COMPARATIVE STUDY ON THE ANTIBACTERIAL ACTIVITIES OF TWO SELECTED MEDICATED SOAPS AND TWO LOCALLY-PRODUCED BLACK SOAPS ON STAPHYLOCOCCUS AUREUS ISOLATED FROM WOUNDS

ABSTRACT

An investigation was carried out on the antibacterial activities of two selected medicated soaps (Tetmosol and Dettol) and two selected locally-produced black soaps (Zee and Royal) against Staphylococcus aureus isolated from wounds using disc diffusion method. The study included the use of a bacterial species isolated from wounds with the aid of a swab stick. The bacterial species was duly identified. The determination of minimal inhibitory concentration and minimum bactericidal activity of the bacterial species was performed by tube and plate methods respectively. Thirteen individuals with wound were sampled, and out of the thirteen (13) individuals sampled, 6(46%) were infected with Staphylococcus aureus, 2 (15%)Pseudomonas aeruginosa, 3 (21%) Staphylococcus epidermis (21%), and 2 (15%) Escherichacoli. The study revealed that Staphylococcus aureus isolated were most susceptible to Tetmosol soaps at the concentration of 10-1 with the highest and the lowest zone of inhibition as 19.66±1.52b and 12.33±1.52a  respectively; and followed by Zee soap at the concentration of 10-1 with the highest and the lowest zone of inhibition as 18.66±0.57b and 11.66±0.57a as compared to the positive control (Levofluxacin) zone of inhibition 24.00±1.00, but were not susceptible to Dettol and Royal soaps. It is obvious that Tetmosol and Zee soaps have the antibacterial agents that can either kill or inhibit the bacterial cells. It was

determined that neither Tetmosol soap nor Zee soap have bacteriocidal effect on Staphylococcus aureus; they all exhibited bacteriostatic effect. Tetmosol and Zee soaps showed significant antibacterial activity greater than those of the Dettol and Royal soaps. It is recommended that Tetmosol and Zee soaps be used for sanitizing the skin.

TABLE OF CONTENTS

Title Page                                                                                                                                           i

Certification                                                                                                                                       ii

Dedication                                                                                                                                          iii

Acknowledgement                                                                                                                               iv

Table of Contents                                                                                                                                v

List of Tables                                                                                                                                      ix

Abstract                                                                                                                                               x

CHAPTER ONE

1.0  Introduction                                                                                                    1

1.1Aims of the Study                                                                                                               5

1.2  Objectives of the Study                                                                                               5

CHAPTER TWO

2.0 Literature Review                                                                                                                         6

2.1 Virulence Factor                                                                                                                           8

2.1.1 Enzymes                                                                                                                                     8

2.1.2 Toxins                                                                                                                                        8

2.1.2.1Superantigens                                                                                                                          8

2.1.2.2Exfolivative Toxins A and B                                                                                                    9

2.1.2.3 Other Toxins                                                                                                                           9

2.2 Antibacterial Activity of Soap                                                                                                       10

2.3 Medical Relevance of Antimicrobial Soap                                                                                   11

2.4 Normal Skin Microbiota and Soap                                                                                               12

2.5 Relationship to Host                                                                                                                     13

CHAPTER THREE

3.0 Materials and Methods                                                                                                                 15

3.1 Material                                                                                                                                        15

3.2 Sample Collection                                                                                                                        15

3.3 Media Preparation                                                                                                                                   15

3.4 Identification of Isolates                                                                                                               16

3.4.1 Isolation of Microorrganisms                                                                                                    16

 3.4.2 Sub-culturing and Preservation of Pure Cultures of Microorrganisms                                16

3.4.3 Gram Staining                                                                                                                           17 

3.4.4 Biochemical Test                                                                                                                       17

3.4.4.1 Catalase Test                                                                                                                          17

3.4.4.2 Coagulase Test                                                                                                                       18

3.5 Bioassay Procedure                                                                                                                      18

3.5.1 Soap Sample Preparation                                                                                                          18

3.5.2 Preparation of Disc                                                                                                                   19

3.5.3 Assay                                                                                                                                          19

3.5.4 Determination of Minimal Inhibitory Concentrations (MICs)                                                  20

3.5.5 Determination of Minimal Bacterial Concentrations (MBCs)                                                  20

CHAPTER FOUR

4.0 Results                                                                                            21

4.1. Morphological and Biochemical Characteristics of Test Isolate                                                                        22

4.2 Zone of Inhibition for Susceptibility Test Results                                                                         23

4.3 Minimal Inhibitory Concentrations (MICs) Results                                                                     24

4.4 Minimal Bacterial Concentrations (MBCs) Results                                                                     25

CHAPTER FIVE

5.0 Discussion, Conclusion and Recommendation                                                                            26

5.1 Discussion                                                                                                                                     26

5.2 Conclusion                                                                                                                        27

5.3 Recommendation                                                                                                                          27

Reference                                                                                                                                            29

LIST OF TABLES

Table 1. Morphological and biochemical characteristics of test isolate                                        22

Table 2. Zone of inhibition in millimeter of S. aureus by the different soap samples               23

Table 3. Minimum inhibitory concentrations                               24

Table 4. Minimum bacteriocidal concentrations                                          25

CHAPTER ONE

1.0 INTRODUCTION

Antibacterial activity is the ability to either destroy bacteria or inhibit their growth. This is significant with respect to the human body in preventing sepsis and skin infections (Black et al, 1981).                                                                                                                                              

Soap may be defined as a chemical compounds resulting from the interaction of fatty acids oil and caustic soda (alkali) possessing the characteristic soap like properties of detergents, surface tension, lowering wetting and emulsifying power and gel formation (Hunt, 1999) as well as acting as both active medication and vehicle for the incorporation of other active substances (Grayson, 1983). It can also be defined as a substance able to mix with both oil and water, used for cleaning, often in the form of a solid or in liquid form, derived from fats or made synthetically (Hunt, 1999). It cleanses by lowering the surface tension of water, by emulsifying grease, and by absorbing dirt into the foam. Scrubbing body or hands, particularly with soaps, is the first line of defense against bacteria and other pathogens that can cause colds, the flu, skin infections and even deadly communicable diseases (Larson, 1988; Kimel, 1996). Conceptually, many people consider that an antimicrobial potion of soaps is effective at preventing communicable diseases.

In the treatment of skin diseases, it causes cooling, drying, hydration, crust and scale removal (Schwartz, 1979). Although bacteria that attack human body are of great importance with reference to health (Fuls et al., 2008) reported the inhibitory potential of antimicrobial and non-antimicrobial soaps in clinical cases (Larson et al.,1987; Toshima et al.,2001) indicated that soaps containing antimicrobial active ingredients could remove more bacteria as compared to plain soap (Osborne and Grube, 1982) had earlier reported that antibacterial containing soaps can remove 65% to 85% bacteria inhabiting human skin. When used properly, washing with soap could reduce Propiobacterium acnes and prevent secondary infections in acne skin (Kuehl et al., 2003) and healthcare-associated transmission of contagious diseases more effectively (Arya et al., 1966).                                                                                                                             

Medicated soaps are industrial produced soaps that have the germicidal substance like chloroxyhlenol potassium mercuric iodide, trichlorocarbanlide etc. incorporated into them, in order to enhance their antibacterial activity (Black et al, 1998). These germicidal substances are normally added in a specified amount and the percentages of the substances used are always stated on the soap-case or inside the leaflet which conveys the information on how to use the soap for various purposes.

Antibacterial soap may include 2, 4, 4-trichloro 2,2- hydroxydiphenyl in an amount of 0.05 to 5% by weight and at least one phosphorus oxyacid (Kimel, 1996). A soap containing phosphorus oxyacid salt phosphoric ester has a wide range of antibacterial activities and marked resistance of discoloration upon exposure to sunlight (Larson, 1988). Medicated soap incorporated in their composition germicidal agents which include hexachlorophene mercuric which tricloran, trichlorocarbaucide. Medicated soaps to a large extend remove dirt and disrupt cytoplasmic membrane to kill microorganisms (Tachibana, 1976). It also works against enveloped virus like human immunodeficiency virus (HIV). Several antimicrobial substances are found in medicated soaps and they have various mode of action on various skin microflora and infections.

Local black soap is an African traditionally manufactured medicated soap, otherwise known as African black soap, is known by different names from various regions. In Ghana, black soap is known as ‘Anago soap’ or ‘Alatasamina’. In Nigeria, it is known by the Hausas as ‘Sabilum-salo’, the Yorubas call it ‘Ose-dudu’ or ‘abuwe’ and the Igbos name it ‘Ncha-Nkota’ (Aliyu et al., 2012; Bella, 2011; Getradeghana, 2000; Summers, 2016). African black soap is a natural source of vitamins A and E and iron (Grieve, 1997) which helps to strengthen the skin and hair. It is made of a combination of water, roasted plantain skin or cocoa pod, palm oil, palm kernel oil or Shea butter. These are common oils used for the production of soap through saponification reactions (Kubmarawa and Atiko, 2000) that make the soap to have antimicrobial properties recognized in the traditional African households (Getradeghana, 2000). Depending on where it is manufactured, black soap contains leaves and bark from plantains, Shea tree, cocoa pods or palm tree leaves. The leaves and bark are sun dried before being roasted slowly in a pot after which different oils including coconut oil, shea butter and palm kernel oil giving antimicrobial properties to the soap are added to the mixture (Getradeghana, 2000). Black soap made with shea butter offers protection against UV rays, whereas black soap made with plantains contains a high concentration of iron along with vitamins A and E (Underwood, 2008). The attribute of the soap includes gentleness on the skin, rich lather, protection against skin disorders (including rashes, eczema, scabies) treatment of skin infection (such as ringworm), protection of even skin toning and smoothness of the skin (Getradeghana, 2000), help relieve acne, oily skin, clear blemishes and various other skin issues. Black soap has been employed to get rid of skin rashes, ringworm, measles and body odors (Adelakun, 1990) and for treating many infections caused by microorganisms as well as for exfoliating and deep cleansing (Underwood, 2008). It also has the ability to emulsify grease and oil that hold dirty particles (Sharma 2006). Having antiseptic properties and being a natural shampoo to avoid dry itchy scalp, it is good for bathing, washing hair and faces and helps keep the skin clear of premature facial lines. Many people prefer this soap because it does not cause resistant bacteria growth. The major fatty acids in palm kernel oil are lauric acid (C12, 48%), meristic acid (C14, 16%) and oleic acid (C18, 15%). Certain fatty acids (medium chain saturates) and their derivations have adverse effects on various microorganisms (Kabara, 1978). The palm kernel oil sample with highest lauric acid value have the highest effect on Staphylococcus aureus, Streptococcus sp. and C. albicans, this confirms that lauric acid is the antimicrobial agent in palm kernel oil (Ugbogu, 2006). S. aureus and Streptococcus sp. which cause skin and wounds infections are inhibited minimally by palm kernel oil. Although the antimicrobial activity observed is low (Kabara, 1978).

According to Paulson (2003) a wound is defined as disruption of cellular and anatomic continuity while its healing is the restoration of continuity.  The biological process can only be accomplished by regeneration, cell proliferation and collage production which can be alleviated by washing the wound surface especially with medicated soap which due to its content of phenolic compounds help in keeping off organisms like Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa from the wound to a certain level. A wound can also result when the operative barrier of the skin is breached by traumatic invasion or whether it is caused by trauma or internationally by surgery. The open area is susceptible to microbial invasion and once a wound has become infected, pus forms in the injured area resulting to wound abscess.

Hence, this study was aimed at comparing the antibacterial activity of two selected medicated soaps and two local black soaps on Staphylococcus aureus isolated from wound infections.

1.1 AIM OF THE STUDY

The aim of this project work is to compare the antibacterial activity of two selected medicated soaps (Tetmosol Medicated and Dettol Medicated soaps) and two local black soaps (Zee and Royal Black soaps) on Staphylococcus aureus isolated from wound infections.

1.2 OBJECTIVES OF THE STUDY       

The objectives of this study include the following:

·       To isolate and identify Staphylococcus aureus from wound infection.

·       To determine the antibacterial activity of two selected medicated soaps and two local black soaps on Staphylococcus aureus isolated from wound infections.

·       To determine the Minimal Inhibitory Concentration (MIC) and Minimal Bacteriocidal Concentration (MBC) of the selected medicated soaps and two locally-produced black soaps used.

·       To compare the antibacterial activity of two selected medicated soap and two local black soaps on Staphylococcus aureus ssisolated from wound infections.

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