COMPARATIVE STUDY OF DISINFECTANT EFFICIENCY OF ETHANOL, BLEACH AND PHENOLICS AGAINST PSEUDOMONAS AERUGINOSA AND STAPHYLOCOCCUS AUREUS

TABLE OF CONTENTS

Title Page                                                                                                                 i

Certification                                                                                      ii

Dedication                                                                                       iii

Acknowledgements                                                                    iv

Table of Contents                                                                    v

List of Tables                                                                          vi

List of Figures                                                                        vii

List of Plates                                                                                      viii

Abstract                                                                                              ix

CHAPTER ONE

1.0       Introduction                                                                        1

1.1       Aims and Objectives                                                   2

CHAPTER TWO

2.0       Literature Review                                                             5

2.1       History of Disinfectants                                             6

2.2       About Disinfectants                                                        7

2.3       Sources of Contamination of Surfaces                        8

2.4       Types of Disinfectants                                               9

2.5       Properties of A Disinfectant                                        18

2.6       General Features of Disinfectant                                 19

2.7       General Features of the Test Organisms                     24

2.9       Mechanism of Actions of Disinfectants against Bacteria  26

2.10    Resistant Action of Bacteria                              27

2.11    Advantages and Disadvantages of Disinfectants           29

2.12    General Guidelines in the Use of Disinfectants             33

CHAPTER THREE

3.0       Material and Method                                    36

3.1       Isolation of Bacteria                                                 36

3.2       Identification of Isolates                                                  37

3.3       Preparation of Disinfectants                                              40

3.4       Antimicrobial Susceptibility Testing (Using Kirby Bauer

Diffusion Assay Well Method)                                      41

CHAPTER FOUR

4.0       RESULTS                                                                           43

CHAPTER FIVE

5.0       Discussion                                                                        57

5.1       Recommendations                                                            60

5.2       Conclusion                                                                           51

References                                                                                      62

Appendix 1                                                                          69

Appendix II                                                                                  71

Appendix III                                                                      74

LIST OF TABLES

Table 1:          Results of diameter of zone of inhibition of Ethanol,

Phenolics and Bleach for Staphylococcus aureus                      44

Table 2:          Results of Diameter of Zone Inhibition of Ethanol,

Phenolics, Bleach for Pseudomonas aeruginosa                                    45

Table 3:          Pseudomonas aeruginosa response to Ethanol, Phenolics

and Bleach                             46

Table 4:          Staphylococcus aureus response to Ethanol, Phenolics

and Bleach                                  47

LIST OF FIGURES

PATTERNS OF THE ANTIMICROBIAL EFFICACIES OF VARYING CONCENTRATIONS OF THE DISINFECTANT ON THE TEST ORGANISMS USING HISTOGRAM

a.                  Fig 1: Pseudomonas aeruginosa disinfectants A test result                48

b.                  Fig 2: Pseudomonas aeruginosa disinfectants B test result                 48

c.                  Fig 3: Pseudomonas aeruginosa disinfectants C test result                 49

d.                  Fig 4: Staphylococcus aureus disinfectants A test result                     49

e.                  Fig 5: Staphylococcus aureus disinfectants B test result                     50

f.                   Fig 6: Staphylococcus aureus disinfectants C test result                     50

PATTERNS OF THE EVALUATION OF THE BACTERIAL PERCENTAGE RESPONSE TO EACH DISINFECTANT USING A PIE CHART

a.                  Fig 7: Disinfectant  A on Pseudomonas aeruginosa     52

b.                  Fig 8: Disinfectant B on Pseudomonas aeruginosa    52

c.                  Fig 9: Disinfectant C on Pseudomonas aeruginosa          53

d.                  Fig 10: Disinfectant A on Staphylococcus aureus       53

e.                  Fig 11: Disinfectant B on Staphylococcus aureus      54

f.                   Fig 12: Disinfectant C on Staphylococcus aureus          54

LIST OF PLATES

Plate 1: Plates showing zones of inhibition                 74

Plate 2: MacConkey media with colonies of Pseudomonas aeruginosa 74

Plate 3: Some of the used plates                          75

Plate 4: Biochemical test for Pseudomonas aeruginosa 75

CHAPTER ONE

1.0       INTRODUCTION

            Microorganisms are minute living things that individually are too small to be seen with the unaided eyes (Tortora et al, 2007).  Though only a minority of microorganisms are pathogenic (disease producing), practical knowledge of microbes is necessary for medicine and related health sciences.  For example hospital workers must be able to protect patients from common microbes that are normally harmless but pose a threat to the sick and injured.  Thousands of people died in devastating epidemics; the cause of which was not understood.  Entire families died because vaccination and antibiotics were not available to fight infection (Johnson and Case, 1995).  This leads to scientific control of microbial growth.  This began only about 100 years ago.  It was Pasteur’s work on microorganism that led scientists to believe that microbes were a possible cause of diseases and need to be eliminated or destroyed.  Some examples off these microbes are; Bacteria, fungi, viruses and protozoa etc (Tortora et al, 2007).

            In the mid 1800s, the Hungarian physician Ignaz Semmeliveis and English physician Joseph Lister used these thoughts to develop some of the first microbial control practice for medical procedures.  These practices include hand washing with microbes killing chloride of lime and use of techniques of aseptic surgery to prevent microbial contamination of surgical wounds (Hamamah, 2004).  Over the last century, scientists have continued to develop a variety of physical methods and chemical agents to control microbial growth.  Control directed at destroying harmful microorganisms is called disinfection.  It usually refers to the destruction of vegetative (non-endospore forming) pathogens example bacteria by using a disinfectant to treat an inert surface or substances (Bhatia and Icchpujani, 2008).

            Bacteria are major causes of disease and even human death.  A disinfectant is one of the diverse groups of chemicals which reduces the number of microorganisms present (normally on an inanimate object).  There are various official definitions of the process of disinfection and disinfectants agents.  It is defined as a chemical that inactivates vegetative microorganism but not necessarily high resistant spores (ISO, 2008).  Cleaning and disinfection of surfaces are essential steps for maintaining the cleanliness of pharmaceutical industries, hospitals and environments (Rollins, 2000).  Disinfectant as effective agents that kill or eliminates bacteria is widely used in various ways; especially in microbial laboratory.  Disinfectant can be mainly divided into five agents; alkylating, sulfhydryl combining, oxidizing, dehydrating and permeable.  The most commonly used disinfectants in laboratories are ethanol, bleach and Isol (Larson and Morton, 1991).  Bleach also known as sodium hypochlorite is a broad spectrum disinfectant, non specific in their action, only action biological material that is present on any surface. They effects by oxidizing the cell of microorganism and attacking essential cell components including lipid, protein and DNA (Ho-Hyuk Jang et al, 2008).    Ethanol, as  a dehydrating agent, lies between the highly specific and broadly based categories.  It is effective against actively growing bacteria and viruses with a lipid based outer surfaces, but is not effective against bacterial spores or viruses that prefer watery environment.  They cause cell membrane damages, rapid denaturalization of proteins with subsequent metabolism interference an cell lyses (Larson and Morton, 1991).  Another surface disinfectant is the compound that contain phenol group, a popular commercial brand of Isol, (a saponated brand of cresol) as a phenolics are intermediate level disinfectant derived from coal tar, that are effective on contaminated surfaces (Bittel and Hughes, 2003).

            However, certain types of viruses and some bacteria are resistant to the killing action of Phenolics compound (ISO, 2008).  Many studies have been done on comparison of disinfectant efficiency, and ethanol and bleach are believed to have immediate effect against most organisms (Carly et al, 2006).  For bacteria species, the effects of ethanol, bleach, phenol on Pseudomonas aeruginosa and Staphylococus aureus are the bedrock of this study.

Pseudomonas aeruginosa is a classical opportunities pathogen with innate resistance to many antibodies and disinfectants.  It is invasive, toxigenic and produces infection in patients with abnormal host deficiencies (Stephen et al, 2004).  Staphylococus aureus occur in 40 – 50% of humans.  Hospitalized patients as well as medical and paramedical staff show higher incidence of carriage of it (Bhatia and Icchpujani, 2008) in this study, disinfectant experiment was conducted using different concentrations of laboratory ethanol as disinfectant A, household bleach (Jik) disinfectant B and saponated brand of cresol (Isol) disinfectant C against Pseudomonas aeruginosa and Staphylococus aureus

1.1       AIMS AND OBJECTIVES

1.         To find out the concentration of disinfectants that will be effective in Gram positive Staphylococcus aureus and Gram negative Pseudomonas areuginosa.

2.         To investigate their differences of sterilizing pattern.

3.         To advise the public on the important of disinfectants and dangers of harmful microorganisms.

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