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

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Product Code: 00001397

No of Pages: 84

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ABSTRACT

Ethanol, Bleach and Phenolics are three kinds of disinfectants which have been widely used in common laboratories.  In this study, a compared experiment on these three disinfectants efficiency was conducted against Staphylococcus aureus and Pseudomonas aeruginosa using agar hole diffusion method.  Different concentrations of bleach (1%, 2%, 3%, 4% and 5%) were used on both organisms.  Also (50%, 60%, 70%, 85% and 95%) of ethanol as well as (5%, 10%, 20%, 25%, and 30%) Phenolics were used. Diffrences in  concentrations tested was because, the original concentrations of the disinfectants differs. After 24 hours of incubation at 370C, the results showed that all the disinfectants inhibited the growth of the test organism in their concentrated forms.  The diameter of zone of inhibitions were measured around each well by using a ruler in millimeters, using different concentrations, their efficacies varied.  The results showed that 30% Phenolics had the best efficiency against both test organisms and 5% bleach had a better effect on Staphylococcus aureus than Pseudomonas aeruginosa, while ethanol showed least sensitivity. 70% concentration gave the highest effect on Staphylococcus aureus as compared with Pseudomonas aeruginosa.

           

 

 


 

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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