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Development of an Experimental Rat Model for Studies on Sub-acute Blood Loss Anaemia

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Development of an Experimental Rat Model for Studies on Sub-acute Blood Loss Anaemia.

ABSTRACT  

Anaemia is one of the most clinically encountered conditions in animals and humans. It continues to be a global public health problem, affecting both developing and developed countries with major consequences for human and animal health as well as social and economic development. The objective of this study was to develop and test a rat model for experimental studies on sub-acute blood loss anaemia. The study design was an experimental research and development protocol, made up of two experiments (1 and 2).

Experiment 1 was for development of the model while experiment 2 was for testing the developed model. A total of 162 albino rats of either sexes, between the ages of 10 and 12 weeks weighing 130 – 250g were used for the study. In experiment 1, 130 rats were assigned at random, into thirteen groups of ten each (5 males & 5 females) labeled 1 to 13. Baseline values of the haematological parameters and serum protein were determined using standard procedures before the commencement of the study.

Rats in group 1 were not bled and served as control. Blood loss was induced at specified periods of time for the remaining 12 groups using the orbital bleeding technique. At the end of the specified period of bleeding for each group, the haematological parameters and serum protein were re-assessed in order to select which group met the target of halving the RBC count, Hb and packed cell volume (PCV) with minimal stress, which is characteristic of sub-acute blood loss anaemia.

TABLE OF CONTENTS

Title page—————————————————— ii
Certification page—————————————————— iii
Dedication—————————————————— iv
Acknowledgement—————————————————— v
Table of contents—————————————————— vii
List of tables—————————————————— xi
Abstract—————————————————— xiv

CHAPTER ONE: INTRODUCTION———————————————– 1
1.1: Background of the study—————————————————- 1
1.2: Statement of Problem—————————————————— 7
1.3: Objective of the Study—————————————————– 8

CHAPTER TWO: REVIEW OF RELATED LITERATURE—————- 9
2.1: Blood – An Overview——————————————————- 9
2.1.1: Blood Cellular Components———————————— 10
2.1.1.1: Erythrocytes (Red Blood Cells)——————– 10
2.1.1.2: Leukocytes (White Blood Cells)——————- 12
2.1.1.3: Platelets (Thrombocytes)—————————- 14
2.2: Anaemia———————————————————————- 15
2.2.1: Symptoms and Signs of Anaemia—————————— 15
2.2.2: Classification and Types of Anaemia————————— 16
2.2.2.1: Morphological Classification of Anaemias——– 17
2.2.2.2: Aetiological Classification of Anaemia————– 18
2.2.2.3: Classification of Anaemia Based on Presence or
Absence of Indicators of Compensatory Erythropoiesis—- 25
2.2.3: Diagnosis of Anaemia—————————————————– 27
2.2.4: Treatment of Anaemia—————————————————— 29
2.2.4.1: Specific Treatment in Iron Deficiency Anaemia————- 29
2.2.4.2: Specific Treatment in Vitamin B12 Deficiency Anaemia— 31
2.2.4.3: Specific Treatment in Aplastic Anaemia———————- 32
2.2.4.4: Specific Treatment in Anaemia of Chronic Renal Failure– 33
2.2.4.5: Specific Treatment in Thalassaemia / Haemoglobinopathies-33
2.2.4.6: Specific Treatment in Immune Haemolytic Anaemia——– 34
2.2.4.7: Specific Treatment in Anaemia Secondary to Leukaemia
and Haematological Malignancies————————————— 34
2.2.4.8: Plants Used in the Treatment of Anaemia——————– 35
2.2.5: Prognosis in a Patient with Anaemia————————————- 36
2.2.6: Consequences of Anaemia————————————————- 37
2.2.7: Anaemia Prevention and Control Strategies—————————– 39
2.3: Models of Anaemia——————————————————————— 41
2.3.1: Haemolytic Anaemia Model———————————————– 41
2.3.2: Iron Deficiency Anaemia Model—————————————— 42
2.3.3: Normovolaemic Acute Anaemia Model——————————— 42
2.3.4: Anaemia of Chronic Disease Model————————————– 43
2.4: Laboratory Rats and Research——————————————————– 43
2.4.1: Cardiac Function and Hypertension————————————– 44
2.4.2: Behavioural and Neuropharmacology of Addiction——————– 44
2.4.3: Arthritis and Related Autoimmune Disorders————————— 45
2.4.4: Learning, Memory and Behaviour—————————————- 45
2.4.5: Endocrinology and Reproductive Biology——————————- 45
2.4.6: Respiratory and Pulmonary Biology————————————— 46
2.4.7: Toxicology and Pharmacology——————————————— 46

CHAPTER THREE: MATERIALS AND METHODS—————————- 47
3.1: Materials——————————————————————————— 47
3.1.1: Study Area and Experimental Animals———————————– 47
3.1.2: Equipment——————————————————————- 48
3.1.3: Laboratory Reagents——————————————————– 49
3.1.4: Experimental Drugs——————————————————— 50
3.2: Experimental Design——————————————————————– 51
3.2.1: Experiment 1: Development of the Model——————————– 51
3.2.2: Experiment 2: Testing the Model—————————————— 53
3.3: Methods———————————————————————————– 54
3.3.1: Blood Removal and Blood Sample Collection————————— 54
3.3.2: Haematological Determinations——————————————- 54
3.3.2.1: Determination of Packed Cell Volume———————— 54
3.3.2.2: Determination of Haemoglobin Concentration————— 55
3.3.2.3: Red Blood Cell (RBC) Count———————————– 55
3.3.2.4: Calculations of Erythrocytic Indices————————— 56
3.3.2.5: Total White Blood Cell (TWBC) Count———————– 56
3.3.2.6: Differential Leukocyte Count———————————– 56
3.3.2.7: Reticulocyte Count———————————————- 57
3.3.3: Determination of Serum Total Proteins———————————- 58
3.4: Data Analysis—————————————————————————- 58

CHAPTER FOUR: RESULTS———————————————————- 59
4.1: Experiment 1: Development of the Model—————————————— 59
4.1.1: Haematology —————————————————————– 59
4.1.2: Total Proteins—————————————————————– 69
4.1.3: Mortality ——————————————————————— 70
4.1.4: Choice of Model ———————————————————— 71
4.2: Experiment 2: Testing the Model —————————————————- 71
4.2.1: Haematology—————————————————————– 71

CHAPTER FIVE: DISCUSSION AND CONCLUSION————————– 100
REFERENCES—————————————————————————— 107

INTRODUCTION  

Anaemia is a reduction below normal in the number of erythrocytes and/or haemoglobin concentration per unit volume of blood for an animal or human of a particular age, species, breed and geographic location (Coles, 1986; Calton and McGavin, 1995; Jones et al., 1997; Ihedioha and Chineme, 2004; Miller, 2009). Haemoglobin is the major carrier of oxygen in the blood and is located within the red blood cells. Its primary function is to deliver oxygen to the tissues of the body, thereby enabling cellular oxidative phosphorylation and generating energy for all cellular works.

Any decrease in the red cell mass reduces the amount of haemoglobin available and therefore reduces the amount and rate at which oxygen can be delivered to the tissues (Cheesbrough, 2006). Anaemia occurs when the rate of red blood cell formation fails to keep pace with the rate of its degradation or loss (Jennings, 1970). Anaemia is usually a sign of another disorder and does not represent a specific diagnosis. It usually develops secondary to another disease, which, more commonly in domestic animals, is a generalized disease process.

The presence of anaemia is therefore an indication of the existence of an underlying disease process (Jones et al., 1997; Mugera, 2000; Ihedioha and Chineme, 2004). With significant anaemia, the patient appears pale. Weakness, malaise and easy fatigability are common complaints or signs seen. The lowered oxygen content of the circulating blood leads to dyspnea on mild exertion. The nails may become brittle and lose their usual convexity to assume a concave spoon shape. Anoxia may cause fatty changes in the liver, myocardium and kidney.

If the fatty changes in the myocardium are sufficiently severe, cardiac failure may develop and compound the respiratory difficulty caused by reduced oxygen transport (Cotran et al., 1999; Mugera, 2000; Ihedioha and Chineme, 2004). 2 The prevalence of anaemia in humans has reached alarming figures in Africa (46%), South east Asia (57%), and in the eastern Mediterranean countries (45%) (Indian Council of Medical Research, 1989).

REFERENCES

Agbor, G. A., Oben, J. E. and Ngogang, J. Y. (2005). Haematinic activity of Hibiscus
cannabinus. African Journal of Biotechnology, 4: 833–837.

Alfrey, C. P. and Lane, M. (1970). The effect of riboflavin deficiency on erythropoiesis.
Seminars in Hematology, 7: 49–90.

American Society of Haematology (2010). Anaemia. www.haematology.org. Accessed 16th
July, 2011.

Andrews, J. S. (1942). Stomach worm (Haemonchus contortus) infection in lambs and its
relation to gastric haemorrhage and general pathology. Journal of Agriculture
Research, 65: 1-18.

Andrews, N. C. (2000). Iron metabolism and absorption. Reviews in Clinical and
Experimental Haematology, 4(4): 283-301.

Aslinia, F., Mazza, J.J, Yale, S.H. (2006). Megaloblastic anaemia and other causes of
macrocytosis. Chinese Medical Research, 4 (3): 236 – 241.

Aster, J. C. (2003). The haematopoietic and lymphoid system. In: Kumar V., Cotran R. S. &
Robbins S. L. (Eds.), Robbins Basic Pathology, 7th ed. Saunders, Philadelphia,
pp. 395-452.

Bain, B. J. (2005). Current concepts: Diagnosis from blood smear. New England Journal of
Medicine, 353: 498-507.

Balarajan, Y., Ramakrishnam, U., Ozaltin, E., Shanker, A. H. and Subramanian, S. V.
(2011). Anaemia in low-income and middle-income countries. Lancet, 378:
2123-2135.

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