1 on the Effects of Varying Energy Intakes on the Adequacy of the Safe Level of Protein Intake in Young Children

  Energy intake (kcal/kg/day)
87 100 118
Number of children 9 9 7
Age (months) 21.6 2.8    
Protein intake (g/kg/day) 1.7 0.01 a 1.7 0.04 1.7 0.`05
Fat intake (% of energy) 11.2 0.6 10 1 0.2 10.1 0.5
Fat absorption (% of intake) 90.2 1.0 91 4 1.2 91.0 1.0
Energy absorption (% of intake) 8.5 0.4 6 7 0.6 7.1 0.6
Weight gain (g/day) 3.5 6.9 20 3 5.9 53.8 7.3
N balance (mg/kg/day)      
N intake 271.2 2.3 276. 4 5.8 277.0 7.4
Urinary N 116.2 6.9 100.56.8 80.2 4.8
Faecal N 111.3 8.9 106.2 10.9 121.6 11.2
Apparent N balance 43.7 6.8 69.6 7.8 75.7 7.6
Urinary creatinine (mg/day) 140.7 11.6 1506 6.8 144 6 8.1
Protein quality      
N absorption (% of intake) 58.9 3.4 61 7 3.6 56 5 3.5
Digestibility (%) 66.7 3.4 69.4 3.7 64 1 3.6
BV (%) 61.3 4.1 71.1 2.7 80 1 2.5
NPU (%) 40.5 2.7 49.2 3.0 51 3 3.1

a Mean SE

TABLE 2. Summary of Blood Constituents in Study Phase 1

  Initial Energy intake (kcal/kg/day)
87 100 118
BUN (mg/dl) 8 7 0 7 a 6 1 0.6 5 4 0 4 3 7 0 5
Albumin (g/dl) 4 2 0.06 4.0 0.08 4 1 0 09 3 9 0 06
Total protein (g/dl) 6 8 0 2 6 6 0.1 6 8 0 1 6.4 0.1
AST (SF units) 355 44 373 43 375 1 5 438 20
ALT (SF units) 14 1 1 3 13.8 2.3 15 7 1.1 22 9 1 3

a Mean + SE

TABLE 3. Initial Characteristic of Six Young Male Children

Subject Age
(months)
Weight
(kg)
Height
(cm)
Weight for height
(% of Thai standard)
a
O.M. 12 8.7 67.8 100
A.B. 12 7.1 68.2 100
N.N. 8 7.3 68.1 90
D.S. 8 8.4 66.7 100
L.S. 9 7.9 67.2 90
K.P. 9 7.3 69.6 94
Mean SE 9.7 0.8 7.8 0.3 67.9 0.4 95.7 2.0

a From P Khanjanasthiti "The Anthropometric Nutritional Classification of the Infants and Preschool Children J Med Ass Thai., vol 60 Suppl 1 (1977)

Objective of Study Phase 2

The specific objective of this study was to determine whether a usual Thai weaning diet would provide adequate protein and energy for the needs of young children over a long-term period.

Experimental Design

Environment

The entire study was conducted in a metabolic ward of Ramathibodi Hospital, Mahidol University. The subjects were under close nursing and medical care and were also provided with adequate play facilities.

Subjects

Six normal, healthy, young male children aged 8-12 months were selected from an orphanage. They were studied after they had been rehabilitated for protein-energy malnutrition for eight weeks or longer and had reached normal weights for height. Their initial characteristics are shown in table 3. The means of age, weight and height were 9.7 months, 7.8 kg, and 67.9 cm, respectively. They were completely reexamined before the beginning of the study to ascertain that they were in good health.

TABLE 4. Proximate Analysis of Usual Thai Weaning Diet

Nutrients per 100 g
Protein (g) 1.70
Fat (g) 0.19
CHO (g) 12.98
Energy (kcal) 69.37
Moisture (g) 85.07
Ash (g) 0.25

Diets

The diets given to the children were largely based on rice and fresh-water fish, supplying protein in a ratio of 70:30 by weight. Fifty grams each of green leafy vegetables and ripe banana were also given daily. Fat intake was provided at a level of 10 per cent of energy intake. Neither vitamin nor mineral supplements were given to the children during the study. Table 4 shows the data for proximate analysis of the diets.

In order to maintain precision in dietary components, diets were prepared in advance for each month of dietary testing. The body weights of the subjects were taken into account for readjustments of protein and energy food contents.

Protein and energy intake levels were 1.7 g/kg/day and 100 kcal/kg/day, respectively. The usual Thai weaning diet is fed in three meals per day. with drinks containing sugar once or twice a day.

Duration

The entire study lasted for 120 days, or four months.

Measurements Taken

Figure 1 shows the experimental design of the study. The entire study was divided into four periods, with each period lasting for 30 days.

1. Diet samples - for each period, a one-day diet sample analysed twice a month for N, energy (bomb calorimetry), and fat.

2. Fasting blood samples were taken initially and at the end of each month for analyses of complete blood count, total serum protein, albumin, and urea N.

FIG. 1 Experimental Design for Long-term Study of Six Infants Given Usual Thai Weaning Food.

TABLE 5. Protein, Fat and Energy Intake of Six Infants Given Usual Thai Weaning Food for Four Months

Period Subject Energy Intake
(kcal/kg/day)
Protein Intake Fat Intake
(% energy) (g/kg/day) (% energy) (g/day)
1 O.M. 98.8 7.2 1.8 9.3 8.7
  A.B. 94.9 7.2 1.8 10 8 8.2
  N.N. 96.9 7.2 1.8 10.0 8.1
  D.S. 103.5 6.8 1.7 11.7 10.7
  L.S. 96.3 7.2 1.8 8.7 7.6
  K.P. 104.9 7.2 1.8 10.9 8.9
  Mean SE 99.2 1.7 7.1 0.1 1.8 0.02 10.2 0.4 8.7 0.4
2 O.M. 91.4 6.8 1.7 9.4 9.7
  A.B. 99.5 7.6 1.9 9.0 7.1
  N.N. 95.2 7.6 1.9 11.5 9.7
  D.S. 100.6 6.8 1.7 9 4 9.1
  L.S. 95.6 6.8 1.8 9.5 8.7
  K.P. 108.6 6.8 1.7 9.8 8.3
  Mean SE 98.5 2.4 7.1 0.6 1.8 0.04 9.8 0.4 8.8 0.4
3 O.M. 87.6 6.0 1.5 8.8 9 3
  A.B. 95.7 7.6 1.9 9.6 7.8
  N.N. 94.0 7.2 1.8 10.5 8.2
  D.S. - - - - -
  L.S. 94.7 6.8 1.7 10.0 9.3
  K.P. 98 7 6.8 1.7 10.9 9.4
  Mean SE 94.1 1.8 6.8 0.3 1.7 0.1 10.0 0.4 8.8 0.3
4 O.M. 98.1 6.4 1.6 8.6 9.4
  A.B. 82.4 6.0 1.7 8.8 7.4
  N.N. 94.0 6.8 1.7 9.5 8.3
  D.S. - - - - -
  L.S. 94.7 6.4 1.6 10.1 9.6
  K.P. 97.2 6.4 1.6 9.9 8.7
  Mean SE 93.2 2.8 6.4 0.1 1.6 0.1 9.4 0.3 8.7 0.4

Statistical analysis:

Period 1 vs. 2 NS NS WS
Period 2 vs. 3 P < 0.025 P<0.05 NS
Period 3 vs. 4 NS NS NS
Period 1 vs. 4 NS P < 0.05 NS

TABLE 6. N Balance in Six Infants Given Usual Thai Weaning Food for Four Months

Period Subject
Number
Energy intake
(kcal/kg/day)
N balance (mg/kg/day)
N intake Urinary N Faecal N Apparent
N balance
1 6 99.21.7 282.8 7.5 92.3 6.2 90.510.0 100.1 8.4
2 6 98.52.4 287.3 9.1 101.2 6.8 101.65.6 84.5 11.4
3 5 94.11.8 273.012.7 111.412.8 90.54.6 71.313.1
4 5 93.22.8 255.4 4.3 123.1 7.1 86.1 3.2 46.1 5.7

Statistical analysis.

Period 1 vs. 2 NS NS NS NS NS
Period 2 vs. 3 P < 0.025 P < 0 05 NS P < 0 05 NS
Period 3 vs 4 NS NS NS NS NS
Period 1 vs 4 NS P < 0.05 P < 0 05 NS P < 0.025

3. 24-hour urine samples were collected during days 7, 14, 21, and 27 to 30 of each period for analyses of total N and creatinine (see fig. 1).

4. Three-day faecal collections were taken at the end of each period during days 27 to 30 of the month for analyses of N, energy, and fat.

5. Body weights were measured daily before breakfast, and other anthropometric measurements, including height, head, and left mid-arm circumference, and tricep skin-fold thickness, were measured every week.

Physical activities

The subjects were allowed normal activities, which included playing on the floor of the metabolic ward. Toys and mobile units were provided. Children were put on metabolic beds during the period of urine collection.

Main Results

continue


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