ABSTRACT

This study was designed to evaluate the effect of co-administration ethanol leaf extract of Ageratum conyzoides Nigerian Bonnylight crude oil on sperm membrane integrity in male Wistar rats. Twenty male Wistar rats (150-180g body weight) were divided into four groups of five rats each. The rats in group I served as the control group and were oral gavaged 3ml/kg of normal saline; group II animals was gavaged 748.33mg/kg body weight of the extract of A. conyzoides, which was 20% of the lethal dose (LD50) (3741.66mg/kg).Group III animals was gavaged 3ml/kg body weight of NBLCO. This dose was calculated as 20% of the LD50 of 14.14 ml/kg. Group IV animals were gavaged 748.33mg/kg body weight of the extract of A. conyzoides, and 3ml/kg body weight of Nigerian Bonnylight crude oil (NBLCO). In all cases, doses were applied daily for 28 days according to animals most recent body weight. The results showed NBLCO significantly reduced parameters of sperm motility as well as sperm  concentration while significantly increases number of immotile sperm and non-progressive sperm compared with groups I and II (control and A. conyzoides) (p<0.05). The co-administration of A. conyzoides with NBLCO to group IV animals significant reversed this trend (p<0.05). It is concluded that NBLCO administration is detrimental to sperm cells as it decreases the sperm motility and concentration which could be prevented by co-administration of ethanol leaf-extract of A. conyzoides.

 

TABLE OF CONTENTS

Pages

Title Page    -        -        -        -        -        -        -        -        -        i

Certification                   -        -        -        -        -        -        -        -        ii

Dedication  -        -        -        -        -        -        -        -        -        iii

Acknowledgements        -        -        -        -        -        -        -        iv

Abstract      -        -        -        -        -        -        -        -        -        v

Table of Contents -        -        -        -        -        -        -        -        vi

List of Tables       -        -        -        -        -        -        -        -        x

List of Figures      -        -        -        -        -        -        -        -        xi

CHAPTER ONE: INTRODUCTION

1.1     Overview of the Study   -        -        -        -        -        -        1

1.2     Objectives of the Study  -        -        -        -        -        -        4

1.2     Research Questions       -        -        -        -        -        -        5

1.3     Justification of Study     -        -        -        -        -        -        5

 

CHAPTER TWO: LITERATURE REVIEW

2.1     Health Implication of Crude Oil        -        -        -        -        -        6

2.2     Antioxidant effect of Ageratum conyzoides Administration     8

2.3     The Role of Oxidative Stress and Antioxidants in Male

Fertility       -        -        -        -        -        -        -        -        9

2.4     Cellular Sources of ROS in Semen    -        -        -        -        13

2.5     The antioxidant system in semen      -        -        -        -        14

2.6     Spermatozoa viability and membrane integrity    -        -        15

CHAPTER THREE: MATERIALS AND METHODS

3.1     Crude Petroleum and Chemical Reagents   -        -        -        20

3.2     Collection of Plant Material    -        -        -        -        -        20

3.2.1  Preparation of Leave Extract  -        -        -        -        -        21

3.2.2  Acute Toxicity Test       -        -        -        -        -        -        22

3.3     Experimental Animals   -        -        -        -        -        -        23

3.3.1  Experimental design and treatment of animals    -        -        23

3.5     6- Seminal Analysis       -        -        -        -        -        -        24

3.8     Statistical analysis         -        -        -        -        -        -        -        26

CHAPTER FOUR: RESULTS

4.1     Results        -        -        -        -        -        -        -        -        27

4.1.1  Sperm concentration following treatment with ethanol leave

extract of Ageratum conyzoides and Nigerian Bonnylight

crude oil      -        -        -        -        -        -        -        -        27

4.2     Discussion  -        -        -        -        -        -        -        -        33

CHAPTER FIVE: SUMMARY, CONCLUSION AND RECOMMENDATIONS

5.1     Summary    -        -        -        -        -        -        -        -        36

5.3     Conclusion -        -        -        -        -        -        -        -        37

5.3     Recommendation -        -        -        -        -        -        -        37

References  -        -        -        -        -        -        -        -        38

 

 

LIST OF TABLES

Pages

Table 4.1:   Comparing the spermatozoa concentration between

the control of ethanol leave extract of Ageratum

conyzoides and Nigerian Bonnylight crude oil      -        31

 

Table 4.2:   Spermatozoa motility for the control, extracts of

A. conyzoides and NBLCO      -        -        -        -        32

 

 

 

LIST OF FIGURES

Pages

Figure 2.1: Showing scheme of harmful effects of ROS                   -        12

Figure 3.1: Diagram of Argeratum conyzoides     -        -        -        21

Figure 4.1: Comparing the total sperm cell concentration in

NBLCO and extract of Ageratum conyzoides       -        30

 

CHAPTER ONE

INTRODUCTION

1.1     Overview of the Study

Crude oil is a naturally occurring, unrefined petroleum product composed of hydrocarbon deposits and other organic materials. A type of fossil fuel crude oil can be refined to produce usable products such as gasoline, diesel and various forms of petrochemicals. It is a non-renewable resource which means that it can’t be replaced naturally at the rate we consume it and is therefore a limited resource.

Crude oil is typically obtained through drilling and is refined and processed into a variety of forms such as gasoline, kerosene and asphalt by the process of fractional distillation.

The chemical components of crude oil include

a.     Light distillates     (i) Naphtha – made into

    gasoline/petrochemicals

(ii) Methane pentane

b.     Middle distillate   (i) Light gas oil

(ii) Heavy gas oil

c.      Residue                  Processed into refinery fuels, waxes, greases.

The effect of Benzene reduces the sperm vitality and sperm motility. The mean acrosin activity, gamma – GT activity and LDH C4 relative activity in the exposed workers were lower and fructose concentration was higher. Benzene can cause abnormal amount of chromosome in their sperm which impacts fertility and fetal development.

Argeratum conyzoides is a common annual herbaceous weed with long history of traditional medicine used in many countries especially in the tropical regions. It is commonly known as “Billy goat weed”.

Extracts and metabolite from this plant is widely utilized in traditional medicine as a purgative, analgesic and as a heart tonic (Tandon et al., 1994). Herbal preparation from the leaves of Argeratum conyzoides has been used in the treatment of high blood pressure, fever, diabetes, pneumonia and numerous infections diseases (Xuan et al., 2004).

Argeratum conyzoides is widely utilized in traditional medicine system wherever it grows, although applications vary by region. Traditional communities in India use this plant as a bacteriacide, antidysenteric and Antilythic (Okunade, 2002), and in Asia, South America and Africa, aqueous extract of this plant is used as bactericide (Okunade, 2002). In central Africa, it is used to treat pneumonia, but the most common use of this plant is to cure wounds and burns (Okunade, 2002).

Argeratum conyzoides leaves are among the forages usually fed to domestic animals in South Eastern Nigeria (Oladejo et al., 2003) but there is dearth of information on its effects on the spermatozoa characteristics and morphology of wistar strain albino rats exposed to arsenic toxicity.

Crude oil discovery minced an important landmark in world industrialization. It is associated with many issues such as exploration, exploitation, pollution (via oil spillage etc) and its                                                                                                                                                                                                                                         attendant environmental impacts. People ingest crude oil via eating contaminated aquatic animals, plants and direct consumption of crude oil itself because people think it is a cure to ailments such as epilepsy, febrile convulsion, malaria (Dienye et al., 2012). Others believe that it protects witchcraft, magic and evil spirits/spells. Environmental pollution do occurs due to continual spillage of crude oil. It is applied to the skin, instilled into orifices, nostrils, vagina, anus, urethra (Dienye et al., 2012). The effects have been shown to be harmful starting from the skin resulting in formation of blisters and epidermolysis (Dienye et al., 2012). Its ingestion may result in nausea, vomiting and if aspirated it could cause chemical pneumonitis. Higher doses have been reported to be associated with renal failure, convulsion, coma etc. (Dienye 2012).

Since the toxicity of crude oil has been shown to be multisystemic (Ogara et al., 2016). It follows that human reproductive system may also be affected by the components of crude oil. Researchers have shown that there is decreasing male reproductive potentials as characterized by falling sperm count, abnormal sperm forms, abnormal morphology, asthenospermia and even azoospermia (Okonofua et al., 2003). Therefore, continuous doses of crude oil ingestion may decrease sperm count, altered morphology viability and mobility. Moreover, the endothelium induced damages from crude oil have been implicated in a wide range of disease such as hypertension, diabetes, myocardial infection etc. (Cines et al., 1998).

1.2     Objectives of the Study

The objective of the study is to evaluate the sperm membrane integrity following crude oil and ethanol leave extract of Ageratum conyzoides administration.

This study will help us to know the effects of crude oil and ethanol leave extract on the sperm.

It will also help to know the antioxidant effect of Ageratum conyzoides and also its radical scavenging on crude oil induced oxidative stress.

1.3     Research Questions

The effect of Argerantum conyzoides on sperm membrane.

Ø Is the sperm membrane lysed?

Ø Is it compromised?

Ø What effect does Argeratum conyzoides has on the sperm membrane?

1.4     Justification of Study

The discovery of crude oil has caused a lot of environmental pollution which has affect humans in diverse way. The environmental products associated with crude oil leads to generation of free radicals in the blood and these are capable of lowering sperm quality causing outright male infertility (Messenger, 2010).

Given that, the underlying pathologies arise from uncontrolled production of free radicals by the crude oil components, it is postulated that anti-oxidants could reverse the actions of the reactive oxygen species (ROS). One of such antioxidant is Argeratum conyzoides. Argeratum conyzoides possessed anti-radical scavenging activity (Singh et al., 2013). This research intends to utilize the anti-oxidant properties of Argeratum conyzoides to alleviate the effects of crude oil on sperm membrane integrity.

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