Incidence of Asymptomatic Bacteriuria Among Pregnant Women In Sabon-Gari Lga – Zaria, Nigeria And The Antimicrobial Susceptibilities of The Isolates.
ABSTRACT
Urinary tract infection (UTI) occurs frequently in pregnancy with a large number of infections being asymptomatic.
This study was therefore conceived to determine theincidence of asymptomatic bacteriuria in pregnant women in Sabon- Gari LGA –Zaria, Nigeria using questionnaire as well as isolate and characterize any bacteriapresent in urine of apparently healthy pregnant women and to determine theantibiogram of such isolates.
Urine samples from two hundred and fifty (250) apparently healthy pregnant women,who were attending ante-natal clinics in Sabon-Gari Local Government Area Zaria,Nigeria, were interviewed and urine samples were collected for isolation of anypossible bacteria present.
Isolation was done by culturing on standard laboratorymedia and identification of isolates were subsequently done using standardbiochemical tests and confirmed using API (bioMerieux, USA) rapid kits.
Theantibiogram of the isolates were determined using selected antibiotics and using discagar diffusion technique. Plasmid isolation using gel electrophoresis was carried outon selected isolates that were multiple antibiotics resistant.
Molecular characterizationusing multiplex Polymerase Chain Reaction (PCR) was carried out to detect nature ofresistance in the resistant isolates.
Primers such as SHV, OXA, TEM, aaclla, gyrAand gyrB, that codes for resistant genes for aminoglycosides, penicillins,cephalosporins and fluoroquinolones were used.
TABLE OF CONTENT
Title Page – – – – – – – – -i
Declaration – – – – – – – – -ii
Certification – – – – – – – – -iii
Dedication – – – – – – – – -iv
Acknowledgement – – – – – – – -v
Special Acknowledgement – – – – – – -vi
Abstract – – – – – – – – -vii
Table of Content – – – – – – – -ix
List of Tables – – – – – – `- – -xiv
List of Figures- – – – – – – – -xv
List of Appendix – – – – – – – -xvi
CHAPTER ONE: INTRODUCTION
1.1 Urinary Tract Infection (UTI)- – – – – – -1
1.2 Asymptomatic Bacteriuria – – – – – – -2
1.3 Statement of Research Problem – – – – – -3
1.4 Justification for the Study – – – – – – -7
1.5 Aim of the Study – – – – – – – -7
1.6 Objectives of the Study – – – – – – -8
1.7 Null Hypothesis – – – – – – – -8
1.8 Limitations – – – – – – – – -9
CHAPTER TWO: LITERATURE REVIEW
2.1 Urinary Tract Infection (UTI)- – – – – -10
2.2 Etiology of Urinary Tract Infection – – – – – -12
2.3 Incidence of Urinary Tract Infection – – – – – -13
2.4 Risk factors for Asymptomatic Bacteriuria – – – – -14
2.5 Prevalence of Asymptomatic Bacteriuria – – – – -15
2.6 Classification of Urinary Tract Infection – – – – -17
2.6.1 Lower UTIs – – – – – – – – -17
2.6.2 Upper UTIs – – – – – – – – -17
2.6.3 Uncomplicated UTIs – – – – – – – -17
2.6.4 Complicated UTIs – – – – – – – -18
2.7 Outcome of Asymptomatic Bacteriuria – – – – -18
2.8 Treatment of Asymptomatic Bacteriuria – – – – -19
2.9 Commonly Implicated Organisms in Asymptomatic Bacteriuria – -22
2.9.1 Escherichia coli – – – – – – – -22
2.9.2 Staphylococci – – – – – – – – -23
2.9.3 Pseudomonas aeruginosa – – – – – – -25
2.9.4 Proteus – – – – – – – – -26
2.9.5 Klebsiella – – – – – – – – -27
2.9.6 Candida – – – – – – – – -28
2.10 Development of Bacterial Drug Resistance – – – – -29
2.10.1 Incidence of Antimicrobial Drug Resistance – – – -29
2.10.2 Non-Genetic Resistance – – – – – – -31
2.11 Types of Bacterial Drug Resistance – – – – – -33
2.11.1 Chromosomal Mediated – – – – – – -33
2.11.2 Extra chromosomal (Plasmid) Mediated – – – – -33
2.12 Acquisition of Antibiotic Resistance by the Transfer of Genetic Information -34
2.13 Biochemical Mechanisms of Acquired Bacterial Antimicrobial Resistance -36
2.13.1 Alteration of Target Receptor – – – – – – -37
2.13.2 Decreased Concentration of the Drug at the Target Site – – -38
2.13.3 Enzyme Inactivation of the Drug – – – – – -40
2.13.4 Synthesis of Resistant Metabolic Pathways – – – – -42
2.13.5 Failure to Metabolize the Drug – – – – – – -43
2.14 Properties of Antibiotics used in UTI – – – – – -43
2.14.1 Penicillins – – – – – – – – -43
2.14.2 Cephalosporins – – – – – – – -43
2.14.3 Monobactam (Aztreonam) – – – – – – -44
2.14.4 Carbapenems – – – – – – – – -45
2.14.5 β-Lactamase inhibitor (Clavulanic acid) – – – – -46
2.14.6 Sulbactam – – – – – – – – -47
2.14.7 Aminoglycosides – – – – – – – -47
2.14.8 Quinoloncs – – – – – – – – -48
2.14.9 Macrolides – – – – – – – – -49
2.14.10 Sulfonamides and Trimethoprim – – – – – -50
CHAPTER THREE: MATERIALS AND METHODS
3.1 Materials – – – – – – – – -51
3.1.1 Media – – – – – – – – – -51
3.1.2 Antibiotic Disks – – – – – – – -51
3.1.3 Sugars – – – – – – – – – -51
3.1.4 API kits – – – – – – – – -51
3.1.5 Reagents and Solutions – – – – – – -52
3.1.6 Equipments – – – – – – – – -52
3.1.7 Primers – – – – – – – – -53
3.2 Method – – – – – – – – -53
3.2.1 Selection of Subjects for Sampling – – – – – -53
3.2.2 Sample Collection – – – – – – – -54
3.2.3 Media Preparations – – – – – – – -54
3.2.4 Determination of Bacteria Count – – – – – -55
3.2.5 Isolation and Identification Tests – – – – – -55
3.2.6 Identification of isolates to specie level (Rapid test using API kits) – -57
3.2.7 Antibiotic Susceptibility Tests – – – – – – -61
3.2.8 Determination of Multiple Antibiotic Resistance (MAR) Index – -61
3.2.9 β-Lactamase Production Test- – – – – – -61
3.2.10 Extended Spectrum β-Lactamase (ESBLs) Production Test – – -62
3.2.11 Plasmid DNA Analysis of Isolates – – – – – -62
3.2.11.1 Preparation and purification of total DNA using Spin-Column Protocol -62
3.2.11.2 Plasmid Extraction – – – – – – – -63
3.2.11.3 Detection of Plasmid (Gel Electrophoresis) – – – – -64
3.2.11.4 Amplification by Multiplex Polymerase Chain Reaction (PCR) – -64
CHAPTER FOUR: RESULTS
4.1 Incidence of Asymptomatic Bacteriuria – – – – -68
4.2 Distribution of Isolates in the Urine Samples – – – -72
4.3 Antibiotic susceptibility Profiles of Isolates – – – – -76
4. 4 Determination of MAR Index- – – – – – -81
4.5 Result of β-Lactamase Production – – – – – -82
4.6 Result of ESBLs production – – – – – – -83
4.7 Result of DNA extraction – – – – – – -85
4.8 Result of the Multiplex PCR amplification – – – – -86
CHAPTER FIVE: DISCUSSION – – – – – – -89
CHAPTER SIX: SUMMARY, CONCLUSION AND RECOMMENDATIONS
6.1 Summary – – – – – – – – -95
6.2 Multiplex PCR – – – – – – – -96
6.3 Recommendations – – – – – – – -96
Reference – – – – – – – – -98
Appendix – – – – – – – – -129
INTRODUCTION
URINARY TRACT INFECTION (UTI)
Urinary tract infection (UTI) occurs frequently in pregnancy with a large number ofinfections being asymptomatic. The frequency of asymptomatic bacteriuria (ASB) inpregnancy accounts for the rationale of urinalysis throughout pregnancy period(Stamm and Hooton, 1993).
Considered clinically unimportant in non-pregnant women, asymptomatic bacteriuriais the most common complications of pregnancy, occurring in 4-7% of normalpregnancies.
It can be associated with varieties of adverse obstetric outcome andmedical conditions such as the development of acute and chronic pyelonephritis, preterm labour, low birth weight, pre-eclampsia, chronic renal disease, and prenatalmortality (Naeye et al., 1986; Stenqvist et al., 1989; Mittendorf et al., 1992).
Many ofthe complications of asymptomatic bacteriuria in pregnancy can be prevented withearly identification and treatment.
Urine is the product of filtration of blood excreted by the kidneys which generallyconsist of water and water soluble waste products which have been removed from theblood stream.
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