ABSTRACT
The present study investigates the prevalence of multi-drug resistant bacteria isolates from selected recreational waters in Umuahia, Abia State Nigeria. The bacteria isolates were identified on the basis of their cultural, morphological, biochemical characteristics and antibiotic susceptibility were tested by disc diffusion method. The percentage occurrence of isolates were Escherichia coli 5(15.2%), Staphylococcus aureus 8(24.2%), Klebsiella sp. 5(15.2%), Enterobacter sp. 9(27.3%) and Pseudomonas aeruginosa 6(18.2%) respectively. In-vitro sensitivity patterns were determined using the disc diffusion method, it revealed that the isolates, Escherichia coli 5(100%) resisted Co- trimoxazole and Amoxicillin, 3(60%) were sensitive to Chloramphenicol, 2(40%)were sensitive to Gentamycin. Staphylococcus aureus, 6(75%) resisted Erythromycin and Gentamycin, 5(62.5%) resisted Septrin and Perfloxacin, 4(50%) resisted Amoxicillin while 3(37.5%) were sensitive to Co- trimoxazole and 5(62.5%) were sensitive to Chloramphenicol. Klebsiella sp., 4(80%) were resitant to Erythromycin, Amoxicillin, Co- trimoxazole and Perfloxacin, 4(40%) were sensitive to Perfloxacin, Gentamycin and 1(20%) were sensitive to Erythromycin and Perfloxacin. Enterobacter sp.8(88.9%) were resistant to Co- trimoxazole, 7(77.8%) resisted Amoxicillin, 5(55.6%) resisted Chloramphenicol, 4(44.4%) resisted Erythromycin, 2(22.2%) resisted Gentamycin while 5(55.6%) were sensitive to Perfloxacin and 3(33.3%) were sensitive to Erythromycin and Gentamycin. Pseudomonas aeruginosa, 5(83.3%) were resistant to Erythromycin and Gentamycin, 4(66.7%) resisted Penicillin and Perfloxacin, 3(50%) resisted Chloramphenicol and Amoxicillin while 2(33.3%) were sensitive to Chloramphenicol and 1(16.7%) proved sensitive to Amoxicillin and Co- trimoxazole. From this study the presence of Multi-drug resistance bacteria in recreational water could present a potential health risk which will contribute to health hazard of poorly managed recreational water sources.
TABLE
OF CONTENT
Title
page i
Certification ii
Dedication iii
Acknowledgement iv
Table
of content v-vii
List
of tables viii
List
of figures ix
Abstract x
Chapter One
1.1
Introduction 1
1.2
Contamination of water sources 1-3
1.3
Aims 3
1.4
Objectives 3
Chapter
Two
1.0 Literature
review 4
2.1
Recreational water 4-5
2.2
Recreational water sanitation 5-6
2.3
Microbiological analysis of recreational water 7-8
2.4
Prevention of disease in recreational water 9
2.5
Multi-drug resistant bacteria 9-10
2.5.1
Antibiotics 10-12
2.5.2
Drug resistance among bacteria 13
2.5.3
Biochemical mechanism of bacteria resistance 14
2.5.3.1
Mutation alteration of target protein 14
2.5.3.2
Enzymatic inactivation of drugs 14
2.5.3.3
By-passing of target 16
2.5.3.4
Ribosome protection 17
2.5.3.5
Target modification 17
2.5.3.6
Acquisition of latest gene 18
2.5.4
Managing the drug mechanism problem 18
Chapter Three
2.0 Materials
and methods 19
3.1
Area of study 19
3.2
Sample collection 19
3.3
Media used 19
3.4
Media preparation 19
3.5
Isolation of bacteria from recreational water samples 20
3.6
Identification and characterization of isolates 21
3.6.1
Colony morphology 21
3.6.2
Microscopic examination 21
3.6.3
Gram staining 21
3.7
Biochemical test 22
3.7.1
Catalase production test 22
3.7.2
Oxidase test 22
3.7.3
Coagulase test 22
3.7.4
Methyl-red test 22
3.7.5
Voges-proskauer test 23
3.7.6
Carbohydrate utilization test 23
3.7.7
Motility test 24
3.8
Test for multi-drug resistance 24
Chapter Four
4.0 Results 25-39
Chapter Five
5.1
Discussion 40-41
5.2
Conclusion 42
5.3
Recommendation 42
References
LIST
OF TABLES
Table Title Page
1
Biochemical
characteristics of isolates obtained 26
2
Percentage occurrence of
isolated organism 27
3
Antibiotic susceptibility
test for various isolated 28-29
organism
4
Antibiotic susceptibility
pattern of Escherichia coli
isolated from selected recreational water
samples 30
5
Antibiotic susceptibility
pattern of Staphylococcus aureus
isolated from selected recreational water
samples 31
6
Antibiotic susceptibility
pattern of Klebsiella sp.
isolated from
selected recreational water samples 32
7
Antibiotic susceptibility
pattern of Enterobacter sp.
isolated from
selected recreational water samples 33
8
Antibiotic susceptibility
pattern of Pseudomonas aeruginoasa
isolated from
selected recreational water samples 34
LIST OF FIGURES
Figure Title Page
1
Anti-biogram of
multi-drug resistance percentage
of Escherichia coli isolated from recreational
water samples. 35
2
Anti-biogram of
multi-drug resistance percentage
of Staphylococcus aureus isolated from recreational
water samples. 36
3
Anti-biogram of
multi-drug resistance percentage
of Klebsiella sp. isolated from recreational
water
samples. 37
4
Anti-biogram of
multi-drug resistance percentage of
Enterobacter
sp. isolated from recreational water samples. 38
5
Anti-biogram of
multi-drug resistance percentage of
Pseudomonas
aeruginosa isolated from recreational
water samples. 39
CHAPTER ONE
1.1
INTRODUCTION
Contamination refers to the presence of
unwanted constituent, contaminants or impurity
In
a material, physical body, natural environment, workplace etc. (IAEA, 2010).
In chemistry, the term
contamination usually describes a single constituent, but specialized fields,
the term could also mean chemical mixtures, even up to the level of cellular
materials. All chemicals contains some level of impurity. Contamination may be
recognized or not and may become an issue if the impure chemical is mixed with
other chemicals or mixtures and causes additional chemical reaction (IAEA,
2007)
In environmental
chemistry, the term in some cases virtually equivalent to pollution, were the
main interest is the harm done on large scale to humans or to organisms or
environments that are important to humans.
In biological sciences,
accidental introduction of foreign materials (contaminations) can seriously
detort the results of experiments were small samples are used. In cases, were
the contaminant is a living microorganism, it can often multiply and take over
the experiment, especially cultures and render them useless.
In environmental
sciences, contamination arises from several potential sources which includes
urban activities, wildlife, and agricultural activities (IAEA, 2007).
1.2 CONTAMINATION OF
WATER SOURCES
Global changes will cause important
perturbation to aquatic ecosystem among these, degradation of microbial quality
of surface water is expected (Bruce et al.,
2006). The climate changes will bring about an increase in the frequency and
intensity of rain events which will be followed by an increasing amount of
water reaching watercourses by runoff. The increased runoff will carry a larger
amount of microorganism which comes to settle on watercourses.
Contamination
of water sources (Bruce and Cutis, 2006) arises from several potential sources
which includes: urban activities, wildlife activities, and agricultural
activities. Water contaminated by both animal and human sources (Marsalek et al., 2006; Palz et al., 2008) of feacal microorganism can present a health risk to
human health for people who consume water from it. Although, it is assumed that
in general animal sources of feacal contamination represent a smaller risk to
human health than human sources. Animals can carry zoonotic pathogens such as Salmonella,Campylobacter, and Escherichia coli which may lead to
gastrointestinal illness and sometimes severe sequelae.
In
urban activities (Bruce et al.,2006),
water sources can be contaminated during rain events; water can runoff in two
major ways: they can reach watercourses via pluvial sewer systems carrying many
pollutants such as organic contaminants and feacal pathogens coming from
domestic waste, domestic fauna etc. They can also reach water sources by
directing to combined sewer system that collects domestic industrial and
pluvial waters. However, during heavy rainfall (Wade and Beach, 2008) and rapid
snow melts, water discharge coming into this system can overwhelm waste
watercourses by overload drains called Combined Sewer Overflows (CSOs).
CSOs
may represent a health risk for people in contact with these waters since the
principle source of bacteria it contains is from human waste, which can contain
a large quantities of feacal pathogen (Jamj, 2008).
Contamination of water sources can occur directly from
human (swimmers and those who bath in water sources) via feacal accidents
mainly were the proportion of children among swimmers or those bathing is high
and if there are babies or toddlers on diapers (Collins and Biemers,2006).
Wide animals can also contribute to feacal pollution
of water sources such as the birds which can be carried of feacal pathogen
including species of Campylobacter, Salmonella, Cryptosporidium (Calderon,
2008). They can contaminate the water by direct deposit of feacal material into
the water sources.
Agricultural sources can contribute to feacal
contamination of water sources in many ways (Calderon and Jamj, 2008). It can
come from animals and manure piles on farm and animal sites. Livestock can
carry and excrete in there feaces, zoonotic agents which may lead to various
health problems from self-limiting gastrointestinal disturbances to severe
disease that are potentially deadly. Zoonotic pathogens most associated with
farm animals includes: Salmonella
enterica,Campylobacter jejuni, Escherichia coli, Giardia sp, and Cryptosporidium
sp. These pathogens can reach water sources following rain events.
1.3 AIM
The aim of this research is to determine the
antibiotic resistant pattern of bacterial isolates from various recreational
waters
1.4 OBJECTIVES
·
To isolate and identify
various bacterial species from recreational waters.
·
To assay for antibiotic
resistance among these isolates
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