PLASMA BIOCHEMISTRY, HAEMATOLOGICAL AND BIOMETRIC PARAMETERS OF PROTOPTERUS ANNECTENS OF ANAMBRA RIVER, NIGERIA

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ABSTRACT

Biochemical, haematological and biometric studies of the African lungfish, Protopterus annectens were carried out in order to establish their mean and reference values which would serve as baseline data for assessment of the health status of the fish as well as reference point for future comparative surveys. The study was carried out between March and August, 2015. In the present study, blood was analyzed using standard techniques, and differences in haematological parameters including haemoglobin concentration, red blood cell count (RBC), white blood cell count (WBC), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were determined. Biochemical parameters such as alkaline phosphate (ALP), aspartate transaminase (AST), alanine transaminase (ALT), total protein, albumin, globuline, glucose, cholesterol level, and urea were determined. Biometric parameters such as condition factor (K), hepatosomatic index, and gonadosomatic index were also determined. These haematological, biochemical and biometric parameters of fish were compared according to sex and seasons. Analysis of variance showed that there were significant differences (P < 0.05) in ALP, AST, ALT, Protein, PCV, RBC, WBC, Hb and MCV, between sex and season. The results indicated that blood parameter level such as PCV, RBC, WBC, Hb, MCV, and MCHC between sexes in dry season were significantly different from those measured in wet seasons. The result also indicated that there were significant (P < 0.05) differences in condition factor (K), hepatosomatic index (HSI) and gonadosomatic index (GSI) between the seasons. There were no significant differences in MCH and MCHC values between the sexes. The values of WBC and PCV were found to be higher in female fish especially in dry season, while the level of haemoglobin and MCV values were higher in male Protopterus annectens. This may be related to the aggressiveness of the male fish. All the biochemistry parameters were higher in dry season except cholesterol and urea, but no significant differences were found among the sexes. The length of the fish varied significantly with season. The highest length was recorded in the March and June, while the highest mean weight frequency was obtained in the month of August. The length-weight relationship showed a negative allometric growth and the condition factor indicated that the fish were in a good condition in the months of April and July. There was increase in Hepatosomatic index (HSI), as the fish length increased. The results of the present study provide useful information for monitoring changes in the health status of fish.

TABLE OF CONTENTS

Title page
Table of Contents
List of Tables
List of Figures
Abstract

CHAPTER ONE: INTRODUCTION AND LITERATURE REVIEW
1.1 Introduction
1.1.2    Statement of the Problem
1.1.3    Justification of the Study
1.1.4    Objectives of the Study
1.2       Literature Review
1.2.1    Plasma
1.3       Haematological Indices in Protopterus annectens
1.3.1    The red blood cells (Erythrocytes)
1.3.2    White blood cell (leucocytes)
1.3.3    Blood Platelets
1.3.4    Packed Cell Volume (PCV)
1.3.5    Haemoglobin (Hb)
1.4       Biochemical Indices in Protopterus annectens
1.4.1    Glucose
1.4.2    Total Protein
1.4.3    Albumins
1.4.4    Glubulines
1.4.5    Blood Urea Nitrogen (BUN)
1.5       Biometric parameters in Protopterus annectens
1.5.1    Condition Factor (K)
1.5.2    Hepatosomatic Index (HSI)
1.5.3    Gonadosomatic index (GSI)
1.6       Description of Protopterus

CHAPTER TWO: MATERIALS AND METHODS
2.1       Study Area
2.2       Fish Sampling
2.3       Haematological Analysis
2.3.1    Packed Cell Volume (PCV)
2.3.2    Haemoglobin Estimation
2.3.3    Red Blood Cell Count (RBC)
2.3.3.1 Erythrocyte Indices
2.3.4    White Blood Cell Count (WBC)
2.3.5    Leukocyte Differential Count
2.4       Biochemical Analysis
2.4.1    Assay for Alkaline Phosphate (ALP)
2.4.2    Assay for Aspartate Aminotransferase (AST)
2.4.3    Assay for Alanine Aminotransferase (ALT)
2.4.4    Determination of Total Plasma Protein
2.4.5    Determination of Plasma albumin
2.4.6    Determination for Plasma Cholesterol
2.5       Biometric Analysis
2.5.1    Calculation of Condition Factor (k)
2.5.2    Calculation of Hepatosomatic Index (HSI)
2.5.3    Calculation of Gonadosomatic index (GSI)
2.6       Statistical Analysis

CHAPTER THREE: RESULTS
3.1       Haematological Parameters
3.2       Seasonal Variations in Haematological Parameters of Protopterus
            annectens of Anambra River, Nigeria
3.3       Leucocyte Differentials
3.4       Seasonal Variations in Leucocyte Differentials of Protopterus annectens of
            Anambra River, Nigeria
3.5       Biochemical Parameters
3.6       Seasonal variations in biochemical parameters of Protopterus annectens of
            Anambra River, Nigeria
3.7       Mean length frequency variation of Protopterus annectens of Anambra
            River, Nigeria
3.8       Mean weight frequency variations of Protopterus annectens of Anambra
            River, Nigeria
3.9       Mean monthly variations in condition factor ‘K’ of Protopterus annectens,
            of Anambra River Nigeria
3.10     Mean Monthly Variations in Hepatosomatic Index (HSI) of Protopterus
            annectens of Anambra River, Nigeria
3.11     Mean Monthly Variations in Gonadosomatic Index (GSI) of Protopterus
            annectens of Anambra River, Nigeria

CHAPTER FOUR: DISCUSSION, CONCLUSION AND RECOMMENDATIONS
4.1       Discussion
4.2       Conclusion
4.3       Recommendation
            References


CHAPTER ONE

INTRODUCTION AND LITERATURE REVIEW 1.1 Introduction


It is recognized that the blood component value exhibit genetic and physiological variations. The genetic variation may be due to site-specific factors within species. Blood comprises 1.3-7% of the total body weight of fish and it represent one of the most active components that contribute to metabolic processes by ensuring gas exchange between the organism and the environment. For this reason, blood parameters are increasingly used as indicators of the physiological condition of sub-lethal stress response in fish to endogenous or exogenous change (Belanger et al., 2001; Mohammadizadeh et al., 2012). In live fish, heamatology and plasma biochemistry provide a minimally invasive tool that can support health of fish, especially in relation to determining potential effects associated with such factors as pollution, disease, age, sex, seasonality and reproduction (Pradhan et al., 2012). Changes in hematological parameters depend upon the aquatic biotope, fish species, age, sexual maturity and health status. The evaluation of physiological condition of fish depends on the availability of reference values. These should be as close as possible to normal values of various blood components considered as reliable descriptors of healthy fish under natural conditions (Pradhan et al., 2012). It is clear that the environment in which fish live influences the metabolic content in blood. Thus, haematological, biochemical and biometric parameters are closely related to the response of the animal to the environment, an indication that the environment where fishes live could exert some influence on the heamatological characteristics (Gabriel et al., 2012). Taking into account the long evolutionary history of fishes and the adaptation to different environment, it is obvious that no species can be used as a representative model for all fishes. One of the difficulties in assessing the state of propagation of natural fish population has been the paucity of reliable reference values in healthy animals under natural habitat (Pradhan et al., 2012).
The African lungfish, Protopterus annectens is a highly prized food fish in Nigeria (Otuogbai, 2001; Otuogbai and Ikhenoba, 2012). It is distributed in shallow parts of rivers and lakes of some West African countries ranging from Senegal to Cameroon where it contributes to a relatively high percentage of artisanal fisheries (Otuogbai, 2001; Otuogbai and Ikhenoba, 2001; Okafor, 2004). Protopterus are omnivores in nature, feeding on fish, shellfish, amphibians and plant matter (Otuogbai, 2001).

The determination of biochemical, haematological and biometric parameters of fishes are carried out for a variety of purposes to;
v    establish a “normal range” of blood parameters (Pradhan et al., 2012);

v    investigate condition that might lead to alterations of some of these values such as sampling methods, temperature, sex, maturity, disease condition or nutrition of the fish (Okafor and Chukwu, 2010; Etim et al., 2014). and
v    ascertain the effects of certain chemical pollutants (e.g insecticide) and sublethal strength of some toxicants (such as heavy metals example, lead) on blood values (Gaafar et al., 2010; Abedi et al., 2013; El-Boshy et al., 2014).
Hematological and plasma biochemistry values of fishes have been studied in many

parts of the world (Njidda and Isidalomen, 2010; Fazio et al., 2013; Okorie et al., 2014). Recent studies disclosed that diseases and environmental conditions are the major constraints of aquaculture especially in the developing counties (Thien et al., 2007; Chi et al., 2008; Phan et al., 2010; FAO, 2012). Another study revealed that plasma biochemistry and hematology of fish enables us to differentiate the normal physiological condition of the animal under research from the eventual pathological modifications....

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