World Journal of Microbiology

Vol. 5(1), pp. 075-081, April, 2017. www.premierpublishers.org, ISSN: 2141-5032x

WJM

Research Article

Asymptomatic urinary tract infection amongst some

Students of Michael Okpara University of agriculture,
Umudike
Nwankwo IU1*, Onwuakor CE2, Ifediora AC3, Azubuike FM4
1*,2,3,4
Department of Microbiology, College of Natural Sciences, Michael Okpara University of Agriculture Umudike, Abia
State, Nigeria.

This work was carried out to determine Asymptomatic Urinary Tract Infection amongst some
students of Michael Okpara University of Agriculture, Umudike and the sensitivity pattern of the
isolates from urine. Using aseptic technique, midstream urine were collected from sixty (60)
students, urinalysis was carried out on the urine samples and was then cultured on CLED and
MacConkey agar using pour plate method. Growth was observed in 26 (87%) of the sample while
there was no growth in 4 (13%) of the sample. Out of the 26 (87%) samples with growth, 14 (47%)
had significant bacteria growth while 12 (40%) had no significant growth. Incidence of
asymptomatic bacteriuria was higher in females 8 (57%) than males 6 (38%). The organisms
isolated were Escherichia coli, Klebsiella species, Staphylococcus saprophyticus,
Staphylococcus aureus, Enterococcus faecalis, Proteus species, and Pseudomonas
aeruginosa. All the Gram positive isolates were sensitive to Gentamycin and all resistance to
Cefuroxime, Ceftazidime, Ceftriaxone, Cloxacillin. The Gram negative isolates were mostly
sensitive to Nitrofurantoin, Gentamycin and Ofloxacin. Therefore, these drugs could be
considered as the first line of drug for the treatment of asymptomatic urinary tract infection.

Keywords: Asymptomatic, urinary tract infection, students, assessment, rate.

INTRODUCTION

Urinary Tract Infection mostly occurs in patients with
anatomically and functionally normal urinary tract and
usually results from spontaneous ascent of bacteria from
the urethra to the bladder. Occasionally, the bacteria
progresses to the kidney and blood stream. However,
since asymptomatic colonization of the urinary tract can
occur, other features such as the presence of
inflammatory markers or follow up cultures are needed to
correctly diagnose a person with urinary tract infection.
Urinary Tract Infection (UTI) represents one of the most
common diseases encountered in medical practice today
and occurring from the Neonate to the geriatric age group
(Kunin, 1994). Despite the widespread availability of
antibiotics, UTI remains the most common bacterial
infection in human body (Tambekar et al, 2006).
Urine is generally considered to be sterile and is believed
to be Germ Free, any source of possible infection occurs
Asymptomatic urinary tract infection amongst some Students of Michael Okpara University of Agriculture, Umudike
through Urethra which initiates the incidence of the
infection. Urinary Tract Infection is caused by the
presence of bacteria in urine, although fungi and viruses
could be involved (Siiri et al, 2009). The predominant
pathogen responsible for Urinary Tract Infection is
Escherichia coli which constitute up to 75 90% and is
followed by Staphylococcus saprophyticus which
accounts for 5 15% of Urinary Tract Infection (Michael et
al, 2007). In addition to the above mentioned bacterial
species, Klebsiella, Proteus, Pseudomonas and
Enterobacter are associated with UTI.
*Correspondence author: Nwankwo I.U., Department of
Microbiology ,College of Natural Sciences, Michael
Okpara University of Agriculture Umudike, Abia State,
Nigeria. Email: immaugo@yahoo.com,
afoma20@yahoo.com
Nwankwo et al 076.
Pathogens like Escherichia coli and Staphylococcus
saprophyticus are associated with population acquired
acute uncomplicated infection where as Klebsiella,
Enterococcus, Proteus species, Enterobacter are known
to confer uncomplicated cystitis and phylonephritis that
are sporadic (Mars, 2002).
Asymptomatic UTI occurs when urinary tract pathogens
enter into the bladder without causing apparent
symptoms. Typically the pathogens are usually
eliminated by host defense factors when they persist only
for a short time in a human host. However, when such
pathogens stay within the urinary system for a long time,
symptomatic urinary tract infection results (Nicolle, 2003).
Asymptomatic bacteriuria is defined by the presence of at
5
least 10 colony forming units (CFU) of organisms per
milliliter in cultures of urine specimens in the absence of
symptoms of infection referable to the Urinary Tract
(Smith, 1994). Asymptomatic bacteriuria occurred reliably
more frequently in females as compared to males and it
is a major criterion of urinary tract infection (Nurullaev
2004). Therefore, it is necessary to investigate the
incidence of Asymptomatic bacteriuria in both apparently
healthy male and female undergraduate students of
Michael Okpara University of Agriculture, Umudike, Abia
State, Nigeria.
MATERIALS AND METHODS
Study Site
Michael Okpara University of Agriculture, which is located
along Ikot-Ekpene road, Umudike, in Ikwuano Local
Government Area, Abia State, Nigeria.
Sample Collection
The participated students were instructed to wash their
external genital with mild toilet soap and rinse thoroughly
with clean water. Early morning mid-stream urine
samples of about 10-15ml were collected using a sterile
universal bottle with air tight screw cap tops. Each urine
sample bottle was labeled with a name, number, sex,
department, and time of collection, etc. Each of the
students was instructed on the mode of collection of the
mid-stream urine and also adequately educated on
precautions to prevent contamination of the specimen.
Inclusion Criteria
All apparently healthy students who indicated the
absence of symptoms from questionnaires issued where
included in the study.
Exclusion Criteria
Students who had taken antibiotics two weeks before or
were currently on antibiotic therapy, or with any
Asymptomatic urinary tract infection amongst some Students of Michael Okpara University of Agriculture, Umudike
symptoms suggestive of UTI or pregnancy as indicated in
the questionnaire were excluded from the study.
PROCESSING OF SAMPLES
Urinalysis
This is used as a pretest to check for Asymptomatic
Urinary Tract Infection or symptomatic urinary tract
infection. A medi-test Combi-9 impregnated with test strip
for urine parameters was dipped into urine samples in a
test tube, the reactions were examined visually
immediately after allowing excess urine to drip off
comparing the colours produced against the colour shade
used for charting and estimating the reactions. The
parameters tested were Nitrite, Proteins H, Ketones,
Glucose, Bilirubin, Urobilinogen, Blood. An Asymptomatic
UTI indicates negative to Nitrite, protein and few pus cells
(leucocytes) while a symptomatic UTI indicates positive
to Nitrite, protein and many pus cells (leucocytes).
Urine Microscopy
This is used as a pretest to check for UTI, a well mixed
urine sample was aseptically transferred into a test tube
and centrifuged at 500-1000g for about 5minutes. The
supernatant fluid was discarded by completely inverting
the tube, the sediment was remixed by tapping the
bottom of the tube, then a drop of the well mixed
sediment was transferred to the glass slide and covered
with a cover slip. It was then examined under the
microscope using x10 objective to focus and x40
objective to view. The leucocytes (pus cell) counts are
usually reported as; Few-up to 10/HPF (High Power
Field) , 11-40/HPF (moderate), and more than 40
WBC/HPF (many). Above 40 indicates Urinary Tract
Infections.
Culturing of Urine Samples
Each of the Urine samples was observed
macroscopically to ascertain if the urine was clear or
cloudy (turbid). Each urine sample was shaken properly
to ensured homogeneity. Ten fold serial dilutions were
made of each specimen and 1ml of the appropriate
dilutions was used as inoculums using the pour plate
method, culturing in cysteine lactose electrolyte
deficient (CLED) agar and Mac Conkey agar. The
o
cultured plates were incubated aerobically at 37 C for
24hours (Cheesbrough, 2000). The plates were
examined macroscopically after 24 hours, the bacterial
colonies were counted and multiplied by 100 to given an
estimate of the number of bacteria present per millimeter
of urine. A significant bacteria count was taken as any
count equal or in excess of 10,000 CFU/ml. Plates with
-5
growth at 10 and above were indicated as significant
-5
bacteriuria while plates that did not show growth at 10

Table 1. Age and gender association with UTI
AGE
Gender
Male Female
16 20 4 5
21-25 7 8
> 26 3 3
Total 14 16
dilution were recorder as negative. Bacterial isolates
were identified based in a combination of cultural,
morphological, biochemical characteristics and Gram
staining.
Antimicrobial Susceptibility Testing
Antimicrobial sensitivity was tested for each isolated
organisms to determine the antibiotic susceptibility
pattern of the different isolates. The agar diffusion disc
techniques described by Bauer et al (1996) was used. A
colony of pure isolate was streaked on sterile Mueller
Hinton agar plates aseptically using sterile inoculating
wire loop. The appropriate multi disc containing minimum
inhibitory concentrations (MIC)of Ciprofloxacin (5g),
Nitrofurantoin (300g), Ceftriaxone (30g), Gentumycin
(10g), Cefuroxime (30g), Ofloxacin (5g), Cefixime
(5g), Ceftazidime (30g), Amoxicillin/Clavulanate
(30g),Erthromycin (5g) and Cloxacillin (5g) was
aseptically placed firmly onto the surface of the dried
plates using a sterile forceps. After 30mins of applying
the disc, the plates were kept at room temperature to
allow diffusion of the different antibiotics from the disc
into the medium. The plates were then inverted and
o
incubated at 37 C for 24hours. The clear zone that
developed around each disc was measured as the zones
of inhibition from underneath each plate with the aid of a
ruler in millimeter (mm). The reaction of the test organism
to each antibiotics was reported, sensitivity and
resistance patterns were recorded in accordance with the
European Committee on antimicrobial susceptibility
testing (EUCAST) Version 6.0 recommendations.
RESULTS
Urine samples from sixty (60) students of Michael Okpara
University of Agriculture, Umudike within the age range of
16 - > 26 years were collected and examined for the
Asymptomatic urinary tract infection amongst some Students of Michael Okpara University of Agriculture, Umudike
World J. Microbiol. 077
Total Percentage (%)
9 30
15 50
6 20
30 100
presence of bacteria using a semi- quantitative method.
The colour of the urine samples ranged from pale yellow
to light yellow, some urine samples were clear and some
turbid in appearance. From the urinalysis leucocytes (pus
cells) were observed in 29 samples, 26 samples were
positive to Nitrite and 13 samples were positive to
protein. Detection of Nitrite, leucocyte and protein is a
rapid method for detecting Asymptomatic Urinary Tract
Infection. The urine samples had the normal pH range of
4.0 8.0.
Table 1 shows the Age and Gender association with UTI.
Out of the 60 urine samples examine, 50% of the positive
cases were observed from those within the age range of
21 25, this was followed by those within the age range
of 16 20 (30%) while the list was from among those
within the age range of > 26 (20%).
The incidence of Bacteriuria among the students is
shown in Table 2. Out of the 30 urine samples cultured
(those that were negative from the preliminary test
urinalysis/microscopy), 14 samples (47%) showed
significant bacteria growth, and 16 samples (53%)
showed no significant bacteria growth.
The characterization and identification test carried out
reveals the bacterial isolates associated with UTI to
belong to the genera Escherichia coli, Klebsiella species,
Staphylococcus saprophyticus, Staphylococcus aureus,
Enterococcus faecalis, Proteus species and
Pseudomonas aeruginosa.
Table 3 shows the frequency of occurrence of the
bacteria isolates from the urine samples with significant
bacteriuria. Escherichia coli had the highest frequency
10(33%) followed by Klebsiella species 5(17%),
Staphylococcus saprophyticus and Staphylococcus
aureus had 4(13%) frequency of occurrence respectively.
The list frequency was observed with Proteus species
and Pseudomonas aeruginosa 2(7%) respectively.
Table 4 (a) and 4(b) depicts the Antibiotic susceptibility
pattern of the isolates. Majority of the Gram positive
bacteria were sensitive to Gentamycin, Erythromycin and
Ofloxacin while some others were resistant to the
Nwankwo et al 078

Table 2. Percentage significant/ non significant bacteriuria among the students

TOTAL No of No of No of
students % students % non students % without
with significant with non significant with no growth
significant bacteriuria significant bacteriuria growth
bacteriuria bacteriuria
Male (16) (16) 6 38 7 44 3 19

Female (14) 8 57 5 36 1 7

Total (30) 14 47 12 40 4 13

Table 3. Frequency of occurrence of bacterial isolates among students with significant bacteriuria

Bacterial isolate No. of samples No. isolated Total number of Percentage

examined isolates occurrence
Male Female
Escherichia coli 14 4 6 10 33

Klebsiella species 14 3 2 5 17

Staphylococcus 14 1 3 4 13
saprophyticus
Staphylococcus 14 4 - 4 13
aureus
Enterococcus 14 2 1 3 10
faecalis
Proteus species 14 - 2 2 7

Pseudomonas 14 2 - 2 7
aeruginosa
Total 14 16 14 30 100

antibiotics. Majority of the Gram negative bacteria were
sensitive to Gentamycin, Ofloxacin and Nitrofurantoin
while some were resistant to the antibiotics.
DISCUSSION
This work was carried out to determine the presence of
Asymptomatic Urinary Tract Infection among sixty (60)
students of Michael Okpara University of Agriculture,
Umudike. 30 urine samples were positive to UTI after a
pretest of urinalysis and urine-microscopy. From the
urinalysis, 27 samples were positive to Nitrite, 12
samples were positive to protein and 29 leucocytes (pus
cells) were observed. Detection of these 3 parameters is
rapid method for detecting Asymptomatic UTI. The report
is in line with (Bent, 2002) that dipstick urinalysis in
practice may provide useful information for the diagnosis
of UTI. The presence of nitrite has predictive value of
Asymptomatic urinary tract infection amongst some Students of Michael Okpara University of Agriculture, Umudike
UTI, since nitrite are formed as a metabolic product from
bactecria that break down nitrate to nitrite e.g. E. coli,
Proteus spp, Klebsiella spp etc. If the dipstick is positive
to either nitrate or leucocyte this increases the probability
of UTI to about 80% while dipstick negative in both nitrate
and leucocyte reduces the probability of UTI to about
20%( Bent,2002).
5
For this study, the significant Asymptomatic UTI is 10
CFU/ml (Cheesbrough, 2000). Asymptomatic bacteriuria
was observed in 47% of the population. The higher
incidence of Asymptomatic UTI rate of female (57%)
compared to that of male (38%) is in agreement with
Prescott,et al (2004) who reported that bacteriuria is
more common in females and is often Asymptomatic with
frequent reoccurrence. Higher cases of Asymptomatic
UTI in female than male has also been observed by other
workers (Frank-Peterside and Oguike, 2006), the higher
incidence in female than male could be due to the shorter
length of the Urethra in females and its closeness to the
 World J. Microbiol. 079

Table 4 (a). Susceptibility pattern of the isolates using gram positive antibiotic disc

Group Isolate No. Antibiotics

Of bacteria Isolated Caz Crx Gen Ctr Ery Cxc Ofl Aug

Staphylococcus 4 4R 4R 4S 4R 4S 4R 4R 1S3R
saprophyticus
Gram
positive
Staphylococcus 4 4R 4R 4S 4R 4S 4R 2S2R 4R
aureus
4 13%
Enterococcus 3 3R 3R 3S 3R 1S2R 3R 3S 2S1R
faecalis
KEY: S = Sensitive
R = Resistance
Caz = Ceftazidime; Crx = Cefuroxime; Gen = Gentamycin; Ctr = Ceftriaxone Ery = Erythromycin ; Cxc = Cloxacillin ; Ofl = Ofloxacin; Aug = Augmentin;

Table 4 (b). Susceptibility pattern of the isolates using gram negative antibiotic disc.

Group Isolate No. Antibiotics

Of bacteria Isolated Caz Crx Gen Cxm Ofl Amx/C Nit Cpr

Escherichia coli 10 10R 9S1R 10S 10R 10S 10R 8S2R 10R
Gram
Negative
Klebsiella species 5 5R 5s 5S 5R 3S2R 5R 5S 5S

Proteus species 2 2R 2R 2R 2R 2S 2R 2S 2S

Pseudomonas 2 2R 2R 2S 2R 1SIR 2R 2S 1SIR

aeruginosa

KEY: S = Sensitive
R = Resistance
Caz = Ceftazidime; Crx = Cefuroxime; Gen = Gentamycin; Cxm = Cefixime; Ofl = Ofloxacin; Amx/C = Amoxicillin/Clavulanate; Nit = Nitrofurantoin; Cpr
= Ciprofloxacin.

external passage that facilitates the entrance of
microorganism into the body. Also the high rate may be
due to the increase in female with poor hygiene practice
and also indiscriminate sexual behavior among the
female students.
Seven (7) bacterial species, Escherichia coli, Klebsiella
species, Staphylococcus saprophyticus, Staphylococcus
aureus, Enterococcus faecalis, Proteus species, and
Pseudomonas aeruginosa were isolated from this study.
These species have also been isolated in similar studies
by different investigators of asymptomatic UTI (Roos et
al, 2006).
The highest frequent occurring organism isolated from
this study was Escherichia coli (33%). The presence of
Escherichia coli as the commonest etiological agent of
UTI has also been reported in previous studies by (De
Francesco et al, 2001). The second most common
organism isolated in this study was Klebsiella species
with (17%), followed by Staphylococcus saprophyticus
and Staphylococcus aureus (13%) respectively. This is
usually found in infection among sexually active young
women and can cause UTI in males of all ages
(Cheesbrough, 2004). The least frequency was observed
with Proteus species and Pseudomonas aeruginosa (7%)

Asymptomatic urinary tract infection amongst some Students of Michael Okpara University of Agriculture, Umudike
Nwankwo et al 080
respectively. Their presence in the urine sample should
however not be overlooked because they have been
reported as agents of urinary tract infection
(Cheesbrough, 2002).
This work has showed that some bacterial has showed
that some bacterial isolates are more susceptible to
treatment with certain antibiotics than others. Majority of
the Gram positive bacteria were sensitive to Gentamycin,
Erythromycin and Ofloxacin while some Gram negative
bacteria were sensitive to Nitrofurantoin, Ofloxacin and
Gentamycin. These drugs could be considered as among
the first line of drugs for treating cases of urinary tract
infection.
The least resistance of the bacterial isolates to
antimicrobial agents was observed to be Caftazidime,
Cefuroxime, Ceftriaxone, Cloxacillin,
Amoxicillin/Clavulanate and Cefixime as seen from this
study. The factors contributing to those resistances may
be due to indiscriminate abuse of antibiotic by students.
Other factors may include poor quality of drugs, poor-
storage and expose drug (Okeke et al, 1999) etc. the
reduction of antibiotic prescription and dispensation have
been associated with reduced antibiotic resistance
(Schiemann et al, 2010).
CONCLUSION
A high incidence of Asymptomatic Urinary Tract Infection
was demonstrated among the females of Michael Okpara
University of Agriculture, Umudike. The most causative
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Asymptomatic urinary tract infection amongst some Students of Michael Okpara University of Agriculture, Umudike
agents that were isolated was the Gram negative
organisms with Escherichia coli being the most frequent
were shown to be sensitive to drugs like Nitro furantoin
and Ofloxacin. The Gram positive isolates e.g.
Staphylococcus aureus were sensitive to Gentamycin.
From this study, it was ascertained that some students
might be healthy showing no signs or symptoms of UTI
but when diagnosed of UTI, they could be having urinary
tract infections. Medical check is needed from time to
time to reduce the risk of UTI among students of the
university. In order to reduce infection from UTI- causing
pathogens, intermediate host of UTI causing parasites
should be eradicated and personal hygiene improved.
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Accepted 14 October, 2016.
Citation: Nwankwo IU, Onwuakor CE, Ifediora AC,
Azubuike FM (2017). Asymptomatic urinary tract infection
amongst some Students of Michael Okpara University of
agriculture, Umudike. World Journal of Microbiology 4(1):
075-081.
Copyright: 2017 Nwankwo et al. This is an open-
access article distributed under the terms of the Creative
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