MOLECULAR CHARACTERIZATION OF MULTIDRUG RESISTANT BACTERIAL UROPATHOGENS FROM PREGNANT WOMEN IN IBADAN METROPOLIS, SOUTH-WEST NIGERIA BY Mojirayo TitilayoDUROWAIYE (Ph.
MOLECULAR CHARACTERIZATION OF MULTIDRUG RESISTANT BACTERIAL UROPATHOGENS FROM PREGNANT WOMEN IN IBADAN METROPOLIS, SOUTH-WEST NIGERIA BY Mojirayo TitilayoDUROWAIYE (Ph.
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Date
2021-03
Authors
DUROWAIYE, Mojirayo Titilayo
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Abstract
Urinary tract infection (UTI) is one of the commonest infectious diseases of women worldwide,
and pregnant women are more predisposed. This study aimed to determine the prevalence of
bacterial infecting agents in UTI during pregnancy, to establish their antimicrobial susceptibility
profiles, identify the socio-economic factors that promote UTI. Virulence factor genes (VFGs) of
the multi-drug resistant bacterial uropathogens were determined in order to establish their
pathotypes. Clean catch mid-stream urine samples were collected, processed and analyzed from a
total of 650 women on their first visit to antenatal clinics of three major health facilities in
Ibadan metropolis. Demographic information was collected using structured questionnaires.
Dip-stick leucocyte esterase test was performed and urinary isolates were identified using
standard conventional biochemical tests and further confirmed to specie level with Microbact®
identification kits (Oxoid). Modified Kirby-Bauer disc diffusion method was used to determine
the antibiotic susceptibility profile and interpreted as specified by the Clinical Laboratory
Standard Institute (CLSI).Genomic DNA extraction and amplification of specific virulence genes
associated with UTI by polymerase chain reaction (PCR) was carried out. Extended spectrum
beta-lactamase production by the most prevalent multi-drug resistant Gram negative isolate was
determined phenotypically and genotypically using universal primers.Molecular phylogenetic
analysis by maximum likelihood method (MLM) was carried out for some sequenced isolates,
and an evolutionary analysis was conducted using MEGA 7®. Significant bacteriuria was
observed in 246(37.8%) antenatal patients, with 149(60.6%) being asymptomatic. The major
bacterial species recovered included Staphylococcus epidermidis 51(21.9%), Escherichia coli
48(19.5%), Staphylococcus aureus 27(10.9%), Streptococcusspp
17(6.0%), Providencia stuartii 12(4.8%), Proteus mirabilis 13(5.2%),Staphyloccocus
saprophyticus 12(4.9%),Enterobacter agglomerans 8(3.2%). Others including Staphylococcus
sppranged from 0.4% to 2.3%. The highest prevalence of bacteriuria with respect to gestational
age, women‟s age group, occupational status and educational level were 53.2% (n=131) third
trimester, 44.2% (n=101) 25-30 years, 35.3% (n=87) housewives, and women with no formal
education 32.1% (n=79) respectively and were statistically significant at P<0.05. Majority
(69.5%) of the isolates were multidrug resistant. The isolates were most resistant to
Cotrimoxazole (66.6%), and most susceptible to Nitrofurantoin (11.9% resistance). Phenotypic
ESBL production was seen in 35(72.9%) of E. coli isolates; the ESBL genes amplified were bla
CTX-M 10(100%), blaTEM 10(100%), bla OXA 10(100%), and bla SHV 7(70%). Virulence
genes amplified in the analyzed MDR ESBL E. coli isolates were fim H 10(100%), pap
10(100%), cnf 5(50%), upa Bc 10(100%), qep 5(50%), Int 1 10(100%), IS 256 8(80%), amp C
8(80%), aac 1(10%), tsp 10(100%), yjaA 9(90%), but chuA gene was absent. Virulent extraintestinal
phylogenetic group B2 strains of uropathogenic E. coli (UPEC) 7(70.0%) were
identified. In the analyzed MDR S. aureus isolates, using simplex PCR, the following were
amplified: mec A 2(20%), amp C 10(100%), ica D 8(80%), ica A 5(50%), Int 1 10(100%), IS 256
7(70%), upa Bc 10(100%), while pvl, hly, aer, qep and upa Cc genes were absent. The
nucleotide sequence of 16SrRNA gene of all the MDR isolates used in the molecular analysis in
comparison with those from GenBank database revealed 100-99% identity.
Description
A DISSERTATION SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES,
AHMADU BELLO UNIVERSITY, ZARIA, NIGERIA
IN PARTIAL FULFILLMENT OF REQUIREMENTS FOR THE AWARD OF THE
DEGREE OF DOCTOR OF PHILOSOPHY (Ph.D) IN PHARMACEUTICAL
MICROBIOLOGY
DEPARTMENT OF PHARMACEUTICAL MICROBIOLOGY,
FACULTY OF PHARMACEUTICAL SCIENCES,
AHMADU BELLO UNIVERSITY,
ZARIA, NIGERIA
Keywords
MOLECULAR CHARACTERIZATION,, MULTIDRUG RESISTANT BACTERIAL UROPATHOGENS,, PREGNANT WOMEN,, IBADAN METROPOLIS,, SOUTH-WEST NIGERIA