Effects of Age and Parity Associated With Protein Energy Malnutrition (PEM) On Some Biochemical Parameters of Some Pregnant Women in Enugu Metropolis of Nigeria

 

1Ikeyi A.*, 2Alumanah E.O. and 2Joshua P.E.

1Dept. of Science Laboratory Technology (Biochemistry Option), Institute of Management and Technology, Enugu, Nigeria.

2Department of Biochemistry, University of Nigeria, Nsukka, Nigeria

 

 

ABSTRACT:

This study aims at investigating the effects of age and parity (number of children had by mother) associated with protein energy malnutrition (PEM) on some biochemical indices in pregnant women in Enugu metropolis of Nigeria. Serum total protein, albumin, urea, total cholesterol, creatinine and calcium were evaluated in three groups of female subjects as part of an investigation on the biochemical changes associated with protein energy malnutrition (PEM) in pregnant women. The first group were 52 pregnant women with low total protein (<52g/l), the second group were 50 pregnant women with normal total protein (>52g/l) while the third group were 50 non-pregnant, non-lactating, apparently healthy women with normal total protein (>63kg). All the subjects were resident in Enugu metropolis and aged between 20 to 40 years. The pregnant subjects were in different gestational stages of pregnancy, having different parity and attending the antenatal clinic of Parklane Specialist Hospital, Enugu. The results show that parity (i.e. the number of children had by mother) correlated negatively (p<0.05) with serum total protein, urea, total cholesterol, creatinine and calcium.

 

KEYWORDS: Age, Parity, Protein Energy Malnutrition (PEM), Pregnancy, Biochemical.

 

 

INTRODUCTION:

Malnutrition continues to be a major health burden in developing countries. It is globally the most important risk factor for illness and death with hundreds of millions of pregnant women and young children particularly affected1. Poor nutrition in pregnancy in combination with infections is a common cause of maternal and infant mortality and morbidity, low birth weight and intrauterine Growth Retardation (IUGR)2. In Nigeria, maternal death per 100,000 births is put at 800 while percentage low birth weight stands at twenty3.

 

It has long been recognised that inadequate food intake produces weight loss and growth retardation and, when severe and prolonged leads to body wasting and emaciation. Nutritional disorders date back as far as the sixteenth century and were recognised under such names as maries, atrophy, atrepsy, etc. In Ireland, William Benette of the society of friends described hunger oedema in March 1847 as that “horrid disease – the result of long continued famine and low living in which limbs and body swell most frightfully”. Specific interest in what we know today as Protein Energy Malnutrition began early in this century4.

Low birth weight babies have increased risk of mortality, morbidity and development of malnutrition. Children who suffer from malnutrition are more likely to have slowed growth, delayed development, difficulty in school and high rates of illness and they may remain malnourished to adulthood5,6. IUGR is associated with poor cognitive and neurological development for the infant and in adulthood, susceptibility to cardiovascular disease, diabetes and renal disease7.

 

Malnutrition remains one of the world’s highest priority health issues not only because its effects are so widespread and long lasting but also because it can be eradicated. Eradication is best carried out at the preventive stage. Hence, the need to identify groups of pregnant women at greater risk of developing PEM. Such high-risk groups can be targeted in any planned intervention programme8.

 

Pregnancy is a normal physiological process associated with major alterations affecting every maternal organ, system and metabolic pathway9. This physiological process results in increased plasma volume and red blood cells, decreased concentration of circulating nutrient-binding proteins and other micronutrients10.

This study aims at investigating the effect of parity (number of children had by mother) associated with protein energy malnutrition (PEM) on some biochemical indices in pregnant women in Enugu metropolis of Nigeria.

 

MATERIALS AND METHODS:

Materials:

Study Subjects:

Three groups of female volunteers were involved in this study. The first group were 52 pregnant women with low total protein (<52g/l), the second group were 50 pregnant women with normal total protein (>52g/l), while the third group were 50 non pregnant, non lactating, apparently healthy women (>52g/l). All the subjects were between 20-40 years of age. All the pregnant subjects were attending antenatal clinic of Parklane Specialist Hospital, were in different gestational stages of pregnancy with different parity. Subjects with complications such as hypertension, diabetes, HIV/AIDS on admission were excluded.

 

Methods:

Collection of Blood Samples and Preparation of Serum:

Blood (2.5mls) was collected from each volunteer by venepuncture and delivered into clean and duely labelled specimen containers. The blood was allowed to clot and then centrifuged at 5000 rpm for 10 minutes. Using a Pasteur pipette serum was separated from the cells and delivered into a clean and dry bottle. It was stored frozen at –200C until it was used.

 

Preparation of Reagents:

The reagents used were high performance enzymatic colorimetric commercial analytical kits (Biosystems Reagents and Instruments, Barcelona, Spain). These commercial kits were purchased and used according to the manufacturer’s direction for all the parameters assayed.

 

Determinations of Sample:

Determination of serum total protein:

Principle: The serum total protein was determined using the Biuret method. The protein in the sample reacted with copper II ion in alkaline mediun forming a coloured complex that was measured spectrophotometrically Determination of serum protein was carried out according to the method of Gornall et al. [11] and Berg et al.12.

 

Determination of serum albumin concentration:

The measurement of serum albumin was by the quantitative method using (B.C.G) Bromocresol green. This method is based on the quantitative binding of albumin in the sample to the indicator 3, 3’, 5, 5’ – tetra bromo-m cresol sulphonephthalein (BCG) (Bromocresol Green). This will form a complex known as the Albumin – BCG complex. This complex absorbs maximally at 578nm, the absorbance (A) being directly proportional to the concentration of Albumin in the sample.

The determination of serum albumin concentration was done according to the method of Doumas et al.13.

 

Determination of serum Globulin concentration:

The determination of serum albumin concentration was done according to the method of Doumas et al.13.

 

Determination of Serum Total Cholesterol:

The total cholesterol concentration of the test individuals was determined using cholesterol enzymatic endpoint method. Cholesterol is determined after enzymatic hydrolysis and oxidation. The indicator quinoneimine is formed from hydrogen peroxide and 4-aminoantipyrine in the presence of phenol and peroxide. The free and esterified cholesterol in the sample originates by means of the coupled reactions described below, a coloured complex is formed that can be measured spectrophotometrically.

 

The concentration of serum total cholesterol was determined according to the methods of Allain et al.14 and Meiattini et al.15.

 

Determination of serum urea concentration:

The urea concentration of the test individuals were determined by the urease – salicylate enzymatic method. This uses the enzyme urease to hydrolyze urea. The ammonia produced reacts with alkaline hypochlorite and phenol in the presence of a catalyst to form indophenol. The coloured complex is measured spectrophotometrically. The determination of serum albumin concentration was done according to the method of Cheestbrough16.

 

Determination of serum creatinine concentration:

Creatinine in the sample reacted with picrate in alkaline medium to form a coloured complex. The coloured complex is measured spectrophotometrically. It is measured within a short period to avoid interference from non creatinine substances.

The serum concentration of serum albumin concentration was done according to the methods of Bartels and Bohmer17 and Fabiny and Ertingshausen18.

 

Determination of serum calcium concentration:

Calcium in the sample reacted with methylthymol blue in alkaline medium to form a coloured complex. The coloured complex was measured spectrophotometrically. Hydroxyquinoline was included in the reagent to avoid magnesium interference.

The determination of serum albumin concentration was done according to the methods of Gindler and King19 and Barnett et al.20.

 

Determination of serum Globulin concentration:

The determination of serum albumin concentration was done according to the methods of Gindler and King [19] and Barnett et al.20.

 

Statistical Analysis:

The data were analysed using the SPSS package of windows version 11.00 (SPSS Corporation, IL). Differences between the means were separated and analysed for statistical difference using the one way ANOVA while correlations between parameters were calculated using the Pearsons correlation coefficient. Difference in means with p values < 0.05 were accepted as significant. Data were presented as means  ± standard deviations.

 

RESULTS:

Three groups of female subjects were involved in this study. The first group represented 52 pregnant women with low total protein, the second group represented 50 pregnant women with normal total protein while the third group represented 50 non pregnant, non lactating apparently healthy women. All the subjects were aged between 20 and 40 years. All the pregnant subjects were in different age ranges and parity. The mean age was 28.90 ± 5.31 for all the subjects studied.

 

Effect of Age on the Different Biochemical Parameters Measured:

Figs. 1 to 4 show the results of all parameters measured for mothers in different age groups divided according to their level of serum total protein. There was no significant difference in the means of the serum total protein of mothers of different age ranges and other parameters measured (p>0.05). Therefore age may not affect the level of serum total protein of a mother and also other parameters measured. A test of correlation showed that age of mother did not correlate significantly with serum total protein. However age of mother correlated positively and significantly with parity only (r = + 0.545) (p<0.05) and no other parameter measured (p>0.05 in each case).

 

Effect of Parity (Number of Children Had By Mother) on the Different Parameters Measured:

Figs. 5 – 7 show results of all parameters measured for mothers with different parity divided according to their level of serum total protein.

A significant difference was seen between the mean serum total protein levels of mothers who have 1 – 2, 3 – 4 and 5 or more children at 95% confidence internal, using analysis of variance test (p<0.05).

A test of correlation revealed that number of children a mother had (parity), correlated negatively with serum total protein. (r = - 0.226, p<0.05).

This suggests that serum total protein levels significantly decreased as number of children increased and vice versa. Age of mother also correlated positively and significantly with parity (r = + 0.545, p<0.05).

 

DISCUSSION

The demand for both energy and nutrient is increased during pregnancy and for well nourished women only a small amount of additional energy is required [21]. Pregnancy is also associated with major alterations in every maternal organ, system and metabolic pathway. Values of biochemical parameters may change as the pregnancy advances from first to third trimester and to parturition and then return towards normal during post partum period. The two major physiological forces driving these changes are:

(1)   The increase in plasma volume, increase in red blood cells and decreased concentrations of circulating nutrient-binding proteins and micronutrients.

(2)   The ever increasing levels of estrogen and progesterone as well as other placental related hormones, which have particular impact on maternal lipids (cholesterol)[9].

 

These two physiological modifications result in two dominant effects: the first reduces levels of biochemical substances such as albumin and haemoglobin which return to normal 8-10 weeks post partum. The second causes lipids to rise during pregnancy and return to normal at post partum. The major consequences of protein energy malnutrition (PEM) are mainly poor weight gain in pregnancy, anaemia leading to high risk delivery and low birth-weight babies that fail to thrive.

 

The result of this investigation also showed there was no significant difference in the mean serum total protein in the different age groups. Therefore age of mother may not be a factor and may not affect the level of serum total protein and other parameters measured. This agrees with the study of Okwu et al. [22], which showed that the lower age groups (below 20 years and 20-24 years) presented higher prevalence of PEM than other age groups, with the effect more prominent in rural areas than in urban areas.

 

The result also showed a negative correlation between parity, serum total protein, urea, total cholesterol, creatinine and calcium as observed in the results. This suggests that serum total protein significantly decreased as number of children increased and vice versa. This implies that parity therefore, may be a contributing factor to PEM. This agrees with the views of McGanity et al. [9] that PEM is manifests in many mothers because their nutritional status has not improved from their last birth and so they are not nutritionally prepared for the next pregnancy.

 

In addition, Lapido10 suggested that many pregnancies in developing countries are unplanned, coupled with inadequate dietary intake due to dietary taboos associated with pregnancy, gender and other cultural beliefs.

 

SUMMARY AND CONCLUSION:

Serum total protein was found to correlate negatively and significantly with parity (i.e. the number of children had by the mother). Also there was no significant difference (p>0.05) between the age of mothers and serum total proteins.

 

The serum levels of urea, creatinine, calcium and cholesterol have been implicated in this study as Biochemical indices or Biochemical Markers of PEM in pregnant women. Serum total cholesterol may however be a useful marker for energy intake and not for protein intake.

 

REFERENCES:

1.        Muller, O. and Krawinkel, M. (2005). Malnutrition and Health in Developing Countries. CMAJ, 171(3):279-293.

2.        Pena, M. and Bacalao, J. (2002). Malnutrition and Poverty. Ann. Rev. Nutr., 22:241-253.

3.        Enwonwu, C. O., Phillips, R. S., Ibrahim, C. D. and Danfillo, I. S. (2004). Nutrition and Oral Health in Africa. International Dental Journal, 54:344-351.

4.        Maclaren, S. D. and Burman, D. (1976). Textbook of Paediatric Nutrition. Churchill Livingstone New York pp 105.

5.        Scrimshaw, N. S. (1998). Malnutrition, Brain Development, Learning and Behaviour. Nutrition Research, 18(2):351-379.

6.        Abidoye, R. O. and Eze, D. I. (2000). Comparative School Performance Through Better Health and Nutrition in Nsukka, Enugu, Nigeria. Nutrition Research, 20(5):609-620.

7.        De Onis, M., Blossner, M. and Villar, J. (1998). Levels and Patterns of Intrauterine Growth Retardation in Developing Countries. European Journal of Clinical Nutrition; 52:583-592.

8.        Torun, B. and Chew, F. (1994). Protein Energy Malnutrition. In Shils,M. E.,Olsen, J. A and Shike, M. eds, Modern Nutrition in Health and Diseases 8th ed. Vol 1 Williams and Wilkins Phil. U.S.A. pp 950 – 974.

9.        McGanity, W.J., Dawson, E.B. and Fogelman, A. (1994). Nutrition in Pregnancy and Lactation. In, Shils, M.E; Olsom, J.A. and Shike, M. (eds.) Modern Nutrition in Health and Disease.  8th ed. Vol. 1 Williams and Wilkins Phil. U.S.A. pp 705 – 726.

10.     Ladipo, O. (2000). Nutrition in Pregnancy: Mineral and Vitamin Supplements. American Journal of Clinical Nutrition, 72(1): 280-290.

11.     Gornall, A. G., Bardawill, C. C. and David, M. N. (1949). Determination of serum proteins by means of the Biuret Reaction. Journal of Biological Chemistry, 177: 751-766.

12.     Berg, B., Petersohn, L., Helm, G. and Tryding, N. (1984). Reference values for serum components in Pregnant Women. Acta Obstetricia et Gynecologica Scandinavica, 63(7): 583-586.

13.     Doumas, B.T., Watson, W.A. and Biggs, H.G. (1971). Amino acids and Proteins. Clinica Chimica Acta, 31: 87.

14.     Allain, C. C., Poon, L. S., Chan, C. S. G., Richmond, W. and Fu, P. C. (1974) Enzymatic determination of total serum cholesterol. Clinical Chemistry, 20:470-475.

15.     Meiattini, F., Prencipe, L., Bardelli, F., Giannini, G. and Tarli, P (1978). The 4-hydroxy benzoate/4-aminophenazone chromogenic system used in the enzymatic determination of serum cholesterol. Clinical Chemistry, 24: 2161 – 21 65.

16.     Cheesbrough, M. (1998). District Laboratory Practice in Tropical Countries. Part 1 Cambridge Univ. Press UK. PP 311-340.

17.     Bartels, H. and Bohmer, M. (1971). Eine mikromethode zur kreatinine bestinmung. Clinica Chimica Acta, 32: 81-85.

18.     Fabiny , D. L. and Ertingshausen, G. (1971). Automated reaction rate method for determination of serum Creatinine with centrifichem.  Clinical Chemistry, 17: 696-700.

19.     Gindler, M and King, J. D. (1972). Rapid colorimetric determination of calcium in biologic fluids with methylmol blue.  American Journal of Clinical Pathology, 58: 376-382.

20.     Barnett, R. N., Sokodon, S. B., and Goldberg, M. H. (1973). Performance kit used for clinical chemical analysis of         calcium in serum. American Journal of Clinical Pathology, 59: 836-843.

21.     World Health Organization (WHO). (1999). Health Situation in the South East Asia Region 1994-1997. New Delhi, WHO Regional Office for South-East Asia

22.     Okwu, G. N., Ukoha, A. Nwachukwu, N. and Agha, N. C. (2007). Studies on the predisposing factors of protein energy malnutrition among pregnant women in a Nigerian community (2007). Online Journal of Health and Allied Science, 3:1.

 

Received on 24.07.2010

Accepted on 12.08.2010        

© A &V Publication all right reserved

Research J.  Science and Tech.  2(2): March –April. 2010: 23-28