1134 The American Journal of Clinical Nutrition 29: OCTOBER 1976, pp. 1134-1141. Printed in U.S.A.
Breast milk composition in Ethiopian and
Swedish mothers. II. Lactose, nitrogen, and protein contents13 Bo L#{246}nnerdal,4 Ph.D., Elisabeih Forsum,5 Ph.D., Mehari Gebre-Medhin,6 M.D., M.P.H. and LeijHarnbraeu.s,7 M .D. ABSTRACT Breast milk from underprivileged and privileged Ethiopian mothers was collected at different stages of lactation and analyzed for total nitrogen, nonprotein nitrogen, lactose, and individual milk proteins (lactoferrin, is-lactalbumin, serum albumin, IgG and 1gM). These values and the milk volume of one meal were compared to corresponding results from well-nourished Swedish mothers. No significant differences between the levels of these constituents or the milk volumes were found between the two groups of Ethiopian mothers. W hen comparison was made be- tween breast milk from these two Ethiopian groups and the Swedish group, the former two showed significantly higher values for the iron-binding protein lactoferrin. Am.J. C/in. Nutr. 29: 1134- 1141, 1976. Human milk is the first food after birth for the majority of children. In many countries, especially the developing ones, it is the only food up to the age of 4 to 6 months. Breast milk is by nature extraordinarily well adapted to the needs ofthe infant and its superiority in feeding infants has been undisputed through- out the years. The estimated requirements of infants for proteins (1) and many other nutrients are, therefore, based on the com- position of breast milk. Breast-fed infants are less liable to infections than bottle-fed infants (2). In areas where nutritionally adequate supplementary foods are scarce breast milk is a valuable supplement during the second year of life. Concomitant with the increased availabil- ity and use of artificial formulae a declining trend in the frequency and duration of breast feeding has been observed in most pants of the world in recent years (3, 4). Although nutri- tionally adequate for the maintenance of growth these artificial substitutes lack some of the unique features of human milk. Breast milk, for example, contains substantial amounts of secretory IgA (5) and the iron- binding protein lactofennin (6) both of which are known to enhance resistance to infection. Breast milk also contains comparatively high levels of the enzyme lysozyme (7), which is capable of degrading bacterial cell wall, and relatively lange amount of a B52-binding pro- tein that has been shown to compete with B12-removing microorganisms thereby acting bacteniostatically (8). Another unique feature is the pattern of essential amino acids and the high content of nonprotein nitrogen (NPN) (9). It has been proposed that the quantity and quality of breast milk are adversely affected by maternal undernutrition but hitherto con- ducted studies are inconclusive. Gopalan (10) showed that the growth pattern of breast-fed infants belonging to undernourished mothers was satisfactory, suggesting adequate milk production. These results have subsequently been confirmed by other authors (Il, 12). In From the Institute of Nutrition, University of Upp- sala, Box 551, S-751 22 Uppsala, Sweden and the Ethiopian Nutrition Institute, Addis Ababa. 2This paper is also listed as no. III in the series The protein content of human milk published by the Insti- tute of Nutrition, Uppsala. Financial support was provided by the Swedish International Development Authority (SIDA) and the Swedish Medical Research Council (MFR) (project no. B76- 19X-4722-Ol. Assistant Professor of Nutrition. Assistant Professor of Nutrition. 6 Director, Ethiopian Nutri- tion Institute. Professor of Nutrition, Institute of Nutrition, Uppsala.
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BREAST MI L K COMPOSI TI ON 1135 contrast, some investigators have shown re- duced lactation performance in malnourished women (13, 14). Of special interest are the observations made by Deb and Cama (15) and of Lindblad and Rahimtoola (16), indi- cating impaired protein quality in the milk from severely malnourished mothers. The present study describes the protein composition of milk from mothers belonging to two different socioeconomic groups in Ethiopia. The results obtained are compared to normal data, reported earlier (17), which are based on milk samples collected from well-nourished Swedish mothers. M aterial Ethiopia The Ethiopian mothers were all recruited from a comprehensive study of the nutritional aspects of preg- nancy and lactation directed by Ethiopian Nutrition Institute (ENI). They represent two different socioeco- nomic groups. The nonprivileged group, comprising 78 mothers, w as selected from a m unicipal antenatal clinic in the Kirkos area of Addis Ababa. All the mothers were illiterate and had a family income of less than Eth $100 (Eth $1.00 = US $0.50). This group consumes a m onotonous diet with an average dietary protein and energy intake less than 60% of that recommended by FAO/W HO. Of the protein 97% is of plant origin. In effect, this diet is inadequate in all respects except for iron and thiamine (18). The privileged group, comprising 26 mothers. were recruited from an antenatal clinic for paying patients in the D epartm ent of O bstetrics and G ynaecology, A ddis Ababa University (formerly Haile Selassie University). Both the mothers and their spouses in this group had com pleted at least secondary school education and their family income was generally above Eth $600 per month. The dietary intake of energy and nutrients in this group was ample (18). The study was conducted during the period April to S eptem ber 1973. Healthy, lactating mothers without glucosuria or albuminuria, with normal blood pressure, and having no clinical symptoms of acute or chronic disease were selected consecutively from both clinics. Further data related to dietary intake, hem atological, and biochem ical characteristics in the third trimester of pregnancy are presented elsew here (18, 19; M . G ebre- M edhin et al., m anuscript in preparation). Sweden The Swedish group of mothers were all healthy and socioeconomically and educationally privileged. In com- mon with practically 100% of contemporary Swedish m others they all gave birth to their babies in a hospital and stayed in the w ard during S to 7 days after the delivery. M ethods Milk collectioti Ethiopia. Samples were collected in the morning at a special clinic at ENI. Mothers were instructed not to breast-feed their children after midnight prior to milk collection. The milk samples were taken after overnight fasting (for the purpose of blood collection). Both breasts were emptied using an electric pump (Sister Majas breast pump; Pump Egnell AB, Trollh#{228}ttan, Sweden), of the type used in Sweden for many years. Pumping was continued until milk no longer appeared in jets but merely in a trickle or drop-wise. All samples were collected by one person and, after registration of volume, were placed in a deep freeze at -20 C within less than I hr after collection. The samples were subsequently transported in a frozen condition to Sweden by direct flight in special containers with dry ice and stored at -70 C. Sweden. Milk samples from the second meal on the 4th and 5th day post partum were collected by a nurse while the mother was still in the maternity ward. Mothers whose children were 3 weeks old or older were visited in their homes by a nurse. The day before the samples were collected, the mother was instructed on how to weigh the child before and after each feeding throughout 24 hr starting early in the morning. On the sampling day. the nurse visited the mother at the second meal of the day when the infant was allowed to nurse on one breast while the other was emptied by pumping. After mixing, a 20-mI aliquot was taken for analysis and the rest of the milk was offered to the child. In some instances samples were kept in an ordinary refrigerator for up to I week before being placed in a deep freeze at -70 C. Further details regarding the clinical material and sampling procedures in both the Ethiopian and Swedish groups are found in the first of these series of investiga- tions (20). A nalvtical procedures Methods for estimation of total nitrogen, NPN, lac- toferrin, serum albumin, a-lactalbumin, lgG, 1gM, and lactose are given in a companion paper. Information regarding the statistical evaluation can also be found in this paper (17). R esults The nitrogen distribution, lactose concen- tration, and milk volume of the first meal of the day for each period of lactation for Ethiopian and Swedish mothers are shown in Table 1 . The individual values of total nitno- gen and nonprotein nitrogen obtained from the Ethiopian mothers are compared with the Swedish normal material in Figure 1. The total nitrogen concentration decreases rapidly during the first 20 days of lactation to reach the level of mature milk. As shown in Table I the nitrogen concentration in one of the
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To t al n i t r o g en #{149}non-pnviteged 0 pr i v i Le ge d *non-priviteged Non-protein nitrogen A priviLeged 8 2 O 1.0 .. S#{ 149} .. 8 #{149} S. 2 I I t . , I . . I I 3 6 L ONNERDAL ET AL . FIG. I . Concentrations of total nitrogen and nonprotein nitrogen in milk from Ethiopian mothers belonging to two socioeconomic groups compared to milk from well-nourished Swedish mothers. S olid line in the middle of striped area = regression line for the Swedish material. S triped area = tolerance interval to the regression line. T ABL E I Nitrogen distribution, lactose concentration, and milk volume (first meal of the day)a Months post pa r t um 0-0.5 0.5 I .5 I .5 3.5 3.5 6.5 6.5 To t al n i t r o g en (mg/mI) Eth. nonprivileged Eth.privileged Swe. privileged 3.14 0.83 3.05 0.59 2.50 O.l7 2.89 0.59 1.93 0.24 1.77 0.33 1.97 0.29 1.61 0.21 1.69 0.30 1.48 0.17 1.74 0.30 NPN (mg/mI) Eth. nonprivileged Eth. privileged Sw e. p r i v i l eg ed 0.46 0.06 0.53 0.09 0.43 0.05 0.46 0.08 0.46 0.03 0.36 0.05 0.41 0.05 0.41 0.04 0.34 0.05 0.38 0.07 0.33 0.05 Lactose ( % w/v) Eth. nonprivileged Eth. privileged Sw e. p r i v i l eg ed 6.46 0.89 5.93 0.58 7.42 0.51 6.60 0.63 7.02 0.56 7.43 0.48 7.64 0.16 7.33 0.47 7.49 0.31 7.64 0.35 7.78 0.48 Mi l k v ol ume ( ml ) Et h. nonpr i v i l e ge d Et h. pr i v i l e ge d Swe . pr i v i l e ge d 1 9 7 94 1 5 7 131 169 91 1 2 4 26 153 66 126 61 145 4 4 188 8 8 2 0 6 7 0 1 0 9 72 n Et h. nonpr i v i l e ge d Et h. pr i v i l e ge d Swe. privileged S 15 3 I S 12 1 4 S 12 2 6 11 4 5 2 Val u es ar e m ean SD. Significantly higher than the corresponding value from Swedish mothers (0.001 < P < 0.01). cSignificantly higher than the corresponding value from Swedish mothers (0.01 < P < 0.05). d5ignificantly higher than the corresponding value from Swedish mothers (P 0.001). periods of lactation was significantly higher in the two Ethiopian groups as compared to the Swedish group. The nitrogen concentra- tion in milk from nonpnivileged Ethiopian mothers seems to increase after l70 days of lactation (Fig. 1). This apparent increase is not significant if the whole period from 20 days is considered. The concentrations of nonprotein nitrogenous substances are more constant throughout the lactation period. The concentrations of the individual human whey proteins are shown in Table 2. The individual values obtained from the Ethi- opian milks as well as the statistical represen-
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tation of the Swedish normal material are shown i n Figures 2 to 5. The milk-specific proteins a-lactalbumin and lactoferrin as well as 1gM exhibit similar variations in concen- TABLE 2 Concentrations of individual human whey proteins tration with time as that of nitrogen. The concentrations of the nonspecific milk pro- teins, serum albumin and IgG, are fairly constant throughout the lactation period. aValues are mean SD. bsignificantly higher than the corresponding value from Swedish mothers (0.001 < P. < 0.01). cSignificantly higher than the corresponding value from Swedish mothers (P 0.001). dSig nificantly higher than the corresponding value from Swedish mothers (0.01 < P < 0.05) Lactose #{149} non-priviLeged 0 privaeged i : : , : : a non-privileged A privileged I - a I a a FIG. 2. Concentrations of lactose and a-lactalbumin in milk from Ethiopian mothers belonging to two socioeco- nomic groups compared to milk from well-nourished Swedish mothers. S o lid lin e in the middle of s trip e d a re a = regression line for the Swedish material. S trip e d a re a = tolerance interval to the regression line. BREAST MI LK COMPOSI TI ON I 137 Monthspost partum 0--0.5 0.5 .5
. 5 3 . 5 3 . 5 -6 . 5 6.5 Lactoferrin Eth. nonprivileged Eth. privileged S w e . p riv ile g e d 3.75 0.93 3.53 0.54 2.64 0.06 3.37 0.71 1.94 0.38 mg/nil 1.67 0.51 1.72 0.67 1.89 0.51 1.65 0.29 1.39 0.26 1.48 0.57 a -La c t a l- bumin Eth. nonprivileged Eth. privileged S w e . p riv ile g e d 3.70 0.31 3.62 0.59 3.58 0.28 3.72 #{216},35d 3.26 0.47 2.76 0.29 2.65 0.35 2.92 0.39 2.78 0.49 2.68 0.59 2.58 0.37 S e ru m albumin Eth. nonprivileged Eth. privileged S w e . p riv ile g e d 0.42 0.09 0.39 0.06 0.43 0.02 0.47 0. I I 0.41 0.07 0.36 0.07 0.36 0.08 0.38 0.06 0.39 0.04 0.38 0.04 0.34 0.07 1 g M Eth. nonprivileged Eth. privileged S w e . p riv ile g e d 0.087 0.070 0.156 # {2 1 6 }, # {2 1 6 }9 5 c 0.094 0.090d 0.045 0.025 0.027 0.023 0.029 0.0 1 3 0.030 0. 180 0.033 0.028 0.021 0.015 0.016 0.012 0.040 0.049 Ig G Eth. nonprivileged Eth. privileged S w e . p riv ile g e d 0.068 0.042 0.032 0.023 0.045 0.012 0.067 0.020 0.027 0.018 0.033 0.01 1 0.048 0.024 0.046 0.012 0.027 0.01 I 0.027 0.014 0.039 0.029
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I L a c t o f e r r i n n o n - p r i v i L e g e d apriviLeged Serum atbumm #{149} non-priviLeged . priviLeged #{149} . a I I #{149}1 I I # {1 4 9 }1 FIG. 3. Concentrations of lactolerrin and serum albumin in milk from Ethiopian mothers belonging to two socio- economic groups compared to milk from well-nourished Swedish mothers. S olid line in the middle of striped area = regression line for the Swedish material. S triped area = tolerance interval to the regression line. IgG non-priviLeged 0 priViLeged FIG. 4. Concentrations of immunoglobulin G in milk from Ethiopian mothers belonging to two socioeconomic groups compared to milk from well-nourished Swedish mothers. S olid line in the middle of striped area = regression line for the Swedish material. S triped area = tolerance interval to the regression line. 1138 LONNERDAL ET AL. Figure 2 shows the concentrations of lactose together with serum albumin. Lactofennin and one of the major human whey proteins, decreases rapidly during the first days of a-bactalbumin. The concentration of lactose lactation to reach a constant level after about increases with the time of lactation while that 20 days. A significantly higher concentration of a-lactalbumin decreases. of lactoferrin was found in Ethiopian breast The concentration of another major whey milk as compared to Swedish milk during the protein, lactofennin, is shown in Figure 3 period 0.5 to 1.5 months (Table 2). This
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#{149} non-priviLeged 1 g M #{149} priviLeged BREAST MI L K COMPOSI TI ON 1139 FIG. S. Concentrations of immunoglobulin M in milk from Ethiopian mothers belonging to two socioeconomic groups compared to milk from well-nourished Swedish mothers. Solid line in the middle of striped area = regression line the Swedish material. Striped area = tolerance interval to the regression line. difference seems to be responsible for the higher nitrogen concentration found in Ethi- opian milk. However, this difference becomes less pronounced as lactation proceeds. The concentration of serum albumin is constant throughout lactation and this is probably explained by a passive transport of serum albumin from the blood to the breast milk. Figures 4 and 5 show the concentrations of the immunoglobulins IgG and 1gM. The concentration of 1gM was significantly higher in the nonprivileged group of Ethiopian mothers than in the Swedish group. In the Ethiopian privileged group the IgG concen- tration was higher than in the other groups. Generally no significant differences in the concentrations of individual proteins between the two Ethiopian socioeconomic groups were obtained, although they all were somewhat higher in the privileged group (Table 2). D iscussion According to earlier studies little diurnal variation in lactose (21) and protein (22) contents have been found in breast milk. Similanily the contents of different nitrogen fractions as casein-N, albumin-N, NPN, and amino acid-N (22) have been reported to remain comparatively constant through the day. These facts seem to justify the compani- son of milk obtained from the second meal of the day from Swedish mothers to milk obtained from the first meal of the day from Ethiopian mothers. However, no data are available regarding diurnal variation of mdi- vidual whey proteins. This is presently under study and a minor reservation should be made until complete information on this subject is obtained. The samples from the three groups differ with respect to the stage of lactation in which they were collected. In the privileged Ethiop- ian group lactation was terminated after only a few months and only samples from early lactation were obtained (Table 1). Current policy regarding maternity care does permit mothers to stay away from work at the most 4 to 6 weeks after delivery. Employed mothers are thus forced to replace breast milk with artificial formulas at an early stage. In the nonpnivileged group, on the other hand, few samples could be obtained from early lacta- tion since mothers traditionally stay indoors during the first 6 to 8 weeks after delivery. Most samples were collected after the second month of lactation (Table I). This has to be considered in the statistical comparisons
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1 1 4 0 L NNERDAL ET AL . betw een the tw o E thiopian groups. M ilk sam ples from the S w edish m others w ere collected during the first 6.5 m onths of lacta- tion m aking com parisons possible betw een the S w edish m aterial and the tw o groups of E thiopian m others. It has been pointed out by M . G ebre-M ed- hin et al. (20) that in this investigation the m ilk volum es of the first m eal of the day w ere not significantly different for S w edish and E thiopian m others (Table 1). H ow ever, this observation should be interpreted w ith reservation. It is reasonable to assum e that the use of a breast-pum p, w hich m ay have m eant a greaten stress to the E thiopian m others than to the S w edish ones, has led to a reduction in m ilk yield in the previous group. O n the other hand, the m ilk volum e of the E thiopian m others m ay also have been influenced in a positive direction as a num - ber of these m others had to w ait several hours after the usual nursing tim e before evacua- tion of the breasts took place. This could pos- sibly provide partial explanation for the fact that the m ilk volum e in the group of non- privileged E thiopian m others w as com parable to that observed in their privileged E thio- pian and S w edish counterparts. A difference in lactofernin concentration of breast m ilk from E thiopian and S w edish m others is evident. The lactofennin concentra- tion of the E thiopian sam ples is also m ark- edly above that of breast m ilk from Japanese m others reported by N agasaw a et al. (6). This finding is of considerable interest and m ay be connected to the very high iron content of the Ethiopian diet. In the highland population daily iron intakes of 300 to 500 m g are com m on because of the consum ption of the iron-rich cereal tefw hich is unique for E thi- opia (23). H ow ever, m ost of the iron is in the form of poorly absorbed com pounds (19, 24). Lactoferrin is know n to act bacteniostatically by retaining iron from iron-dem anding m i- croorganism s (25), although the m echanism s involved are not know n. Further investiga- tions are required to elucidate the relationship betw een iron intake and the lactofennin con- tent of breast m ilk. The high 1gM levels of the nonpnivileged group could probably be explained by the constant infections to w hich this group is exposed. H ow ever, since accurate m ethods are not currently available for determ ining the concentrations of the m ajor breast m ilk im m unoglobulins, secretory IgA and IgA , a com plete evaluation of the im m uno-defensive m echanism s involved cannot be perform ed. No overall differences w ere found betw een the E thiopian groups on the one hand and the S w edish group on the other w ith respect to the constituents analysed in the study. A s m others from such different incom e groups are know n to have m arkedly divergent die- tary intakes, it is rem arkable that the present investigation does not provide evidence that undernutrition has any pronounced effects on the quality of breast m ilk, at least in so far as protein and lactose are concerned. Lindblad and R ahim toola (16) studied the am ino acid com position of breast m ilk from undernour- ished m others from P akistan and E thiopia (selected from the sam e low socioeconom ic group as in this study). A m ong the P akistani m others the am ino acid pattern w as changed in a w ay that m ay indicate a change in protein com position (16). This change w as not ob- served in E thiopian m others (B . S . Lindblad, personal com m unication). H ow ever, it is con- ceivable that the P akistani m others w ere under m ore severe nutritional stress than the low socioeconom ic E thiopian group. In conclusion, this report seem s to lend support to the other studies that have show n that the quality and quantity of breast m ilk are not affected by m aternal m alnutrition. H ow ever, published inform ation in this field is lim ited and inconclusive. A fter all, it is the ability of the child under the prevailing condi- tions to optim ally utilize its m others m ilk that is of im portance. In view of this, further inform ation regarding the volum e of breast m ilk produced, as w ell as the quantity con- sum ed and utilized by the child is needed. Furtherm ore, these param eters need to be considered in relation to various ecological conditions and determ inants of child health and developm ent. W e are very grateful to D rs. A nders V ahlquist, Y ngve H ofvander, Francis K am w endo, S am R oss, and P rof. B o V ahlquist for the arrangem ent and realization of this joint study. The collaboration of the E N I M etabolic U nit S taff, in particular, S isters lngrid Tesfatsion and A m sale G ebre-Y ohannes and M iss B ent B org is gratefully acknow ledged. W e also w ish to thank M rs. E w a Johans- son for collecting the S w edish m aterial. For skillful
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BREAST MI L K COMPOSI TI ON 1 1 4 1 technical assistance we would like to express our grati- tude to Mrs. Annika JUgare and Mrs. Maivor Sj#{246}strand. The data program for the statistical evaluation was developed by Drs. Hans Falk and Torgny Groth. Drs. B Johansson and N. Harboe are thanked for gifts of lactoferrin standards. R eferences I. FAO/W HO Ad Hoc Expert Committee. In: Energy and Protein requirements. W orld Health Organiza- tion. Technical report series. No 522, pp. 56-58. WHO Ge n e v a , 1 9 7 3 . 2. MATA, 1. J., J. J. URRUTIA AND A. LEcIiTIG. Infection and nutrition of children of a low socio- economic rural community. Am. J. Clin. Nutr. 24: 249, 1971. 3. VAHLQUIST, B. Evolution of Breast Feeding in Europe. J. Trop. Paediat. Environ. Child Health I: 11, 1975. 4. BERG, A. The Nutrition Factor, W ashington: The Brookings Institution, 1973, p. 91. 5 . HANSON, L.-A., AND B. G. JOHANSSON. Immuno- logical studies of milk. In: Milk proteins. New York: Academic Press, 1970, pp. 45-123, vol. 1. 6. NAGASAWA, T., 1. KIVOSAWA AND K. KUW AI1ARA. Amounts of lactoferrin in human colostrum and milk. J. Dairy Sci. 12: 1651, 1972. 7. CHANDAN, R. C., R. M. PARR\ AND K. M. SHAHA- RI. Lysozyme, lipase and nibonuclease in milk of various species. J. Dairy Sci. 4: 606, 1968. 8. GULLBERG, R. Possible Influence of Vitamin B,,- Binding Protein in Milk on the Intestinal Flora in Breast-Fed Infants. Scand. J. Gastroenterol. 8: 497, 1973. 9. LONNERDAL, B., E. FORSUM AND L. HAMBRAEUS. The protein content of human milk I. A transversal study of Swedish normal material. Nutr. Rept.. Intern. 13: 125, 1976. 1 0 . GOPALAN, C. Protein intake of breast-fed poor In- dian infants. J. Trop. Pediat. 2: 89, 1956. 11. GOPALAN, C., AND B. BELAVADY. Nutrition and Lactation. Fed. Proc. 20 suppl. 7: 177, 1961. 12. VENKATACHALAM, P. 5. A Study of the Diet, Nutri- tion and Health of the People of the Chimbut Area. New Guinea; Monograph 4, Dept. Public Health, Govt. of Papua, 1962. 13. KARMARKAR, M. G. , AND C. V. RAMAKRISHNAN. Studies on human lactation. Acta Paediat. Scand. 49: 599, 1960. 14. SUKHATME, P. V., ANI) D. BASU. The present pattern of production and availability of foods in Asia. In: Proceedings of the First Asian Congress of Nutri- tion. Hyderabad: Nutrition Soc. India, 1972, p. 19. 15. DEB, A. K., AND H. R. CAMA. Studies on human lactation. Dietary nitrogen utilization during lacta- tion and distribution of nitrogen in mothers milk. Brit. J. Nutr. 16: 65, 1962. 16. LINDBLAD, B. S., ANt) R. J. RAHIMTOOLA. A pilot study of the quality of human milk in a lower socio-economic group in Karachi, Pakistan, Acta Paediat. Scand. 63: 125, 1974. 17. LONNERDAL, B., E. FORSUM AND L. HAMBRAEUS. A longitudinal study of the protein, nitrogen and lactose contents of human milk from well-nourished Swedish mothers. Am. J. Clin. Nutr. 29: 1 127, 1976. 18. GEBRE-MEDHIN, M., ANI) A. GOBEZIF. Dietary intake in the third trimester of pregnancy and birth weight of offspring among nonpnivileged and pnivi- leged women. Am. J. Clin. Nutr. 28: 1322, 1975. 19. GEBRE-MEDHIN, M., A. KILLANDER, B. VAHLQUIST AND E. W UHIB. Rarity of anemia of pregnancy in Ethiopia. Scand. J. Haematol. 16: 168, 1976. 20. GEBRE-MEDIIIN, M., A. VAIJLQUIST, Y. H0FvANu- ER, L. UPPSXLL ANt) B. VAHLQLIST. Breast milk composition in Ethiopian and Swedish mothers. 1. Vitamin A and fl-carotene. Am. J, Clin. Nutr. 29: 441, 1976. 21. MORRISON, S. D. Lactose. In: Human Milk; Yield Proximate Principles and Inorganic Constituents, Aberdeen: University Press, 1952, pp. 63-67. 22. MACY, I. G., B. Nisis, M. BROW N ANI) H. A. HUNSCHER. Human milk studies. VII. Chemical analysis of milk representative of the entire first and last halves of the nursing period. Amer. J. Diseases Childhood 42: 569, 1931. 23. ALMGXRD, G. High content of iron in tef, Eragrostis Abyssinica Linn and some other crop species from Ethiopia-a result of contamination. Ann. Agni. CoIl. Sweden 29: 215, 1963. 24. HOFVANDER, Y. Hematological investigations in Ethiopia with special reference to high iron intake. Acta Med. Scand. Suppl. 494, 1968. 25. BULLEN, J . J., H. J. ROGERS AND L. LEIGH. Iron-binding proteins in milk and resistance to Eschenichia coli infection in infants. Bnit. Med. J. 1: 69, 1972.
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The Conflict With Slavery and Others, Complete, Volume VII, The Works of Whittier: The Conflict With Slavery, Politicsand Reform, The Inner Life and Criticism by Whittier, John Greenleaf, 1807-1892