Antimicrobial Resistant Escherichia Coli Genes in Children Aged Below Five Years Presenting with Diarrhoea at Thika Level 5 Hospital, Kiambu County, Kenya
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Date
2015-10
Authors
Kang'ethe, Kamwati Stanley
Journal Title
Journal ISSN
Volume Title
Publisher
Kenyatta University
Abstract
Diarrhoea is one of the leading causes of illness and death among children in
developing countries, where an estimated 1.3 billion episodes and 4 to 10 million
deaths occur each year in children less than 5 years of age. The study envisioned to
determine the genetic basis of extended spectrum β-lactam (ESBL) resistance in E.
coli isolated from children aged below 5 years attending Thika Level 5 Hospital
Kiambu County in Kenya. Faecal samples were obtained from 384 children
presenting with diarrhoea. A questionnaire was used to collect data concerning the
age of the child, gender and additional information recorded included on history of
hospitalization and antibiotic use. Samples were sub-cultured on MacConkey agar
and suspect E. coli isolates identified. Antimicrobial susceptibility profiles were
determined for the most common antimicrobials against Gram-negative bacteria.
The types and prevalence of Extended-Spectrum βeta-Lactamases were also
determined. Polymerase Chain Reaction methods were used for detection of the
Extended-Spectrum βeta-Lactamases genes and carriage of integrons. Conjugation
experiments were performed to determine the potential of horizontal transfer of
resistance genes. The plasmid content for defining diversity of plasmid replicon
types was also determined among donors and transconjugants. Approximately
25% of all isolates were resistant to ampicillin, tetracycline, chloramphenicol,
streptomycin, amikacin sulphamethoxazole-trimethoprim and amoxicillinclavulanic
acid (p=0.0051; OR=2.27; 95% CI: 2.01-2.33). Isolates (50%) had a
Minimum Inhibitory Concentration mode50 value within the resistance range
except for ceftazidime, Gentamicin, chloramphenicol, Nalidixic acid, Ciproflocin
and Kanamycin. Factors identified to predispose children to carry highly resistant
isolates included recent history of hospitalization and antimicrobial use and having
acute diarrhoea (p=0.0026; OR=4.21; 95% CI: 3.76-4.98 ) There was no
significant difference in antimicrobial resistance patterns for isolates from rural
and urban children (p=0.049). Carriage of a combination of blaCTX-M+ blaTEM or a
combination of blaCTX-M+blaOXA genes was highly associated with resistance to
multiple β-lactams (p=0.00015; OR=2.31; 95% CI: 1.99-2.65). Isolates with
multiple β-lactamases were also likely to carry integrons encoding resistance to
other antimicrobials. Plasmids that mediated resistance to most antimicrobials
belonged to incL/M, HI2 and F-type. In conclusion, most of isolates recovered
from these children were resistant to first line treatment for diarrhoea and therefore
a need to revise the existing empiric treatment of E. coli infections. Resistance was
transferrable via conjugation and integrons were significantly implicated in
resistance dispersion hence increase in the prevalence of multidrug resistant
isolates (p=0.0017; OR: 4.03; 95% CI: 3.81- 4.04). There is a need to encourage
parents and doctors to follow the treatment guidelines for diarrhoea in which case,
the oral re-hydration therapy zinc and vitamin C rather than antibiotics should be
administered in cases of watery bacillary diarrhoea, implement proper
antimicrobial use polices and launch national surveillance programs to monitor
emergence and spread of MDR isolates.
Description
A Thesis Submitted in Fulfilment for the Award of the Degree of Doctor of Philosophy (Medical Microbiology) in the School of Medicine of Kenyatta University, October, 2015