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1 HAEMATOLOGICAL PROFILE AND FERRITIN LEVELS IN CHILDREN WITH PROTEIN ENERGY MALNUTRITION IN UNIVERSITY OF ILORIN TEACHING HOSPITAL ILORIN A DISSERTATION SUBMITTED TO THE NATIONAL POSTGRADUATE MEDICAL COLLEGE OF NIGERIA IN PART FULFILLMENT OF THE REQUIREMENT FOR THE FELLOWSHIP OF THE COLLEGE in PAEDIATRICS BY DR AISHAT OLUWATOYIN SAKA NOVEMBER, 2009

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HAEMATOLOGICAL PROFILE AND FERRITIN LEVELS IN CHILDREN

WITH PROTEIN ENERGY MALNUTRITION IN UNIVERSITY OF ILORIN

TEACHING HOSPITAL ILORIN

A DISSERTATION SUBMITTED TO THE NATIONAL POSTGRADUATE

MEDICAL COLLEGE OF NIGERIA IN PART FULFILLMENT OF THE

REQUIREMENT FOR THE FELLOWSHIP OF THE COLLEGE in

PAEDIATRICS

BY

DR AISHAT OLUWATOYIN SAKA

NOVEMBER, 2009

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Declaration

I, DR. AISHAT OLUWATOYIN SAKA, hereby declare that this dissertation is

original unless otherwise acknowledged. The dissertation has not been presented to

any College for Fellowship examination nor has it been submitted elsewhere for

publication.

---------------------------------------- -------------------------

Dr. Aishat Oluwatoyin Saka Date;

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Certification

We hereby certify that Dr. Aishat Oluwatoyin Saka of the Department of Peadiatrics

and Child Health, University of Ilorin Teaching Hospital, Ilorin, prepared this

dissertation under our close supervision.

First Supervisor Prof. Ayodele Ojuawo -------------------------------------------

Signature

Second Supervisor Dr Aishat Abdulkarim ---------------------------------------

Signature

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Dedication

This dissertation is dedicated to Almighty Allah, the author and giver of knowledge

and to the millions of children in the developing world struggling to survive

malnutrition.

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Acknowledgements

I am sincerely grateful to my teacher and supervisor, PROFESSOR AYODELE

OJUAWO, who did not only supervise this project work but ensured that I had a

sound and firm training as a Resident in Paediatrics, thank you sir, may God bless and

reward you.

To my second supervisor Dr Aishat Abdulkarim, thank you for taking time to

supervise the work. I am especially grateful to DR O.T ADEDOYIN, I appreciate

your mentorship and critical review of this project.

I wish to wholeheartedly show profound gratitude to Prof Adeoye Adeniyi (Baba) for

his advices and encouragement at each stage of my residency training.

To all consultants in Department of Paediatrics, PROF WBR JOHNSON, DR OA

MOKUOLU, DR SK ERNEST, DR ADEGBOYE, DR O ADESIYUN, DR

ADEBOYE MAN, DR JK AFOLABI and DR O V ADEBARA, I appreciate you all.

I am grateful to the authorities of University of Ilorin Teaching Hospital, Ilorin for

giving the approval for the study. Many thanks to my colleagues, the nursing staff

and all staff in the department of Paediatrics and Child Health, UITH Ilorin, thanks

for your support.

DR OLAWUMI (HOD) and MR LUQMAN LATUBOSUN of Haematology

Department UITH, Ilorin, DR BILAMIN of the Chemical Pathology Department

UITH, Ilorin, DR FOWOTADE of the Department of Microbiology, UITH Ilorin. I

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am grateful for your kind and immense assistance in the laboratory component of this

work.

Many thanks to Dr Aderibigbe of the Department of Community Health and

Epidemiology UITH Ilorin for your assistance in statistical analysis.

Special thanks and regards to my husband DR M J SAKA who has displayed real

friendship, sacrifice and support in ensuring I attain a comfortable height

academically. May the good lord bless and reward you. To my children, thanks for

your little prayers and enormous support. I love you all.

Above all, my sincere appreciation goes to the Almighty God for seeing me through

in the pursuit of my career so far.

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Table of Contents

Title i

Declaration ii

Certification iii

Dedication iv

Acknowledgement v-vi

Table of Contents vii-viii

Appendices ix

List of abbreviations x-xi

List of Tables xii-xiii

List of Figures xiv

Summary xv-xvii

Introduction 1-6

Literature Review 7-48

Justification 49

Aims and Objectives 50

Materials and Methods 51-67

Results 68-85

Discussion 86-90

Conclusion 91

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Recommendations 92

Limitations of the study 93

References 94-110

Appendices 111-121

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Appendices

i Information Sheet

ii Informed Consent Form

iii Study Proforma

iv Socio Economic Classification Scheme

v Serum Ferritin Absorbance Curve

vi Ethical approval, U.I.T.H Ilorin

vii National Postgraduate Medical College approval

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Abbreviations

EDTA Ethylene diamine tetra acetate

ELISA Enzyme linked immunosorbent assay

EPU Emergency Paediatric Unit

FBC Full Blood Count

Hb Haemoglobin

Hct Haematocrit

MCH Mean Corpuscular Haemoglobin

MCV Mean Cell Volume

MCHC Mean Corpuscular Haemoglobin Concentration

nm nanometer

Nos Numbers

PEM Protein Energy Malnutrition

PCV Packed Cell Volume

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RBC Red blood cell

SEC Socio Economic Class

UITH University of Ilorin Teaching Hospital

WBC White blood cell

RDW Red blood cell distribution width

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List of Tables

I : The Wellcome Classification of PEM

II: Modified Wellcome Classification of PEM

III: Gomez classification of PEM.

IV: Gomez and modified Gomez weight-for-age classification of

malnutrition

V: Mclaren and Read classification of nutritional status in early Childhood.

VI: The World Health Organization Classification of PEM.

VII: The socio-demographic characteristics of the subject and controls.

VIII: Anthropometric measurements of the study population

IX: Distribution of types of protein energy malnutrition according to the age.

X: Haematologic profile of the study population

XI: Haematolgic Profile of children according to the types of PEM and

controls.

XII: Red Cell Morphology and Prevalence Of Anaemia In Children With

PEM.

XIII: Stool RBC in PEM and Controls.

X IV: Relationship between stool RBC and serum ferritin in PEM .

XV: Serum Ferritin Level according to the Age and Sex in the total Study

population

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XVI: The Levels of Serum Ferritin according to the types of PEM and

Controls

XVII: Prevalence of Iron Deficiency And Iron Deficiency Anaemia Amongst

PEM.

XVIII: Relationship between clinical features and serum ferritin levels in PEM

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List of Figures

Figure 1: Picture of the peripheral blood smear prepared.

Figure 2: Comparisons of normal peripheral blood smear with smear of

microcytic, hypochromic anaemia.

.

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Summary

This is a case control study carried out at the University of Ilorin Teaching Hospital

to determine the haematologic profile and serum ferritin levels of children with

Protein Energy Malnutrition compared to that of controls. It also determined the

prevalence of anaemia and iron deficiency anaemia in children with Protein Energy

Malnutrition

The socio demographic characteristics, anthropometry, clinical and laboratory

parameters of children with Protein Energy Malnutrition were studied. The

haematologic profile was analyzed by the auto analyzer, serum ferritin was analyzed

by Nova Path TM Ferritin kit. Stool was analyzed for red blood cells. Data entry and

analysis were carried out with a micro-computer using the Epi info version 3.5 (2008)

software packages.

A total of 180 children 90 with Protein Energy Malnutrition and 90 controls were

studied. Fifty Nine(65%) were males and 31 (34.4%) were females with a male to

female ratio of 1.9:1.0. The mean age of the children with Protein Energy

Malnutrition was 22.7 + 14.4 months (Range of 9-59months) compared to 29.3 + 16.9

months (Range 6-59 months) for the controls and the difference was not significant

(p=0.08).

The children with Protein Energy Malnutrition had lower mean values for

haemoglobin, haematocrit and mean corpuscular haemoglobin (p<0.05). The subjects

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also had higher mean values of total white cell count, neutrophil and lower

lymphocytes counts (p<0.05). The highest lymphocyte count was seen in children

with Kwashiokor. Platelet counts were comparable in the two groups.

No significant differences were observed in the haematologic profile of the various

types of PEM, except for the mean corpuscular haemoglobin concentration which was

higher in children with Marasmic-Kwashiokor. Children with Kwashiokor had the

highest mean for haemoglobin, (31.6±1.6g/dl) and haematocrit (10.7±0.4%), while

subjects with marasmus had the lowest mean for haematocrit (27.6±5.8%),

haemoglobin (9.1±2.1g/dl) and mean corpuscular haemoglobin (22.9±2.3pg/cell).

The subjects had a lower mean serum ferritin compared to the controls. Serum ferritin

increased with age and females had a higher serum ferritin levels compared to males

(p=0.00001).

The prevalence of anaemia among the children with PEM was 74.4% of which 75%

had microcytic hypochromic anaemia. The prevalence of iron deficiency among

subjects was 24.4%, while 16.6% of these subjects had iron deficiency anaemia.

In conclusion, children with Protein Energy Malnutrition had lower red cell indices

and higher white cell count the controls. Serum ferritin level was lower in the

subjects than in the controls. A high prevalence of anaemia and iron deficiency

anaemia with microcytic hypochromic anaemia was observed in children with PEM.

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It is recommended that anaemia and iron deficiency should be sought for in children

with Protein Energy Malnutrition and such should be treated. There is the need to

improve nutrition education among mothers and health workers. Food fortification as

well as supplementation of the important micronutrient especially iron should be

enhanced, as a good nutritional status remains the bedrock for good health and

prevention of diseases.

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Introduction

Malnutrition refers to a group of pathological disorders resulting from imbalance

between intake of essential nutrients and body requirement for these nutrients.1

In other words, malnutrition results from not eating enough food or from intake

of food that although adequate in amount, lack the essential nutrients necessary

for normal growth and development.1

Protein Energy Malnutrition (PEM), is defined as a spectrum of diseases arising

as a result of an absolute, or relative deficiency of calories and or protein in the

diet.2,3 It is globally the most important risk factor for illness and death, with

hundreds of millions of young children affected.4

Malnutrition, with its two constituents of protein–energy malnutrition and

micronutrient deficiencies, continues to be a major health burden in developing

countries.4Apart from marasmus and kwashiorkor (the 2 forms of protein–

energy malnutrition), deficiencies in iron, iodine, vitamin A and zinc are the

main manifestations of malnutrition in developing countries.4Also,nutritional

deficiency contributes to a high rate of morbidity and mortality in Nigeria

especially among infant and young children.5

According to UNICEF in 2005, malnutrition was associated with approximately

50% of child deaths worldwide6. Protein Energy Malnutrition is estimated to

affect every fourth child in the developing world,6 with the regional prevalence

for the severe forms ranging from 1-7%.7 PEM is associated with 49% of the 10

million deaths occurring in children in the developing world and 52% of all

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under five deaths in Nigeria.8 with 24% and 16% of the total under-5 Nigerian

population estimated to have suffered from mild-moderate and severe

malnutrition respectively from 1973 to 19839,10 .The hospital based incidence of

severe Protein Energy Malnutrition in Nigeria varies from 3.18% in Ilorin,11

4.39% in ibadan10 and 4.5% in Ife.12

Many classifications of malnutrition have been used over the past 30 years. The

choice of classification depends on the purpose or the type of study for which it

is to be used.13 There are classification for clinical studies and for community

survey. Classifications for clinical studies are based on the presence or absence

of edema, and are the severe forms. These severe forms of PEM are Marasmus,

Kwashiorkor and Marasmic Kwashiorkor.2,3 Community survey classification is

based on deficit in weight for age and 90% of Harvard standard is taken as cut

off point for separating normal from malnourished children.7 WHO published a

new classification for PEM called the Z-score or standard deviation classification

of malnutrition. 14 In this classification, protein–energy malnutrition is defined

by measurements that fall below 2 standard deviations. Z-scores classification

include height for age (HAZ) Z-score, weight-for-age Z-scores (WAZ), weight-

for-height Z-scores (WHZ) and body-mass- index for- age Z-scores (BMIZ).

Malnutrition is then defined as value below the normal weight for age

(underweight), height for age (stunting) and weight for height (wasting).15

Protein Energy Malnutrition is caused by low intake of food, which contains

macro and micronutrients. PEM results in various changes in the body including

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changes in haematologic profile of the body. Anaemia has always been a

constant feature of protein energy malnutrition and may be normochromic

normocytic, microcytic hypochromic, or, macrocytic.20, 21.

The anaemia of malnutrition may be attributable to various factors such as iron

deficiency, and /or reduced red cell production in adaptation to a smaller lean

body mass.3,21 Erythropoietin deficiency, deficiencies of vitamins (folic acid,

B12,) or trace elements (copper, zinc), infection and chronic diseases may also

be implicated as causes of anaemia in malnutrition.3,21-22

Iron deficiency is the most widespread nutrition problem in the world.16-18 Iron is

a nutritionally important mineral whose function includes structure of

haemoglobin and myoglobin, which are important for oxygen and carbon

dioxide transport and as an oxidative enzyme in cytochrome c and catalase. Lack

of iron impairs the normal cognitive development of 40 % to 60 % of the

developing world’s young children.7

Ferritin is an iron storage protein, the level provides a relatively accurate

estimate of the body‘s iron stores in the absence of inflammatory disease and so

it very useful in assessing body iron status .19

Despite the extensive knowledge of iron metabolism, diagnosis of iron

deficiency often remains a difficult problem, whatever methods are used,

especially in children with protein malnutrition.3.112 Estimation of haematologic

changes such as haemoglobin alone is not completely representative of iron

deficiency. Other methods such as the use of serum ferritin , serum iron, Total

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iron binding capacity and Transferrin receptors all have their limitations.

Diminished stainable iron in a bone marrow aspirate is generally accepted as the

reference standard for iron status. 3, 58,112

Several stidies12, 24-27 have attempted to evaluate iron status and the haematologic

changes in children with protein energy malnutrition.

Alemnji G A, Thomas K D, Durosinmi M A e tal24 in Ife studied twenty

malnourished children’s haemotogram and iron status in the acute phase and

during rehabilitation and concluded that the iron status was low in malnutrition

and that none of the haematological parameters was significantly different after

rehabilitation for all the four types of PEM. However, detailed changes in the

blood profile of the PEM children studied were not emphasized in the study and

the study population was small to make any logic conclusion.

Said A, El-Hawary M F, Sakr R et al25 in another study used haematological as

well as biochemical parameter to study anaemia in PEM found and out that

hypochromic anaemia was predominant in 83.3% of moderate Kwashiorkor with

50% being normocytic and 33.3% microcytic. In severe kwashiorkor, anaemia

was normocytic in 51.7% and microcytic in 19.2%. Macrocytosis was found in

38.8% in severe kwashiokor. In marasmus, normocytic anaemia was revealed in

74% and microcytic in 40%. Macrocytic anaemia was detected in 26%. In the

mild cases, normocytic normochromic anaemia occurred in 70% of the cases

while macrocytosis was encountered among 30% of the cases. Serum iron and

transferrin level dropped in both kwashiokor and marasmus, the extent of

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decrease was greater in the former. The percent transferrin saturation was

elevated in severe cases, particularly severe kwashiokor and marasmic

kwashiokor .In this study serum ferritin which is the best indicator of iron stores

was not utilized and the other details about the haematologic changes in

malnutrition was not addressed by the study .24,25

Another study by Wickramasinghe S N, Gill D S, Broon D N et al26 described

the limited value of serum ferritin in evaluating iron status in PEM. Twenty five

underweight and stunted infants and young children, during recovery from

malnutrition were studied. Many were iron-deficient and, as judged by ESR

values, all but four had evidence of infection or inflammation, or conditions

known to be associated with elevated values for plasma ferritin concentration

and erythrocyte protoporphyrin content and for decrease in the iron saturation of

transferrin .The finding that a number of the plasma ferritin concentrations were

quite high (11 of 48 children had more than 100g/L) was perhaps not surprising

because even in presumably normal subjects in the age range of the

malnourished subjects, concentrations greater than 100g/L was sometimes found.

The study could not conclude whether this instability of serum ferritin in these

malnourished children is related to inflammation or to other factors.The study

did not include haematologic changes in malnutrition.

In another study that looked at the haematologic profile as well as serum ferritin

level of PEM children matched with controls, red cell count, mean corpuscular

haemoglobin and reticulocyte index were significantly lower in PEM than in

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controls. While serum ferritin was significantly higher and total iron-binding

capacity was significantly lower in PEM children than in controls. The anaemia

was attributed to mixed deficiencies resulting in ineffective erythropoiesis. 27The

contribution of iron deficiency to the anaemia seen in PEM was not addressed in

this study.

Laditan in another study on PEM children reported that pack cell values were

within the lower limits of normal values for healthy Nigerian children. This

study however did not highlight other haematologic parameters in PEM.12

Relevance Of The Study To The Practice Of The Discipline

The outcome of this study would provide evidence for the magnitude of iron-

ferritin depletion in children with PEM. It would also provide information about

the contribution of malnutrition to the prevalence of iron deficiency anaemia in

children. It would also provide baseline information on the prevalence of iron

deficiency anaemia in malnourished children in the North-central zone of

Nigeria and thus a template for future research in the future. Clinical finding

related to iron deficiency would provide information for early identification of

iron deficiency so that early identification and treatment would be instituted.

This would go a long way to decrease morbidity and mortality from iron

deficiency especially in the malnourished child.

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Literature Review

Introduction

Malnutrition refers to a group of pathological disorders resulting from imbalance

between intake of essential nutrients and body requirement for these nutrients. In

other words, malnutrition results from not eating enough food or from intake of

food that although adequate in amount, lack the essential nutrients necessary for

normal growth and development. Protein Energy Malnutrition (PEM), is defined

as a spectrum of diseases arising as a result of an absolute, or relative deficiency

of calories and or protein in the diet.28-30

Malnutrition continues to be a major public health problem throughout the

developing world, particularly in southern Asia and sub-Saharan Africa.28-32

Diets in populations are frequently deficient in macronutrients (protein,

carbohydrates and fat, leading to protein–energy malnutrition), micronutrients

(electrolytes, minerals and vitamins, leading to specific micronutrient

deficiencies) or both.30, 33, 34, 35

In children, protein–energy malnutrition is defined by measurements that fall

below 2 standard deviations under the normal weight for age (underweight),

height for age (stunting) and weight for height (wasting).33 Wasting indicates

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recent weight loss, whereas stunting usually results from chronic malnutrition.33

Of all children under the age of 5 years in developing countries, about 31% are

underweight, 38% have stunted growth and 9% show wasting.30 Protein– energy

malnutrition usually manifests early, in children between 6 months and 2 years of

age and is associated with early weaning, delayed introduction of complementary

foods, a low-protein diet and severe or frequent infections.33,36,37

Historical Perspective

Severe malnutrition used to be known by several names such as infantile

atrophy, arthrepsia, inanition and Marasmus (Marasmos in Greek means to

“wither away” or “wasting away”), 2,38the later term being the one that has

survived common usage.

Kwashiorkor was recognized and described early in the turn of this century. In

Germany,39 where it was noticed in German children weaned on a diet of cereal

flour, the condition was called Mehlnarschaden (German for flour

dystrophy).40,41 In 1908 the Spanish literature described it as Syndrome

Policarencial Infantil42 .In that same year it was described as Culebrilla in

Yucatan,43 Mexico. The French literature described it in 1913 as Boufissure d’

Annam44 (meaning the swelling disease of Vietnam). More reports were made in

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temperate regions in 191345 and 192146while a report was made in Kenya in

1926.47

Cicely Williams, in 1933 while working in the Gold Coast (present day Ghana)

among the Ga people described the same disease entity using the native Ga word

“kwashiorkor” (Ga for disease of the weanling).2,47 It was with this report that

this disease in children acquired its present name.31

In 1956 Fredico Gomez and his colleagues described the clinical picture

preceding death and the apparent cause of death in malnourished children

admitted to the nutrition department of the children’s hospital in Mexico.47, 48.

His paper was a landmark of achievement because it used a single anthropometry

measurement (weight) to develop an indicator. Weight for age was related to the

severity of PEM and the risk for mortality.4

Prevalence of PEM

PEM is highly prevalent in almost all developing countries.2,13 It is estimated to

affect every fourth child in the developing world6,8 and approximately 6.6

million of the 12.2 million under -5 deaths occurring annually in 3rd world

countries are attributed to malnutrition.6,9 About 1-7% of under -5 children

suffer from the severe forms of malnutrition like kwashiorkor and marasmus.6,7

This small percentage of severe forms identified is an indication that larger

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number of mild and moderate malnutrition occurred unrecognized. The

prevalence range from 0.5 - 20% for severe form and 3.5 to 40.4% for moderate

forms world wide. In Africa, the prevalence for the severe form range from 1.7 -

9.8% and 5.4- 44.9% for moderate forms .7,54

Geographical variation is seen in the frequency of the different forms of PEM

.7,54 Breast feeding and weaning practices have been shown to play an important

role in the determination of the age distribution and types of malnutrition

observed.13 In Africa kwashiorkor was more common but a changing pattern is

now being experienced, marasmus is becoming more prevalent. This change has

been attributed to changes in feeding practices. Weaning is a common

precipitating factors in Latin America where infantile marasmus is common.

Kwashiorkor and marasmus are common in Asia where prolong breastfeeding

has been implicated as a major contributor to PEM.7-8,12-13.

Aetiology

PEM is an outcome of interplay of several factors that affect a child. These

diverse factors range from inadequate diet, infections, socio-cultural and

economic factors. Infectious causes and inadequate diet have been identified as

the most important.7,78

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Diet

PEM was being thought to result from in adequate protien calorie ratio in the

habitual diet. Kwashiorkor resulted from deficiency in protein predominately

while marasmus was from a deficiency in calorie in the diet with adequate

protein. Weaning with diluted milk formulae resulted in infantile marasmus

while kwashiorkor was common in young children fed on diet low in protein. In

some situation where protein intake is adequate but calorie intake is not, some

protein will be used for energy leading to continue protein deficiency. Hence

protein-energy malnutrition is used to describe the condition.3,7

Some studies have found no difference in dietary pattern of children who

develop kwashiorkor and those with marasmus.2,3,35 Other studies suggest that

PEM does not result from isolated deficiency of protein or calorie ,but rather

PEM results from the body’s adaptation to nutritional stress.18,54Successful

adaptation results in marasmus while mis adaptation results in kwashiorkor.2

Infection

Infection such as diarrhea, measles and acute respiratory tract infection have

been known to be associated with increase incidence of PEM. Kwashiorkor is

often preceded by an episode of infection with diarrhea or respiration tract

infection. Measles is another common infectious disease affecting children in

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developing world. Its impact is more because of the secondary complications and

prolonged illness associated with it. Mechanism by which infection leads to

malnutrition include, reduced food intake, malabsorption of nutrient and

metabolic losses during infection.2, 54

Socioeconomic Factors

Several socioeconomic factors have been implicated in the etiology of PEM.13,60

Inadequate diet, poor housing, increase incidence of infection all result from

poverty. Poverty, infection and malnutrition are related by a vicious cycle.

Insufficient house hold food security, inappropriate care for mother and child

and inadequate health care services provision by the government are some of the

numerous causes of PEM in the developing world. Cultural belief, superstition

and taboos about food belief also influence nutrient intake.13,60,77

Types of Malnutrition

There are various classifications of malnutrition

On the basis of the particular nutrient(s) implicated in the balance between

supply and body requirement, malnutrition can be broadly classified into.1,30:

A Macronutrient malnutrition (relating to energy and protein)

1 Protein – Energy malnutrition

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2 Over nutrition and obesity (effect of nutrient excess)

B. Micro nutrient malnutrition

1 Vitamin nutritional disorder

2 Minerals and trace elements nutritional disorders

Classification of Protein Energy Malnutrition

As a nutritional deficiency disorder, PEM results in growth failure, poor

nutritional status and, in due course, may also result in distinctive clinical

manifestations. These characteristic changes produced by PEM constitute the

main features for classifying the disorder into the various types and severity.51

In all cases, anthropometric indicators of nutritional status (weight-for-age,

height-for-age, and weight-for-height) usually form the basis for the

determination of the extent and severity of the nutritional deprivation and when

combined with certain clinical signs that are of high prognostic significance, for

example nutritional edema further allows differentiation into various types of

severe forms.51

There are classifications for clinical studies and for community survey.

Classification for clinical studies is based on the presence or absence of edema,

and are the severe forms. 2,3 These severe forms of PEM are Marasmus,

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Kwashiorkor and Marasmic Kwashiorkor. Community survey classification is

based on deficit in weight for age and weight for height.2,30-33

Clinical Classification

An International Working Party convened in 1969 agreed to a simple

classification of PEM which was published in 1970 as the Wellcome

classification2,50 in an attempt to bring orderliness to the classification of the

disorder.

In the Wellcome classification only two criteria were used to classify the

malnourished child; these are presence or absence of edema and the deficit in the

body weight for age.

The initial classification gave rise to four groups. Children with edema whose

weight was between 60 & 80% of expected weight for age were classified as

kwashiorkor, while those without edema in this weight range were classified as

underweight. Children without edema whose weight was below 60 % of

expected for age were classified marasmus or as Marasmic Kwashiokor in the

presence of edema.

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Table I : The Wellcome Classification of PEM2,50

Weight (% of standard) Edema Present Edema Absent

60-80 Kwashiorkor Underweight

<60 Marasmic-

kwashiorkor

Marasmus

Modified Wellcome Classification of PEM

It was noticed that the Wellcome classification excluded children who either had

weights above 80 per cent of the standard weight -for –age or even those with

normal weight-for –age and who still had overt kwashiokor .This exclusion

promoted the misconception that kwashiokor was invariably associated with

overt weight loss, thus implying dietary deficiency. This conception is not true as

kwashiokor can and sometimes arise in children who by any anthropometric

criteria cannot be classified as malnourished. This further explains the fact that

dietary deficiency is not a prerequisite for the development of kwashiorkor.2,50

Thus this called for the modification of the classification which takes account of

this and seem more appropriate.

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Table II: Modified Wellcome Classification of PEM 2,50

Weight (% of standard)* Edema Present Edema Absent

>80 Kwashiorkor Normal nutrition

60-80 Underweight-kwashiorkor Underweight

<60 Marasmic-kwashiorkor Marasmus

Classification for Community Survey

Classification on the basis of weight for age is used to assess the magnitude of

the problem in the community. This classification does not indicate the duration

or type of malnutrition.54 The joint FAO/WHO32 expert committee on nutrition

emphasized that weight deficit in relation with age may be negative as a measure

of the duration of malnutrition. Weight in relation to height gives rise to a

measure of current nutritional status, while height for age gives rise to a picture

of past nutritional history. The former is referred to as wasting while the latter is

stunting. Most community surveys used 80% of weight for age as level below

which malnutrition is defined and 90% height for age as a level below which

stunting is defined.54

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Gomez classification is the most widely used. It is based on deficit in weight for

age. In this classification, malnutrition is classified as first, second and third

degree.47

Table III: Gomez classification of PEM.

Malnutrition Body weight(% of standard)

First degree 75-90

Second degree 60-75

Third degree <60

The classification developed by Gomez F, Galvan R R, Frenk S et al47 was based

on three prior selections, an anthropometric indicator, a reference population

with which to compare the index child or community41 and cut off points to

classify children according to varying degrees of malnutrition. All classifications

developed after Gomez have all relied on these elements.48

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Table IV: Gomez and modified Gomez weight-for-age classification of

malnutrition2,47,48

Nutritional status Gomez classification

(%)

Modified Gomez

classification

(%)

‘Normal’ nutrition > 90 weight for age > 80 weight for age

Mild or grade I PEM 75-90 weight for age 70-80 weight for age

Moderate of grade II

PEM

60-75 weight for age 60-70 weight for age

Severe or grade III PEM < 60 weight for age <60 weight for age

Nutritional edema

Following the publication by Gomez in 1956, 1st degree, 2nd degree and 3rd

degree malnutrition became common place terminology among nutritionist’s and

medical personnel. Within the spectrum of PEM, three broad clinical types were

recognized, namely kwashiorkor, Marasmus and marasmic-kwashiorkor.

However, the criteria to distinguish these three conditions were based on clinical

observations, which are subjective, and ill-defined.51

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In, the same year, Waterlow, while trying to establish guidelines for rational

action or intervention, recommended a classification based on weight and height

to determine the nutritional status of children in a community. This was based on

the fair relationship that existed between weight and height.53

In 1972, Mclaren and Read proposed a classification of nutritional status in early

childhood based on observed weight as a percentage of ideal weight.51,52

Table V:Mclaren and Read classification of nutritional status in early

Childhood52

Classification observed weight as % of ideal

Obesity 120%

Overweight 110-120%

Normal range 90-110%

Mild PEM 85-90%

Moderate PEM 75-85%

Severe PEM <75%

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The World Health Organization Classification of PEM

In 1990 the WHO proposed a new classification of Protein Energy Malnutrition

based on the weight for height and the height for age Z-scores and the presence

or absence of symmetrical edema of the feet.55

Table VI: The World Health Organization Classification of PEM

Mild

Malnutrition

Moderate

Malnutrition

Severe Malnutrition

Weight-for-

Height

<-1 to >-2 Z-score

(90%-80%)

<-2 to >-3 Z-Score

(70%-79%)

<-3 Z-Score (70%)

Height-for-

Age

<-1 to >-2 Z-score

(90%-95%)

<-2 to >-3 Z-Score

(85%-89%)

<-3 Z-Score (<85%)

Symmetrical

Edema

No No Yes(Edematous

Malnutrition)

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Pathological Changes

Pathological changes have been noted in the body of a child with PEM. These

changes can be categorized as 3,7,54

(i) The organs.

(ii) Haematology.

(iii) Biochemical.

Changes in the organs

Liver- Fatty infiltration, seen in kwashiorkor first noticed in the periphery then

spread towards the portal tract and cortical level until it affects the parenchyma.

Changes can be reversed with treatment.3,7

Pancreas-children with PEM show marked atrophy of the pancreas. Pancreatic

calcification may ensue. Changes are reversible with treatment.3,7

Kidney-kidney size is reduced in severe PEM; Pyelonephritics changes with

scaring of the medulla and focal areas of calcification are common. Glomerular

changes, thickening of basement membrane and increased cellularity of capillary

tuft also is present. Changes are reversible with treatment.3,7

Gastro intestinal tract- Mucosal atrophy of the small intestine. The villous

atrophy decreases the absorptive surface. Villous shortens, crypts /villous ratio

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increase. Histology of gastrointestinal mucosa changes from columnar shapes to

cuboidal shapes and brush borders are lost. Inflammation cell infiltrations are

seen in laminar propria.3,7.

Thymolymphatic system-Thymus is greatly reduced in children during severe

PEM. Degree of thymic atrophy correlates closely with depletion of lymphocytes

.Tonsil sizes are significantly smaller with PEM, size of spleen is also decreased.

There is decrease in thymic dependent lymphocyte associated with impaired cell

Mediated immunity. However, B –lymphocytes responsible for humoral

immunity are not affected.3,7.

Muscle- There is marked reduction both in bundle and fibers of the muscle.3

Heart;- Heart size is usually reduced. ECG changes suggest non specific

myocardial damage. Reversal of changes occurs following full treatment.3,7

Haematological changes;

Some degree of anaemia is always found in children with PEM. Moderate

anemia with peripheral blood film showing normocytic or microcytic, hypo

chromic cell occur in PEM. Haemoglobin levels are reduced as a consequence of

low levels of plasma protein. Reticulocytes increase in response to anaemia with

increase erythroid activity. Anaemia may be due to associated deficiency of iron,

folate riboflavin and vitamin E.3,7,59.

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Biochemical changes

Low levels of serum sodium, potassium, magnesium copper and zinc have been

reported in PEM.7

TYPES OF PEM

There are various forms of protein energy malnutrition , various distinctions

have been made between the various forms based on the clinical presentation,

such as the presence or absence of edema and weight loss . Some workers feel

that this distinction between the various forms of severe PEM need not be

emphasized as they are different facets of the same disease. 2,7 58For the purpose

of classification and management , the following are the clinical presentation and

features of the severe forms of protein energy malnutrition.

Marasmus

It is characterized by progressive wasting of the subcutaneous tissue and the

muscle. Marasmus is diagnosed when subcutaneous fat and muscle are lost

because of endogenous mobilization of all available energy and nutrients.28 This

condition is a consequence of inadequate food intake. It is often precipitated or

preceded by failure of breast feeding, infection or congenital abnormalities.

When an infant develops this condition, it is described as infantile Marasmus and

when seen in older children it is called late marasmus.28 Marasmus is defined as

a weight of <60%of the expected weight for age without edema.2,28

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There are many mechanisms responsible for marasmus but in global terms,

marasmus results from a negative energy balance.3,7 It can result from decreased

intake, increased energy expenditure, or both as it may occur in either acute or

chronic disease states.56 Reasons why a continuous nutritional food deficit leads

to Marasmus rather than kwashiorkor are unclear but theories have shown that

Marasmus is a successful adaptation to continuous nutritional deficits, while

kwashiorkor is a dysadaptation. 2,3. The adaptations to energy deficit include

reduced activity, lethargy, decrease in basal energy metabolism slowing of

growth and then weight loss.56

Clinical presentation of marasmus include prominent bones and joint, hanging

skin folds over the buttocks and thighs, disappearance of buccal fat pads and

distended abdomen. There are also changes in the skin and in the hair. The skin

becomes loose and may eventually hang in folds; this is common in the buttocks

and thighs. It may also be wrinkled, dry or scaly.2-3 The buccal fats in the cheeks

of young infants may persist long after the subcutaneous tissue of other part of

the body have disappeared. Its disappearance is a poor prognostic sign. The

abdomen may be scaphoid or distended. Anal or rectal prolapse (from loss of

perianal fat) can also occur. Diarrhea is sometimes present with accompanying

dehydration. Anaemia is usually present.2-3,7.

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Marasmic children are usually unhappy and irritable but usually respond

normally to attention and feed more hungrily. Edema is usually absent. There

are no constant biomedical derangements associated with marasmus.3,28, 56

Kwashiokor

Kwashiorkor usually manifests with edema, changes to hair and skin color,

anaemia, hepatomegaly, lethargy, severe immune deficiency and early death. It is

common in children between one and three years old. The physical findings in

kwashiorkor include poor growth, edema, muscle wasting, and mental changes.

These are the four cardinal signs which are always present in kwashiorkor. Other

features include skin changes, hair changes, and moon face appearance, signs of

micronutrient deficiency, hepatomegaly and intractable diarrhea. Anaemia is a

usual feature. Anaemia is usually moderate and when severe is usually due to

iron and/or folate deficiency.

Marasmic- Kwashiorkor

They are children that have both clinical features of kwashiorkor and marasmus.

In this condition, edema appears in a clinically undernourished child who has not

been growing for weeks or months. The degree of stunting is significantly

greater in them suggesting that the duration of illness is longer in marasmic–

kwashiorkor than in kwashiokor . Features like psychosocial, skin and hair

changes as well as other feature of kwashiorkor are also present.56

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Complications

Infection

Infections are common complications of severe PEM. It is often severe and

reflects impaired immunological status of these children. It might be difficult to

diagnose because the typical features such as fever and leucocytosis might be

absent. Children with kwashiorkor are susceptible to gram negative organisms

such as Escherichi. coli, Proteus species and Pseudomonas Aeruginosa. Viruses

like measles, herpes simplex may cause serious infections and HIV/AID is also a

common infections. Severe candidaisis are frequently seen. Tuberculosis being

aggressive are usually difficult to identify in these children. Intestinal parasitic

infections (especially ascaris) are high in PEM. Hookworm infection is less

common in the malnourished children because they are too weak to walk around

to contract the infection. Malaria is common and it does appear to be less severe

in the malnourished.3,7,47

Hypoglycemia and Hypothermia

Fasting blood sugar may be decreased but symptomatic hypoglycemia is

uncommon and may develop if feeding is infrequent. Hypoglycemia is common

in marasmus while energy store are depleted often in association with

septicemia. It can present as twisting convulsion and unconsciousness.7,60

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Hypothermia is manifestation of impairment of thermoregulatory center and

energy stores. It occurs often in severely wasted children.

Cardiac failure

Cardiac failure is commonly seen in the treatment phase and sometimes

associated with anaemia.3,7 During recovery, the heart size increases rapidly

owing to chamber enlargement, and left ventricle increase in proportion to

increasing weight. However if dietary repletion is rapid, especially with a high

sodium load, heart failure may develop and sudden death may occur.3

Investigation and Treatment of PEM

There are some investigations required for the diagnosis and treatment of PEM

these include1-3,7,.

1 Complete blood count-Haemoglobin and packed cell volume usually are

lower than normal .There might be leucocytosis or leucopenia in the

prescence of infection. Platelet count is usually within normal limits.2

2 Random blood sugar-blood sugar might be low or normal. Children with

PEM have an increase susceptibility to low blood sugar because of the

increase cortisol level which occurs in response to the severe malnutrition

and concomitant infection present in these children.1,3

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3 Blood electrolyte, urea and creatinine –evidence of derangement of these

parameters can be present. High sodium as well as low potassium is

typical of malnutrition because of active transport of sodium out of the

cell as a result of increase permeability of the cells.3-4

4 Serum lipids and protein levels –changes seen in serum protein in

malnutrition include low serum albumin, transferin and apolipoprotien B .

PEM. This is as a result of changes in the liver where these proteins are

synthesized in severe malnutrition. 3 Plasma free fatty acids are raised

while the level of triglycerides and phospholipids are reduced with a low

ratio of esterified fatty acids to free cholesterols.7

5 Microbiological cultures -culture of blood and other body fluids such as

urine and cerebrospinal fluid can be done to know the organisms

responsible for any infection and their sensitivity patterns to antibiotics.

6 Stool for ova of parasite and occult blood to identify parasite if present.

7 Tuberculin skin test - for diagnosing tuberculosis even though it might be

negative because of immune suppression present in severe PEM despite

the presence of an active tuberculosis infection. Clinical presentation

along side with other investigations would be useful in making a diagnosis

in suspected cases.

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8 Radiological chest x ray this is important because of the higher association

of malnutrition with tuberculosis.

9 HIV screening – this is important because of the common association of

PEM with HIV/AIDS.

10 Hormone; - increased serum levels of corticosteroid , cortisol and growth

hormones are common manifestation of PEM.

Treatment

Treatment is divided into phases;

(i) Stabilization phase (2-6 wks)

(i) Rehabilitation phase

(ii) Follow up between -6 months

Main stay of treatment has been elucidated by several workers 65,77. This includes

A Dietary management . These involve improving the nutritional level of the

child as soon as possible. Dietary management should follow stated protocol

such as the one provided by WHO.14 This should be adapted for the particular

locality where it is to be used. The protocol entails that what ever diet is chosen,

the energy density should be between 75-100kcal /100ml, the osmolarity should

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be lower than 350-400mOsmol/L and 6-12% of the calories should come from

protein.3,14. Vitamin and other micronutrients should also be provided.

B Management of complication

Anaemia-Iron deficiency is the commonest cause of anaemia. It can be treated

with oral forms of iron. Associated folate deficiency can be treated with oral

dose of folic acid. Severe anaemia can be managed with blood transfusion of

packed cells to prevent heart failure.

Hypoglycemia; Treated with intravenous glucose and early monitored and

quantified feeding.

Infection; Gastro intestinal respiratory and urinary tract infection are common in

children with severe PEM. Rationale antibiotics use including the use of broad

spectrum antibiotics initially such as a combination of an aminoglycoside and a

penicillin compound to cover for both such gram positive and gram negative

organism. This can be changed to the sensitive antibiotic as soon a result of

sensitivity is out.

Parasitic infection, such as malaria and other intestinal infestation should be

managed appropriately.

Hypothermia; Child should be kept warm and temperature monitored regularly.

Shock and electrolyte derangements; Prompt and proper fluid and electrolyte

management should be instituted especially when diarrhea and vomiting are

present.

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Long Term Care

Follow up care for nutritional rehabilitation and growth monitoring is required

for children with PEM. This can be done at health centers or domiciliary visit if

the situations permit. Nutritional education, immunization and growth

monitoring should be offered60,83.

Risk Factors for Protein Energy Malnutrition in Nigeria

Protein-energy malnutrition is still highly prevalent in Nigeria as a result of

many factors which include faulty weaning practices, poverty, poor sanitary

conditions, minimal medical attention, and endemic childhood infections.60

Inadequate production or shortage of food is no longer thought to be the usual

cause of malnutrition. Instead the problem develops from, and are maintained by,

a complex network of socio-economic and health determinants.3,7 Studies have

shown that in developing countries, the nutritional status of children has a

significant direct relationship with household income. 12,13,75 Other socio-

economic factors such as educational level of parents, distribution of food in the

family, demographic data, immunization status, childhood illness, intestinal

parasitoses, and childhood nutrition also have significant association with the

nutritional status of children.83,98-100.

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Prevention of Malnutrition

Protein Energy Malnutrition can be prevented by improving maternal diet and

health, and providing good antenatal care. Other modalities of prevention include

appropriate infant feeding practice which include, practice of exclusive breast

feeding and good complementary feeding practice. Prevention of infections and

diseases by immunization, diarrheal management, growth monitoring and

sustenance of other child survival strategies will also prevent malnutrition.8,14,60.

Haematologic Changes In Protein Energy Malnutrition

In children with protein energy malnutrition, the haemoglobin concentration is

usually low ,it may fall to 8g/dl of blood.57,58 However, some PEM children with

features of acute complication who are admitted to the hospital, a normal

haemoglobin levels may initially be present . This can be attributed to a

reduction in plasma volume present at this stage of the illness.

The mechanism of anaemia in PEM has been attributed to various causes such as

protein deficiency which leads to an increased oxygen consumption and

erytropoitein production. This invariably results in a drop in erythropoiesis and

reticulocyte counts.59 Red cell maturation is blocked at the erythrocyte level

and the erythropoietin-sensitive stem cell pool is slightly decreased57-59. The

production of red blood cells (RBCs) is governed by various growth factors. Of

all the factors stimulating erythropoiesis has a more important regulatory role

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than erythropoietin. Erythropoietin binds to specific receptors on the erythroid

bone marrow precursors [colony forming unit-erythroid (CFU-E) and burst

forming unit-erythroid (BFU-E)], thus stimulating their differentiation and clonal

maturation into mature RBCs 59,76. Other requirements for normal erythropoiesis

are protein, iron for haemoglobin synthesis, vitamin B12 and folate for DNA

synthesis, as well as other vitamins such as pyridoxine, riboflavin, thiamine,

vitamins C and E, and trace elements such as cobalt 59,77.

Some investigators, who have also found normal erythropoietin production in

PEM, have suggested that the impairment of erythropoiesis in PEM could be due

to an abnormality in the erythroid progenitor cell pool or ineffective

erythropoiesis82. It has also been shown that in anaemia of some chronic diseases

there could be a blunted serum erythropoietin response to anaemia83, which was

not the situation in the anaemia of PEM infants in a study27, where the

erythropoietin response to anaemia was preserved.

Another possible etiology of anaemia in PEM is a reduction in the plasma

volume to a variable degree seen especially in children with Kwashiokor, the

total red cell mass also decreases in proportion to the decrease in the lean body

mass as a result of protein deficiency which reduces metabolic needs of the

body.57

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The anaemia in PEM is usually normocytic normochromic, but there is a

considerable variation in size and shape of the red cell on the blood film.58

The white cell count and platelets counts are usually normal.3 The marrow is

most often normally cellular or slightly hypocellular, with a reduced erythriod-

myeloid ratio.3 Erythroblastopenia, reticulocytopenia, and a marrow containing a

few giant pronormoblast may be seen in the presence of infection.58

During repletion in the recovery phase an increase in plasma volume may occur

before an increase in red cell mass, anaemia may then become more severe

despite reticulocytosis. Also during this stage of repletion, occult deficiencies of

iron, folic acid, riboflavin, vitamins E and B12 may occur and thus worsen the

anaemia in these children .57-59

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Nutritional Anaemia

Anaemia is defined as a reduction of the red blood cell volume or haemoglobin

concentration below the range of values occurring in healthy persons.61 It results

in a reduction in circulating red cells, haemoglobin and haematocrit. A reduction

in haemoglobin concentration usually reduces oxygen carrying capacity of the

blood but this is usually apparent when the haemoglobin Concentration fall

below 7-8g/dl.84 When anaemia occurs as a consequence of a nutritional

deficiency, any of these pathologic processes may be involved either singly or

combined . This is referred to as nutritional anaemia.

Nutritional anaemia may be due to iron deficiency (inadequate iron intake, poor

iron absorption, or excess iron losses), vitamin deficiency such as vitamin A,

vitamin B group(pyridoxine B6, riboflavin B2, folate B9,and cyanocobalamin

B12), vitamin C and vitamin E deficiencies.58,62 Nutritional disturbances such as

protein and calorie deficiency states can also result in anaemia.

Prevalence of iron deficiency anaemia in PEM

Nutritional anaemia is a common problem in Nigeria among children with iron

deficiency being the commonest micronutrient deficiency, affecting more than 2

billion people world wide. Iron deficiency anaemia (IDA) is reported to be the

most common nutritional anaemia in the world. It affects 20% to 50% of the

world's population and it is common in young children.5 In the United states ,

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data from the National Health and Nutrition Examination survey found the

prevalence of iron deficiency to be 9% among children 1-2 years. Despite this

fact the prevalence of iron in vulnerable groups such as children with protein

energy malnutrition is extremely variable.25-26 In Nigeria, the Prevalence of iron

deficiency anaemia has been reported to be as high as 20-25% among children, 5

while the prevalence of iron deficiency anaemia in children with PEM in Nigeria

was 10% .75

Normal term infants are born with sufficient iron stores to prevent iron

deficiency for the first 4-5 months of life. Thereafter, enough iron needs to be

absorbed to keep pace with the demands of rapid growth. For this reason,

nutritional iron deficiency is most common between 6 and 24 months of life. A

deficiency earlier than age 6 months may occur if iron stores at birth are reduced

by prematurity, neonatal anaemia, small birth weight, or perinatal blood loss.65A

significant body of evidence indicates that iron deficiency, in addition to causing

anaemia, has adverse effect on multiple organ systems. Thus, the importance of

identifying and treating iron deficiency extends past the resolution of any

symptom directly attributable to a decreased hemoglobin concentration.64,66

Anaemia is classified according to the red cell on peripheral blood film and the

mean corpuscular volume (MCV). The primary classification includes;84

Hypochromic microcytic (small pale red cells and low MCV).

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Macrocytic (large red cell with large MCV).

Normocytic normochromic (cells of normal size and normal MCV).

History of Iron deficiency Anaemia

A disease believed to be iron deficiency anaemia was described in about 1500

B.C. in the Egyptian Ebers papyrus, it was termed chlorosis or green sickness in

Medieval Europe, and iron salts were used for its treatment in France by the mid-

17th century.68 Thomas Sydenham recommended iron salts as treatment for

chlorosis, but treatment with iron was controversial until the 20th century, when

its mechanism of action was more fully elucidated.68

Etiology of Iron Deficiency Anaemia In PEM

Iron deficiency anaemia is the most common type of anaemia, and is also known

as sideropenic anaemia. It is the most common cause of microcytic anaemia. The

causes of iron deficiency anaemia are numerous. 69Dietary deficiency,

malnutrition, infection and infestation are the leading cause of iron deficiency

ananemia.68,84

Iron deficiency anaemia occurs when the dietary intake or absorption of iron is

insufficient, and haemoglobin, which contains iron, cannot be formed.68 Also,

the popular complementary food given to infants in developing countries like

Nigeria such as corn pap which are very low in protein, energy, and other vital

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nutrients such as iron that support optimal growth can contribute to iron

deficiency.101 Besides, the traditional method of preparing corn pap permits

nutrients losses especially iron loss.102 This popularly used Corn pap (gruel)

contains 3.8 to 7.9% protein, grossly deficient vitamins and minerals and fat

content less than 2% .

Iron deficiency anaemia in PEM can be caused by parasitic infections, such as

hookworms. Intestinal bleeding caused by hookworms can lead to fecal blood

loss and haemoglobin/iron deficiency. Chronic inflammation caused by parasitic

infections as well as diversion of iron to fetal erythropoiesis, intravascular

haemolysis and haemoglobinuria contributes to anaemia in most developing

countries.

Iron deficiency occurs in three stages and this include.69-70,112

Stage 1 characterized by a reduction in serum iron and mobilization of storage

iron. Levels of transport iron and haemoglobin are reduced.

Stage 2 characterized by iron-deficient erthropoiesis involving a decrease in

transport iron (low serum ferritin, serum Transferrin receptor and free

erythrocyte protophyrin). This stage is referred to as iron deficiency without

anaemia as haemoglobin concentration is within normal limit.

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Stage 3 Iron deficiency anaemia is the final stage of iron deficiency. It develops

when the supply of transport iron decreases to the point where haemoglobin

synthesis is restricted. Thus there is low haemoglobin and low iron stores

Clinical features of iron deficiency

Iron deficiency anaemia is characterized by pallor (reduced amount of

oxyhaemoglobin in skin or mucous membrane), fatigue and weakness. Because

it tends to develop slowly, adaptation occurs and the disease often goes

unrecognized for some time. In severe cases, dyspnea can occur. Unusual

obsessive food cravings, known as pica can occur. Geophagia, and pagophagia

may also develop. Hair loss and lightheadedness can also be associated with iron

deficiency anaemia. Other symptoms include sore tongue, fainting and

breathlessness. Nails may become weak, brittle and koilonychia occurs with

severe anaemia.68

Effect of Iron deficiency anaemia in PEM

Iron deficiency is a condition which there is a lack of cellular iron to the tissues.

It represent a spectrum of mild to severe forms characterized by the presence or

absence of physiologic, biochemical, haematologic and functional changes in the

body.65,112

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Clinical consequences of iron deficiency anaemia include poor cognitive

performance, anaemia, poor immune function and decreased work output. 112

Iron deficiency anaemia for infants in their earlier stages of development may

have significantly greater consequences than it does for adults. It affects the

neurological development by decreasing learning ability, altering motor

functions, and permanently reducing the number of dopamine receptors and

serotonin levels. Iron deficiency occurring during development can lead to

reduced myelination of the spinal cord, as well as a change in myelin

composition. Several studies have proven these facts.65,69,112 Additionally, iron

deficiency anaemia has a negative effect on physical growth. Growth hormone

secretion is related to serum transferrin levels, suggesting a positive correlation

between iron-transferrin levels and an increase in height and weight.71

Diagnosis

Anaemia may be diagnosed from symptoms and signs, but when anaemia is mild

it may not be diagnosed from mild non-specific symptoms. Anaemia is often

first shown by routine blood tests, which generally include a complete blood

count (CBC). A sufficiently low haemoglobin or haematocrit value is

characteristic of anaemia, and further studies will be undertaken to determine its

cause and the exact diagnosis.19,56.

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In progressive iron deficiency, sequences of biochemical and haematologic

events occur. First, the tissue iron stores represented by bone marrow

haemosiderin disappear. The level of serum ferritn, an iron storage protein,

provides a relatively accurate estimate of the body iron stores in the absence of

inflammatory disease. Normal ranges are age dependent, and decreased levels

accompany iron deficiency. Next the serum iron level decreases, iron binding

capacity (TIBC), serum transferrin increases and the percentage saturation falls

below normal and free erythrocyte protoporphyrins accumulate. As deficiency

progresses, the red blood cells(RBC) becomes smaller than normal and their

hemoglobin content decreases .This can be determined by changes in the mean

corpuscular hemoglobin (MCH) and mean corpuscular volume(MCV). With

increasing deficiency the red blood cells become deformed and misshapened and

present with characteristic microcytosis, hypochromic poikilocytosis, and

increased red cell distribution width. The reticulocyte may be normal or

moderately elevated, nucleated red blood cells are seen in the peripheral blood if

anaemia is severe. White blood cell counts are normal, sometimes

thrombocytosis occurs though thrombocytopenia may occur in severe forms.

There will be a high red blood cell distribution width (RDW), reflecting a varied

size distribution of red blood cells. In about a third of cases, occult blood can be

detected in the stools19,90.

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Treatment of Iron Deficiency Anaemia

If the cause is dietary iron deficiency, iron supplements, usually with iron (II)

sulfate, ferrous gluconate, or iron amino acid chelate NaFeEDTA, will usually

correct the anaemia. The recommended daily oral dose of elemental iron is 3-

6mg/kg/day. Mild cases may be treated with 3 mg/kg/day before break fast.

Parental administration is rarely required. Iron therapy results in an increased

reticulocyte count within 3-5 days and maximal between 5 and 7 days. The

hemoglobin level begins to increase thereafter. The rate of haemoglobin

increases is inversely related to the haemoglobin level at diagnosis. In moderate

cases an elevated reticulocyte count 1 week after initiation of therapy confirms

the diagnosis and documents compliance. When iron deficiency is the only cause

of anaemia, adequate treatment usually results in resolution of anaemia in 4-6

weeks. Treatment is continued for a few months to replenish stores. While

adequate medication is given, family must be educated about child’s diet, which

should contain iron fortified diet.65,68.

Assessment Of Iron Status

There is a wide variety of laboratory methods that can be used to assess iron

status in the body. These methods include ; Haemoglobin , haematocrit, mean

corpuscular volume(MCV), mean corpuscular haemoglobin concentration

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(MCHC); and mean corpuscular haemoglobin(MCH).Others include serum Iron,

total iron binding capacity, serum Ferritin and serum Transferrin receptor.65

Haemoglobin Concentration; The amount of hemoglobin in the blood. The

hemoglobin concentration is measured in grams per deciliter (g/dL) of whole

blood. It measures the presence of anaemia but can not distinguish between

anaemia of iron deficiency and other causes. It is interpreted according to age

and sex-specific references.

Haematocrit; The haematocrit is a test that measures the volume of blood in

percentage that is comprised of the red blood cells.

Mean Corpuscular Volume (MCV); This is haematocrit divided by the red

blood cell count. It is a measurement of the volume occupied by a single red

blood cell and it is an indication of individual red cell size. Normal value is

between 73-93fl. Decreases values indicates microcytic anaemia while raised

value indicates microcytic anaemia.

Mean Corpuscular Haemoglobin Concentration (MCHC); This is

haemoglobin concentration divided by haematocrit. It represents the average

concentration of haemoglobin in the red blood cell. It is expressed as a

percentage. Increased values indicate spherocytosis while decreased value

indicates iron deficiency. Normal value is between 13-31g/dl.

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Mean corpuscular Haemoglobin (MCH); This is haemoglobin concentration

divided by the red cell count. It represents the average haemoglobin contained in

each red cell. It is influenced by the size of the cell and haemoglobinization.

Normal values are between 27-34pg. increased values occur in microcytic

anaemia while decreased values are seen in microcytic anaemia.

Serum Ferritin

Structure: Ferritin is a large protein shell (MW 450,000) comprised of 24

subunits, covering an iron core containing up to 4000 atoms of iron. In normal

condition it may represent 25% of total iron found in the body.73

Function: Ferritin acts as the soluble storage form of iron in tissue . It may serve

other functions as well although these are controversial. It is found in most cells

of the body, especially macrophages, hepatocytes and erythrocytes. Its synthesis

occurs in the liver and the rate correlates directly with the cellular iron content.

Control of ferritin synthesis occurs post-transcriptionally (at the mRNA level).

There are iron- and cytokine-responsive elements in ferritin mRNA. Increased

iron or cytokines (such as IL-1, IL-6) promotes ferritin translation, resulting in

increased iron storage. This is one of the causes of iron "sequestration" that

occurs in animals with chronic or inflammatory disease and will lead to reduced

serum iron values. The function of serum ferritin is not known, but the

concentration correlates well with the amount of stored iron in normal (and most

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diseased) subjects. Serum ferritin concentrations are quite stable from day-to-

day, in contrast to serum iron73,74.

Measurement: Sensitive methods are needed, since serum levels are very low.

Immunologic assays requiring species-specific reagents, such as RIA and

ELISA, have been employed. Canine and feline-specific ferritin assays are also

available74.

Variation in disease: Low serum ferritin: A decrease in the amount of stored

iron is the only known cause for low serum ferritin result. This is the strong point

of this test. Increased stored iron (overload) is associated with raised serum

ferritin levels. Unfortunately, ferritin is an acute phase reactant protein, so liver

disease, infection, inflammation, or malignancy will result in high serum ferritin

levels. The concurrent presence of these conditions in an iron-deficient subject

may raise serum ferritin concentration to a level not clearly indicative of iron

deficiency; or raise the level in a subject with normal iron stores to an extent

suggestive of iron overload.74

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Overview of PEM and Iron (Serum Ferritin)

PEM is a generalized disorder affecting the structure and function of the entire

body. According to various reports the incidence of iron deficiency in PEM is

extremely variable.5,79 Iron deficiency in PEM might arise from dietary

deficiencies. This can arises from reduced dietary iron and/or low bioavailability

of dietary iron. Despite the high bioavailability of iron in human milk, it contains

a low iron store. This might result in iron deficiency anaemia in children

exclusively breast fed after 6 months. The choices of complementary foods

usually further increase the risk of iron deficiency as most complementary foods

in the developing world are poor sources of iron. 3,99,101

Despite the fact that dietary deficiency of iron is prevalent among children with

PEM, hepatic iron and bone marrow free iron might also be elevated in severe

PEM, as there is a reduction in the concentration of serum protein especially the

transport protein. Iron is usually tightly bound to proteins during storage and

transport to prevent free iron forming the extremely damaging hydroxyl radical.

Reduced serum transport proteins such as transferrin will result in a reduction in

total iron binding capacity and an increase in concentration of free iron.3

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Low serum transferin in PEM has been associated with increase mortality in

severe PEM131, 132 while free iron has been implicated in the etiology of PEM

especially kwashiokor.131, 132

Several studies on haematologic and biochemical parameters have revealed low

iron stores in children with Protein Energy Malnutrition25, 26, 97. Some of these

studies demonstrated high serum ferritin levels despite the low iron stores.

Intestinal absorption of iron is maintained in PEM children with high serum

ferritin while those with low ferritin levels have impaired absorption.134 In the

acute phase of PEM ferritin level might be elevated as ferritin serves as an acute

phase reactant26 and iron supplements at this stage has been found to be

associated with increased mortality135. However during the recovery phase of

PEM, with accompanying increase in the lean body and red cell mass, iron is

required, and stores are quickly exhausted so that classic iron deficiency features

occur.136 At this stage iron supplements would be required.

Prevention and control of iron deficiency anaemia in PEM

Interventions to prevent protein– energy malnutrition range from promoting

breast-feeding to food supplementation schemes. Micronutrient deficiencies

would best be addressed through food-based strategies such as dietary

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diversification through home gardens and small livestock. To be effective, all

such interventions require accompanying nutrition-education campaigns and

health education interventions. To achieve the hunger- and malnutrition-related

Millennium Development Goals, we need to address poverty, which is clearly

associated with the insecure supply of food and nutrition22,4

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Justification

Protein-energy malnutrition is still highly prevalent in Nigeria. Malnutrition can

be due to macro or /and micronutrient deficiency. Micro nutrient deficiency is

becoming increasingly common with iron deficiency being the commonest. The

impact of iron deficiency is enormous especially on the developing child’s

growth, developmental, cognitive and intellectual abilities. Despite this fact the

prevalence of iron in vulnerable groups such as children with protein energy

malnutrition is extremely variable.25-26 Iron deficiency results in nutritional

anaemia thus there is a need to assess the haematologic profile of the group in

question(PEM) to properly asses this important trace element.

Generally and especially in these part of the country there a paucity of literatures

on the hematological profile and ferritin levels of children with Protein Energy

Malnutrition. Majority of the studies carried out on hematological profile in

malnutrition were done several decades ago, many of such were not in this

environment. Thus, there is paucity of data on this subject and a hence the need

for this study. This will enable us to predict the status of this essential

micronutrient in this group of children, as well as their haematological profile.

Iron deficiency and its attendants consequences would thus be prevented in this

already compromised group of children. This would in turn reduce childhood

mortality, and further enhance child survival in this part of the world.

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Aims and Objectives

General objectives

To assess the haematological profile and serum ferritin levels in children with

PEM in UITH, Ilorin.

Specific Objectives

To determine;

1. The haematological profile of children with PEM

2. The serum ferritin levels of children with PEM.

3. The prevalence of anaemia in children with PEM

4. The prevalence of iron deficiency anaemia in children with PEM in Ilorin

5. The relationship between the clinical feature and serum ferritin levels in

PEM.

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Materials and Methods

Study Design

The study was a case control study in which subjects were children diagnosed

for PEM and controls were children with normal nutritional status without

haematological or infectious conditions.

Background of study site

The University of Ilorin Teaching hospital is located in Ilorin West Local

Government Area of Kwara State. This lies in the North Central zone and serves

as a gateway between the Northern and Southern parts of the country. The state

population stands at 2.4million,85 Ilorin west local government area is populated

by 36,466.86 people and the vegetations is largely Guinea Savannah.25 – 28 The

hospital serves as a referral center for Kogi, Niger, Osun, and Ekiti states. It has

an Emergency Pediatrics Unit that treats children on referral and also renders

consultation to outpatients. The Emergency Pediatrics Unit is a 30 bedded unit

with a functional side laboratory. The Hospital also offers efficient laboratory

services.

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Study population

This survey was conducted in the Emergency Paediatric ward of UITH Ilorin

among children (6 -59 months old) with malnutrition.

Sampling techniques

All consecutive admissions into the Emergency Paediatric Unit with a diagnosis

of PEM based on the Wellcome’s classification that fulfilled the inclusion

criteria were enrolled. Controls were well children attending the routine clinic

without haematologic or infectious condition.

Inclusion criteria

1. Children between the ages 6 months and 59 months diagnosed with PEM

according to Wellcome classification.

2. Patients whose parents consent.

Exclusion criteria

1. Patients on drugs containing iron

2. History of blood transfusion in the last 3 months

3. History suggestive of on going haemolysis, such as haematuria, and other

indices of chronic blood loss, malignancy and peptic ulcer disease.

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4. Children with haemoglobinopathies such as Sickle cell anaemia,

thalassemia, and patients with HIVs Positive status.

Research Instruments

Interviewer administered semi-structured questionnaire.

Stadiometer, tape rule, and weighing machine.

Proforma (Questionnaire)

A semi–structured questionnaire (proforma) was used to collect information

from the subjects using interview method. The questionnaire was translated into

other languages such as Yoruba, and Nupe and used for respondents (mothers

and care givers) who do not understand the English language. Relevant

information on the child’s socio-demographic characteristics, nutritional indices

and laboratory findings were obtained.

Pre- Testing of instrument

The research instrument was pre-tested among mothers of children with Protein

Energy Malnutrition in the children’s ward at children specialist hospital Ilorin.

This was to ensure validity and reliability of the instrument. It also gave an idea

of the level of difficulty and complexity, which could hinder the administration

of the instrument. The pre-tested instrument was then analyzed and necessary

modification was effected.

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Validity of the profoma (questionnaire)

Senior Laboratory Scientist and Lecturers in the Department of Pediatrics,

Chemical Pathology and Hematology were consulted for a careful vetting of the

initial items in the proforma. The correction and suggestions on the tool lead to

the modification of some of the items of the instrument all of which were

incorporated into the final copy of the proforma.

Institution/College approval

The study was approved by the Ethics and Research Committee of the University

of Ilorin Teaching Hospital (Appendix vi), and also by the National Post

Graduate Medical College of Nigeria (Appendix vii)

Consent

The caregiver were interacted with and were provided with adequate

explanations about the study as contained in the information sheet (Appendix i)

with a view to obtaining their consent by signing or thumb printing on the form

for that purpose.

All the patients received standard treatment for the primary conditions. The

study pro forma used is as displayed in Appendix ii.

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Method of data collection

The researcher along side with research assistants made up of Paediatrics

residents that were trained, were involved in sample collections and

administration of questionnaires. The researcher and the Laboratory Scientist in

hematology, chemical pathology and microbiology were involved in sample

analysis and interpretation of laboratory results.

Sample size determination

The minimum sample size was determined using the formula (29,30)

N=2z2pq

d2

Where

N= desire sample size

Z = the standard normal deviate usually set at 1.96 (or simply at 2.0) which

corresponds to 95% confidence interval.

P = the proportion in the target population estimated to have a particular

characteristics (malnutrition) The prevalence from a previous study is put at 10

%( 0.10)75

q=1-p

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= 1.0 – 0.10=0.90

d =observed difference of 10% or more taken as being significant.

N = 2 x (1.96)2 x 0.10 x0.90

(0.10)2

= 69

The minimum calculated sample size was 69; making allowance for at least 10%

attrition rate, the calculated sample size was 76. To obtain an equal distribution

for statistical analysis, 90 subjects each were selected for the study and control

groups with a total of 180 participants for the study.

Data Collection /analysis

Structured questionnaire was used to obtain information’s on;

1. Socio-demographic characteristic.

2. Clinical profile of sign and symptoms.

3. Anthropometric measurements.

4. Laboratory parameters.

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Socio-Economic Index Score

Socio-economic index scores were awarded to the occupations and educational

attainments of their parents or caregivers using the Oyedeji socio –economic

classification scheme13 (Appendix IV). The mean of four scores (two for the

father and two for the mother) approximated to the nearest whole number was

the social class assigned to the child as proposed by oyedeji.13 For example, if

the mother was a junior school teacher (score = 3) and father a senior teacher

(score=2) and the educational attainment of the mother was primary six

(score=4), and the father was a school certificate holder(score=2), the socio-

economic index score for this child

was; 3+2+4+2/4=2.75 to the nearest whole number 3.Socio economic class I

and II are the upper socio-economic class, class III is the middle class while

socio economic class IV and V are the lower socio-economic class.13

Clinical Examination; Clinical examination and anthropometric measurements

such as weight and height were carried out among the subjects by the

investigator. The weight was measured to the nearest 0.1kg using standardized

scales while the length was measured to an accuracy of 0.1m using an

infantometer for children less than 2 years and a stadiometer for height among

children older than 2 years children. The body mass index (BMI) were calculated

using the WT(kg)/HT(m)2 formula.6 The Z scores for height/age, weight/age

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were calculated for each child by comparing the height and weight of the

subjects to the reference values 15 for the age and sex expressed as a ratio of the

standard.

Blood specimen collection; Under strict aseptic conditions, after cleaning the

blood collection site thoroughly with 70% alcohol, 5ml of venous blood was

collected by venepucture using a fixed hypodermic needle. Three milliliters of

the withdrawn blood was decanted into a sample bottle containing ethylene

diamine tetra acetate (EDTA) and gently mixed to prevent clotting while the

remaining 2ml was decanted into a heparinized bottle which was left to stand for

2 hours and the serum was separated by centrifuging and serum was decanted

into another bottle and frozen at -20˚C.

Stool collection and analysis; Freshly passed stool was collected using pre

coded specimen bottles. Some of the stool samples were analyzed by the

researcher under the supervision of a Senior Laboratory Technologist within an

hour of collection or preserved using 10% formol saline when immediate

analysis was not possible. Each stool sample was subjected to macro and

microscopic examinations using saline and iodine preparation to assess red blood

cells.90,91

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Laboratory Analysis;

Full blood count

Under close supervision of senior lab scientists, pathologists and other members

of staff in the laboratory an automated blood analyzer model/Symax KX 21®

was used to analyse the full blood count. This works on the principle of

electronic gating. The passage of cells through charged orifices result in voltage

changes whose magnitude and pulse frequency suggests cell size and count

respectively.

The machine was used to analyse composition and concentration of the cellular

components of blood such as: red blood cell (RBC) count, hemoglobin,

haematocrit and red blood cell indices (mean corpuscular volume (MCV), mean

corpuscular haemoglobin (MCH), and mean corpuscular haemaoglobin

concentration (MCHC)); white blood cell (WBC) count, classification of White

Blood Cells (WBC differential);and Platelet Count. The limitation of the use of

this procedure is its inability to differentiate between monocyte, eosinophils and

basophils.

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Peripheral blood film preparation for cell morphology

A small drop of venous blood was placed on a glass microscope slide, using a

glass capillary pipette. A spreader slide was positioned at an angle 45˚and slowly

drawn toward the drop of blood. The spreader slide was brought in contact with

the drop of blood and was drawn away. The spreader slide was further pulled

out, leaving a thin layer of blood behind glass slide with a well-formed blood

film. After drying for about 10 minutes, the slide was stained manually using the

Wrights’ stain.

A low power examination of a peripheral blood smear, the 50X or 100X

objective of the microscope was selected and the area of morphology was

examined in a consistent scanning pattern to avoid counting the same cell(s)

twice. A differential count of at least 100 red blood cells (200, 500, or 1000) was

performed, and any abnormal morphology of RBCs observed during the

differential count was recorded. Each morphologic abnormality observed was

quantified ("graded") separately as to severity ("slight to marked" or "1+ to 4+").

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Figure 1: Picture of the peripheral blood smear prepared. The arrow points

to the zone of morphology.

Serum ferritin assay

Serum Ferritin concentrations was analyzed by enzyme—linked immuno

adsorbant assay using the Nova Path TM Ferritin kit.93 Serum was prepared from

whole blood specimen collected under aseptic condition into an EDTA bottle.

Serum was capped and stored at -20 oC.95

Principle of the assay; The ferritin quantitative test is based on the principle of

a solid phase enzyme linked immunosorbent assay. The assay system utilizes

rabbit anti-ferritin for the solid phase (micro titer wells) immobilization and

mouse monoclonal anti-ferritin in the antibody–enzyme (horseradish peroxidase)

conjugate solution. The serum was allowed to react simultaneously with anti

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bodies, resulting in the ferritin molecules being sandwiched between the solid

phase and enzyme linked antibodies. After 45 minutes incubation at room

temperature, the wells were washed with water to remove the unbound labeled

antibodies. A solution of 3, 3, 5, 5’- Tetramethylbenzidine (TMB) was added and

incubated for 20 minutes, resulting in the development of a blue color. The color

development was stopped with the addition of 1N HCl, and the resulting yellow

colors measured spectrophotometrically at 450nm. The concentration of ferritin

is directly proportional to the color intensity of the test sample.95, 96

Standardization and quality control; The reference standards are calibrated

against the international committee for standardization in Hematology (ICSH)

Expert panel on iron, human liver standard (NIBSC_WHO80/602). Good

laboratory practice requires that quality control specimens (controls) be run with

each calibration curve to verify assay performance. To assure proper

performance, a statistically significant number of controls were assayed to

establish mean values and acceptable ranges.96, 96

Calculation of Serum ferritin level

In constructing the standard curve, the ferritin standard value was checked on

each vial. The absorbance for ferritin standard (vertical line) was plotted against

the Ferritin standard concentration (horizontal) line on a linear graph paper. The

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absorbance for the controls and each unknown sample was read from the

curve.(Appendix V)

Normal mean serum ferritin level for children in this age group was 31ng/ml65,

low serum ferritin was defined as levels below 31ng/ml. Iron deficiency was

defined as serum ferritin levels below 31ng/ml, while iron deficiency anaemia

was defined as haemoglobin less than 11g/dl and serum ferritin < 31ng/ml.64,75,97

Methods for assessing iron deficiency

Haemoglobin; The level falls in the third stage of iron deficiency and the degree

of fall reflects the severity of the iron deficiency. Anaemia was defined as

haemoglobin <11g/dl.

Haematocrit; The estimation of packed cell volume has the advantages of

simplicity, accuracy, few technical errors and can be done in capillary samples.

Mild anaemia was haematocrit =26-29%, moderate anaemia was haematocrit

=21-25% and Severe anaemia, was haematocrit of <20%. 110

Red blood cell count; the estimation of the red cell count is a simple and

accurate test if an electronic counter is available.

Mean corpuscular Volume (MCV) This is the haematocrit divided by the red

blood cell count. It is a measurement of the spacies occupied by a single red

blood cell and it is an indication of individual cell size. Decreased values

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indicate microcytic anaemia (as seen in iron deficiency anaemia, thalasemia ),

while increased values indicate differentials that encompass macrocytic anaemia

(for example Vitamin B12 deficiency, folate deficiency). MCV 70-74fl was

taken as normal range. Values less than 70fl was taken as microcytosis and

>74fl as macrocytosis.

Mean corpuscular haemoglobin (MCH).The MCH is the haemoglobin

concentration per cell (haemoglobin mass/red blood cell), expressed in

picograms per cell (pg, 10-12 g). The MCH is calculated from the haemoglobin

and RBC by the following formula:

The MCH is decreased in patients with anaemia caused by impaired

haemoglobin synthesis. The MCH may be falsely elevated in blood specimens

with turbid plasma (usually caused by hyperlipidemia) or severe leukocytosis.

Mean corpuscular haemoglobin concentration (MCHC); The MCHC is the

average haemoglobin concentration per total red blood cell volume (ratio of

haemoglobin mass to RBC volume), as determined from the following equation:

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The MCHC is decreased in microcytic anaemias where the decrease in

haemoglobin mass exceeds the decrease in the size of the red blood cell. It is

increased in hereditary spherocytosis and in patients with haemoglobin variants,

such as sickle cell disease and hemoglobin C disease).

Red cell morphology

Normocytic normochromic Normal red blood cells ("normocytes,"

"discocytes") are round to very slightly ovoid cells with a mean diameter of

approximately 7 mm and a central pale area ("area of central pallor")

approximately 1/3 the diameter of the cell that gradually fades towards the more

deeply stained periphery.

Hypochromia

Hypochromia is a decreased amount (MCH) and concentration (MCHC) of

haemoglobin in red blood cells. In the peripheral blood smear, hypochromic cells

have an expanded central zone of pallor. Small hypochromic red blood cells

(microcytes) are usually present, and the mean corpuscular volume (MCV) is

decreased. Microcytosis and hypochromia are characteristic of iron deficiency

anaemia and other microcytic, hypochromic anaemias [anaemia of chronic

disease, hereditary haemoglobinopathies with diminished globin synthesis

(thalassemias, haemoglobin E, haemoglobin H), red blood cell enzyme

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deficiencies (sideroblastic anaemias, lead poisoning, and pyridoxine

deficiency)].

Figure 2 : Comparisons of normal peripheral blood smear with smear of

microcytic, hypochromic anaemia.

Microcytes

Microcytes are small red blood cells (MCV < 70 fL) with decreased amounts of

haemoglobin formed as a result of iron deficiency and defective haemoglobin

synthesis, imbalance of globin chains, or defective porphyrin synthesis. Possible

clinical causes of microcytosis include iron deficiency anaemia, thalassemia, the

anaemia of chronic disease, lead poisoning, and sideroblastic anaemia.

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Benefit of study to participants.

The children that participated benefited from the study by knowing their blood

profile and ferritin level. Children with anaemia depending on its severity were

treated. Those with iron deficiency also had treatment commenced. Nutritional

education was given to the parents or care giver. The children were followed up

in the nutrition clinic.

Data analysis

Data entry and analysis were carried out with a micro-computer using the Epi

info version 3.5 (2008) software packages.94 Chi-square test and student t-test

were used to test for statistical significance of the difference for discreet and

continuous variable respectively. A p value of < 0.05 was regarded as

significant. Analysis of variance (ANOVA) was used for some comparisons

when indicated. Kruskal-Wallis One Way Analysis Of variance – a non

parametric test was used for some variables when indicated.

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Results

The Socio-Demographic Characteristics of The Subject and Controls

A total of 90 children with Protein Energy Malnutrition (PEM) admitted into the

Emergency Paediatiric Unit were studied, 90 apparently well children attending

the Well-Child Clinic served as controls.

Table VII shows that of the 90 children with PEM, 59 (65.6%) were males and

31 (34.4%) were females, with a male to female ratio of 1.9:1.0. The mean age

of the children with Protein Energy Malnutrition was 22.7 + 14.4 months (Range

of 9-59months) compared to the mean age of 29.3 + 16.9 months (Range 6-59

months) in the controls and the difference was not significant (p=0.08).

Forty two percent(38) of the children with Protein Energy Malnutrition were in

socio-economic class (SEC) IV, 26 (28.8%) in SEC III, 18(20%) in SEC V and

only 2(2.2%) in SEC I. The subjects were of a lower socioeconomic class

compared to the controls (p=0.00001).

Of the 90 mothers interviewed, 29(32.2%) had primary education, 25(27.8%)

had no form of education, while 21(23.3%) and 15(16.7%) had secondary and

tertiary education respectively. The educational status of mothers of children

with PEM were lower compared to that of controls (p=0.0002).

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TABLE VII: The socio-demographic characteristics of the subject and

controls

Variable PEM

(n=90)

Controls

(n=90)

χ2 p

Age (month)

Range

Mean S.D

Gender

Male

Female

Social Economic Class

I

II

III

IV

V

Maternal Educational Status

None

Primary

Secondary

Post secondary

Exclusive Breast Feeding

Practices

Yes

No

Age of introduction of

complementary food (Months)

MeanS.D

Type of complementary feeds

Guinea corn gruel

Maize gruel

Infant formula

Fortified gruel

Milk

9.0-59.0

22.714.4

59(65.6%)

31(34.4%)

2(2.2)

6(6.8)

26(28.8)

38(42.2)

18(20.0)

25(27.8)

29(32.2)

21(23.3)

15(16.7)

25(27.8%)

65(72.2%)

5.11.93

68(75.6%)

9(10%)

2(2.2%)

10(11.1%)

1(1.1%)

6.0-59.0

29.316.9

53(58.9%)

37(41.1%)

4(4.4)

27(30.0)

35(38.9)

16(17.8)

8(8.9)

9(10)

20(22.2)

23(25.6)

38(42.2)

33(36.7%)

57(63.4%)

5.62.4

56(62.2%)

11(2.2%)

2(2.2%)

16(17.8%)

5(5.6%)

t=7.95

0.85

28.17

19.025

1.63

t=2.35

12.93

0.08

0.356

0.000012

0.0002

0.2019

0.26

0.024

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The percentage of PEM children exclusively breast fed (27.8%) was lower than

36.7% found in the controls. However the difference did not reach statistical

significant difference (p=0.02).The mean age for the commencement of

complementary feeds was 5.1±1.93 months in children with Protein Energy

Malnutrition compared to 5.6±2.4 months in the controls, and the difference was not

significant (p=0.126) .

Over 68% of the PEM cases were fed with guinea corn gruel, 11.1% with

fortified gruel, 10% with maize gruel and only 1.1% added milk to the feeds.

The children with PEM were fed more with non nutritive feeds when compared

to the controls (p=0.02).

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Anthropometric Measurements Of the Study Population

Table VIII shows that the mean weight of PEM children was 7.9 + 3.2 kg while

that of the controls was 13.9 + 13.6kg and the difference was significant

(p=0.0001). The mean height was 0.75 + 0.11m for PEM and 0.94±25.3m for

controls, the difference was significant (p=0.0001).

The mean for the mid upper arm circumference was 11.9 + 3.4cm and 16.4 +

2.6cm for the subjects and controls respectively. The mean BMI for the controls

(15.2±3.4kg/m2) was significantly higher than the BMI for the subjects

(13.3±4.5kg/m2), (p=0.001). Occipito frontal circumference was similar in both

groups (p=0.03). The mean Z score of weight for age was -3.2±1.8 for PEM and

-5.03± 1.0 for controls, there was a statistically significant difference ( p=0.001).

The mean weight for height Z score was -2.0±3.45 and 0.01±4.2 in subjects and

controls respectively, there was a statistically significant difference in these

values.

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TABLE VIII: Anthropometric measurements of the study population

Anthropometry PEM

(n=90)

Controls

(n=90)

t p

Weight(kg)

Range

Means ± S.D

Height (m)

Range

Means ±S.D

Body Mass Index(kg/m2)

Range

Means± S.D

Mid Arm

Circumference(cm)

Range

Mean± S.D

Occipitofrontal

circumference(cm)

Range

Mean±

Weight For Age Z score

Mean± S.D

Weight For Height Z score

Mean± S.D

3.2-13.2

7.9±3.2

0.5-0.80

0.75±111

7.0-23.7

13.3±4.5

9.2-16.1

13.2±8.6

40.0-51.0

45.6±2.1

-3.2±1.8

-2.01±3.45

4.2-23.9

13.9±13.6

0.6-0.99

0.94±25.3

8.2-29.1

15.2±3.4

12.0-19.0

15.1±1.6

41.0-56.0

47.7±8.2

-5.03±1.0

-0.01±4.2

192.86

12.86

9.99

4.22

4.82

303.98

17699.4

0.0001

0.0001

0.001

0.04

0.03

0.001

0.001

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Distribution of Types of Protein Energy Malnutrition According to the Age

in the Study Population

Table IX Shows that 50 (55.6%) children with PEM were Underweight,

21(23.3%) had Marasmus, 11 (12.2%) had Marasmic-Kwashiokor and 8 (8.8%)

had Kwashiorkor. Over a half of the children were in the 10 - 19 months age

bracket.

Table IX: Distribution of types of protein energy malnutrition according to

the age in the study population

Age range

(months)

Marasmus

(%)

Kwashiorkor(%) Marasmic-

kwashiokor(%)

Underweight Total

(%) (%)

0-9

10-19

20-29

30-39

40-49

50-59

60-69

Total

0

10

4

0

4

1

2

21(23.3)

0

8

0

0

0

0

0

8(8.8)

0

6

5

0

0

0

0

11(12.2)

4 4

26 50

4 13

10 10

2 6

2 3

2 4

50(55.6)90(100)

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Haematologic Profile of the Study Population

Table X shows that the mean haematocrit values for the subjects and controls

were 30.4 ± 6.3% and 32.0 ± 6.1% respectively while the mean haemoglobin

values for the subject and controls were 10.1±2.1g/dl and 10.9±15.0g/dl

respectively and the difference was statistically significant (p=0.019 and 0.003

respectively) with the svalue for PEM being lower.

The mean values of the mean corpuscular volume were 72.4 ± 10.9fl and 72.6 ±

13.6fl in the subjects and controls respectively and the values were similar (p =

0.913).

The mean values for mean corpuscular haemoglobin concentration and mean

corpuscular haemoglobin in the subjects were 30.4±2.8g/dl RBC and

24.3±10.5fl.These values in the controls were30.3±1.8 and25.6±1.6,and were

similar in both the subjects and controls (p>0.05).

The mean value of platelets count were 291.8 ±131.7 and 326.4 ±133.9 for the

subjects and controls respectively and the difference was statistically significant

(p=0.0001) with PEM children having a lower platelet count compared to the

controls.

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TABLE X Haematologic profile of the study population

Haematological

Parameters

PEM

mean±S.D

Controls

mean±S.D

t P

RBC (x100 cell/mm3)

Haemoglobin (gldl)

Haematrocrit (%)

MCV(fl)

MCH (pg/cell)

MCHC(gHbldl RBC)

Platelet count(x103

cell/mm3)

White cell count(x103

cell/mm3)

Neutrophils (%)

Lymphocyte (%)

Total protein(g/l)

Albumin(g/l)

4.0±0.9

10.1±2.1

30.4±6.3

72.4±10.9

24.3±10.5

30.4±2.8

291.8±131.7

12.8±11.6

49.5±12.3

52.7±12.3

3.24±1.36

2.11±0.87

4.2±0.9

10.9±15.0

32.0±6.1

72.6±13.6

25.6±10.6

30.3±1.8

326.4±133.9

5.9±8.7

43.8±5.9

59.4±7.5

3.90±1.03

2.94±0.65

2.39

18.58

2.97

0.01

0.68

0.68

180.18

20.38

15.64

23.80

12.83

4.50

0.123

0.019

0.003

0.091

0.41

0.411

0.0001

0.001

0.001

0.00002

0.0044

0.0352

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The mean total white cell count of 12.8 ± 11.6×109 was higher in PEM than 5.9

± 8.7×109 in controls and the difference is significant (p=0.001) with the subject

having a higher white cell counts than controls. The Neutrophil value was 49.5 ±

12.3 % and 43.8 ± 5.9 % for PEM and controls respectively, the difference was

statistically significant. (p = 0.0001). The mean value for lymphocyte was 52.7 ±

12.3 % for PEM and 59.4 ± 7.5% for controls. It was statistically significant. (p

= 0.00002) with a lower count in the subjects compared with controls.

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Haematolgic Profile According to the Types of PEM

Table XI shows that the mean values of red cell count, haemoglobin, and

haematocrit, MCV, MCH, and MCHC were similar in all the types of

malnutrition (p>0.05), however, the mean MCHC was significantly high in all

the types of PEM, with Marasmic- Kwashiokor having the highest mean.

Children with Kwashiokor had the highest levels of haemoglobin, (31.6±1.6g/dl)

and haematocrit (10.7±0.4%), while subjects with Marasmus had the lowest

values of haematocrit (27.6±5.8%), haemoglobin (9.1±2.1g/dl) and mean

corpuscular haemoglobin (22.9±2.3pg/cell).

Underweight children had the highest mean of white cell count (13.8±14.5×103

cell/mm3) while Marasmic –Kwashiokor had the lowest mean count, however,

the difference is not statistically significant (p=0.750). The neutrophils counts

were similar in all the types of Protein Energy Malnutrition (p=0.438).

Children with Kwashiokor had the highest lymphocyte count while underweight

had the lowest lymphocyte count with a statistical significant difference

(p=0.05).

The children with Marasmic-Kwashiokor had the lowest level of platelets

(226.5±48.7×103cell/mm3) though, there was no significant difference in the

platelets count in all the various types of Protein Energy Malnutrition (p=0.171).

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Table XI: Haematolgic Profile Of Children According To The Types Of

PEM

Haematologic

Parameters

Marasmus

n=21

Mean±S.D

Kwashiokor

n=8

Mean±S.D

Marasmic-

kwashikor

n=11

Mean±S.D

Underweight

n=50

Mean±S.D

t p

RBC(×10 6

cell/mm3)

Haemoglobin

(g/dl)

Haematocrit (%)

MCH(pg/cell)

MCHC(hb/dlRBC)

MCV(fl)

Total WBC (x103

cell mm3)

Neutrophils%

Lymphocyte%

Platelet (x103 cell

mm3)

3.82±0.91

9.1±2.1

27.6±5.8

22.9±2.3

31.2±2.9

68.1±11.9

12.9±8.8

47.2±17.9

51.8±13.9

273.9±156.2

4.28±0.05

10.7±0.4

31.6±1.6

22.4±0.5

31.5±2.1

73.0±3.6

11.2±0.7

52.3±13.4

67±4.1

270.0±5852

4.28±0.05

10.4±1.7

31.1±5.1

23.5±1.3

33.1±1.5

74.4±6.2

9.9±1.3

44.7±7.4

59.9±4.2

226.5±48.7

4.08±9.5

10.3±2.4

31.08±21

25.2±13.9

29.4±2.9

73.3±2.1

13.8±14.5

51.2±9.8

49.1±7.1

316.8±137.6

0.85

1.89

1.77

0.36

4.43

1.231

0.405

0.915

11.62

1.7022

0.47

0.1575

0.157

0.7815

0.006

0.3015

0.750

0.438

0.0001

0.171

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Red Cell Changes In Children With Protein Energy Malnutrition.

Table XII shows that all the children with Kwashiorkor had mild anaemia

8(100.0%). Moderate anaemia was found in 9(42.0%) children with Marasmus,

3(27.3%) with Marasmic-Kwashiorkor and 10(20.0%) children with

underweight .Severe anaemia was found in only 2(9.5%) of the Marasmic and

4(8.0%) of the underweight children.

All the subjects with Kwashiorkor and Marasmic –Kwashiokor had microcytic

cells whereas microcytosis and macrocytosis was seen in 66.0% and 23.8% of

subjects with marasmus. All the subjects with kwashiorkor and Marsmic –

Kwashiokor had microcytic hypochromic anaemia while it was observed in

90.5% and 76% of marasmus and Underweight children respectively.

The prevalence of anaemia in PEM was 74.4% as a total of 67 out of the 90

subjects had haemoglobin level below 11g/dl.

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Table XII: Red Cell Morphology and Prevalence Of Anaemia In Children

With Protein Energy Malnutrition.

Red cell

Changes

marasmus

n=21

Kwashiokor

n =8

Marasmic-

kwashiokor

n=11

Underweight

n=50

Anaemia

Mild

Moderate

Severe

RBC size

Normocytic

Microcytic

Macrocytic

PBF*

appearance

Normochromic

Microcytic-

hypochromic

Prevalence of

Anaemia

Hb˚<11g/dl

2(9.5%)

9(42.9%)

2(9.5%)

2(9.5%)

14(66%)

5(23.8%)

2(9.5%)

19(90.5%)

19(21.1%)

8(100%)

Nil

Nil

Nil

8(100%)

Nil

Nil

8(100%)

8(8.9%)

3(27.3%)

3(27.3%)

Nil

Nil

6(54.6%)

Nil

Nil

11(100%)

6(6.6%)

6(12%)

10(20%)

4(8%)

12(24%)

12(24%)

26(52%)

12(24%)

38(76%)

34(37.8%)

*PBF -Peripheral blood film

˚Hb - Haemoglobin

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Table XIII shows that only 3 of the PEM had RBC in their stool and there was

no statistical significant difference in stool RBC in subjects and

controls(p>0.05). There was also no significant difference in the serum ferritin

levels of PEM who had RBC in their stool and those who did not have RBC in

their stool. (Table XIV)

Table XIII : Stool RBC in PEM and Controls.

Stool RBC Present Stool RBC Absent χ2 p

PEM PEM 3 87

Controls Nil 90 2.48 0.478

Table XIV : Relationship between stool RBC and serum ferritin in PEM.

Stool RBC n Serum ferritin( Mean±SD) t p

Present 3 310.1±286.7

Absent 87 496.7±320.5 0.821 0.512

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Serum Ferritin Levels In The Study Population

Table XV shows that the serum ferritin levels increased as age advanced in the

subjects, however, the mean ferritin levels were higher in the controls compared

to PEM cases (p=0.0001). The mean serum ferritin was significantly higher in

females (p=0.00001).

Table XV: Serum Ferritin Level (ng/ml)according to the Age and Sex in the

total Study Population.

Age

(months)

PEM

Serum ferritin

Mean

Controls

Mean

t p

Age(months)

0-19

20-39

40-59

Sex

Males

Females

Total

332.±26.5

496.5±36.7

630.±49.2

349±316.4

534±376.7

386.1±39.2

459.6±1.4

536.9±27.3

355.7±42.9

428±343.3

486.1±373.5

450.7±23.7

345.70 0.004

455.66 0.003

597.14 0.0001

46.06 0.0001

48.05 0.0002

41.60 0.003

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Serum Ferritin Distribution Of The Various Classes Of PEM

Children with Marasmus had the highest mean serum ferritin level of

600.5±392.6ng/ml, while those with kwashiorkor had the lowest level of

165.5±6.2ng/ml and the difference was statistically significant (p=0.009).

Table XVI: The Levels Of Serum Ferritin According To The Types Of

PEM and Controls

Serum Marasmus Kwashiokor Marasmic- Under- Controls t p

Ferritin Kwashiokor weight

Range 4.5-1000 15.8-170 11.2-780 1.0-1000 67-1000

ng/ml

Mean±SD 600.5±392.6 165±6.2 320±252.7 395.2±340 450.7±23.7 4.010 0.09

Prevalence Of Iron Deficiency And Iron Deficiency Anaemia Amongst PEM

Table XVII shows that, the prevalence of iron deficiency was 24.4% in the

children with Protein Energy Malnutrition. It also shows that the prevalence of

iron deficiency anaemia was 16.6% in these children.

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Table XVII: Prevalence of Iron Deficiency And Iron Deficiency Anaemia

Amongst PEM.

Clinical Profile and Serum Ferritin Levels

Table XVIII shows that those with fever had the highest mean value of serum

ferritin 455.22±365.42 and a significant relationship with serum ferritin

(p=0.03). while those with glossitis had lowest mean serum ferritin value but

was not statistically significant (p>0.05).

Iron Status Marasmus Kwashiokor

n=21 n=8

Marasmic-

kwashiokor

n=11

Underweight Total

n=50 Total (%)

Iron

deficiency

Iron

deficiency

anaemia

4(4.4%) 4(4.4%)

4(4.4%) 3(3.3%)

3(3.3%)

3(3.3%)

11(12.2%) 22(24.4%)

5(5.5%) 15(16.7%)

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Table XVIII: Relationship between clinical features and serum ferritin

levels in PEM.

Clinical

feature

Present

Serum ferritin

N (mean±S.D)

Absent

Serum Ferritin

n(mean±S.D)

t p

Vomiting

Fever

Pallor

Diahorea

Cough

Failure to

gain

Edema

Skin

changes

Stomatitis

Glositis

54(408.44342.56)

71(455.22365.42)

25(408.44342.56)

60 (369.5330.4)

49 (429.1367.3)

63 (437.3356.4)

29 (384.1319.9)

28 (320.9301.4)

15 (431.4311.6)

2 (225.5317.5)

36(406.34347.18)

19(257.68±214.19)

64(406.3±347.18)

30(501.5±369.9)

41(394.9±326.20)

27(357.9±325.9)

59(422.5±357.4)

62(455.3±361.2)

75(409.9±356.3)

88(417.8±348.8)

0.026

2.249

0.026

1.715

0.046

0.992

0.441

1.716

0.217

0.7

0.980

0.027

0.980

0.090

0.644

0.323

0.660

0.089

0.828

0.4424

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DISCUSSION

Majority of the children in all the four classes of Protein Energy Malnutrition

studied were in the age range of 10-19 months and Sixty five percent of the

subject were males. Sixty two percent of the subjects belong to families of low

socio economic class and mothers with low level of education. These are major

risk factors for the development of malnutrition and nutritional anaemia in PEM

as found in other studies. 99,101

In this study, haematologic changes observed include a significantly lower mean

values for MCH, MCV and RBC count in children with PEM as compared to

the controls. A lower mean values was also observed in the haematocrit and

haemoglobin values of children with PEM as compared to controls a finding

similar to previous studies.12, 27 Red cell changes observed among the various

classes of PEM in this study showed that there were no significant differences in

the red blood cells in the various types of PEM except for the MCHC. The

lowest red blood cell counts, haemoglobin and haematocrit values were observed

in marasmus while underweight had the highest of these values. These red cell

changes can be attributed to adaptation to lower metabolic oxygen requirements

ands decrease in lean body mass seen in PEM.78

This study found a significant leucocytosis and neutrophilia among children with

PEM as compared to controls, this is similar to a previous study where there was

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a significant rise in leukocyte count in the patients with PEM compared to the

controls. despite the fact that children enrolled in that study had no apparent

infection.97 Leucocytosis in these children can be a result of infection which is

seen commonly in PEM:both PEM and infection, either clinical or subclinical

have been reported to act synergistically .81 This has been an important factor in

determining morbidity and mortality attributed to PEM.27 However, a lower

lymphocyte count was observed in the malnourished children compared to

controls. The lower lymphocyte count can be attributed to changes in the

Thymus which is greatly reduced in children during severe Protein Energy

Malnutrition. The degree of Thymic atrophy correlates closely with depletion of

lymphocytes and a decrease in the thymic dependent lymphocyte is also

associated with impaired immunity.74

The study also shows that the underweight children had the highest white cell

count; this can be attributed to their immune response to infection that is not so

severely compromised as compared to those of the more severe forms of PEM. A

significant difference in the lymphocyte count was seen in the various classes of

Protein Energy Malnutrition, this is in keeping with a previous study.24 No

significant changes was observed in the platelet of the various classes of PEM

but there was a significant difference in the controls compared to PEM. Children

with PEM had a significantly lower platelet count.

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In this study, a lower mean serum ferritin level was observed in the

malnourished compared to controls. This is in contrast to previous studies where

a higher serum ferritin value was observed in PEM as compared to controls, 26

though the subjects studied were in the recovery phase of PEM. However, this

study is in conformity with other studies that assessed the serum ferritin levels in

PEM in the acute phases and found that serum ferritin was lower in the children

with malnutrition.24, 97 This low serum ferritin level is a true reflection of iron

deficiency in these children, as serum ferritin is the best indicator for iron

deficiency because a decrease in the amount of stored iron is the only known

cause for low serum ferritin result.74

There was a statistical significant difference in the serum ferritin level of the

various classes of malnutrition. The lowest mean value was seen in children with

kwashiorkor. Marasmus had the highest mean serum ferritin and this is not

surprising as the low serum protein seen in marasmus allows an increased free

unbound iron. Serum protein is necessary for the binding and transport of iron.66

The number of subjects that had red blood cell in the stool was not significant

and there was no significant difference in the serum ferritin of those that had

RBC in the stool compared to those who did not. This finding shows that iron

losses through stool was not common in these group of children studied. Thus

iron deficiency can be attributed to dietary deficiencies majorly in this study.

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A high prevalence of anaemia (74.4%) among children with Protein Energy

Malnutrition found in this study, is in consonance with previous studies where

anaemia was said to be a common feature of Protein Energy Malnutrition.12,25-27

Mild to moderate anaemia with Microcytic hypochromic cells was the most

commonly observed in PEM in this study, this is in contrast with previous

studies where anaemia seen in Protein Energy Malnutrition was normocytic

normochromic.25, 27, 75 Several factors have been implicated in the aetiology of

anaemia in PEM and these include deficiencies of iron, folate, Vitamin B12 ,

infections, blood loss, haemolysis, erythroid hypoplasia and adaptation to lower

metabolic oxygen requirements and decrease in lean body mass.78 Microcytic

hypochromic anaemia found to be the commonest in this study can be attributed

to deficiency of iron which is a common cause of microcytic hypochromic

anaemia. This finding strongly support the fact that iron deficiency was

commonest cause of nutritional anaemia in this part of the world.57 Nutritional

anaemia among children in this study can be attributed to the type of food which

was eaten by majority of the PEM children in this study. Eighty five percent of

PEM children in this study were fed with common staple food such as maize

gruel with iron content of 0.02mg, guinea corn and millet, which do not contain

any iron at all. 101 These provisions are below the recommended daily

requirement of elemental iron which is 3-6mg/kg/day.

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The prevalence of iron deficiency anaemia is 16.6% in this study, this finding is

higher than values documented in previous studies .27, 75 This higher prevalence

of iron deficiency anaemia in this study can result from a low practice of

exclusive breastfeeding (27.8%) reported in this study as, early age of

introduction of complementary feeds, and nutritionally poor complementary

feeds given to the children.

This study also shows that serum ferritin was highest in the children with PEM

that presented with fever, and there was a significant relationship between fever

and serum ferritin level.

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Conclusion

It is concluded from this study that;

1. The subjects have a lower haemoglobin, heamtocrit, platelet count and

lymphocyte and a higher total white cell count and Neutrophils than the

controls.

2. Malnourished children had a lower serum ferritin levels than the controls.

3. The prevalence of anaemia was 74.4% in children with Protein Energy

Malnutrition

5. Microcytic hypochromic anaemia was found to be the commonest form of

anaemia in Children with Protein Energy Malnutrition.

6. The prevalence of iron deficiency anaemia in children with Protein Energy

Malnutrition is 16.6%

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Recommendations

From this study it therefore recommended that,

1. Anaemia should be sought for and treated in children with Protein Energy

Malnutrition. Iron deficiency should be considered as a most likely cause

of anaemia in this group of children in this part of the world.

2. Efforts should be increased by health workers to promote exclusive breast

feeding in the first six months of life.

3. Adequate enlightenment of mothers and health workers on appropriate

energy dense and fortified complementary foods for children.

4. Food fortification as well as supplementation of the important

micronutrient especially iron should be enhanced as a good nutrition status

remains the bedrock for good health and prevention of diseases.

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Limitation of study

1. The limitation in the use of the auto analyzer makes it impossible

to differentiate between monocyte, eosinophils and basophils.

2. Other biochemical tests of assessing the iron status of the

subjects and their relationship with serum ferritin could not be

carried out on account of inaccessibility of the reagents kits.

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References

1. Ulasi TO, Ebenebe J. Nutritional disorders in children. In: Azubuike JC,

Nkanginieme KEO eds Paediatrics and Child health in the tropical region.

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Appendix I

Information Sheet

What Is The Study About

The study aims at documenting the hematological profile and ferritin levels of

children with PEM in UITH Ilorin.

What Is Expected Of You If You Agree To Participate

You will be expected to provide answers to simple questions like your child’s

age, sex, your level of education and occupation. Your child will then be

examined in detail and certain laboratory investigations shall be performed after

collection of blood and stool samples

Your Participation Is Voluntary

Your child’s participation is voluntary and you may withdraw him or her at any

time of the study without any repercussion.

Confidentiality

The information obtained will be treated in absolute confidence. No part or

whole of such information shall be divulged to anyone except the investigators.

We owe it a duty to keep your child’s records absolutely secret.

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Benefit From Participation

Your child will benefit by knowing his or her blood profile and iron status and

knowing if he or she has occult blood in the stool in which case adequate

treatment instituted. In case of any abnormal incidental finding, adequate referral

shall be done to appropriate specialist.

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Appendix ii

Informed Consent Form

Mr/Mrs/Chief/Alhaja/Dr.----------------------------------------------

Whose address is ----------------------------------------------------

Hereby give consent to---------------------------------------------

To carry out research on my child/ward as explained to me. I am aware the test

to be carried out will not harm my child/ward.

I have the right to withdraw at any point of the study if I so wish.

All the terms of this consent have been explained to me in a language that I

understand. I am also aware that the information on my child on my child shall

be kept in strict confidence.

Sign---------------------- Sign-----------------

Child’s Parent/Guardian Interviewer

Date-----------------------

Time----------------------

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Appendix iii

Study Profoma

HEAMATOLOGICAL PROFILE AND FERRRTIN LEVELS IN CHILDREN

WITH PROTEIN ENERGY MALNUTRITIONIN UITH ILORIN

Hospital No------- Serial No -----

A) Sociodemographic data

1. Age---------

2. Sex 1)Male 2)Female

3. Mother’s Education level 1) Post 30 2) 30 3) 20 4)10 5

None

4. Father’s Education Level 1)Post 30 2) 30 3)20 4) 10 5) None .

5. Mother’s Occupation: ---------- 6. Father’s Occupation:--------

7. Family type 1)Monogamy 2)Polygamy

8. No of sibling(s) 1)1-2 2) 3-4 (3) ≥5

B) Feeding pattern;

9. Was the child Exclusively breastfeed yes No

10. At what age was complementary feeding commenced----------

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11. What type of food is given---------?

12. How is the child given the food (1) cup & spoon (2) feeding bottle?

13. Who feeds the child (1) mother (2) care giver

14. How many times is the child fed in a day --------------------

(C) Any Associated illness Yes No

15. If yes what illness (1) Pneumonia (2) Tuberculosis (3) Diarrheal disease

(4) Others (specify) ------

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D) Clinical Profile symptoms and Signs

S/N Symptoms and Signs Yes =1 No=2

16. Fever

17. Vomiting

18. Diarrhea

19. Cough

20. Failure to gain weight

21. Edema

22. Pallor

23 Skin changes dry peeling or hyper pigmentation

24. Angular stomatis

25. Glositis

26. Fissured nail

27. Papillary atrophy

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(E) GROWTH MEASURE:

S/N Anthropometry Indicators Value

29. Weight for age

30. Height for age

31. Weight for height

32. BMI=W(kg)/HT(m)2

33. Mid Arm Circumference

34. Occipital frontal

circumference OFC

35. Percentage of expected

weight

(F) LABORATORY; HEMATOLOGICAL PARAMETERS

RBC (cells X l06.ccı) -----------------------

Hb (g/dl) --------------------------------------

Hct(%)-----------------------------------------

MCV (fl) --------------------------------------

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MCH (pg/cell) -------------------------------

MCHC (%) -----------------------------------

PLATELET COUNT (cell/mm3)

WBC (per cells/ mm3)

Serum Protein (Total and albumin)

Peripheral blood film PBF---------------------------------------------

(G) Iron Status

Serum ferritin(ng/mL)

Stool examination for Red blood cell Present Absent

(H) Outcome Of The Patient

Well and discharge--------Yes Died

If yes, what is number of days in hospital-----------------

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Appendix Iv

Socio-Economic Classification Scheme By Oyedeji

For Occupation

Class Occupation

I Senior Public Servants, Professionals, Managers,

large scale traders, businessmen and contractors.

II Intermediate grade public servants and senior school

teachers.

III Junior school teachers, drivers, artisans.

IV Petty traders, labourers, messengers .

V Unemployed, full-time housewife, students and

subsistence farmers.

For Educational Status

Class Educational Attainment

I University graduates or equivalents

II School certificate holders ordinary level (GCE) who

also had teaching or other professional training.

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III School certificate or grade II teachers certificate

holders.

IV Modern three and primary six certificate holders.

V Those who could either just reads and write or were

illiterate.

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Appendix v

Serum Ferritin Absorbance Curve

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Table XI: Haematolgic Profile Of Children According To The Types Of

PEM and Controls

Haematologic

Parameters

Marasmus

n=21

Mean±S.D

Kwashiokor

n=8

Mean±S.D

Marasmic-

kwashikor

n=11

Mean±S.D

Underweight

n=50

Mean±S.D

T p Control

RBC(×10 6 cell/mm3)

3.82±0.91

4.28±0.05

4.28±0.05

4.08 ±9.5 0.85 0.47 42. ±0.9

Haemoglobin (g/dl)

9.1±2.1 10.7±0.4 10.4±1.7 10.3±2.4 1.89 0.1575 10.9±15.0

Haematocrit (%)

27.6±5.8 31.6±1.6 31.1±5.1 31.08±21 1.77 0.157 32.0±6.1

MCH (pg/cell) 22.9±2.3 22.4±0.5 23.5±1.3 25.2±13.9 0.36 0.7815 25.6±10.6

MCHC (hb/dlRBC) 31.2±2.9 31.5±2.1 33.1±1.5 29.4±2.9 4.43 0.006 30.3±1.8

MCV (fl) 68.1±11.9 73.0±3.6 74.4±6.2 73.3±2.1 1.231 0.3015 72.6±13.6

Total WBC (x103 cell

mm3)

12.9±8.8 11.2±0.7 9.9±1.3 13.8±14.5 0.405 0.750 5.9±8.7

Neutrophils% 47.2±17.9 52.3±13.4 44.7±7.4 51.2±9.8 0.915 0.438 43.8±5.9

Lymphocyte% 51.8±13.9 67±4.1 59.9±4.2 49.1±7.1 11.62 0.0001 59±7.5

Platelet (x103 cell mm3) 273.9±156.2 270.0±5852 226.5±48.7 316.8±137.6 1.7022 0.171 326.4±133