ISOLATION AND IDENTIFICATION OF BACTERIAL CONTAMINANTS FROM MICROSCOPES

ABSTRACT

Some parts of the microscope frequently touched with hands can act as vehicles for transmission of infections. The study analyzed forty (40) swab samples from microscopes routinely used in hospitals, clinics and medical laboratories within Aba and Umuahia for bacterial contamination. The samples were cultured on MacConkey, Blood and Mannitol Salt agar and incubated aerobically at 37oC for 24 hours. Isolation, identification and Antibiogram of isolates were done by standard microbiological techniques. A total of 17 bacterial organisms comprising five (5) bacterial genera were recovered from the microscopes in this study. Streptococcus species 7(41.2%), was the most frequently isolated organism followed by Staphylococcus aureus 4(23.5%), Pseudomnas aeruginosa 4(23.9%) and Bacillus species 1(5.9%) and Serretia species 1(5.9%). The fine and coarse adjustment knobs harbored more bacterial contaminants (69.2%) than other parts of the microscope samples - eye piece and stage condenser (30.8%). Results of the antibiotic susceptibility test of the isolates presents a general pattern of mild resistance to easily available, across counter drugs. All the isolates showed 100% sensitivity to Streptomycin. The level of contamination varied depending on the exposure of the environment; hence the low incidence of bacterial contamination could be due to regular cleaning of the microscope and use of hand gloves by the microscope users. Microscopes used in medical facilities should be cleaned regularly.    

TABLE OF CONTENTS

Cover page                                                                                                                              i

Title page                                                                                                                                ii

Dedication                                                                                                                              iii

Certification                                                                                                                            iv

Acknowledgement                                                                                                                  v

Table of contents                                                                                                                    vi

List of tables                                                                                                                           viii

Abstract                                                                                                                                  ix

CHAPTER ONE

1.0           Introduction                                                                                                              1

1.1       Aim of study                                                                                                               3

1.2       Objectives of Study                                                                                                    3

1.3       Significance of study                                                                                                  3

CHAPTER TWO

2.0           Literature review                                                                                                      4

2.1       Bacteria contamination and Health Dangers                                                              4

2.2       Fomites and infection transmission                                                                            4

2.3       The Microscope                                                                                                          9

2.4       Types of microscope                                                                                                  11

2.4.1    Optical microscope                                                                                                     11       

2.4.2    X-ray microscope                                                                                                       12

2.4.3    Electron microscope                                                                                                   12

2.4.4    Fluorescence microscope                                                                                           14

2.4.5    Scanning probe microscope                                                                                       15

2.5       Components of the microscope                                                                                  16

2.6       Care for the microscope                                                                                             17

2.7       Common bacterial contaminants of fomites/microscopes                                         18       

CHAPTER THREE

3.0           Materials and Methods                                                                                            20

3.1           Study Area                                                                                                                  20

3.2           Collection of Samples                                                                                                20

3.3           Preparation of Culture Media                                                                                     20

3.4           Bacterial inoculation and isolation                                                                             21

3.5           Antibiotic Susceptibility Testing                                                                                21

3.6           Characterization and Identification of Organisms                                                      21

3.7           Gram Stain                                                                                                                  21

3.8           Biochemical Tests                                                                                                      22

3.8.1    Catalase Test                                                                                                               22

3.8.2    Coagulase Test                                                                                                           22

3.8.3    Citrate Test                                                                                                                 22

3.8.4    Motility, Indole, Urease Test (MIU)                                                                          23

3.8.5    Triple Sugar Iron Test                                                                                                23

3.8.6    Oxidase Test                                                                                                               24

3.9       Data Analysis                                                                                                              24

CHAPTER FOUR

4.0           Results                                                                                                                        25

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendation                                                      33

5.1       Discussion                                                                                                                   33

5.2       Conclusion                                                                                                                  35

5.3       Recommendation                                                                                                       35

References                                                                                                                             36                   

CHAPTER ONE

1.0       INTRODUCTION

Countless reasons have been shown to influence bacterial transfer between surfaces; including the bacterial species involved, moisture levels, pressure and friction between contact surface and lastly, inoculum sizes on surfaces (Chen et al., 2001; Rusin et al., 2002). Studies have also shown that hospital/clinical surfaces can easily be contaminated with bacteria and that viruses can easily be transferred to hands from hands to mouth (Rusin et al., 2002). Fomite is any objects or substances capable of carrying infectious organisms, such as germs or parasites and hence transferring them from one individual to another. Door handles, skin cells, hair clothing and bedding are common hospital sources of contamination (Barrie, 2009). Fomites are associated particularly with nosocomial infections, commonly known as hospital acquired infections (HAI), as they are possible routes to pass pathogens between patients.  Stethoscope and neckties are two such fomites associated with health care providers, basic hospital equipment, such as IV drip tubes, catheters and life support equipment can also be carriers, when the pathogens form biofilms on the surfaces. Careful utilization of such objects prevents cross-infection. A preliminary study suggests that aerosols assist viruses in travelling distances increasing the likelihood of transmission.

Fomites consist of both porous and non porous surfaces or objects that can become contaminated with pathogenic microorganisms and serve as vehicles in transmission. During and after illness, pathogenic and non pathogenic organisms are shed in large number in body secretions including blood, feces, urine, saliva, and nasal fluids. Fomites become contaminated with fungi, bacteria and viruses by direct contact with soiled hands, contact with aerosolized virus (large droplet spread) generated via talking, sneezing, coughing, or vomiting or contact with airborne organisms that settles after disturbance of a contaminated fomites (i.e shaking a contaminated blanket).

Once a fomite is contaminated, the transfer of infectious organisms may readily occur between inanimate and animate objects, or vice versa and between two separate fomites (if brought together) (Talaro, 2011). According to a study conducted by Talaro (2011), otherwise called pancic study recovered 3 to 1,800 PUF (plague forming unit) of rhinovirus from fingertips of volunteers who handled contaminated doorknobs or faucets. Using celiphage PRD-1 as a model, demonstrated that 65% of virus could be transferred to uncontaminated hands and 34% to the mouth. He also reported that the nature and frequency of contact with contaminated surfaces vary for each person depending on age, personal habits, type of activities, personal mobility, and the level of cleanliness in the surrounding. Viral transfer and disease transmission is further complicated by viriations in virus survival on fomites is influenced by intrinsic factors which include fomites properties or virus characteristics and extrinsic factors, including environmental temperature, humility etc. if viruses remain viable on surfaces long enough to come in contact with a host, the virus may only need to be preserve in small number to infect the host.

After contact with the host is achieved, viruses can gain entry into the host systems through portals of entry or contact with the mouth, nasopharynx, and eyes. Host susceptibility to viruses in influenced by previous contact with the virus and the condition of the host immune system at the time of infection (Rhame, 2012). Nosocomial infections constitute over 25% of infection.

1.1       AIM:

This aim of this study is to evaluate the bacterial contamination of microscopes and to determine the antibiogram of the isolates.

1.2       OBJECTIVES OF THE STDUY

The specific objectives of the study include;

i.               To isolate and identify bacteria from various parts of microscopes routinely used in hospitals, clinics and laboratories.

ii.              To determine the antibiotic sensitivity pattern of the isolates.

1.3       SIGNIFICANCE OF THE STUDY

The significance of this study is to establish the possible presence of some pathogenic organisms on microscopes in hospitals, and for laboratory technicians, health workers or medical practitioners in general to know the need and importance of appropriate aseptic and enhanced hygienic practices.

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