1. Virulence of uropathogenic Escherichia coli isolates and their potential to be diarrheagenic
Marium Khaleque1, Selina Akter2, Humaira Akhter1, Sirajul Islam Khan1, Anowara Begum1
Affiliation :
1 Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh.
2 Department of Microbiology, Jessore University of Science and Technology, Jessore- 7408,
Bangladesh.
Presented By: Department of Microbiology
Marium Khaleque University of Dhaka

2. Introduction

3. Escherichia coli UTI UPEC Virulence Factors of DEC
Family: Enterobacteriaceae
common inhabitant of the gastrointestinal tract of humans and animals.
Some strains of E. coli can diverge from their commensal cohorts, taking on a more pathogenic nature.
These strains acquire specific virulence factors which confer an increased ability to adapt to new niches and allow them to cause a broad spectrum of disease
The sub-set of genes involved in the subversion of host responses and attacking of host cell machinery makes each pathovar distinct

4. Virulence Factors of DEC
UTI
UPEC
Virulence Factors of DEC
Escherichia coli
E. coli can be classified into following groups:
UPEC (Uropathogenic E. coli) collected from UTI patient
Commensal
Pathogenic
The commensal E. coli is the nonpathogenic and often considered to be the normal inhabitant of the gastrointestinal tract of warm-blooded animals
Diarrheagenic
Extraintestinal
UPEC
NMEC
can cause infection in the gastrointestinal tract
EPEC
EHEC
ETEC
EIEC
EAEC
DAEC
is the group capable of infecting sites other than intestine

5. Virulence Factors of DEC
UTI
UPEC
Virulence Factors of DEC
Escherichia coli
A urinary tract infection (UTI) is an infection in any part of the urinary system including kidneys, ureters, bladder and urethra.
E. coli is responsible for more than 80% of the UTI cases
Causative agents
E. coli
Klebsiella spp
Enterobacter spp
Proteus
Pseudomonas
Staphylococcus saprophyticus

6. Aims and Objectives
Assessment of virulence properties of uropathogenic E. coli (UPEC), isolated from urine samples of UTI patients.
Whether these strains share the virulence properties with diarrheagenic E. coli by monitoring the presence of five different diarrheagenic E. coli virulence traits in these UPEC strains.

7. Virulence Factors of DEC autotransporter toxin
Escherichia coli
UTI
UPEC
Virulence Factors of DEC
E. coli that can enter, colonize urinary tract and cause infection are known as uropathogenic E. coli
Virulence factors of UPEC
Type 1 fimbriae
virulence factors associated with bacterial cell surface
α-haemolysin
secreted and exported to the site of action
P fimbriae
CNF1
S fimbriae
CDT
afimbrial adhesins
autotransporter toxin

8. Virulence Factors of DEC
Escherichia coli
UTI
Virulence Factors of DEC
UPEC
vt2
pCVD
eltB
vt1
estA
EHEC
EPEC
ETEC
EIEC
EAEC
eaeA
bfpA
ial

9. Identification of isolates
Work plan
Microscopy
Identification of isolates
Use of selective and differential media
Biochemical profiling
Preliminary screening
Biochemical screening
Identification using the fluorogen MUG
Biochemical Identification
Observation of UTI - associated virulence properties
Observation of genes associated with diarrheagenic E. coli virotypes by multiplex PCR assay
hemolysis activity
Biofilm formation
UTI associated
adhesin genes by
multiplex PCR assay

10. Result

11. Identification of isolates
Among 71 isolates 56 (78.8%) isolates were identified as E. coli
Biochemical
Screening
Microscopy
selective and differential media
MUG test

12. Observation of UTI - associated virulence properties
Hemolysis activity
Among 56 isolates 5 isolates gave clear zone of β-hemolysis around the colony.
No α-haemolysis was found among the E. coli isolates.
β-hemolysis
γ- hemolysis

13. Observation of UTI - associated virulence properties
Biofilm formation assay
Among 56 isolates 45% isolate showed the capability of biofilm formation
16 gave significant biofilm formation ability and
9 gave moderate level of biofilm formation ability in-vitro.
Naves et al., 2008

14. Observation of UTI - associated virulence properties
Detection of UTI associated adhesin genes by multiplex PCR assay
Type I fimbriae
Among 56 E. coli isolates
27% was fim1 (+)ve
P fimbriae
43% was papC (+)ve
11% was afa (+)ve
No isoletes found (+)ve for sfa adhesin genes
Fig: Agarose gel elctrophoresis showing PCR amplification product of papC (328 bp) gene and afa (750 bp), fim1 (700 bp) gene and No band appeared for sfa adhesin genes

15. Potentiality of UPEC isolates to be diarrheagenic
Detection of diarrheagenic virulence genes by multiplex PCR assay
All the 56 UPEC strains were subjected to multiplex PCR for the detection of
heat stable toxin (estA)
Only 7 isolates was carrying DEC virulence gene
heat labile toxin (eltB)
bundle forming pilli (bfpA),
EA (pCVD) gene
Verotoxin 2 (vt2)
Verotoxin 1 (vt1),
Eae (eaeA)
Invasive associated locus (ial)

16. Detection of diarrheagenic virulence genes by multiplex PCR assay
Potentiality of UPEC isolates to be diarrheagenic
Detection of diarrheagenic virulence genes by multiplex PCR assay
All the 56 UPEC strains were subjected to multiplex PCR for the detection of 2
ETEC
heat stable toxin (estA)
Only 7 isolates was carrying DEC virulence gene
heat labile toxin (eltB) 1
ETEC
EPEC
bundle forming pilli (bfpA), 1
EA (pCVD) gene
EAEC 1
Verotoxin 2 (vt2)
EHEC 1
Verotoxin 1 (vt1),
Eae (eaeA)
EIEC
Invasive associated locus (ial) 1

17. Comparison with other study
Similar study in Brazil, Iran, Denmark, Germany 16
out of 225 UPEC strain
Conclusion
In Bangladesh, isolates of UPEC have been found sharing virulence property with Diarrheagebic E. coli
But these UPEC strains were found to share with different DEC virulence properties ….NOT limited to EAEC
In our study, we have found almost equal
no. of UPEC strains carrying virulence gene of ETEC, EHEC, EPEC, EIEC or EAEC.
Among these ETEC property was found in higher no. of UPEC strains.
Brazil aat seq of EAEC
2007 13
out of 138 UPEC strain
Iran EAEC property
2014
increased adherence to human bladder epithelial cell
EAEC virulence gene in UPEC
In the presence of EAEC gene
OUTBREAK
Copehegan, Denmark
in 1991
10.8% of 265 UPEC strains
Germany EAEC property

18. THANK YOU ALL

19. Patient information
Cultures were collected from 71 patient . Among the patients 47 (66.2%) were female and the rest 24 (33.8%) were male, indicating higher occurrence of UTI in females in a random sample
Fig. Age and sex distribution pattern of 71 UTI patients of the study.