ABSTRACT
Globally, human health is under threat due to the presence of diseases caused by microorganisms such as bacteria and viruses. Bacteria are found on almost all surfaces with which people come into contact and therefore pose a threat to human health. Antimicrobial resistance is a major health concern as these microorganisms can cause untreatable diseases. I tested the surfaces found in three buildings on the campus of the American University of Nigeria in Yola, Adamawa State, for the presence of harmful bacteria. Swabs collected were streaked on lysogeny broth agar to determine if bacteria were present. Selective and differential media were used to identify the type of bacteria found. Also, the samples were subjected to an antimicrobial susceptibility test using replica plating. The result showed the surfaces were contaminated with Escherichia coli, Enterobacter aerogenes, and Enterococcus faecalis, with E. coli and E. aerogenes being resistant to ampicillin. This study showed that AUN is not only contaminated with pathogens, but also with bacteria resistant to ampicillin. The institution should provide hand sanitizers around campus and also create awareness
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programs on the importance of hand washing. Also, the efficiency of cleaning can be increased by making use of stronger agents and increasing the frequency of cleaning.
Keywords
American University of Nigeria, bacteria, antimicrobial resistance, microbial contamination, surfaces, students and staff
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TABLE OF CONTENTS
CERTIFICATION…………………………………………………………………………………………. ii
READERS’APPROVAL………………………………………………………………………………. iii
DEDICATION ……………………………………………………………………………………………… iv
ACKNOWLEDGMENTS ………………………………………………………………………………. v
ABSTRACT …………………………………………………………………………………………………. vi
LIST OF TABLES …………………………………………………………………. ix
LIST OF FIGURES ………………………………………………………………… x
CHAPTER 1 …………………………………………………………………………………………………. 1
INTRODUCTION …………………………………………………………………………………………. 1
Antibiotic resistance ……………………………………………………………………………………….. 2
Resistance in bacteria …………………………………………………………………………………….. 3
Increasing health risks due to microorganisms ………………………………………………….. 4
Public places and bacteria ………………………………………………………………………………. 5
Bacteria in universities …………………………………………………………………………………… 7
HYPOTHESIS …………………………………………………………………………………………….. 12
AIMS AND OBJECTIVES …………………………………………………………………………… 12
CHAPTER 2 ……………………………………………………………………………………………….. 13
MATERIALS & METHODS ………………………………………………………………………… 13
Study Site …………………………………………………………………………………………………….. 13
Sampling ……………………………………………………………………………………………………… 13
Materials …………………………………………………………………………………………………….. 14
Lab analysis ………………………………………………………………………………………………… 14
CHAPTER 3 ……………………………………………………………………………………………….. 22
RESULTS …………………………………………………………………………………………………… 22
Surface analysis …………………………………………………………………………………………… 22
Microbial Identification ………………………………………………………………………………… 23
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Antimicrobial susceptibility analysis ………………………………………………………………. 26
CHAPTER 4 ……………………………………………………………………………………………….. 30
DISCUSSION ……………………………………………………………………………………………… 30
CHAPTER 5 ……………………………………………………………………………………………….. 35
CONCLUSION ……………………………………………………………………………………………. 35
References …………………………………………………………………………………………………… 36
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LIST OF TABLES
Table 1. Appearance of microorganisms on differential media …………………………… 18
Table 2. Where bacteria were found ……………………………………………………………….. 23
Table 3. Differential and Selective Media used and the microorganisms detected … 24
Table 4. Susceptibility test of bacterial colonies against antibiotics …………………….. 26
Table 5. Amount of DNA in each sample ………………………………………………………… 28
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LIST OF FIGURES
Figure 1. Projections of deaths that could be attributable to antimicrobial resistance yearly by 2050 ……………………………………………………………………………………………….. 3
Figure 2. Structure of Staphylococcus aureus …………………………………………………… 10
Figure 3. Gel electrophoresis under UV light …………………………………………………… 28
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CHAPTER 1
INTRODUCTION
Human health is under serious threat globally by microorganisms, especially viral and bacterial diseases. Bacteria and viruses can be found on surfaces and could potentially cause harm. Infectious diseases are emerging at an alarming rate and have contributed to a good number of deaths globally. Although over the past decade the mortality rates of these diseases have declined, the impact these diseases have on the world remains substantial. Infectious diseases are the second leading cause of death worldwide, with 57 million deaths occurring each year, as reported by the Center for Disease Control and Prevention (CDC) (Fauci, Touchette, & Folkers, 2005).
There are increasing global concerns about certain issues such as antibiotic-resistant bacteria, Zika virus, HIV/AIDS, Ebola virus, and SARS. These diseases have a high mortality rate and have been identified by the World Health Organization as diseases likely to cause epidemics (WHO, 2015). In Africa, emerging diseases are a public health concern not only due to their high mortality rate, but also due to the fact that developing countries lack good health facilities. Despite countermeasures that have been developed due to advanced technology (therapeutics and vaccines), infectious diseases that affect both human health and the economic stability of societies have not been contained (Morens & Fauci, 2013). Bacteria and viruses that have developed resistance to antimicrobial agents are more life threatening and are now a serious public health concern.
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Antibiotic resistance
Antimicrobial resistance has become a major global concern. Aside from bacteria and viruses, other microorganisms are rapidly developing a resistance to antimicrobial agents being used to kill them. This evolution makes the treatment of infectious diseases less effective and, in the long run, may cause death. Though sometimes viewed as an apocalyptic fantasy, antibiotic resistance, which might allow even minor injuries to kill, is indeed a reality in the 21st Century (WHO, 2014). Antimicrobial resistance is a global concern because it will make the treatment of infectious diseases less effective and prolong illnesses (WHO, 2016). Additionally, organ transplantation, C-section delivery, and other medical procedures may become more risky due to the lack of effective antimicrobials.
The increasing global concern for antimicrobial resistance has urged scientists to further research this issue. Projections made by scientists showed deaths that could be attributable to antimicrobial resistance yearly by 2050 will be high (Fig.1). Most deaths will most likely occur in Asia and Africa. Although antimicrobial resistance is increasing, less researches have been carried out on new drugs to curtail these resistant pathogens.
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Figure 1. Projections of deaths that could be attributable to antimicrobial resistance yearly by 2050
Resistance in bacteria
Various bacteria have developed a resistance to particular antibiotics. To name a few, bacteria such as Mycobacterium tuberculosis and Staphylococcus aureus have developed a resistance to certain antibiotics (Shanks & Peteroy-Kelly, 2009). Klebsiella pneumoiae, a bacterium known for causing intestinal infections, is resistant to carbapenem antibiotics and has spread globally, leading to several deaths (WHO, 2016). Additionally, Escherichia coli which causes intestinal infections, is resistant to fluoroquinolone antibiotics. Currently, many countries around the world have no effective treatment for these bacteria. Other bacteria, such as S. aureus (Fig.2) and members of the family Enterobacteriaceae, which are resistant to methicillin and carbapenem, have also contributed to many untreatable conditions in different countries (WHO, 2016). The more resistant a strain of bacteria is, the greater the threat to human health.
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Increasing health risks due to microorganisms
Pathogens can be found on almost all surfaces. Bacteria have the ability to grow anywhere, even in narrow surfaces, and have the ability to move (Männik et al., 2009). They can also survive on surfaces for an extended period. Some diseases caused by bacteria and viruses are life threatening and require immediate attention (Morris, 2016). Prophylaxis such as vaccinations may help lower the probability of getting infected. Also, good hygiene behaviors and good cleaning practices could lower the risk of getting infected. Globally, pandemics are rapidly spreading and have left people concerned about their health. People are more likely to catch diseases caused by bacteria and viruses in places where people congregate and where they make frequent contact with non-living objects.
Before assessing the level of microbial contamination on surfaces, hygiene standards must be considered which are set by regulatory bodies (White, Dancer, Robertson, & McDonald, 2008). Such standards for pathogens were set by the World Health Organization (WHO) at <1cfu/cm2, which means colony forming unit should not exceed 1 per cm2.
People in congregated areas are increasingly exposed to health risks daily. Health risks associated with public places vary; however, bacteria and viruses are the most easily contracted microorganisms. Microorganisms have several routes through which they can infect humans, including the mouth and from the hand to the mouth (Butz, Fosarelli, Dick, Cusack, & Yolken, 1993). Exposure routes of bacteria could be from shopping carts, offices, restrooms (especially public restrooms), and even on university campuses.
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Public places and bacteria
The human skin is the main organ of contact with microbes. The skin on its own houses many bacteria which could be beneficial or harmless. Even though most bacteria found on the human skin are harmless, S. aureus (Kloos & Musselwhite, 1975), Corynebacterium minutissimum, and Pseudomonas aeruginosa could cause skin diseases, such as atopic dermatitis and erythrasma (Ross & Neufeld, 2015).
Studies have shown that bacteria, such as Acinetobacter calcoaceticus and S. aureus, commonly occur on hospital surfaces. “Hospital” bacteria lead to nosocomial infections which are infections acquired in hospital settings. In this research, the life span of the bacteria found on dry hospital surfaces was found to be 13 days, which could be an additional factor in transmission of nosocomial infections (Getchell-White, Donowitz, & Groschel, 1989). In the past years, much attention has been paid to nosocomial infections as the pathogens causing these infections became resistant to antibiotics. In comparison, there are few studies focusing on infections originating on public surfaces, such as public telephones, shopping carts, buses, shopping carts, office furniture, and even on surfaces in university campuses.
Public telephones, which are commonly used in some parts of the world, have been investigated and are found to harbor pathogens. A particular study carried out in Melbourne, Australia, identified certain potentially pathogenic bacteria on 20 telephones, such as Acinetobacter anitratus, Enterobacter cloacae, Pantoea agglomerans, and S. aureus (Ferdinandus, Hensckhe, & Palombo, 2001). From this study, it was ascertained that telephones could be labelled as potential reservoirs for pathogens.
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Similarly, transportation systems such as buses and trains harbor pathogens. Transport systems in Portland, Oregon, USA, were investigated for the presence of pathogens on seats, floors, and railings (Yeh, Simon, Millar, Alexander, & Franklin, 2011). The research further analyzed the pathogens for any resistance in certain antibiotics. Bacteria were found to be more abundant on floors than railings, handles, and seats. Various species of Staphylococcus, such as S. epidermidis, S. haemolyticus, S. saprophyticus, S. warneri, and S. xylosus, were found in the buses. Even though there was no high level of multi-drug resistance, some of the species such as S. xylosus and S. haemolytics were found to be resistant to gentamycin and erythromycin and could engage in horizontal transfer of resistance to other species (Yeh et al., 2011).
Subsequently, pathogens are also commonly found on shopping carts, which may be touched by a large number of people each day. A study carried out across the United States in 5 grocery stores found E. coli, Yersinia pseudotuberculosis, Klebsiella oxytoca, and E. cloacae on shopping carts (Gerber & Maxwell, 2012). In offices, pathogens may be present on surfaces and passed around from one surface to another. Office equipment and furniture, such as printers and computers, carry these pathogens, and since they may be not be regularly cleaned, people may have a high chance of getting infected (Reynolds, Watt, Boone, & Gerba, 2005).
Another “public” place, university campuses, may have high population densities, which makes it easier for bacteria to spread. Students may be exposed to many health risks on campuses, which can be contracted from unclean surfaces, cafeteria food, or bathrooms. Communicable diseases spread easily in places with many people as is the case with universities. The presence of harmful bacteria in a university can pose a
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threat to the health of students, staff, and faculty (Morris, 2016). These bacteria can be easily passed around because as people interact in congregated areas, they may pick up bacteria which could potentially infect them as well as people around them (Shanks & Peteroy-Kelly, 2009). Students in a university normally make use of computers, library study tables, cafeteria tables, and restrooms. All these surfaces may harbor bacteria and could potentially harm people. Unless students are frequently washing their hands or making use of hand sanitizers, they are more likely to get infected.
Bacteria in universities
Previous studies on bacteria in university campuses have confirmed the presence of bacteria on surfaces. For example, pathogens such as E. coli have been found on university tables in clinics, laboratories, and libraries (Burnham, Peterson, Vavrek, & Haas, 2009). Disinfection protocols may have an impact on the level of contamination. However, pathogens could still be present regardless of the cleaning being done on surfaces.
Health risks to students on university campuses could result from the use of public computers, ingestion of cafeteria food, use of bathrooms, or contact with railings, and door handles. Cafeterias may be cleaned frequently, but may still harbor bacteria. Food particles could fall off on the tables in a cafeteria, and if the tables are not probably cleaned, could be the cause of microbes being present. Contact between students and these tables could transfer pathogens to people. Bathrooms are generally known to harbor bacteria. Public restrooms are visited by people with different hygiene behaviors, which can influence the type of bacteria found on restroom surfaces (Flores et al., 2011). Urinary Tract Infections (UTIs) may be contracted from the use of public
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toilets, and these are often caused by E. coli, which is commonly found on restroom surfaces. Depending on the cleaning schedule of a facility, the level of contamination can be minimal.
In addition to restrooms, computer keyboards in computer labs could also be infected with bacteria. Campus computers are often accessed by many people daily, and frequent contact could contribute to high microbial activity. Keyboards may be not routinely cleaned, as well (Anderson & Palombo, 2008). Microorganisms, such as S. aureus, Enterobacteriaceae, Enterococcus faecalis, and Bacillus cereus, were found on computer keyboards in a university computer lab with S. aureus having the highest colony growth on computers used by multiple people.
Another study conducted in a university in New York City sampled different surfaces in lecture halls, restrooms, libraries, and cafeterias (Shanks & Peteroy-Kelly, 2009). The findings revealed that the bacteria found on these surfaces, including E. coli, Salmonella typhi, S. aureus, were resistant strains. This poses a greater risk to people on campuses as the diseases caused by these pathogens will be harder to treat (Shanks & Peteroy-Kelly, 2009). This shows that students may potentially become infected and that these infections might be hard to curtail due to the resistant strains.
The health risk to students may be even higher if bacteria found on campuses have developed a resistance. Such resistance is often due to a mutation or frequent use of an antibacterial agent to clean the surfaces. The bacteria could adapt to the substance being used to clean and natural selection will take place. The only way to curtail diseases caused by bacteria resistant to antibiotics is by prescribing the right dosage of
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antibiotics. This is a pressing public health issue.
In addition, the rise in antimicrobial resistance has also been attributed to the frequent use of antimicrobial agents while cleaning. Some bacteria found on public surfaces that have resistant strains include E. coli, Salmonella, and Enterococcus species (Conly, 2002). The bacteria mentioned are often found on campuses. The different types of bacteria found on surfaces depends on seasons and the different uses of the buildings (Dunn, Fierer, Henley, Leff, & Menninger, 2013).
The level of microbial contamination in a university depends on several factors. These include the cleaning schedule practiced by the Facilities Management and, cleaning materials and products being used (e.g., types of chemicals). The surfaces of a university campus can be seen as non-critical since the surfaces come in contact mostly with human skin and is not necessarily contaminated by blood and bodily fluids (Rutala & Weber, 2001). The U.S.-based Center for Disease Control and Prevention (CDC) has emphasized the need to disinfect surfaces even though they have not been contaminated with blood or body fluids (Rutala & Weber, 2001). If the surfaces get cleaned frequently with a strong disinfectant, the bacterial load will be minimal or negligible.
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Figure 2. Structure of Staphylococcus aureus
Pathogens could be the cause of the high prevalence of bacterial infections on campuses. Numerous infections such as typhoid and flu recorded by clinics on university campuses are most likely transmitted in the dormitories or academic buildings due to the crowded lifestyle on campuses and contact with surfaces (Pechter, 2011). Inadequate personal hygiene increases the level of contamination and also puts students at risk of contracting diseases. Some of these diseases are life threatening and require immediate attention.
The literature on health risks to university students on campuses appears sparse, particularly for universities in developing countries where standards may be lower. Therefore, I investigated facilities on the American University of Nigeria (AUN) campus in Yola, north-eastern Nigeria, for the presence of bacteria on surfaces commonly touched by students. The aim of my study was to determine which strains of bacteria occur in these public places and whether any strains were resistance to antibiotics. The outcome of this work helps raise awareness regarding potential
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harmful pathogens on campus. Findings will be shared with Facilities Management of the university.
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RESEARCH QUESTION
Are students and staff exposed to harmful bacteria when they use the AUN facilities?
HYPOTHESES
Null hypothesis (H0):
Bacteria found on surfaces in AUN facilities pose no health risks to students and staff.
Research Hypothesis (H1):
Bacteria found on surfaces in AUN facilities pose health risks to students and staff.
AIMS & OBJECTIVES
Aim:
To determine potential health risks to students and staff who use the AUN facilities.
Objectives
To swab surfaces in AUN and check for bacteria.
To identify the types of bacteria found on surfaces in AUN facilities.
To determine if the bacteria found are potentially harmful to health.
To examine antimicrobial resistance in the bacteria found in AUN.
To perform molecular analysis so as to determine the cause of antimicrobial resistance
Based on my findings, to recommend potential changes in cleaning approaches to AUN Facilities management and clinic.
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