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This document presents the results of a study comparing the plasma osmolarity and levels of various biochemical parameters in healthy breastfed infants versus non-breastfed infants. The study found significantly higher levels of urea, sodium, potassium, total proteins and calculated plasma osmolarity in infants fed cow's milk or formula compared to breastfed infants. These differences indicate that non-breastfed infants have a higher renal solute load from alternative milks compared to the optimized nutrient balance provided by human breastmilk.
Thesis mphil
- 2. DIFFERENCES OF PLASMA OSMOLARITY IN HEALTHY BREASTFED AND NON BREASTFED INFANTS THESIS FOR M.Phil BIOCHEMISTRY BY DR. MUHAMMAD MUSTANSAR SUPERVISOR DR. ZAMIR AHMED PROFESSOR OF BIOCHEMISTRY SERVICES INSTITUTE OF MEDICAL SCIENCES, LAHROE INTRODUCTION Breastfeeding is the most natural and safest way to feed an infant. Breastfeeding provides a unique combination of proteins, lipids, carbohydrates, and minerals in quite appropriate amount. Human milk is a complex species-specific biological fluid for an infant with all nutritional needs for growth, development and biochemical equilibrium. The total protein content of human milk is the lowest among all mammals due to relatively slow growth of baby in human beings as compared to others. The concentration of minerals in human milk is about 1/3rd lower than in cow’s milk. This fact together with reduced protein content leads to a lower solute load, which is adequate for the infants developing kidney. (Lawrence 1994)
- 3. Devies and Saunders (1973) described that the renal solute load of cow‟s milk is considerably higher than that of breast milk. This is shown by high urea level in non-breast fed infants. The blood urea level in breast fed infants was 22mg/dl and that of non-breast infants was 47mg/dl calculated by „‟t‟‟ test for a sample of 61 infants. The sodium level in cow‟s milk is 3.6 times more than in human milk. For cows milk it is 22meq/L and for human milk is 7meq/L. Hyper natermia is associated with cows milk feeding. Experiments with newborn rats on high salt in takes have shown that hypertension can develop. (Ruth 1989) SOLUTE LOAD Composition of Human, Cow and Buffalo milk (100ml) NUTRIENT HUMAN COW BUFFALO Water g 85.7 88.0 84.0 Energy Kcal 70.0 61.0 97.0 Protein g 1.0 3.2 3.7 Fat g 4.4 3.4 6.9 Lactose g 6.9 4.7 5.2 Minerals g 0.20 0.72 0.79
- 4. Human milk provides optimal nutrition to infants and the appropriate balance of nutrients. The relatively low contents of minerals and protein in breast milk are adequate and present comfortable load to immature infant kidney. Human milk has relatively low sodium content allowing the fluid requirement of the infant to be met while not over taxing the kidneys. The osmolarity of human milk approximates to that of human serum and is 286m osmol/L, where as that for cows milk is 350m osmol/L. (Dale 1975) OPTIMAL NUTRITION Lonnerdal and Chen (1990) reported that formula-fed infants experience much higher plasma levels of most amino acids and consistently elevated blood urea levels. This implies a decreased efficiency of utilization of the protein in the formula. Different formulas with similar nitrogen concentration may contain dissimilar levels of true protein and non-protein nitrogen depending on their method of preparation. FORMULA FEEDING
- 5. AIMS AND OBJECTIVES The aim of the present study is to highlight the importance of mother’s milk and to discourage the usage of infant formula or cows/buffalos milk to an infant. The objectives are 1) To find out Serum Sodium, Potassium, Urea, Glucose, Total Serum proteins, Albumin, Globulin and A/G ratio (Albumin/Globulin) in breastfed and non-breastfed infants. 2) To determine plasma osmolarity and its comparison in breastfed and non-breastfed infants. In our society due to some social and cultural believes many mothers deprive their infants from the benefits of breastfeeding. The infants are fed by cow milk/buffalos milk purchased from the market. The infant formula milk is very costly and it is not possible for an average family to afford formula milk for an infant. The milk of cows and buffalos is having three times more proteins and 3-4 times more minerals as compared to the human milk. By feeding the milk of cow or buffalo/infant formula the infant is being loaded with more proteins and minerals, which can lead to high osmolarity, sodium, potassium, urea and serum proteins. The effects of these may not be visible immediately but may have consequences in later life. IMPORTANCE OF STUDY
- 6. MATERIALS AND METHODS STUDY DESIGN This was a descriptive cross-sectional study, which conducted on infants up to the age of 6 months being fed either mother’s milk, infant formula or cow/buffalo milk. SAMPLE SIZE A total sample of one hundred and eighty infants up to the age of 6 months were taken included. The sample were divided into three groups of sixty infants in each group. Group I:- Infants on mother’s milk. Group II:- Infants on formula milk. Group III:- Infants on cow/buffalo milk. SELECTION CRITERIA Infants up to the age of 06 months Either sex The Infants being fed milk. No weaning diets. Taking the particular milk for at least one month. Appropriate mile stone EXCLUSION CRITERIA Infants of less than one month or more than six months age Low birth weight infants Premature delivered infants Infants having gross congenital anomaly of GIT, Urinary tract and Cardiovascular System The infants having history of recurrent chest infection Infants on weaning diets Infants on mixed feeding
- 7. SAMPLE COLLECTION LABORATORY ANALYSIS Serum Sodium Level (Gowenlock, 1988) Serum Potassium Level (Gowenlock, 1988) Serum Urea Level (Marsh et al, 1965) Serum Glucose Level (Trinder, 1969) Total Serum proteins Level (Reinhold, 1953) Serum Albumin Level (Doumas et al, 1971) Serum Globulin Level (Total proteins – serum albumin level) A/G ratio Serum Albumin Level (Sood, 1994) Serum Globulin Level Plasma osmolarity (estimated) (Vapro Osmometer) Plasma osmolarity (calculated) (Robert, 1999) CALCULATION OF OSMOLARITY Osmolarity mosm/l = 2{Na+meq/l+K+ meq/l}+ (glucose) mg/dl + (BUN) mg/d 18 2.8
- 8. Appendix I PROFORMA Name of Child:Name of Father:Age:Address: Feeding Practice for the last one month: Only on Mother’s Milk On Buffalos Milk On Cows Milk On infant formula Name: ________________ Preparation: ___________Dilution or without DilutionDilution or without DilutionEXAMINATION OF INFANT LABORATORY REPORT Serum Sodium Level _______________meq/l Serum Potassium Level _______________meq/l Serum Urea Level _______________mg/dl Serum Glucose Level _______________mg/dl Total Serum proteins Level _______________g/dl Serum Albumin Level _______________g/dl Serum Globulin Level _______________g/dl A/G ratio _______________ Plasma osmolarity (estimated) ___________m osml/l Plasma osmolarity (calculated) __________m osml/l STATISTICAL METHODS STATISTICAL METHODS (Danial WW 2005) For the purpose of statistical analysis the information recorded on the proforma was stored in spreadsheet of SPSS software (Statistical Program for Scientific Studies) version 13. The data were analyzed as follows. 1. Description 2. Analysis ANOVA One Way. 3. Inference Post HOC Test.
- 9. AGE DISTRIBUTION OF BRESTFED AND NON BREASTFED INFANTS Mode of Feeding No. of Cases (n) Mean Age in Months Standard Deviation (SD) Formula Milk 60 4.03 1.657 Mother Milk 60 3.54 1.650 Fresh Milk 60 4.55 1.489 WEIGHT DISTRIBUTION OF BREASTFED AND NON BREAST FED INFANTS Mode of Feeding No. of Cases (n) Mean Weight in Kg Standard Deviation (SD) Formula Milk 60 4.58 0.86 Mother Milk 60 4.86 1.16 Fresh Milk 60 4.84 0.80
- 10. BLOOD GLUCOSE LEVEL IN BREASTFED AND NON-BREAST FED INFANTS Mode of Feeding No. of Cases (n) Mean Glucose Level in mg/dl Standard Deviation (SD) Formula Milk 60 90.47 7.0 Mother Milk 60 95.30 9.2 Fresh Milk 60 95.43 9.3 BUN AND BLOOD UREA LEVEL IN BREASTFED AND NON BREAST FED INFANTS Mode of Feeding No. of Cases (n) Urea mg/dl BUN mg/dl Mean SD Mean SD Formula Milk 60 22.81 4.02 10.66 1.88 Mother Milk 60 10.45 3.84 6.14 1.40 Fresh Milk 60 15.35 7.22 7.14 3.36
- 11. SERUM SODIUM AND POTASIUM IN BREASTFED AND NON-BREAST FED INFANTS Mode of Feeding No. of Cases (n) Na+ m mol/L K+ m mol/L Mean SD Mean SD Formula Milk 60 147.43 3.87 5.15 0.49 Mother Milk 60 138.43 4.20 4.83 0.34 Fresh Milk 60 146.5 5.17 5.27 0.55 COMPARISION OF CALCULATED AND ESTIMATED PLASMA OSMOLARITY IN BREASTFED AND NON BREAST FED INFANTS T TEST DISTRIBUTION Mode of Feeding T d.f Sig (2Tailed) Mean Difference 95 % CI of the differences Lower Bound Upper Bound Formula Milk 5.137 118 0.000 7.43 4.568 10.299 Mother Milk 5.158 118 0.000 5.23 3.214 7.220 Fresh Milk 4.296 118 0.000 6.90 3.764 10.202
- 12. CALCULATED AND ESTIMATED PLASMA OSMOLARITY IN BREASTFED AND NON BREAST FED INFANTS Mode of Feeding No. of Cases (n) Calculated Osmolarity m osm/L Estimated Osmolarity m osm/L Mean SD Mean SD Formula Milk 60 314.05 8.48 306.62 7.32 Mother Milk 60 291.92 6.31 286.70 4.63 Fresh Milk 60 310.30 9.09 303.32 8.71 COMPARISION OF CALCULATED AND ESTIMATED PLASMA OSMOLARITY IN BREASTFED AND NON BREASTFED INFANTS (ANOVA) Variable s Differen ce Sum of Squares d.f Mean Squares VR P Value Calculate d Osmolarit y Between Groups 16837.87 8 2 8418.939 129.872 0.000 Within Groups 11474.03 3 177 64.825 Total 28311.91 1 179 Estimate d Osmolarit y Between Groups 13673.54 4 2 135.788 13.847 0.000 Within Groups 8911.767 177 50.349 Total 22585.31 1 179
- 13. COMPARISION OF CALCULATED AND ESTIMATED PLASMA OSMOLARITY IN BREASTFED AND NON BREASTFED INFANTS POST HOC TEST Variable Group Code Group Code Mean Difference * P Value 95 % Confidence Interval Lower Bound Upper Bound Calculated Osmolarity 1 2 22.1333* 0.000 19.2324 25.0343 3 3.7500* 0.012 0.8491 6.6509 2 1 - 22.1333* 0.000 - 25.0343 - 19.2324 3 - 18.3833* 0.000 - 21.2843 - 15.4824 3 1 - 3.7500* 0.012 - 6.6509 - 0.8491 2 18.3833* 0.000 15.4824 21.2843 Estimated Osmolarity 1 2 19.9167* 0.000 17.3601 22.4733 3 3.3000* 0.012 0.7434 5. 8566 2 1 - 19.9167* 0.000 - 22.4733 - 17.3601 3 - 16.6167* 0.000 - 19.1733 - 14.0601 3 1 - 3.3000* 0.012 - 5.8566 - 037434 2 16.6167* 0.000 14.0601 19.1733 *Mean Difference is significant at the 0.05 level. Code Group 1 Formula Milk Code Group 2 Mother Milk Code Group 3 Fresh Milk CONCLUSION The present study has clearly highlighted that “Mother‟s Milk is the best milk”. The serum levels of BUN, urea, total proteins, albumin, sodium, potassium and plasma osmolarity are all significantly low as compared to the infants on bottle feeding taking infant formula or fresh milk. The bottle feeding either by fresh milk or by the cheaper partially modified formula predispose the infants developing systems to high solute loads and osmolarity which may not have immediate consequences but may have in later life like childhood obesity and raised blood pressure.