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Le Infezioni in Medicina, n. 2, 98-100, 2001
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articles
Expression of P fimbriae
of uropathogenic
Escherichia coli strains
Espressione delle fimbrie P in ceppi di E. coli uropatogeni
Güner Cos‚ar1, Mine Hos‚gör1, Onur Özgenç2, Süleyha Hilmioǧlu3,
Hüseyin Tas‚li1
1
Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Ege University,
Department of Infectious Diseases and Clinical Microbiology, Social Security Bozyaka
Hospital,
3
Department of Microbiology and Clinical Microbiology, Faculty of Medicine, Ege University, Izmir, Turkey
2
■ INTRODUCTION
with cystitis of the Social Security Bozyaka
Hospital and 24 E. coli strains were isolated
from the urine of children patients with
pyelonephritis at the Ege University Hospital.
A total of 222 E. coli strains were collected in
two years. The E. coli isolates were identified by
standard laboratory identification techniques
and then tested for P fimbriae.
Bacterial adherence is an important initial step in
the pathogenesis of urinary tract infections
(UTIs). In a number of cases, fimbriae have been
shown to mediate this adherence. Fimbriae enable the bacteria to bind to specific receptor
structures and thereby to colonize specific surfaces [1-5]. Fimbriae of uropathogenic Escherichia
coli (UPEC) can be divided into three categories
on the basis of their hemagglutinating activities:
1) Type 1 fimbriae that cause mannose-sensitive
hemagglutination of guinea pig erythrocytes;
2) MR fimbriae that cause mannose-resistant
hemagglutination (MRHA) of human erythrocytes. This group can be subdivided on
the basis of receptor specificities into P, M, S
and X fimbriae;
3) FIC fimbriae that cause no hemagglutination
(HA) of any erythrocytes [6-10]. P fimbriae
differ in serotype and the molecular weight
of the fimbrial subunit [11, 12].
Detection of P fimbriated E. coli: The occurrence of P fimbriae on uropathogenic E. coli
strains was determined by using a P-specific particle agglutination kit (Orion Diagnostica, Espoo,
Finland) [18]. The E. coli strains to be tested were
cultivated on Tryptic Soy Agar (TSA) overnight
at 37 °C. One drop of phosphate buffer solution
(PBS) from the dropper was dispensed on the
test and control areas of the test card. A mixing
stick was used to harvest 2 to 4 bacterial colonies
from TSA, and they were suspended in the PBS
droplets on the test and control areas. The bacterial suspension mixed with the reagents. The
test card were tilted for two minutes and observed for the agglutination reaction. The result
was positively interpreted, if agglutination was
visible by the naked eye in the PF test reagent
within two minutes, but not seen in the control
reagent. A positive control card was also used.
P fimbriae, which are subdivided into serologically distinct groups (F7-F13), recognize the αD-Gal-(1R4)-β-D-Gal region of globosides [11,
13]. Vaccination with purified P fimbriae prevents experimental pyelonephritis in monkeys
[14]. P fimbriae are known to occur in association with a limited number of E. coli serotypes
[15-17]. The aim of this study was to investigate
the prevalence of P fimbriae of E. coli causing
human urinary tract infection.
Electron microscopy: Bacterial suspensions of
positive P-specific particle agglutination were
prepared for electron microscopy after being
washed in sterile distilled water. One drop of
the suspension was placed on a film-coated
coppergrid, and excess fluid was removed with
filter paper strips. The bacteria were negatively
stained for 30 s with 2% (w / v) phosphotungstic acid (pH 7). Grids were kept at ambient temperature for one night and examined
■ MATERIALS AND METHODS
Bacteria: One hundred and ninety eight E. coli
strains were isolated from the urine of patients
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ogenic E. coli strains 34 (97%) were fimbriated in
children with pyelonephritis, and of 26 E. coli
strains five (19%) were fimbriated in patients
with cystitis.
Väisänen-Rhen et al [26], reported that of 35
uropathogenic E. coli strains, 33 (94%) with P
fimbriae occurred in children with clinical
pyelonephritis and out of 26 E. coli strains, five
(19%) occurred with cystitis. In another study,
Väisänen-Rhen et al [16] found that the prevalence of P fimbriae of E. coli strains was 76%
with pyelonephritis and 23% with cystitis.
Latham et al [27] reported that, out of 23
pyelonephritogenic E. coli strains, 13 (57%) were
fimbriated, and of 116 E. coli strains isolated in
patients with cystitis, 22 (19%) were fimbriated.
Siegfried et al [28], suggested that P fimbriae
production was significantly more frequent in
pyelonephritogenic E. coli strains than in lower
urinary tract infection E. coli strains in children.
Lo Bue et al [29], showed that P fimbriated E.
coli was the most prevalent microorganism in
acute urinary tract infections (34%).
According to the findings of the above investigations, P fimbriated E. coli strains are more frequent in patients with clinical pyelonephritis
than with cystitis.
In the present study, of 24 uropathogenic E. coli
strains three (13%) P fimbriae-positive strains
occurred in children with clinical pyelonephritis and of 198 E. coli strains 29 (15%) occurred
with cystitis.
In this study, the prevalence of P fimbriae of
uropathogenic E. coli strains in patients with
cystitis is quite similar to the above findings.
By contrast, in this study the prevalence of P
fimbriae of pyelonephritogenic strains is lower
than the findings reported in the literature.
The results of this study suggest that P fimbriae
occur with a limited number of E. coli strains and
indicate that many cases of pyelonephritis may
be caused by non-P fimbriated E. coli strains.
Figura 1 - Electron micrograph of fimbriated Escherichia coli strain that is P-specific particle agglutination positive x 33,000.
with a Joel 100 transmission electron-microscope. The micrograph was taken of fimbriae
observed on bacteria from preparation.
■ RESULTS
P fimbriae of a total of 222 uropathogenic E. coli
strains were examined. Of the total, 31 (14%)
were found with P fimbriae. Out of 198 E. coli
strains, 29 (15%) were P fimbriated in patients
with cystitis. Of 24 pyelonephritic isolates, three
(13%) had P fimbriae. Prevalence of P-fimbriae
of E. coli strains were found to be quite similar in
patients with cystitis and pyelonephritis. The
fimbriated E. coli strain is shown in figure 1.
■ DISCUSSION
One of the various virulence factors of uropathogenic E. coli strains is adherence to uroepithelial
cells [17, 19-21]. P fimbriae adhere to glycolipids
on mucous membrane and urothelial cells [22,
23]. There is an important role of pyelonephritis
associated pili (PAP) protein adhesins in the development of acute pyelonephritis in children
[24].
Möllby et al [25], showed that out of 35 uropath-
Key words: P fimbriae, uropathogenic Escherichia
coli, urinary tract infection
SUMMARY
curred in children with clinical pyelonephritis,
and of 198 E. coli strains 29 (15%) occurred in
children with cystitis. Prevalence of P fimbriae
of E. coli strains was found to be quite similar
in patients with cystitis and pyelonephritis.
The occurrence of P fimbriae in a total of 222
uropathogenic Escherichia coli (UPEC) strains
was investigated. Out of the total, 31 (14%)
were P fimbriated. Of 24 pyelonephritogenic
E. coli strains, three (13%) with P fimbriae oc-
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RIASSUNTO
minazione delle fimbrie P il 13% (3/24) dei ceppi
pielonefritogeni isolati in bambini affetti da pielonefrite e il 15% (29/198) dei ceppi isolati da bambini con cistite. La prevalenza dei ceppi di E. coli
provvisti di fimbrie P è risultata simile sia nei pazienti con cistite sia in quelli con pielonefrite.
Nel presente studio è stata valutata l’incidenza di
ceppi di Escherichia coli uropatogeni (UPEC)
dotati di fimbrie P. Complessivamente, dei 222
ceppi di Escherichia coli testati, sono risultati
provvisti di fimbrie P 31 ceppi (14%).
In particolare, sono risultati positivi per la deter-
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