COMPARATIVE STUDY OF ANTIFUNGAL ACTIVITIES OF AQUEOUS AND ETHANOLIC EXTRACTS OF SEED AND LEAF OF GARCINIA KOLA (HECKEL) AND LEAF OF MITRACARPUS HIRTUS (L.) DC.

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ABSTRACT

A comparative study of the antifungal activities of the leaf extract of Mitracarpus hirtus, and the seed and leaf extracts of Garcinia kola was carried out using the Agar well diffusion method on five fungal isolates (Candida albicansPenicillium marneffei, Microsporum canis, Rhizomucor pusillus and Aspergillus niger). Different concentrations of the plant extracts (50,100 and 200mg/ml) were used against the five pathogenic fungi. Data collected was subjected to t-test and Analysis of Variance (ANOVA). Means were separated by Duncan Multiple Range Test (DMRT) at p≤0.05. The results show that ethanol used as solvent was effective in extracting the active compounds. Results of the qualitative phytochemical compositions of seeds and leaves of G. kola and the leaves of M. hirtus showed the presence of bioactive compounds of interest such as alkaloids, tannins, flavonoids, saponins, phenols, anthraquinones, terpenoids, phlobatannins, steroids and glycosides. Results show that the extracts from leaf of Garcinia kola contained higher percentage of alkaloids (10.05±0.21), phenols (9.54±0.01) and flavonoids (4.80±0.14), than the leaf of M. hirtus (2.50±0.14, 8.10±0.02 and 3.97±0.03), and the seed of G. kola (0.83±0.05, 0.44±0.06 and 0.36±0.04) respectively. The leaf of M. hirtus contained higher percentage of tannins (7.61±0.01) than the leaf and seed of G. kola (2.83±0.03 and 1.34±0.05) respectively. The seed of G. kola contained higher percentage of saponins (5.52±0.05) than the leaves of M. hirtus and G. kola (2.81±0.05 and 2.25±0.35) respectively. The M. hirtus ethanol extract exhibited the best inhibitory activities against most of the test fungi, producing inhibition zones ranging from 12.05±0.07-17.99±0.01mm. There was resistance from Aspergillus niger. The G. kola seed extract also inhibited most of the test fungi. The inhibition zone observed ranged from 10.08±0.11-15.08±0.11mm. There was also resistance from Rhizomucor pusillus. M. hirtus leaf ethanolic extract significantly inhibited Candida albicans, while G. kola seed ethanolic extract also significantly inhibited Microsporum canis and Penicillium marneffei respectively. The activity index of ethanol extract of M. hirtus leaf (0.86 and 0.90 mm) was higher than that of G. kola seed (0.83 and 0.88 mm) when both fluconazole and ketoconazole were used as standard drugs. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of M. hirtus ethanolic leaf extract on the fungal isolates ranged from 6.25-12.5 mg/ml and 12.5-25 mg/ml respectively. The G. kola ethanolic seed extract has minimum inhibitory concentration (MIC) of 50 to 100 mg/ml and minimum fungicidal concentration (MFC) of 100 to 200 mg/ml respectively. Gas chromatography-mass spectrometry (GC-MS) analysis of the leaf and seed extracts of M. hirtus and G. kola was performed. The major constituents identified in ethanolic leaf extract of M. hirtus are 2,4-Di-tert-butylphenol (3.791%), 2-Octylcyclopropene-1-heptanol (6.230%), cis-11-Hexadecenal (2.179%), Cholestan-3-one,4,4-dimethyl-(5α) (4.154%). The major constituents identified in the ethanolic seed extract of G. kola are Hexadecanoic acid (11.276%), Octadecanoic acid (13.569%), n-Hexadecanoic acid (5.038%), l-(+)-Ascorbic acid 2,6-dihexadecanoate (2.444%), 4,7-Octadecadiynoic acid (2.996%), Falcarinol (4.419%) and 6-Octadecenoic acid (2.595%), which might be responsible for antifungal activity of ethanolic leaf and seed extracts of M. hirtus and G. kola. The implication of these results are discussed in relation to the antimicrobial activities of medicinal plants and the use of Garcinia kola seed and Mitracarpus hirtus leaf in treating some fungal infections affecting man.  






TABLE OF CONTENTS

Title Page                                                                                                                    i

Declaration                                                                                                                 ii

Certification                                                                                                               iii

Dedication                                                                                                                  iv

Acknowledgements                                                                                                    v

Table of Contents                                                                                                       vi

List of Tables                                                                                                              x

List of Figures                                                                                                             xi

List of Plates                                                                                                               xii

Abstract                                                                                                                      xiii

 

CHAPTER 1: INTRODUCTION

1.1                   Background of the Study                                                                    1

1.2                   Ethno-medical History in Nigeria                                                      4

1.3                   Plants of Study                                                                                    5

1.3.1                Garcinia kola (Heckel)                                                                       5

1.3.2                Mitracarpus hirtus (L.) DC                                                                6

1.4                   Some Medicinally Important Phytochemicals                                   7

1.4.1                Phenolic compounds                                                                           8

1.4.1.1             Flavonoids                                                                                          8

1.4.1.2             Phenolic acids                                                                                     10

1.4.1.3             Tannins                                                                                               12

1.4.2                Terpenoids                                                                                          12

1.4.3                Alkaloids                                                                                            14

1.4.4                Saponins                                                                                             15

1.4.5               Glycosides                                                                                          16

1.4.6                Sterols                                                                                                 17

1.5                   Some infections Caused by Fungi                                                      18

1.6                  Justification of Study                                                                           22

1.7                   Statement of the Problem                                                                   23

1.8                   Aims and Objectives                                                                          24

 

CHAPTER 2: LITERATURE REVIEW                                                               26

2.1                   Antimicrobial Activities                                                                     26

2.2                   Phytochemical Screening                                                                   31

2.3                   Antioxidant Activity                                                                           33

2.4                   Hepato-protective Activity                                                                 34

2.5                   Anti-inflammatory Activity                                                               35

2.6                   Anti-infertility Activity                                                                      36

2.7                   Anti-cancer Activity                                                                           37

2.8                   Anti-diabetic Activity                                                                         37

2.9                   Central Nervous System (CNS) Activity                                           38

2.10                 Gas Chromatography Mass Spectrometry (GC-MS) Analysis          38

 

CHAPTER 3: MATERIALS AND METHODS                                                    41

3.1                   Study Area                                                                                          41

3.2                   Experimental Design                                                                          41

3.3                   Source of Plant Materials Used                                                          41

3.4                   Test Microorganisms Used and their Sources                                    42

3.5                   Macroscopic and Microscopic Features of Fungal Isolates Used    42

3.6                   Extraction Method                                                                              44

3.7                   Preparation of the Media and Inoculation                                          44

3.8                   Reactivation of Test Microorganisms (Stock Cultured)                        45

3.9                   Determination of Zone of Inhibition                                                  45

3.10                 Determination of Minimum Inhibitory Concentration (MIC)  46

3.11                 Determination of Minimum Fungicidal Concentration (MFC) 47

3.12                 Determination of the Activity Indices of Plant Extract and

Antibiotics                                                                                          47

3.13                 Qualitative and Quantitative Phytochemical Screening of Plants

Extract                                                                                                47

3.13.1              Qualitative phytochemical screening                                                 47

3.13.1.1           Test for alkaloids                                                                                47

3.13.1.2           Test for saponins                                                                                 48

3.13.1.3           Test for tannins                                                                                   48

3.13.1.4           Test for flavonoids                                                                              48

3.13.1.5           Test for phenols                                                                                  48

3.13.1.6           Test for anthraquinones                                                                      48

3.13.1.7           Test for terpenoids                                                                              48

3.13.1.8           Test for steroids                                                                                  49

3.13.1.9           Test for phlobatannins                                                                                    49

3.13.1.10         Test for glycoside                                                                               49

3.13.2              Quantitative determination of the chemical constituents                        49

3.13.2.1           Alkaloid determination                                                                      49

3.13.2.2           Tannin determination                                                                         49

3.13.2.3           Flavonoid determination                                                                    30

3.13.2.4           Saponins determination                                                                      50

3.13.2.5           Phenols determination                                                           

3.14                 GC-MS Analysis of Ethanol Fraction of Mitracarpus hirtus

Leaves and Garcinia kola Seeds                                                        51

 

3.14.1              Extraction process                                                                              51

3.14.2              The gas chromatography-mass spectrometry analysis (GC-MS)     51

3.14.3              Identification of chemical constituents                                              52

3.15                 Statistical Analysis                                                                             52

 

CHAPTER 4: RESULTS AND DISCUSSION                                                      53

4.1       Results                                                                                                            53

4.2       Discussion                                                                                                       149

4.2.1    Presence of bioactive compounds in seeds and leaves of G. kola and

the leaves of M. hirtus                                                                                    149

4.2.2    Percentage of bioactive compounds in the plant materials studied                        150

4.2.3    Diameter of zone of inhibition                                                                       151

4.2.4    Activity indices of M. hirtus and G. kola extracts in comparison

with fluconazole and ketoconazole                                                                153

4.2.5    Minimum inhibitory concentration (MIC) and minimum fungicidal

concentration (MFC)                                                                                     154

4.2.6    Phytochemical components of the ethanol leaf extract of M. hirtus

and the seed extract of G. kola using GC-MS                                                155

 

4.2.7    GC–MS profile and Retention time of crude leaf ethanol extract of

Mitracarpus hirtus and seed extract of Garcinia kola                                   156

 

CHAPTER 5: CONCLUSION AND RECOMMENDATIONS                           158

5.1       Conclusion                                                                                                      158

5.2       Recommendations                                                                                          159

References                                                                                                     160

Appendices                                                                                                    186

 

 

 

 

LIST OF TABLES

                                      

4.1       Qualitative phytochemical screening of the seeds and leaves of

G. kola, and the leaves of M. hirtus.                                                               53

 

4.2       Comparative Phytochemical composition of Garcinia kola seed and leaf           55

4.3       Comparative Phytochemical composition of Mitracarpus hirtus leaf

and Garcinia kola seed                                                                                   57

 

4.4       Comparative Phytochemical composition of Mitracarpus hirtus leaf

and Garcinia kola leaf                                                                                    59

4.5       Diameter of zone of inhibition (mm) of ethanol extract of M. hirtus

leaf against some pathogenic fungi                                                                61

 

4.6       Diameter of zone of inhibition (mm) of ethanol extract of G. kola

seed against some pathogenic fungi                                                               63

 

4.7       The activity index of the extract of both plants in comparison with

standard drug (Fluconazole)                                                                           65

 

4.8       The activity index of the extract of the both plants in comparison with

standard drug (Ketoconazole)                                                                        67

 

4.9       MIC and MFC (mg/ml) of M. hirtus leaf ethanol extract on fungal

isolates                                                                                                                        69

 

4.10     MIC and MFC (mg/ml) of G. kola seed ethanol extract on fungal isolates            71

 

4.11     Phytochemical components of M. hirtus leaf ethanol fraction

using GC-MS                                                                                                  73

 

4.12     Phytochemical components of G. kola seed ethanol fraction

using GC-MS                                                                                                  121

 

 

 

 

 

 

 

LIST OF FIGURES

 

4.1       Chromatogram of different chemical compounds identified in the

ethanol extract of Mitracarpus hirtus leaf using GC-MS.                              75

4.2       Generalized GC–MS profile of crude leaf ethanol extract of

Mitracarpus hirtus                                                                                          119

4.3       Chromatogram of different chemical compounds identified in the

ethanol extract of Garcinia kola seed                                                             123

 

4.4       Generalized GC–MS profile of crude seed ethanol extract of

Garcinia kola                                                                                                  147

 

 

 

 

 

LIST OF PLATES

 

1          Zones of inhibition of aqueous and ethanol extracts of

Mitracarpus hirtus leaf against Penicillium marneffei                                  180

2          Zones of inhibition of aqueous and ethanol extracts of G. kola

seed against Penicillium marneffei                                                                 181

3          Zones of inhibition of aqueous and ethanol extracts of G. kola

seed against Penicillium marneffei                                                                 182

4          Zones of inhibition of aqueous and ethanol extracts of G. kola

seed against Microsporum canis                                                                    182

5          Garcinia kola leaves                                                                                       183

6          Garcinia kola seeds                                                                                        184

7          Mitracarpus hirtus leaves                                                                               185

 

 

 

 


 

CHAPTER 1

INTRODUCTION

 

1.1       BACKGROUND OF THE STUDY

The use of healing powers of plants is as old as man. Till date, natural plants of various types are used in traditional African medicine for providing healing to various ailments even before and after the spread of modern and scientific medicine.  Ethno-medicine is the oldest medicinal system and often traditionally referred to as the Cradle of Mankind (Van Vuuren, 2008). Traditional herbal drugs have been used to cure infectious diseases for so many years in various parts of the world (Adebayo and Krettli, 2011). There has been a developed interest in local medicine the whole world because traditional medicine is not used by all people (Adebayo and Krettli, 2011).  In developing countries, the health care has been maintained by other practices based on traditional values attached to them (Adebayo and Krettli, 2011).

In many developing countries, including Nigeria, 80% of patients use indigenous herbal remedies to treat infectious diseases (Nasir et al., 2015). Despite the availability of modern medicine in some communities, herbal medicines (use of medicinal plants) have continued to maintain popularity for historical and cultural reasons, in addition to their efficacy and cheaper cost (Lifongo et al., 2014). They serve as essential sources of new pharmacological bioactive substances since the entire parts of a plant, from roots to seed heads and flowers, are used in traditional treatments and can, therefore, act as major compounds used for drug production (Lifongo et al., 2014). Moreover, molecules from natural products have represented about 80% of drugs that have been put into the market (Ntie-Kang et al., 2013). The use of plant remedies has steadily grown globally in recent times as well as the look for new bioactive compounds that eventually could be produced as important drugs for the cure of infectious diseases (Nasir et al., 2015).

Plants contain bioactive compounds which are considered the source of active ingredients for modern and traditional medicine. Plants produce primary metabolites like proteins, lipids, nucleic acids and starch from substances such as water, carbon dioxide, nitrogen and numerous nonorganic salts in appreciable amounts. These base metabolites are changed into secondary metabolites (alkaloids, steroids, terpenoids, saponins, flavonoids) that are used as drugs (Akerle-Heywood and Synge, 1991). Long-term history of indigenous plants has offered good crude materials for many factories such as drug manufacturing companies, cosmetic, perfumery and dietry (Sathiya et al., 2008). The presence of many life enhancing components in plants have stimulated scientists to examine various plants with hope of determining their potential on antimicrobial effects (Nayak and Lekley, 2006). It is generally believed that most of the steadily used antimicrobials which are mainly chemical drugs may not be efficient and some of these agents exhibit dangerous effects to users of the dugs (Udegbunam et al., 2014). For example, Stevens-Johnson syndrome and toxic epidermal necrolysis, and hypersensitivity reactions are associated with the administration of antimicrobials such as sulfonamide and fluoroquinolones, and penicillin, respectively (Ferrandiz-Pulido and Garcia-Patos, 2013). All the professionals/scientists that formulated strategies to tackle the antimicrobial resistance situation knew that production of new and safe antimicrobials is more important than any other suggested solutions/strategies (Roca et al., 2015). Many initiatives and programs have been set up by many countries/organizations with the aim of developing new, effective, and safe antimicrobials (Roca et al., 2015). Thus, researchers/scientists are now looking everywhere in the environment including animal, plant, soil, and marine for potentially new and safe antimicrobial agent (Nasir et al., 2015). Unfortunately, the speed at which microbes develop antimicrobial resistance outpaces the rate of discovery/development of new drugs (Huttner et al., 2013). For instance, the 10x’20 initiative proposed in 2010 is aimed at developing ten new, safe, and more active antibiotics by 2020 (Infectious Diseases Society of America, 2010).

Currently, antimicrobial resistance is primarily the major challenge to world health and factors such as global climatic change, globalization (increased international travel and food importation/exportation), and change in demographics are worsening the crisis (Cheng et al., 2016; WHO, 2014). The rising and continuous spread (as a result of moving genetic elements) of multiple drug- and pan-drug-resistant microbes (such as carbapenem-, quinolone-, extended-spectrum β-lactam-, vancomycin- , methicillin- and colistin-resistant bacteria) which show resistance to almost all antimicrobial agents presently known to man, have put the globe in confusion (http://www.amr-review.org/Publications) .  The economic and health conditions of antimicrobial resistance on a worldwide scale is great and dreadful. A current review on “global crisis of antimicrobial resistance” organized by Jim O’Neill, estimated that about 700,000 human beings die globally every year due to antimicrobial-resistant diseases (http://www.amr-review.org/Publications). The review also projected that by 2050, the societal and monetary cost of not handling the antimicrobial resistance challenges could be US$100 trillion (Piddock, 2016). Other current researches estimated population decrease of between 11 million and 444 million people and a decrease in the size of the worldwide economic profile by 0.1-3.1% by 2050, if more active antimicrobial drugs are not developed (Fitchett and Atun, 2016). The sum of money use in producin a new antimicrobial drugs has been projected to be US$1 billion (Huttner et al., 2013) and projection of US$30 billion is require to handle antimicrobial resistance crisis now before it gets out of control (Piddock, 2016). The effect of antimicrobial resistance is devastating in poor countries, including Nigeria, where the money spent in managing resistant infections and resultant deaths are not put into consideration (Huynh et al., 2015).


1.2       ETHNO-MEDICAL HISTORY IN NIGERIA

In spite of the well-compiled ethnobotanical journals, very limited scientific information (e.g., efficacy, phyto-chemistry) is available on traditional medicinally used plants in Nigeria (Adebayo and Krettli, 2011; Conrad et al., 2014). From journals obtained so far, the oldest documents on antimicrobial properties of Nigerian plants are those of Dalziel (Dalziel,1956) in 1937 and 1957, respectively. Twenty years later, few other journals on antimicrobial activities and chemistry of Nigerian plants appeared in the literature (Odebiyi and Sofowora, 1979). Around 1980s, a handful of studies on antimicrobial potential of Nigerian plants became obtainable in the journal (Wolinsky and Sote, 1984; Aladesanmi et al., 1986). However, from 1990 till date, there has been overwhelming number of articles in the literature on the antimicrobial activities and chemistry of Nigerian medicinal plants. This current development in the scientific authentication of antimicrobial properties of Nigerian medicinal plants may be as a result of rise in public enlightenment, technological advancements and a number of publications in local books supporting the need for such studies (Iwu, 1993; Dalziel, 1955).

Further reasons for advancement of work on Nigerian medicinal plant include sourcing for new pharmacological active compounds to be produced as drugs or as templates for analog production and the evaluation of ethno-medicine and herbal medicinal products/mixtures (Yuan et al., 2016; Nasir et al., 2015). Medicinal properties of Nigerian plants are associated to synergetic effects of phytocompounds (like phenols, flavonoids, alkaloids, saponins, tannins, and essential oils) and phytochemical compounds contained in their tissues (Okigbo et al., 2009). Scientific documentations of studies on antimicrobial properties of Nigerian plants would give clearer understanding of the level of research that has been done to explain the antimicrobial potentials of these plants.


1.3       PLANTS OF STUDY

1.3.1    Garcinia kola (Heckel)

Garcinia kola (Heckel) (bitter kola) is a species of flowering plant in the Clusiaceae or Guttiferae family. Bitter kola is found in Benin Republic, Cameroon, Republic of  Congo, Ivory Coast, Gabon, Ghana, Liberia, Nigeria, Senegal and Sierra Leone. Its natural habitat is subtropical or tropical moist lowland forests. The nuts, fruit, bark and seeds fruit of the plant have been helpful for hundreds of years in folk medicine to cure ailments from coughs to fever (Iwu, 1993). Garcinia kola trade is still important to the tribes and villages in Nigeria.

Seeds of the plant are mainly used in Southern Nigeria and parts of West Africa as masticatory regardless of its bitter taste (Iwu et al., 1987). It is also used for traditional and social ceremonies, traditional way of welcoming guests and worship of deities among traditionalists in some part of West Africa (Iwu, 1993). Bitter kola is one of those few herbal plant referred to as ‘wonder plant’ because of its’ wide use in fighting numerous infectious diseases in Africa ethno-medicine. Garcinia kola is eaten raw as adjuvant of true kola and quoted as Guinea worm remedy and as a vermifuge (Sofowora, 1982). It prevents colics and particularly known to improve singing voice, due to its effectiveness for bronchitis and throat troubles (Iwu, 1993). The uses of other parts of Garcinia kola tree for traditional medicine have been documented (Adaramoye et al., 2005a; Sofowora, 1982).

In spite of the socio-economic value of Bitter kola, planting of the species is not common. Factors that have negatively affected farmers from cultivating Garcinia kola include seed dormancy or germination problems that reduces seedling steady existence. Most viable trees were left in the forest when farmers create plots out of the forest (Adebisi, 2004). Scientists have studied the germination difficulties/challenges of G. kola seeds and proposed many ways of breaking seed dormancy (Gyimah, 2000; Anegbeh et al., 2006; Kanmegne and Ndoumou, 2007; Oboho and Ogana, 2011; Oboho and Urughu, 2010). However, there is much need to investigate easy and practicable strategies that farmers could easily adopt with low professional input. G. kola seeds has both seeds coat dormancy and physiological dormancy probably imposed by the chemicals in the seed (Oboho and Urughu 2010). Seed coat dormancy of bitter kola can be minimized by removing the seed protective cover before planting, whereas physiological dormancy or endogenous dormancy can be minimized by soaking in distilled water for 3 days (Yakubu et al., 2014). Removal of the seed protective cover, soaking in distilled water for 3 days, putting inside air tight transparent polythene bag and watering the seeds when required for steady moisture gives an early germination period of two weeks (Yakubu et al., 2014).


1.3.2    Mitracarpus hirtus (L.) DC:

Mitracarpus hirtus belongs to the Rubiaceae family, and is commonly distributed throughout gardens, farms and fields in tropical and subtropical regions; United States of America, India, Malaysia, west and east African countries (Alqasim et al., 2013). In different parts of tropical Africa, it is traditionally used for treatment of sore throat. It is simply referred to as tropical girdlepod. In Nigeria, the liquid extract from the upper parts of the plant above soil is rubbed on the body against skin diseases and on wounds (Abere et al., 2007). Orally, it is employed as an antidote to arrow poison, anti-dysentery and anti-diarrhea (Abere et al., 2007). Ethno botanical surveys showed wide use of M. hirtus for treating fungal diseases such as ringworm, rashes, itching, eczema, toothache, venereal diseases, by rubbing leaves on skin or taken orally (Alqasim et al., 2013).

Mitracarpus hirtus is an erect plant. The leaves are opposite and decussate, elliptical and slightly stalked, with a serrated stipular collar. The flowers are white, sessile, grouped in axillary glomerules. They have a persistent calyx, fused to the ovary and surmounted by 4 lobes of which 2 are reduced and 2 are developed, 4 petals welded into a tube topped by four lobes (Akobundu and Agyakwa, 1989). The fruit is a dehiscent capsule, globular in shape. It is 2 mm long and surmounted by parts of the calyx. The line of dehiscence is located at the equator of the capsule. The capsule contains a seed in each loculus. The seeds are elongate, forming a cross with branches bent towards the center. They are 1 mm long and 0.5 mm wide. The protective seed coat is light brown and finely tuberculous (Grard et al., 2010).


1.4    SOME MEDICINALLY IMPORTANT PHYTOCHEMICALS

Phytochemicals are biological metabolites, which are natural chemical compounds seen in plants, which shade plant cells from ecological disturbances like stress, pollution, drought, UV light exposure and pathogenic infections (Ali and Alqurainy, 2006). These substances are referred to as secondary plant metabolites and offer health importance to human beings. They are known to act as synergistic agents, allowing food materials to be used more effectively by the body. Some of the important roles of phytocompounds are reduced toxicity, easy availability, reduced cost and their biological properties like antimicrobial activities, antioxidant properties, modulation of detoxification enzymes, lowering of platelet aggregation, enhancement of the immune system and modulation of hormone metabolism and antineoplastic properties (Andre et al., 2010). Phytochemicals are not major nutrients and are not needed by the human body for maintaining life, but have beneficial effects to stop or to fight some common infections (Holst and Williamson, 2008). The pharmarcological activities of some medicinal plants can justifiably be considered to, among others, the bioactive compounds in them especially the saponins, alkaloids, sterols, phenolic acids, tannins, terpenoids and flavonoids.

 

1.4.1    Phenolic compounds

Phenolic compounds are phytochemicals that have one or more than one aromatic rings with at least one hydroxyl group. In plants, they play a protective role by minimizing the effect of aggression by predators, parasites and also protect plants from ultraviolet radiation. Phenolics and tepernoids are ubiquitous in fruits, cereals, legumes and vegetables. Plant phenolics include flavonoids, phenolic acids and tannins (Piero et al., 2015).


1.4.1.1    Flavonoids

Flavonoids are low molecular mass polyphenolic antioxidants that are found to be present in vegetables, fruits and beverages such as astea and wine (Nyamai et al., 2015). Flavonoids are thought to have many medicinal values. Flavonoids have been revealed to have antihyperglycemic property (Muriithi et al., 2015). Flavonoids are thought to enhance cardiac function, reduce anginas and decreases cholesterol levels. These substances act by regulating inflammation mediators (Sánchez et al., 2008). Flavonoids have also been found to decrease production of pathogenic thrombosis in mice models (Jiang et al., 2010). A substrate of sea buckthorn which contains high level of flavonoids has been found to restore cardiac function and enhance blood circulation in patients with coronary heart disease. Flavonoids have been employed in the management of hypertension and chronic cardiac insufficiency as they obstruct the enhancement of necrosis factor kappa-B (Lim et al., 2006). Flavonoids like flavone C-glycoside, kakonein and caesalpin P enhance the function of pancreatic islet cells and have diabetic properties (Mohammad et al., 2006). It has been reported that the flavonoid glycosides cause pancreatic beta cell regranulation and have been employed in clinical management of diabetes as a result of increased sensitivity of insulin (Seeram et al., 2006).

Anthocyanins and proanthocyanins are flavonoids. Anthocyanins are believed to inhibit formation of free radicals thus protecting cardiomyocytes after ischemic episodes (Benkhayal et al., 2009). These compounds have also been known to reduce the quantity of nitric oxide by preventing the effect of nitric oxide synthase (Ngugi et al., 2012). Anthocyanins have anti-inflammatory property as they block cyclooxygenase enzyme. These flavonoids block the impact of VCAM molecules thereby inhibiting expression and attachment of endothelial cells with leucocytes. These phytocompounds are also said to reduce the amount of interferon necrosis factor-gamma, interleukin-2 and prevention of mast cell degranulation and tumor necrosis factor-alpha (Joseph et al., 2011). Proanthocyanins and anthocyanins have antibacterial activities and block the sticking of bacteria to the mucous membrane of the urinogenital passage (Pataki et al., 2002). Studies have revealed that anthocyanins have hepatoprotective effect towards hepatocytes of hepatitis B and A patients and paracetamol-induced hepatotoxicity (Lin et al., 2002). They reduce prostaglandin levels by blocking COX-2 thereby acting as anti-inflammatory substance in inflammated connective tissue and joints and enhance type II collagen production (Howell, 2002). Proanthocyanins are known to better clinical manifestation of pancreatitis such as abdominal pain, nausea and vomiting and to reduce the pathological developments that take place (Knox et al., 2000). These chemical compounds hinder sensitivity of intestinal cells to insulin and inhibits α-glucosidase enzyme in the intestinal lumen thereby reducing sugar levels (Ali et al., 2003). Proanthocyanins and anthocyanins inhibit the effect of enzymes that stimulate apoptosis thereby conferring protective property on cardiomyocytes after ischemic injury (Vinson et al., 2002). Anthocyanins have been employed in treatment of Epstein-Barr virus induced lymphoma, gastric adenocarcinoma, ovarian carcinoma and pulmonary carcinomas (Bagchi et al., 2002).

 

1.4.1.2   Phenolic acids

Phenolic acid compounds are aromatic secondary plant metabolites evenly distributed in plants. Phenolic acids that are found in in the environment can be divided into two major groups; benzoic acid derivatives and cinnammic acid derivatives such as ferulic acid and caffeic acid. Ferulic acid, a phenolic acid is believed to have a broad spectrum of therapeutic properties against diseases such as cardiovascular, diabetes, neurodegenerative, inflammatory diseases and cancer (Uraji et al., 2013). These pharmacological properties are said to be due to the antioxidant properties of the phenolic acids (Joshi et al., 2001). Ferulic acid inhibits lipid peroxidation and destroys superoxide free ion radical. The structural features of phenolic acids make them to have the antioxidant activities. These chemical compounds posses a phenolic nucleus and an unsaturated side chain that can produce a resonance stabilized phenoxy group. Reactive radicals collide with these substances/elements gaining a hydrogen atom and producing a phenoxy radical (Iwase et al., 2000). Phenolic acids and its ester derivatives decrease the amount of inflammatory mediators such as prostaglandin E2, tumor necrosis factor-alpha (Appendino et al., 2006). Ferulic acid derivatives have been said to hinder the effect of cyclooxygenase-2 promoter property in human colon cancer DLD-1 cells enroute the β-galactosidase reporter gene assay system (Ronchetti et al., 2006). Diabetes, an endocrine defect is characterized by hyperglycemia leading to oxidative stress because of the high synthesis of free radicals. Phenolic acids lower the harmful effect of streptozotocin by stopping the effect of free radicals synthesize in the pancreas by streptozotocin (Uttara et al., 2009). The reduction in harmful effect and oxidative stress in the pancreatic cells make beta cells to grow vigorously and produce more insulin. Increased insulin production leads to decrease in glucose levels due to increased glucose utilization by extra hepatic tissues. Phenolic acids are also said to protect lipids, DNA and proteins from oxidative stress thereby exerting anticancer activities (Delmas et al., 2006). These phytocompounds also act on channels that control induction to apoptosis, regulation of proliferation and reaction to oxidative stress. Phenolic acids have been revealed to hinder occurrence of pulmonary cancers in mice, inhibit mutagenesis and lower urinary N-nitrosoproline amount in humans. Phenolic acids restore normal homeostasis by stimulating apoptosis in cancer cells (Kang et al., 2011). Phenolic compounds absorb UV light, producing a stable phenoxyl radical radiation thereby ending free radical chain reactions. These compounds sustain the physiological properties of cells by scavenging deleterious radicals and chain reactions and hinder radiation-induced oxidative reactions (Uttara et al., 2006).

Phenolic compounds sustain the properties of cells exposed to alcohol stress by stopping the lipid peroxidative chain and scavenging free radicals. The mode of action is known to be by abstraction of H+ by hydroperoxyl and hydroxyl radicals through a free phenolic substrate to produce a phenoxyl radical which then forms products that are passed out in bile (Delmas et al., 2006). Phenolic acids are said to stop oxidative modification of proteins by lowering the possibility of oxidative attack on them (Delmas et al., 2006). Phenolic acids improve the endogenous antioxidant defense system, restores the damage caused by nicotine and prevents cells from oxidative destruction (Uttara et al., 2006). These compounds protect cell membrane by stopping the free radicals, increase the antioxidant level and block the leakage of marker enzymes into circulation.


1.4.1.3 Tannins

Tannins are polyphenols that are produced from different parts of varying plants belonging to so many species. It is produced in large quantities in the tree fruit pod, leaves, bark, wood, fruit and roots and also in plant gall. Tannins can be grouped into two broad categories – condensed tannins and hydrolysable tannins. The tannin epigallo-catechin-3-gallate is believed to exhibits anti-diabetic properties (Kang et al., 2011). In medical terms, all types of tannins may involve in the control of glucose level in the blood. Tannin has been indicated to enhance the receptor cells to make use of carbohydrate. It has been revealed that tannins act as an antifungal substance at increased concentrations by coagulating the protoplasm of the microbes (Adekunle and Ikumapayi, 2006). Consequently, the possible mode of action of tannins have been attributed to interference with energy production by uncoupling oxidative phosphorylation or interference with glycoprotein of cell surface (Harekrishna et al., 2010). Ellagic acid and quercetin work in combination to lower viability, proliferation and trigger apoptosis of MOLT-4 human leukemia cells (Gao et al., 2001). Ellagic acid and resveratrol are believed to completely inhibit skin tumorgenesis in mice (Cassidy et al., 2000).


1.4.2     Terpenoids

Terpenoids are chemical substances produced from five carbon isoprene units majorly isopentenyl pyrophosphate and its isomerdimethylallyl pyrophosphate by terpene synthases. Terpenoids have antioxidant potential and also work with most regulatory proteins. Plant liquid extracts have been employed both culturally and in modern medicine in the fight of cancer and inflammatory diseases. Terpenes are important inhibitors of NF-kB in modern medicine (Piero et al., 2015). NF-kB system is a cytoplasmic sensor that responds to so many internal and external signals such as genotoxic stress and hypoxia as well as challenges in the immune system. NF-kB also plays active role in the production of cellular resistance against anti-apoptotic signalling and apoptosis. Most of the terpenes in plants are in the form of terpene derivatives (terpenoids). Sesquiterpenoids are the major tepernes and are said to have NF-kB signaling inhibitory activity whereas diterpenoids and triterpenoids are also confirmed to have several active inhibitors of NF-kB signaling system (Mertens-Talcott et al., 2003). Aucubin, a monoterpenoid that are seen in plants as glycoside derivative stops the nuclear translocation of P65 subunit of NF-kB complex in enhanced mast cells and also block the degradation of IkBa protein (Mertens-Talcott et al., 2003). Limonene and their derivative perillyl alcohol are said to have inhibitory potential on mammary and pancreatic tumors (Yang et al., 2003). These two phytocompounds are also said to block proliferation and metastasis of gastric cancer.

Artemisinin, a lactone extracted from Artemisia annua is mainly used as an anti-malarial drug but it is also employed as an antifungal, anticancer, immunosuppressive and antiangiogenesis properties (Salminen et al., 2008). Terpenoids also enhance the skin tone, improves the concentration of antioxidants in wounds, and ressucitate inflammed tissues by increasing blood flow (Lyss et al., 1998). Terpenoids also increase lung function (Mujoo et al., 2001). The seeds and leaves of S. spectabilis are employed in the management of diabetes because of the presence of phytochemicals including terpenoids (Grace et al., 2013). Terpenoids have been confirmed to lower diastolic blood pressure and reduce the amount of sugar in blood in diabetic and hypertensive patients respectively (Grace et al., 2013). The anthraquionone in the plant liquid extracts of Polygonum multiflorum have been employed in the management of peripheral neuropathy, a complication that is connected with diabetes mellitus (McPartland et al., 2001).


1.4.3   Alkaloids

Alkaloids are biochemical compounds that have nitrogen and are derived from so many amino acids. Alkaloids are said to have blood glucose lowering property. Alkaloids tetrandine and berberine have been indicated to exhibit antioxidant property responsible for many biological potentials connected with this plant including antidiabetic effect (Yang et al., 2003). Alkaloid fractions have exhibited hypoglycemic effect in mice (Cassidy et al., 2000). The alkaloids l-ephedrine of Ephedra distachya herbs have demonstrated hypoglycemic property in diabetic mice because of restoration and regeneration of atrophied pancreatic islets that encourages the production of insulin (Piero et al., 2015). Alkaloids with therapeutic properties usually act by affecting biochemical transmitters of the nervous system such as γ aminobutyric acid, acetylcholine, dopamine and serotonin. Alkaloids are also confirmed to be anti-arrythmic properties, antihypertensive properties, anticancer and anti-malarial potential (Abdirahman et al., 2015; Verma et al., 2013). Alkaloids are said to have neuro-protective, cholinergic and antioxidant properties in Alzheimer’s disease (O’Brien et al., 2006). These chemical compounds have retentive-memory and cognitive-enhancing properties on Alzheimer’s patients. The therapeutic importance of these phytocompounds is known to be by restricting inflammatory reactions and oxidative stress, inducing cholinergic transmission, increasing estrogen and other neurotropic substances and inhibiting β-amyloid toxicity- formation (O’Brien et al., 2006). These chemical compounds block acetylcholinesterase enzyme. Inhibition of this enzyme enhances acetylcholine activity which is one of the main methods in the fight of Alzheimer’s disease. Tetramethylpyrazine, an amide alkaloid is believed to show hypotensive potentials by preventing platelet aggregation and vasoconstriction (Guruvayoorappan et al., 2014). This alkaloid has been confirmed to induce inotropic and chronotropic reactions on isolated atria. Tetramethylpyrazine is employed in the fight of occlusive cerebral arteriolar diseases as a result of its vasodilatory potentials. Alkaloids have also been revealed to have cytotoxic, trypanocidal and antimicrobial properties. These chemical compounds act by intercalating DNA thereby impairing transcription and replication causing frame-shift mutations (Guruvayoorappan et al., 2014). Alkaloids are also said to show antimicrobial and trypanocidal property by blocking of protein biosynthesis and by association with neuroreceptors (Francis et al., 2002).


1.4.4    Saponins

Saponins are plant compounds that occur either as steroid alkaloids, glycosides of triterpenoids or steroids. These phytocompounds are said to have immunostimulant, hypocholesterolaemic, hypoglycemic potential and anticarcinogenic activities (Ros, 2000). The hypoglycemic potential of saponins is associated to enhancement of pancreatic β-cells, prevention of glucose transport across the brush border cells of the small intestines and hinder the transfer of glucose from the stomach to the small intestines. Saponins are also revealed to prevent gastric emptying in a dose dependent manner (Tan and Vanitha, 2004). Saponins reduce cholesterol level by forming large micelles that are then removed in bile. These chemical compounds are believed to decrease serum levels of low density lipoproteins-cholesterol and lower intake of cholesterol in the intestines (Chung, 2004). Saponins are known to act as adjuvants in improving antibody synthesis and in the stimulation of cell mediated immune system. These chemical compounds are known to interact with antigen-presenting cells and enhance interferon and interleukin production thereby mediating immune-stimulant properties (Guruvayoorappan et al., 2014). Saponins block tumor cell development by apoptosis in leukemia cell line and by cell cycle arrest in breast cancer cell line. They also exert anti-proliferative active to prostate carcinoma cells by inducing apoptosis and cell cycle arrest at G1 phase. Saponins enhance apoptosis by promoting cytochrome c-caspase pathway. The nature and position of the sugar portion in saponins promotes the tumor specificity of cytotoxic action. Saponins are known to reduce the risk of cancer and other chronic infections. These chemical compounds are necessary and active for both hormone dependent cancer and non-hormone dependent cancer (Hostanska et al., 2005). Saponins are also thought to have antifungal and hypocholesterolemic properties. These properties are said to be as a result of combination with bile acids to produce micellar aggregates. Saponins hinder liver injury and hyperlipemia caused by lipid peroxidation (Lee et al., 2004).

The mode of action of these phytocompounds in this case is by blocking of lipid peroxide peroxidation and prevention of lipid peroxide formation. Saponins are also said to prevent HIV infection in vitro in addition to having antitumor potentials. This property can be associated to the preventive ability of HIV-induced cell fusion but have no direct action on reverse transcriptase potential of the virus (Jassim and Naji, 2003). Saponins have been revealed to have superoxide scavenging property on oxygen radicals that are implicated in the formation and initiation of several infections (Zhu et al., 2004). This pro-oxidative effect makes saponins to behave as hydrogen abstractor leading to initial reaction of lipid oxidation. Saponins have been known to exhibit strong antifungal properties (George et al., 2002). The main mechanism attributed to the antifungal potentials of saponins is their assocition with membrane sterols.


1.4.5    Glycosides

Glycosides are secondary metabolites found in plants that have a glycoside unit and act on the contractile behaviour of the cardiac muscle. These chemical compounds have been employed traditionally for the management of cardiac arrhythmias and congestive heart failure as they improve contractile force (Liu et al., 2000). Digitalis is the commonest cardiac glycoside used both traditionally and in modern medicine. This glycoside has two glycosides; digitoxin and digoxin whose structures vary only by an extra hydroxyl group on digoxin. Cardiac glycosides work by prevention of Na+, K+-ATPase resulting to reduced intracellular K+ions and increased intracellular Ca2+ and Na+ ions. Digitalis directly prevents proliferation of androgen dependent and androgen independent prostate cancer cell lines by forming apoptosis and enhancing intracellular Ca2+ (Newman et al., 2008). Digitoxin suppresses hyper-secretion of IL8, a protein linked to lung inflammation thereby blocking the activation of the NF-B signaling pathway in cystic fibrosis (Newman et al., 2007). These phytocompounds have been confirmed to exert cytotoxic effects in both cell lines derived in advanced cancer and normal prostate epithelial cells. Oleandrin, a glycoside obtained from oleander, causes apoptosis by sustaining Ca2+ rise that follows release of cytochrome c from mitochondrion and caspase activation. Oleandrin is also known to induce cell arrest at G2-M phase of the cell cycle in a dose dependent treatment (Hartmann, 1998). Oleandrin potential to prevent cell development and tumor cell multiplication is known to be due to suppression of the up-regulation of pERK and pAkt production (Hartmann, 1998).


1.4.6    Sterols

Phytosterols are subgroup of steroids that have structures and functions similar to cholesterol. Phytosterols in plants act as nutrient for the production of secondary metabolites control fluidity and permeability of cell membranes and also work as biogenic precursors of growth factors (Patel and Thompson, 2006). Phytosterols appear either as sterols or stanols; the saturated types of sterols. The intake of stanols in the intestines is lower than that of sterols resulting to decrease concentrations in blood serum. Phytosterols prevent intake of cholesterol in the intestines. Phytosterols and cholesterol need Niemann-Pick C1-like protein for their movement in the intestine cells. Cholesterol is esterified in the enterocytes by acetyl-coenzyme A, acetyltransferase-2 enzyme and are put into chylomicrons and taken to the lymphatic system. Medical studies have revealed that phytosterol absorption leads to up to 15% decrease of LDL-cholesterol (O’Neill et al., 2005). Intake of plant stanols lowers both plant cholesterol and sterol concentrations in the serum (Vanhanen et al., 1993). β-Sitosterol is the main phytosterol in plants and it is also obtained in human serum in combination with its glycoside at reduced concentrations. β-sitosterol and β-sitosterol glycoside have been indicated to lower incidences of inflammatory diseases and carcinogen-induced cancer (Ivorra et al., 1987). These phytocompounds are also confirmed to have insulin releasing property, anti-complement and antipyretic effect (Ivorra et al., 1987). β-sitosterol and their glycoside have immune modulating properties on non-infectious conditions such as allergies and rheumatoid arthritis and chronic infectious diseases such as Human Papilloma Virus and tuberculosis (Harshal and Prakash, 2014). The mixture is also revealed to increase the lytic effect of natural killer cells to cancer cell lines in vitro. β-sitosterol and its glycoside have anti-inflammatory property as they prevent both tumor necrosis factor alpha and interleukin-6 in a dose dependent manner.


1.5       SOME INFECTIONS CAUSED BY FUNGI

Mycoses are fungal infections of animal, including humans. Fungi are members of the class of eukaryotic living organisms that includes microbes like yeasts and moulds, as well as the macroscopic mushrooms. Fungi belong to a large group of organisms that includes yeasts, moulds and dermatophytes (fungi that cause parasitic skin disease in humans) (Wilson, 2006). Some species grow as unicellular yeasts that reproduce asexually by budding or binary fission.  There are so many important fungal diseases in man caused by a number of fungi. These infections range from superficial skin infections like dermatomycosis to generalized mycoses such as histoplasmosis and coccidiodomycosis (Kwon-Chung & Bennett, 1992). The normal commensal yeast may become pathogenic whenever alteration of the skin layer microclimate or host defense response occurs (Ashbee, 2007). Some fungi are dimorphic in nature. Dimorphic fungi can change between a hyphal phase and a yeast phase in relation to environmental conditions (Alexopoulos et al., 1996). The fungal outer cell wall is made up of chitin and glucans whereas glucans are also seen in plants and chitin found in the exoskeleton of arthropods (Bowman, 2006). Fungi are the known organisms that have these two structural molecules/compounds in their cell wall.

 Fungi are found everywhere in nature and live as free-living saprobes that has no known gain from parasitizing humans or animals. Since fungi are found everywhere in the society and are sometimes cultured from diseased body surfaces, it may be hard to evaluate whether a fungus found during disease is a disease causing organism or a transient environmental contaminant.  

Infection is penetration of pathogen into body tissues which is preceded by rapid growth of the organism. Infection may be clinically unnoticed or may result in disease because of cellular wound from competitive metabolism, elaboration of harmful metabolites, replication of the fungus, or an immune reaction. Successful infection may lead to disease, which is a departure from or interruption of the normal structure or function of body parts, tissues, organs, or systems that is seen as a characteristic set of signs and symptoms and whose etiology, epidemiology, pathology, and prognosis are known or unknown.

Rhizomucor pusillus: Mucormycosis (sometimes referred to as zygomycosis) is a dangerous but rare fungal infection caused by some species of moulds called mucormycetes.  These organisms live throughout the environment, mainly in the soil and in decaying plant remains, such as leaves, compost piles, or decayed wood (Richardson, 2009).

Infection is established as soon as susceptible host come in contact with the fungal spores in the environment. For instance, the lung or sinus forms of the infection can occur after someone inhales the spores. These types of mucormycosis mainly occur in human beings with health challenges or take drugs that reduce the body’s capacity to combat germs and sickness (Petrikkos et al., 2012; Ribes et al., 2000).   Mucormycosis can also form on the skin after the fungus penetrates the skin through a wound, scrape, burn, or other type of skin trauma.

Aspergillus niger: Aspergillosis is the name used to identify so many types of diseases caused by fungi of the genus Aspergillus. Aspergillosis affects humans, birds and other animals. Aspergillosis exist in chronic or acute forms which are clinically very different.  Major cases of acute aspergillosis are found in patients that have seriously compromised immune systems, e.g. those with underlying respiratory tract infections, like asthma (Denning, 2014).  Most commonly, aspergillosis occurs in the form of chronic pulmonary aspergillosis (CPA), aspergilloma, or allergic bronchopulmonary aspergillosis (ABPA) (Goel, 2015). Aspergillosis occurs when the body defense system fails to stop Aspergillus spores from gaining entrance to the bloodstream through the lungs. Without the body showing strong immune response, fungal cells are free to spread across the entire body and can infect major organs such as the kidneys and heart. Most human beings are thought to breathe in thousands of Aspergillus spores every day, but they do not cause harm to most people’s health because of strong immune responses.

Penicillium marneffei: Penicilliosis is a disease normally caused by the fungus Penicillium marneffei. The name of the organism and the name of the infection have been renamed. P. marneffei is now called Talaromyces marneffei, and penicilliosis now called talaromycosis (Yilmaz et al., 2014). Most people who contract talaromycosis have a medical condition that weakens their immune system, such as HIV/AIDS, or another condition that lowers the body’s capacity to combat germs and sickness (Vanittanakom et al., 2006). People contract talaromycosis after breathing in T. marneffei from the environment. However, the exact environmental source is unknown. T. marneffei has been found in bamboo rats and their burrows, but people who touch or eat these rats are not more likely to get sick from T. marneffei. The fungus can make people sick weeks to years after they come in contact with it (Bulterys et al., 2013). Bumps on the epidermal layer are a common symptom. These bumps are usually small and painless. The bumps usually appear on the face and neck but can also appear in other places on the body. Other symptoms include: (Wong and Wong, 2011) fever, general discomfort, weight loss, cough, swollen lymph nodes, difficulty in breathing, swelling of the liver and spleen, diarrhea and abdominal pain.

Candida albicans: Candidiasis is a disease usually caused by yeasts that are seen in the genus Candida. (CDC, 2014).  Some of the hundreds of Candida species can cause disease in humans; notably Candida albicansCandida usually stays inside the body (in an environment like the vagina, mouth, gut, and throat) and on the skin surface without causing any disease. Candida yeasts can cause diseases once they grow extensively over the body or once they penetrate deep into the body (for instance, the bloodstream or internal organs such as the kidney, heart and brain).

Candidiasis that are found in the mouth or throat is called “thrush” or oropharyngeal candidiasis. Candidiasis in the vagina is usually called a “yeast infection.” Invasive candidiasis occurs when Candida species penetrate the bloodstream and cause disease to the internal organs such as the kidney, heart and brain. Very uncommon, yeast infections can be invasive, affecting other parts of the body (CDC, 2014). This can lead to fever together with other signs depending on the parts attacked.

 More than 20 species of Candida are causing infection with Candida albicans being the most predominant (CDC, 2014). Infections of the mouth are most common among children less than one month old, the elderly, and those with weak immune systems.  Conditions that result in a weak immune system include HIV/AIDS, the medications used after organ transplantationdiabetes, and the use of corticosteroids. Other risks include dentures, following antibiotic therapy, and breastfeeding (Walker, 2008).

Microsporum canis: Tinea capitis (usually referred as "herpes tonsurans", "ringworm of the hair" (Rapini et al., 2007) "ringworm of the scalp", and "tinea tonsurans’’) is a cutaneous fungal infection (dermatophytosis) of the scalp (Freedberg, 2003). The infection is usually caused by dermatophytes in the  Microsporum and Trichophyton genera that attack the hair shaft. The clinical manifestation is usually single or multiple spots of hair loss, often with a 'black dot' appearance (often with cut-off hairs), that may be connected with inflammation, scaling, itching and pustules. Rarely in adults, tinea capitis is commonly seen in pre-pubertal children, mainly in boys than girls and is caused by Microsporum canis.

Up to eight species of dermatophytes are connected with tinea capitis.  Diseases from Microsporum species are common in Africa, Southern and Central Europe, South America and the Middle East. The disease is infectious and can be spread by human beings, animals, or objects where the fungus inhabits. The organism can also exist in a carrier state on the scalp, without clinical manifestation.


1.6       JUSTIFICATION OF STUDY

Plants contain bioactive compounds which are considered the source of modern and traditional medicine (Nasir et al., 2015). Medicinal plants are important sources of unique antimicrobial molecules.

They are also major sources of strongly important unique pharmaceutical compounds since every parts of a plant, from roots to seed heads and flowers, are used in ethno-medicine (Lifongo et al., 2014).

In many developing countries, including Nigeria, 80% of patients use indigenous herbal remedies to treat infectious diseases (Nasir et al., 2015). Nigeria have alvalanche of medicinal plants and there are alot of studies that have been conducted to screen antimicrobial properties of these plants.

Bitter kola has anti-cancer properties and is use in folklore remedies for the management of ailment such as liver disorders, hepatitis, diarrhea, laryngitis, bronchitis and gonorrhea (Adaramoye et al., 2005; Ezeifeka et al., 2004; Okojie et al., 2009).

M. hirtus has been employed for the management of fungal diseases like ringworm, rashes, itching, eczema, toothache, venereal diseases, by rubbing leaves on skin or taken orally (Alqasim et al., 2013).

In recent times, the cost and availability of various synthetic drugs have hampered the effort made in the management of mycotic infections as the emergence of drug resistant microbes is gradually incapacitating current antibiotics.

This ugly trend therefore calls for the need for cheaper and more available alternatives such as the use of chemotherapeutics and chemoprophylaxis made from plants; to combat the disease causing organisms in humans.


1.7         STATEMENT OF THE PROBLEM

Infectious disease has continued to be a problem militating against the health of man including farm animals. Despite the progress made in the treatment and control of the etiologic agents, fungal diseases have remained serious and hidden causes of illness and death in the world.

The incidence of diseases because of drug resistant microorganisms and the emergence of some unknown disease causing microbes pose enormous public health concern (Idu et al., 2006).

The use of synthetic medicine is usually expensive and not readily available and the emergence of drug resistant microbes is gradually affecting current antibiotics. It is largely noticed that most of the readily used antimicrobials which are mostly synthetic fall short of combating the enormous existing and emerging diseases. These existing agents (drugs) are also capable of eliciting terrible effects to the recipients (Udegbunam et al., 2014).

According to Huttner et al. (2013), the frequency level in which microbes develop antimicrobial resistance outpaces the rate of discovery/development of new drugs, whereas many plants possess unexploited potentials capable of handling the drug resistance by many disease-causing microorganisms.

 

1.8             AIMS AND OBJECTIVES

The aims of this study are:

1. To compare the antifungal potentials of aqueous and ethanol extracts of the seeds and leaves of Garcinia kola, and the leaves of Mitracarpus hirtus

2. To evaluate the effectiveness of the plant extracts in treating human fungal infections.

while the objectives include to:

i. Determine the qualitative and quantitative phytochemical components of the plants extracts.

ii.  Determine the diameter of zone of inhibition of the plant extracts

iii. Determine minimum inhibitory concentration value/level of the plant extracts

iv. Determine minimum fungicidal concentration of the plant extracts

v. Compare the activity indices of both plants extracts with standard drugs (Fluconazole and Ketoconazole)

vi. Carryout Gas Chromatography and Mass Spectrometry Analysis of the plants extract

 

 

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    I am thoroughly impressed with Projectshelve.com! The project material was of outstanding quality, well-researched, and highly detailed. What amazed me most was their instant delivery to both my email and WhatsApp, ensuring I got what I needed immediately. Highly reliable and professional—I'll definitely recommend them to anyone seeking quality project materials!

  • Anonymous

    3 months ago

    Its amazing transacting with Projectshelve. They are sincere, got material delivered within few minutes in my email and whatsApp.

  • TJ

    5 months ago

    ProjectShelve is highly reliable. Got the project delivered instantly after payment. Quality of the work.also excellent. Thank you