Isolation and Identification of Bacteria from Urinary Tract Infections - Open Journal Systems
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2432 Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3
Isolation and Identification of Bacteria from Urinary Tract
Infections
Noor Hamid Abbas Al- Marzoog1 , Heba Saleh Shaheed2, Alaa Yousif Oudah1
1
Assist. Lecturer, 2Lecturer, Dr. Medical Laboratory Techniques Department / AL-Mustaqbal University College,
51001 Hillah, Babil, Iraq
Abstract
A urinary tract infection (UTI) is an infection in any part of urinary system (kidneys, ureters, bladder and
urethra). Most infections involve the lower urinary tract, the bladder and the urethra. Women are at greater
risk of developing a UTI than are men. In this study, (50) urine sample were collected from (50) patients
with suspected urinary tract infection, during the period from December (2017) to March (2018). The results
of this study revealed that (70%) of samples was positive for bacterial culture, while (30%) of sample have
no growth. The bacterial culture shown that (18) (51.43%) of bacterial isolates was E. coli followed by
Staphylococcus aureus (8) (22.86%) and Klebsiella spp (17.14%) (6) , Proteus spp .(8.57%) (3). In-vitro,
the susceptibility of bacteria towards some antibiotics included in this study revealed that the Ciprofloxacin,
Gentamicin, and Chloramphenicol are the most effective antibiotics to different types of pathogenic bacteria.
Key Words: Urinary Tract Infection, Bacteria, Antibiotics.
Introduction of the isolates8.
Urinary Tract Infection is an infection that affects part Other organisms that can be responsible for UTIs
of the urinary tract. It is classified as the most common include Gram positive cocci, such as Enterococcus
and occurring nosocomial bacterial infection in human faecalis, Staphylococcus aureus and coagulase negative
populations around the world 6 . Urinary tract infection staphylococci (CoNS). Other Gram negative organisms
is classified into three types: acute pyelonephritis, lower responsible for causing UTIs include Klebsiella
UTI and asymptomatic bacteruria 2. UTI is a condition species, Proteus species, Pseudomonas aeruginosa and
caused by pathogenic invasion of the epithelium, Enterobacter species (Baraboutis et al., 2011).
which lines the urinary tract from the minor calyx to
prostatic urethra. The proliferation of bacteria in the Material and Methods
urothelial can be asymptomatic or symptomatic, which Patient
causes inflammatory response and symptomatic case
characterized by a wide range of symptoms including, This study includes 50 patients with suspected
fever, lethargy, anorexia and vomiting7. urinary tract infection during the period from December
(2017) to March (2018), urine sample was taken. The
However, both genders are susceptible to this type clinical diagnosis of urinary tract infection was carried
of infection, but women are more, as their reproductive out with the help of physician.
anatomy and physiology are more sensitive. Half of
all women by 32 years age had experienced at least an Preparation of Culture Media:
infection history 10.
A group of general culture media were prepared
Many different microorganisms can cause UTIs according to the instructions of the company and
though the most common pathogens causing the simple sterilized by autoclaving at121oC for 15 minutes. Such
ones in the community are Escherichia coli and other as Blood Agar Medium, MacConkey Agar Medium,
Enterobacteriacae, which accounts approximately 75% Nutrient agar medium, Muller-Hinton agar medium,Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3 2433
Simmons’ citrate medium, MR –VP Medium and Antibiotic Susceptibility Testing:
Peptone water medium.
Disk Diffusion Test
Identification of isolated bacteria
It was performed by using a pure culture of previously
Identification of isolated bacteria was carried out identified bacterial isolate. The most effective antibiotic for
by using culture methods and biochemical tests. The each bacterial isolate was determined as recommended by
samples was first culture on Blood agar and MacConkey CLSI (2016) .
agar then incubated for (24 h. at 37 C) after that the
A -The inoculums to be used in this test was prepared
bacterial growth was diagnosed by using specific culture
by adding 5 isolated colonies grown on blood agar plate
and biochemical tests.
to 5 ml of nutrient broth and incubated at 35oC for 18
Microscopic Examination and Colonial hours and compared with (0.5) MacFarland standard
Morphology: tube.
A single colony was taken from each primary B - A sterile swab was used to obtain an inoculum
positive culture and its identification depended on the from the bacterial suspension. These inoculums were
morphological properties (colony size, shape, color and streaked on a Mueller-Hinton agar plate and left to dry.
nature of pigments, translucency, edge, elevation and
C - The antibiotic discs were placed on the surface
texture). Bacterial smear stained with Gram stain was
of the medium at evenly spaced intervals with flamed
used to check the cellular morphological properties
forceps or a disc applicator and incubated for 18 hours.
of bacterial cells, including Gram reaction, shape,
arrangement, capsules, spores, etc. D -Inhibition zones were measured using a ruler and
compared with the zones of inhibition determined by
CLSI (2016).
Results and Discussion
Bacterial growth of urine samples
In this study, fifteen urine sample of suspected patients with UTI according to diagnosis made by physician were
collected, 35(70%) of specimens was confirmed bacterial infection with positive urine culture, while 15(30%) of
specimen was confirmed bacterial infection with negative urine culture.
Table (1) Bacterial growth of urine samples
Isolates
Result of urine culture
No. %
Urine sample with positive bacterial culture 35 70%
Urine sample with negative bacterial culture 15 30%
Total 50 100%
In the present study 35 isolates of Gram negative bacteria, while 27(77.14%) of specimens were Gram
and Gram positive bacteria detected in urine specimens negative. As summarized in Table (2).
from study groups, out of 35(70%) specimens positive
bacterial culture, 8(22.86%) were Gram positive2434 Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3
Table (2) Isolated bacteria from urinary tract infections
Isolated bacteria Number Percent %
1 E. coli 18 51.43
2 Klebsiella spp. 6 17.14 77.14
3 Proteus spp. 3 8.57
4 Staphylococcus aureus 8 22.86
Table 3. The isolated bacteria diagnosed according to biochemical tests.
Biochemical test
Isolated bacteria
Indole Methyl red Voges-Proskauer Citrate Catalase
1 E. coli + + _ _
2 Klebsiella spp _ _ + +
3 Proteus spp. + + _ _
4 Staphylococcus aureus +
In this study the highest prevalence of specific The isolation rate of bacteria in this study is
bacteria was found to be E. coli recorded among all the comparable to rates were reported in previous studies
UTI patients to be (51.43%) followed by Staphylococcus done by Getenet and Wondewosen. (2011) 4 who found
aureus with percent (22.86%), Klebsiella spp. (17.14%), that (50 %) of the isolates were E. coli. Also Ouno et
Proteus spp. (8.57%). al. (2013) reported that E.coli was found to be the most
predominant uropathogen isolated from the patients
Uropathogenic E. coli from the gut is the cause of
with UTI followed by P. aeruginosa, K. pneumoniae, S.
80–85% of community-acquired urinary tract infections
1
aureus, and P. mirabilis.
.
Effect of antibiotics on bacterial growth, which
The results of this study are in agreement with
isolated from urinary tract infection:
Shahab et al. (2017) who find that of positive urine
culture, (22.5%) were Gram positive bacteria, while Some antibiotics were used to show the effect on
(77.5%) specimens were Gram negative bacteria, and different types of bacteria isolated from patient with
Getenet and Wondewosen. (2011) 4 was in agreement urinary tract infection which include E. coli, Klebsiella,
with this study who showed that Gram positive and Proteus and Staphylococcus aureus. It has been found
Gram negative bacterial proportion was (19.1%) and that there is a clear variation in resistance, and all isolates
(80.9%), respectively. Also Karzan et al. (2017) 5 also shown resistance to most of these antibiotics.
find that Gram negative bacteria were the most isolates
than Gram positive bacteria.Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3 2435
Table (4) bacterial susceptibility to antibiotics
Antibiotics
Bacterial isolates
CN CIP C AM K AMX
E. coli S S S S R S
Klebsiella S S S R S R
Proteus S S S R S R
Staphylococcus aureus S S S S S S
CN: Gentamicin, CIP: Ciprofloxacin, , AM: Ampicillin, C: Chloramphenicol, K: Kanamycin, AC: Amoxicillin
The isolated uropathogens were sensitive to the 2. Buonsenso, D, Cataldi, L. Urinary tract infections
various antibiotics. Among the isolated pathogens all in children: a review. Minerva Pediatr 2012;
of them shows 100% sensitivity towards Ciprofloxacin, 64:145-57.
Gentamicin, and Chloramphenicol. While bacteria show 3. Clinical and Laboratory Standards Institute
different resistance rate to Kanamycin, Ampicillin, and (CLSI). Performance standards for Antimicrobial
Amoxicillin show. Staphylococcus aureus was sensitive susceptibility testing. Approveds tandard
to all the antibiotics. M100-S20. 2016; 32 (3).
4. Getenet, B, Wondewosen, T. Bacterial Uropathgens
The results of this study was in agreement with in Urinary Tract Infection and Antibiotic
study done by Ouno et al., (2013). Susceptibility Pattern in Jimma Univversity
Specialized Hospital, Southwest Ethiopia. Ethiop J
Recently there is a wide spread distribution of UTI all
Health Sci. 2011; 21: 141-146.
over the world due to multi-drug resistant uropathogens
5. Karzan, MK, Faeza, BO, Shahida N. Isolation and
caused by Resistance plasmid. The effects of urinary
Identification of Urinary Tract Infectious Bacteria
tract infectious pathogens on urinary tract organs are:
and Exploring their Anti-drug Potential against
cystitis, urethritis and pyelonephritis.
Some Common Antibiotics. J Microb Biochem
Financial Disclosure: There is no financial Technol. 2017; 9(6): 285-289 – 285.
disclosure. 6. Najar, M, Saldanha, C, Banday, KA. Approach to
urinary tract infections. Indian J Nephrol. 2009; 19:
Conflict of Interest: None to declare. 129-139.
Ethical Clearance: “All experimental protocols 7. Onuo, GA, Korir, SC, Cheruiyot, JC. Isolation,
were approved under the Department of Medical identification and characterization of urinary
tract infections bacteria and the effect of different
Laboratories Techniques were carried out in accordance
antibiotics. J Nat Sci Res. 2013; 3: 150-159.
with approved guidelines”.
8. Sefton, AM. The impact of resistance on the
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