ANTIBIOGRAM AND MICROBIAL CARRIAGE OF CAMPUS SHUTTLE DOOR HANDLES

(A STUDY OF FEDERAL UNIVERSITY OF TECHNOLOGY, AKURE)

By

Author

Presented To

Department of Microbiology

ABSTRACT

The transmission of infection via fomites constitutes a major threat to public health especially in the developing countries. This study was carried out to investigate the microbial carriage and antibiotics pattern of bacteria on the door handles of campus shuttle operating in the Federal University of Technology Akure. Samples were randomly collected from a total of one hundred and fifty (150) door handles following standard laboratory techniques. Enumeration of the bacterial counts was carried out using pour plating technique while the bacteria isolates were identified using cultural, morphological and biochemical characteristics. The results show that the drivers’ door handles, front passengers’ door handles and back passengers’ door handles had overall mean aerobic bacterial counts of 195.56 ± 2.05 (x104cfu/ml), 228.08 ± 1.56 (104cfu/ml) and 176.92 ± 2.70 (x104cfu/ml) respectively. Bacteria isolated and identified wereEnterobacter aerogenes (17.1%), Escherichia coli (12.4%),Klebsiella pneumoniae (10.1%), Proteus spp. (4.7%), Pseudomonas aeruginosa (4.7%), Salmonella enteritidis (2.3%), Staphylococcus aureus (17.1),Staphyloccoccus epidermidis (22.5) andStreptococcus pneumoniae (9.3%) while the fungal isolates wereAspergillus fumigatus (29.3%), Aspergillus niger(19.5%),Candida spp. (6.1%), Fusarium spp. (14.6%),Mucor spp. (9%), Penicillium spp. (15.9%), and Rhizopus spp. (3.7%). The antibiotics pattern of the bacteria indicated that all the bacteria isolated were susceptible to pefloxacin (19.25 ± 4.031-26.33 ± 4.041mm) and gentamycin (15.00 ± 2.685-25.00 ± 3.916mm) while all were resistant to amoxicillin (3.67 ± 0.082-13.75 ± 2.217mm) except Enterobacter aerogenes. These campus shuttles aside their usage as a means of transportation could also serve as a means of transmission of both pathogenic and non-pathogenic microorganisms which pose public health risks. Personal hygiene and sanitation such as hand washing and the use of antimicrobial wipe to clean the hands could serve as a means of reducing the incidence of microbial transmission to drivers and passengers.

TABLE OF CONTENTS

Content Page

Title I

Certification II

Dedication III

Acknowledgements IV

Table of contents V

List of tables IX

List of plate X

Abstract XI

CHAPTER ONE

1.0 Introduction 1

1.1 The objectives of the study 4

1.2 Justification 4

CHAPTER TWO

2.0 Literature review 5

2.1 Background 5

2.2 Roles of different surfaces in the transmission of pathogens 8

Content Page

2.2.1 Mobile phones 8

2.2.2 Tooth brushes 9

2.2.3 Toilet door handles 10

2.2.4 Currency and coins 10

2.3 Factors that favours transmission of infection to humans from fomites 11

2.3.1 Individual susceptibility 11

2.3.2 Environmental factors 11

2.4 Possible preventive measures against transmission of infection via fomites 11

2.5 Antimicrobial activities of antibiotics 12

2.6 Antibiotics resistance 13

CHAPTER THREE

3.0 Materials and methods 15

3.1 Ethical clearance 15

3.2 Sample collection 15

3.3 Media preparation and isolation of microorganism 15

3.3.1 Nutrient agar 15

3.3.2 Potato dextrose agar 16

3.3.3 Eosin methylene blue agar 16

Content Page

3.3.4 Blood agar 16

3.3.5 Salmonella shigella agar 17

3.4 Total viable count 17

3.5 Purification of bacterial isolates 18

3.6 Cultural and morphological identification of bacteria 18

3.7 Identification and biochemical characterization of bacterial isolates 19

3.7.1 Gram staining 19

3.7.2 Indole test 19

3.7.3 Citrate test 20

3.7.4 Oxidase test 20

3.7.5 Urease test 21

3.7.6 Catalase test 21

3.7.7 Coagulase test 21

3.8 Identification and characterization of fungal isolates 22

3.8.1 Isolation of pure fungal isolates 22

3.9 Antibiotics sensitivity test 22

CHAPTER FOUR

4.0 Results 25

Content Page

4.1 Total bacterial load 25

4.2 Frequency of bacterial isolates associated with shuttle door handles 25

4.3 Identity of the bacterial isolates 33

4.4 Frequency of fungal isolates associated with the shuttle door handles 36

4.5 Antibiotics susceptibility pattern of bacterial from the door handles 41

CHAPTER FIVE

5.0 Discussion 48

CHAPTER SIX

6.0 Conclusion 54

6.1 Contribution to knowledge 54

6.2 Recommendation 54

References 56

LIST OF TABLES

Table Title Page

1. Total viable count of bacteria on the door handles 26

2. Frequency of total bacteria isolated from the shuttle door handles 27

3. Frequency of the bacterial species isolated from the entire door handles 28

4. Frequency of bacterial species from the drivers’ door handles 30

5. Frequency of bacterial species from the front passengers’ door handles 31

6. Frequency of bacterial species from the back passengers’ door handles 32

7. Biochemical characteristics of bacterial isolate 34

8. Morphological characteristics of bacterial isolate 35

9. Frequency of fungal isolates obtained from the entire door handles 37

10. Frequency of fungal species on the entire door handles 38

11. Frequency of fungal species on the drivers’ door handles 39

12. Frequency of fungal species on the front passengers’ door handles 40

13. Frequency of fungal species on the back passengers’ door handles 43

14. Morphological characteristics of fungal species 44

15a. Zones of inhibition of the bacterial isolates measured in millimeter 45

15b. Zones of inhibition of the bacterial isolates measured in millimeter 46

LIST OF PLATE

Plate Title Page

1. Antibiotics pattern of Streptococcus pneumoniae 47

CHAPTER ONE

1.0 Introduction

The spread of infectious disease through hand contact has been an area of major public health concern because of the frequent contact of the hand with fomites which are potential carriers of pathogenic organisms may lead to an alarming rate of outbreaks of infections transmitted by the fomites. Worldwide annually there are 1.7 million deaths from diarrhoeal diseases and 1.5 million deaths from respiratory infections (Pruss-Ustun and Covahan, 2006), these are examples of diseases which could be contracted by humans via fomites. According to (Itah, 2004), Gram positiveStaphylococcus aureus, and Gram negative bacteria such as Escherichia coli, Klebsiella species, Pseudomonas species, were found to contaminate various contact surfaces including chairs, tables, windows, door handles and many other common household fixtures. The presence of these pathogenic bacteria on environmental surfaces poses a potential risk to vulnerable, immune-compromised individuals (Pruss-Ustun and Covahan, 2006).

It has been shown that hard, non-porous surfaces such as door handles have the highest bacterial transfer rates to hands (Rusin et al., 2002). In recent past, a lot of effort has been invested in emphasized hand hygiene through hand wipes and hand sanitizers (Hota, 2004). Hand wipes with higher ethanol content are more effective in not only antimicrobial activities but also remover of endospores via mechanical action (Stout et al., 2010). Even though people are commonly aware of such practices, the possibility of inaccessibility or lack of use of these practices do exist. According to Hansen and Knochel (2010) up to 60% 0f adults do not wash their hands when appropriate. People believe that microbes are only present in research laboratories, hospitals or clinics and thus they have a misleading feeling of security in other places or while touching other surfaces such as door handles (Stout et al., 2010). Lack of knowledge of the roles of micro-habitat such as door handles in dissemination of microorganisms is a threat to public health. In fact 80% of infections are spread through hand contact with hands or other objects (Reynold and Hurst, 2005). Reynold and Hurst (2005) used an invisible fluorescent tracer for artificial contamination of public surfaces. They found that contamination from outside surfaces was transferred to 86% of exposed individual’s and 82% tracked the tracer to their home or personal belongings hours later. The viability of Gram positive and Gram negative organism under various environment conditions have been described (Noskin et al., 1995). Some microbes are infectious at very low doses and can survive for hours to weeks on nonporous surfaces, such as countertops, telephone and door handles (Reynold and Hurst, 2005). Enterococci have been found to survive in dry conditions and on various fabrics utilized in the health care environment. Infectious doses of pathogen may be transferred to the mouth after handling everyday contaminated objects such as the door handles. Door handles are contaminated with various types of microorganisms including varying species of bacteria and fungi and this may be a source of nocosomial infections in hospitals. Scientific information about the occurrence of microorganisms on various objects outside the health care facilities is very little and needs to be enriched in order to educate people on the necessity of improving the habit of hand washing to reduce microbial transmission.

In the past 60 years, antibiotics have been critical in achieving a dramatic rise in life expectancy and significant improvements in public health (El-Astal, 2005). However, disease-causing microbes have become increasingly resistant to the antibiotics commonly in use (El-Astal, 2005). It has been clearly shown that the use of antimicrobials leads to selection of resistant strains both in the individual and in the community, and overuse or inappropriate use only increases this risk (Reynold and Hurst, 2005). History suggests that microbes will never run out of ways of developing resistance, but we may run out of effective antimicrobials (Reynold and Hurst, 2005). The Gram negative enteric bacilli are common causes of a wide variety of infections involving diverse anatomic sites in both healthy and compromised hosts (El-Astal, 2005). In general, among adults, the incidence of infection due to these agents increases with age. Thus, as the mean age of the population increases, so will the number of these infections (Reynold and Hurst, 2005). Drug resistance is a serious medical problem. Progressive increase in resistance to commonly used antibiotics with many gram-negative bacilli being multidrug-resistant has been noticed (Reynold and Hurst, 2005). The emergence of antibiotic resistance in the management of infections is a serious public health issue, particularly in the developing world where apart from high level of poverty and ignorance, there is also high prevalence of fake and spurious drugs of questionable quality in circulation (El-Astal, 2005). This has led to a significant increase in morbidity.

There also appears to be a significant lack of studies highlighting the susceptibility patterns of locally prevalent microorganisms (El-Astal, 2005). Knowledge of etiological agents of infections and their sensitivities to available drugs is of immense value to the rational selection and use of antimicrobial agents and to the development of appropriate prescribing policies (El-Astal, 2005). Microbial transmission via several surfaces such as automated teller machines, mobile phones, door handles and currency has been extensively studied but the role of commercial buses door handles in the dissemination of pathogenic microorganisms have not been properly investigated. Therefore this research project will be centered on the role of commercial buses door handle in the transmission of pathogenic microorganism to human via hand contact and the antibiotics pattern of the potential isolates.

1.1 The objectives of the study

The aim and objectives of this research are to:

(a) assess the role of campus shuttle door handles in dissemination of pathogenic microorganisms; and

(b) determine the antibiotics susceptibility and resistance pattern of the potential isolates.

1.2 Justification

Over the years, different researches has been conducted to examine the role of various surfaces, such as tables, computer key boards and mobile phones on the carriage and dissemination of pathogenic infection, but the role of door handles of commercial shuttle buses as a route of microbial transmission has not been reported. Hence the need to assess the carriage and transmission of microorganism by shuttle door handles. The knowledge of this is expected to broaden our understanding of the microbial carriage of the door handles and their antibiotics pattern. This will help the University management to implement public health preventive and control measure to forestall future outbreak of infection that may result from this pathogenic microorganisms associated with the shuttle door handles.

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