MULTI-DRUG RESISTANT (MDR) BACTERIA ISOLATED FROM SELECTED RECREATIONAL WATERS IN UMUAHIA, ABIA STATE.

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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