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Product Code: 00007159

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Urinary tract infection (UTI) constitutes a major health problem in pregnant women. It is more common in this group due to the anatomical and physiological changes that occur during pregnancy. This study was undertaken to characterize antibiotic resistant Escherichia coli isolated from pregnant women in Umuahia. A total of 250 mid stream urine samples from pregnant women in different trimesters were analyzed and cultured on cystein lactose electrolyte deficient (CLED) agar and MacConkey agar. Isolation and identification of Escherichia coli was done by Gram staining and biochemical analysis. Antibiotic susceptibility test was done by Kirby-Bauer disc diffusion method. The prevalence rate of UTI in the study population was observed to be 32%. The prevalence of   Escherichia coli among positive isolates was (59) 73.7%. Majority (117) of those with Escherichia coli present in their urine presented at the second trimester (52.5%) while 23 (39.9%) presented at the third. Only 17 (6.8%) of the studied subjects were in their first trimester and 5 (8.4%) of those had Escherichia coli present in their urine. Among all those that had   Escherichia coli, 64.4% of them were aged 25-34 years. However, there was no significant relationship (P>0.05) between age or trimester and presence of Escherichia coli, which signify that bacteriuria is independent on age and trimester in the study population. The relationship between those that had previous case of UTI and presently had Escherichia coli was found to be statistically significant at (P<0.05). Escherichia coli isolated from the pregnant women showed 100% resistance to amoxicillin and tetracycline. Imipenem and gentamicin should be the antibiotic of choice for treating UTI in the study area because they showed sensitivity of over 70%. Thirty, 30 (50.8%) of the Escherichia coli isolates showed potential to produce biofilm. Extended spectrum beta-lactamase (ESBL) production among Escherichia coli isolates was found to be 12 (20.3%) while 2 (3.4%) isolates produced chromosomal ambler C (iAmpC). Out of 10 highly resistant Escherichia coli isolates, 4 had plasmid. Screening of pregnant women for possible UTI at early stage of pregnancy should be considered an essential care in the study environment to avoid complications in pregnancy.


Title Page                                                                                                        i

Declaration                                                                                                      ii

Dedication                                                                                                      iii

Certification                                                                                                    iv

Acknowledgement                                                                                          v

Table of Content                                                                                             vi

List of Tables                                                                                                  viii

List of Plate                                                                                                    ix

Abstract                                                                                                          x


CHAPTER 1: INTRODUCTION                                                              1

1.1        Background of Study                                                                         1

1.2        Problem Statement                                                                              3

1.3        Significance of Study                                                                         4

1.4        Justification                                                                                         4

1.5        Scope of Study                                                                                   4

1.6        Aim and Objectives                                                                            5

1.6.1    Aim                                                                                                     5

1.6.2    Specific objectives                                                                              5

CHAPTER 2: LITERATURE REVIEW                                                  6

2.1       Escherichia coli                                                                                   6

2.2       Urinary Tract Infections                                                                     7

2.2.1    Causes and risk factors associated with urinary tract infections        8

2.2.2    Diagnosis of urinary tract infections                                                   9

2.2.3        Specimen collection and processing                                                   9

2.2.4    Epidemiology  of urinary tract infections                                           10

2.2.5    Pathogenesis of uropathogenic Escherichia coli                                 10

2.2.6    Symptoms of urinary tract infections                                                 12

2.3       Urinary Tract Infection and Pregnancy                                              12

2.4       Vaginal Ecology and UTI                                                                   13

2.5       Bacteriuria                                                                                           14

2.6       Antibiotic Resistance                                                                          15

2.6.1    Causes of antibiotic resistance                                                            16

2.6.2    Prevention of antibiotic resistance                                                      17

2.6.3    Mechanisms of antibiotic resistance                                                   18


CHAPTER 3: MATERIALS AND METHODS                                       20

3.1       Collection of Specimen                                                                       20

3.2       Microscopy                                                                                         20

3.3       Preparation of Culture Media                                                             20

3.4       Isolation, Characterization and Identification of Isolates                  21

3.4.1    Gram staining                                                                                      21

 3.4.2   Biochemical tests                                                                                22 Indole test                                                                                           22 Oxidase test                                                                                        22 Kligler iron agar reaction                                                                    22 Methyl red-Voges Proskauer test                                                       23

3.5       Antibiotic Susceptibility Testing                                                        23

3.6       ESBL Producers Screening and Confirmation                                   24

3.7       Biofilm Production                                                                             24

3.8       Detection of AMPC                                                                           24

3.9       Plasmid Analysis                                                                                 25

3.10     Statistical Analysis                                                                              26

CHAPTER 4: RESULTS AND DISCUSSION                                        27

4.1       Results                                                                                                27

4.2       Discussion                                                                                           40


5.1       Conclusion                                                                                          44

5.2       Recommendation                                                                                44

References                                                                                          46

Appendix                                                                                            55




4.1:      Frequency of occurrence of bacterial isolates                                                 28

4.2:      Distribution and frequency of Escherichia coli isolates per trimester of       

            pregnancy                                                                                                        29

4.3:      Age Distribution and frequency of Escherichia coli isolated from study      31

4:         Antibiotic susceptibility pattern of Escherichia coli isolates                          32

4.5:      Biofilm, extended spectrum Beta-lactamase (ESBL) and chromosomal      

            ambler C (iAmpC) production among isolates                                               33

4.6:      Presence of plasmid among Escherichia coli isolates                         34

4.7:      Distribution of cured plasmids                                                                       37

4.8:      Frequency of previous case of UTI and its relationship with presence of     

            Escherichia coli in urine                                                                                  38

4.9:      Cumulative responses of 250 women to different signs and symptoms of

            UTI and it’s relationship with Escherichia coli                                               39












4.1:      Plasmid analysis of E. coli isolates showing presence of plasmid      36









Pregnancy causes numerous changes in a woman’s body. These changes along with an already short urethra which allow bacteria quick access to the bladder and difficulty with hygiene due to a distended pregnant belly increase the frequency of urinary tract infection (UTI) in pregnant women. Indeed, UTI is among the most common bacterial infections that complicate pregnancy (Johnson and Kim, 2015).

UTI refer to the presence of microbial pathogens within the urinary tract and it is usually classified by the infection site: bladder (cystitis), kidney (pyelonephritis) or urethra (bacteriuria) (Gonzalez and Schaeffer, 1999). UTI has been defined as the commonest bacterial infection (Demilie et al., 2012; Parveen et al., 2011). It is regarded as the most common hospital acquired infection (Koffour et al., 2012; Kolawale et al., 2009). Human urine can support bacterial growth due to its favourable chemical composition (Asscher and Sussman, 1969). According to Awaness et al., (2000), UTI affects all age group but are relatively common in females as compared to males due to the following reasons; pregnancy, urethra in women is shorter, women tend not to empty their bladder as completely as men, bacteria enter the bladder during intercourse. Also in women, the lower third of the urethra is continually contaminated with pathogens from the vagina and the rectum and there is absence of prostatic secretion which is believed to contain bactericidal substance.

Bacteriuria during pregnancy may be classified as asymptomatic bacteriuria, infections of the lower urinary tract (cystitis), or infections of the upper urinary tract (pyelonephritis). Lower tract bacteriuria is associated with an increased risk of developing pyelonephritis in pregnancy, which is itself associated with adverse maternal and fetal outcomes (Glaser and Schaeffer, 2015). UTI is an independent risk factor for renal carcinoma and bladder cancer. It is also a risk factor for premature delivery, foetal loss and kidney infections (Linhares et al., 2013). In pregnancy, complications such as hypertensive disease of pregnancy, anemia, chronic renal failure, premature delivery and foetal mortality may occur as a result of UTI (Delzell and Lefevre, 2000; Foxman, 2002). Some symptoms of UTIs include; pain or burning sensation during urination, the need to urinate more often than usual, blood or pus in urine, cramps or pain in the lower abdomen, strong smelling urine, pain during sexual intercourse, nausea, vomiting and malaise (Lane and Takhar, 2011). Gilstrap and Ramin (2001) recommend that all pregnant women be screened for the presence of bacteriuria at their first prenatal visit. This is because failure to treat bacteriuria during pregnancy may result in as many as 25% of women experiencing acute pyelonephritis, which has been implicated in preterm labor, transient renal failure, acute respiratory distress syndrome (ARDS), sepsis and shock, and hematologic abnormalities.

Ordinarily, UTIs are caused by a variety of Gram-negative and Gram-positive bacteria. The Gram-positive bacteria are Staphylococcus spp, Streptococcus spp and Enterococcus spp. The Gram-negative bacteria are Escherichia spp, Klebsiella spp, Enterobacter spp, Citrobacter spp, Proteus spp, Serratia spp, Salmonella spp and Pseudomonas spp (Ouno et al., 2013). Among these, 80-90% of UTI is caused by Escherichia coli (E. coli).

The World Health Organization (WHO) defines antimicrobial resistance as a microorganism’s resistance to an antimicrobial drug that was once able to treat an infection caused by that microorganism (WHO, 2014a). As a result, standard treatments become ineffective, infections persist and may spread.  Resistance is a property of the microbe, not a person or other organism infected by a microbe. Certain features such as production of extended-spectrum beta-lactamases (ESBLs), presence of plasmid, generation of biofilm, can confer resistance to an organism. ESBLs are beta-lactamases equipped for giving bacterial protection from the penicillin, cephalosporins (first, second and third ages) and aztreonam, by hydrolysis of these anti-infection agents. These anti-toxins are hindered by lactamase inhibitors, for example, clavulanic acid. As of late, there has been worry that a few strains of E. coli can create ESBL. These enzymes are significant because when they are produced by bacteria, they can make the bacteria resistant to certain commonly used antibiotic medicines. This implies that organisms that produce them can keep on duplicating, causing increasingly serious contamination and ending up progressively hard to treat.

All bacteriuria in pregnancy need to be dealt with and antimicrobial decision in pregnancy need to reflect wellbeing for both the mother and the foetus (Glaser and Schaeffer, 2015). Pregnant women with urinary tract contaminations ought to be followed up firmly after treatment on the grounds that up to one third will encounter a repeat.


UTI in pregnancy contributes significantly to maternal and perinatal morbidity and mortality. This, coupled with a steady rise in drug resistant pathogens attributed to the excessive or inappropriate use of antimicrobial therapy and the indiscriminate use of broad-spectrum antibiotics, indicates the need for a more proactive approach in the use of antibiotics especially for pregnant women. Hence, an attempt to characterize E. coli a major cause of UTI.


This study sets out to analyse and characterize antibiotic resistant pathogens from pregnant women in Umuahia, with interest in E. coli known to be the most common cause of UTI. It’s findings will provide insight into the drug susceptibility pattern of E. coli associated with UTI in the study area. As the safety of mother and child during pregnancy continues to be an issue of great concern to all across the globe, it is expected that the findings of this research will guide physicians and healthcare providers who attend to these women in drug prescription in the case of UTI even before culture and sensitivity is carried out.


Bacterial infection during pregnancy remains a relevant complication to healthcare even in developed countries. Due to widespread multi-resistance to antibiotics coupled with the physiologic changes associated with pregnancy, these women cannot use any type of drug even after culture and sensitivity. As a result of this, treatment options are becoming very narrow. Hence to ensure appropriate therapy, current knowledge of E. coli which is one of the most common organisms involved in UTI in pregnancy and the antibiotic susceptibility pattern is very important.

1.5                      SCOPE OF STUDY

This study will characterize antibiotic resistant E. coli isolates from pregnant women in Umuahia. Urine specimen of these women will be analysed for significant bacterial growth.



1.6.1   Aim

The aim of this research is to characterize antibiotic resistant E. coli isolated from urine samples of pregnant women in Umuahia and also to establish baseline data from antibiotic susceptibility tests that could guide prescriptions in our health facilities.

1.6.2        Specific objectives

1.      To determine prevalence of Escherichia coli isolates causing UTI in pregnant women in Umuahia.

2.      To determine the resistance pattern of Escherichia coli from UTI in pregnant women to various antimicrobials.

3.      To determine the potential to produce biofilm in the isolates.

4.      To determine the presence of ESBL production in the isolates.

5.      To carry out plasmid analysis on highly resistant isolates.



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