Journal of Applied Pharmaceutical Science 02 (04); 2012: 90-98

ISSN: 2231-3354
Received on: 12-04-2012
Susceptibility of Bacteria Infecting Urinary Tract to
Revised on: 19-04-2012
Accepted on: 26-04-2012
Some Antibiotics and Essential Oils
DOI: 10.7324/JAPS.2012.2422

Mohamed T. Shaaban, Hanan A. Ghozlan and Marwa M. El Maghraby

ABSTRACT

This study investigates the most common urinary tract infection bacteria and their
sensitivity to antibiotics and some essential oils from Egyptian plants. The Urinary tract bacteria
Mohamed T. Shaaban, were sampled from patients expressing any symptoms of urinary tract infection except those
Marwa M. El Maghraby taking antibiotics. The bacteria were isolated, cultured, and identified. 64 bacterial isolates were
Botany department, identified as E.coli (28 isolates), Klebsiella pneumonia (9 isolates) Pseudomonas aeruginosa (6
Faculty of Science,
isolates), Proteus mirabilis (6 isolates), Staph.aureus (5 isolates) , Enterococcus faecalis (4
Menofiya University, Egypt.
isolates), Morganella morganii ( 4 isolates ) and Pseudomonas fluorescens (2 isolates). The
isolates showed different degrees of sensitivity to different antibiotics . Among the essential oils
of five medical plants known for their application in folk medicine in Egypt , oil of Dill
(Anethum graveolens ) showed the highest effect , affecting more than 50 % of both gram +ve
Hanan A. Ghozlan
Botany& Microbiology Department, and gram –ve bacteria ,followed by Parsley (Petroselinum hortense) and Celery (Apium
Faculty of Science, Alexandria graveolens) affecting 48% & 41% of the isolates respectively . The oil of Thyme (Thymus
University, Egypt. valgare) was effective against Gram –ve bacteria only. The lowest effects were recorded to the
oil of Chamomile (Marticaria recutita) affecting only 5% of the tested isolates.

Keywords: urinary tract infection, antibiotics, bacteria, essential oils.

INTRODUCTION

The urinary tract is the body's filtering system for removing waste liquid, or urine; it
comprises the kidneys, ureters, bladder and urethra (Ramadan, 2003). A urinary tract infection is
caused by bacteria that enter the urinary tract; women are more likely than men to get UTI because
of their urinary tract's design, men have a larger urethra, so it is more difficult for bacteria to enter
the urinary tract. Nearly half of all women will have a UTI at some point in their lives (Marild et
al., 1998; Craig, 2001; Foxman, 2003; Parlak et al., 2007). Urinary tract infection are categorized
into either lower tract infection, located in the bladder and/or urethra (cystitis and urethritis) and
upper tract infection, located in ureters, collecting system, and parenchyma (pyelonephritis). It is
For Correspondence necessary to understand the difference between both types to make an accurate diagnosis. Cystitis
Mohamed T. Shaaban,
Email: is defined as an inflammatory condition of the urinary bladder, whereas pyelonephritis is defined
[email protected] as a diffuse pyogenic infection of the pelvis and parenchyma of kidney.
 Journal of Applied Pharmaceutical Science 02 (04); 2012: 90-98

Signs and symptoms of cystitis include dysuria, performed, The presence of more than 5 WBC/HPF indicated
frequency, urgency, malodorous urine, enuresis, hematuria and pyuria (Hindler & Munro, 2007).
suprapubic pain. On the other hand, the signs and symptoms of
pyelonephritis include; fever over 38.5C, chills along with cost Microbiological investigation
vertebral angle or flank pain and tenderness with pyuria and All samples positive for one or both leukocyte and nitrite
positive urine culture (Dulczak & Kirk, 2005). Most of UTI are and positive for one or both WBCs, and bacteria were inoculated
caused by gram-negative bacteria like Escherichia coli, Proteus on chromogenic media and incubated at 37°C for 24 hours. After
mirabilis, Proteus vulgaris Klebsiella sp, Pseudomonas the incubation period, the plates were examined for growing
aeruginosa, Acinetobacter, Serrati, and Morganella morganii. bacterial colonies. The isolated colonies were purified and sub
Also UTI are caused by Gram positive bacteria include cultured for characterization. Streaking technique was used; an
Enterococcus, Staphylococcus especially coagulase-negative isolated colony can then be used as a source of inoculums for a
staphylococci, and Streptococcus agalacticae (Tangho & pure culture. The agar media were used are, MacConkey's (Oxid
Mcaninch, 2004). At least 80% of the uncomplicated cystitis and ®) Agar selective for: gram-negative bacteria, the growth of gram-
pyelonephritis are due to E.coli. Whereas Proteus mirabilis and positive bacteria was inhibited by the crystal violet dye and bile
Klebsiella pneumoniae infection accounts 10% and 6% salts in the media (Schlager, 2001).
respectively. Adherence properties of some organisms prevent the Hemolysis with Blood Agar this medium contains 5%
normal washout of these organisms by bladder emptying and sheep's blood differential for hemolysis, based on the ability to
mucosal host defense mechanisms (Ashkenazi et al., 1991; Tangho break down hemoglobin or red blood cells, 3 groups of
& Mcaninch, 2004). Treatment of UTI with the appropriate microorganisms can be described alpha-hemolysis a green to light-
antibiotic can minimize mortality, morbidity and any renal damage brown halo is seen around the colonies, Beta-hemolysis a clearing
from acute UTI. Choosing the appropriate antimicrobial agents is seen around the colonies, and Gamma-hemolysis no hemolysis is
sounds difficult, but advances in the understanding of the observed (Dulczak, & Kirk, 2005).
pathogenesis of UTI, the development of new diagnostic tests, and
the introduction of new antimicrobial agents have allowed
physicians to appropriately tailor specific treatment for each patient Isolates maintenance
(Schlager, 2001). Down the ages essential oils and other extracts of Isolates were maintained on nutrient agar plates and slants
plants have evoked interest as source of natural products. They (Magdigan & Martinko, 2005). Subculture were made Bimonthly
have been screened for their potential uses as alternative remedies transfer, and kept in the refrigerator for further investigation
for the treatment of many infectious diseases (Tepe et al.,
2004).The present work aims at the isolation and identification of Characterization of bacterial isolates
bacteria infecting urinary tract and testing the susceptibility of the Colony characteristics the isolates colonies were
isolates to some antibiotics and essential oils from some plants of examined for appearance, smell, and pigmentation (Cowan &
medicinal uses in Egypt. Steel, 1965; Black, 1996).

MATERIALS AND METHODS Gram stain and morphology characters

Gram staining is an empirical method of differentiating
Sample collection
bacterial species into two large groups (Gram-positive and Gram-
A total of 100 clinical samples were collected equally
negative) based on the chemical, primarily the presence of high
from Al-Seguiny Hospital, Alexandria and El-Miry Hospital, Kafr
levels of peptidoglycan, and physical properties of their cell walls.
Eldwar (50% females and 50% males). Samples were obtained
A Gram positive results in a purple/blue color while a Gram
from patients in different ages, expressing any symptoms of
negative results in a pink/red color. Smears of isolated bacterial
urinary tract infections excluding those treated with antibiotics
were stained with Gram technique and examined microscopically
prior to sampling. Patients were sampled by clean catch midstream
for gram stain response, cell shape, and aggregation (Bergey et al.,
urine. All samples were coded and processed at the time of
1994).
collection according to the standard method. Samples were then
transferred the lab of microbiology for further investigation using a
portable cooler to avoid any bacterial multiplication (Robert et al., Biochemical characterization test
1991). The bacterial isolates were tested for their biochemical
characteristics using Oxidase test; to detect the presence of the
Urinalysis, Microscopy intracellular cytochrome oxidase enzyme which allows the
Urinalysis was performed on all collected specimens organism to use oxygen as part of respiration. Organisms that are
using. Dipstick screening technique for leukocyte esterase and oxidase positive called oxidative fermentative, will cause the
nitrite using multisticks of Medi-Test Combi 10 ® SGL, a routine reagent to turn purple within 10-20 seconds. 24 h colonies were
direct microscopy of a centrifuged sample was performed for the used for this test. This test was performed as follows: a drop of the
examination of white blood cells and bacteria examination was oxidase reagent (tetramethyl-p-phenylene diamine hydrochloride)
 Journal of Applied Pharmaceutical Science 02 (04); 2012: 90-98

was placed on a disk of filter paper. A small amount of the sterilized filter paper saturated with reach natural oil separately.
bacterial colony was rubbed on the filter paper using a toothpick. Discs were placed on Muller- Hinton agar plates inoculated with
Blue-black development was observed within 10-20 seconds for the bacterial isolates. Plates were then incubated at 37˚C for 24
positive results. Any changes after 20 seconds were disregarded. A hours. The diameter of growth inhibition zones were measured
positive test indicates a non-fermenting bacteria and vice versa across the disk and data were recorded to the nearest millimeter
(MacFaddin, 2000). (Milhau et al., 1997).
Catalase test was carried out by placing a drop of
hydrogen peroxide on the bacterial culture. A positive test is RESULTS
indicated by froth formation. A positive test indicates production of
Sampling design and handling
catalase enzyme to facilitate cellular detoxification (Taylor &
Hundred patients were selected from Kafr El-Dawar (a
Achanzar, 1972).
public hospital lab situated in a marginal area) and from
Indole test was carried out by culturing pure bacterial
Alexandria (a public hospital lab situated in one of the most
strain in sterile peptone broth for 24-48 h before dipping a strip
populated commercial area) according to clinical investigations
wetted with Kovac's reagent. A positive result is shown by the
and symptoms. Gender represented 50% of each. Half of the
presence of a red or violet color indicating the presence of
patients were above 40 years old, and the other half were under 40
tryptophanase system, which splits indole from tryptophan
years old. Only 55 samples showed positive results 63.6% females
(MacFaddin, 2000).
and 36.4% males. Among them 66% were above 40 years old and
44% were under 40 years old table (1) fig (1).
Antibiotic sensitivity tests
All purified isolated strains were inoculated on Muller- Table. 1: Distribution of UTI in the sampling areas.
Hinton Media (Oxid ®) and then incubated at 37˚C for 24 hours in Number of
an incubator. Antimicrobial susceptibility of isolates was tested by Number of infected % of
infected samples /
samples / Kafr infected
the disk diffusion method using antibiotic disc (Oxid®) with the Alexandria
Eldwar patients samples
patients
minimum inhibitory concentration (MIC) (Alan et al., 2006). Females 22 / 25 13 / 25 63.6%
Agents tested were Amoxicillin/ Clavulanate, Pipracillin/ Males 14 / 25 6 / 25 36.4%
>40 years old 20/ 25 13 / 25 66%
Tazobactam, Cefotaxim, Imipenem, Amikacin, Norfloxacin, <40 years old 16 / 25 6 / 25 44%
Trimethoprim/ Salfamethoxazole (oxoid®). These antibiotics were
chosen as they are the antibiotics of choice in the treatment of UTI
(Wood & Washington, 1995).
B

Cluster analysis and isolates identification

A
For cluster analysis, positive and negative readings were
coded as 1/0, respectively. The data obtained in this study
subjected to statistical analysis and a matrix Euclidean distance
among the isolates was formed for the construction of dendogram.
The statistical calculations were done by the System of Statitics
SYSTAT 12 software. Cluster analysis was used to investigate
similarities in bacterial profiles of the samples (Sharma, 1996).

Identification of isolates
All bacterial isolates were identified using highly Fig 1: Distribution of UTI in the sampling areas (a): gender, (b): age.

automated VITEK®2 System that depends on supplemental tests

and provides same-day right identification (Bosshard et al., 2006). URINALYSIS RESULTS
Three identification cards GN ID (Gram negative Investigation of leucocytes, nitrites, WBC and pyuria
identification), GP ID (Gram positive identification cards) and Leucocytes and nitrites were investigated using the
YST ID (yeast and yeast like organisms) were used (Wallet et al., dipstick technique. This technique is a routine lab test. Phase
2005). contrast light microscopy was used to test the samples for pyuria
and bacterurea.
Effect of natural oils on isolated strains Results in figure (2) show 26 cases (24%) of samples
The crude natural oils of five Egyptian plants namely, were positive for leukocytes, 14 cases (13%) of samples were
Celery (Apium graveolens), Chamomile (Matricaria recutita) , Dill positive for nitrite, 15 cases (14%) of samples were positive for
(Anethum graveolenes), Parsley (Petroselinum hortense) and both leukocyte and nitrite of the dipstick test, and 17 cases (15%)
Thyme (Thymus vulgare) were tested for their antibacterial effects. of samples were positive for pyuria, and 38 cases (34%) of samples
The tests were performed by the disk diffusion method, using were positive for bacteriuria of microscopy test.
.
 Journal of Applied Pharmaceutical Science 02 (04); 2012: 90-98

They all showed non-haemolytic reaction on blood agar

medium. Although all strains seemed to be identical, they showed
differences in antibiotic sensitivity pattern.

Gram- negative bacilli

This group comprises of twenty-seven strains, all growing
on MacConkey agar. 33% of them were of entire medium white
colonies. Cells were long straight rods. They were of γ-haemolytic
type on blood agar medium. 22% of the strains of them were of
irregular small green colonies. Cells were short rods and of β-
Fig. 2: Urine analysis (%). haemolytic type on blood agar medium. Other 22% of the strains
were of entire large yellow colonies. Their cell shape was long
UTI causative organisms
straight rods of gamma γ-hemolytic type on blood agar medium.
Samples of positive leucoscytosis and bacteriuria were
15% of them were of entire small white colonies. Cells were of
selected for further microbiological investigations. One ml of
long straight rods and of beta (β) hemolytic type on blood agar
bacterial suspension was plated on MacConkey and Blood agar
medium. The last 8% strains were of entire small green colonies of
plates. Samples were characterized for single or multiple infections
alpha α-hemolytic. Members of this group showed differences in
according to the morphological description of the plates.
antibiotic sensitivity.
Developed colonies were coded and purified using the streaking
For studying the antibiotic sensitivity pattern of members of
plate method technique. 64 bacterial strains and 4 fungal strains
this group, strains 5, 42, 36a and 50b were selected (Fig. 4). Strain
were obtained from this process. Bacterial strains were examined
5 was sensitive to all tested antibiotics except Norfloxacin and
phenotypically for Gram reaction, cell shape and culture
trimethoprin. Strain 42 was sensitive to all tested antibiotics except
description. According to Gram reaction and cell shapes, isolates
Pipracillin, Norfloxacin and Trimethoprin. Strain 36a was sensitive
were grouped into 3 categories: Gram- positive cocci, Gram-
to all tested antibiotics except Imipenem and Amikacin while strain
negative cocco bacilli, and Gram- negative bacilli.
50b was sensitive to all tested antibiotics except Amoxicillin,
Pipracillin and Trimethoprin.
Phenotypic characterization of the isolates
Gram- positive cocci
Nine strains were included in this group. All of the nine 6
strains were not able to grow on MacConkey agar. They were 5
6
oxidase negative and non-indole producers. 60% were of entire,
large, yellow colonies. Cells were forming irregular grape-like 7 4 3
4 3 7
group. They were all of β-hemolytic type on blood agar medium.
The other 40% were of entire small white colonies. They were all 2 5
of γ-hemolytic type on blood agar medium.. 1 1

2
Strain 5 Strain 42

7
6
5
5 6 7
4 4 4 6
7 6 5
5
3 3
3
7 3 2
2 1 2 1 2 4
1 1
Strain 8a Strain 11

Fig. 3: Antibiotic sensitivity test of strains 8a and 11 representing the Gram- Strain 36a Strain 50b
positive cocci, where 1: Amoxicellin, 2: Pipracillin; 3: Cefotaxin 4: Imipenem, 5: Fig. 4: Antibiotic sensitivity test of representative strains of the Gram- negative bacilli where 1:
Amoxicillin, 2: Pipracillin, 3: Cefotaxin, 4: Imipenem, 5: Amikacin, 6: Norfloxacin, and 7:
Amikacin, 6: Norfloxacin, and 7: Trimethoprin.
Trimethoprin..

Gram- negative coccobacili

Other strains
It comprises of twenty-eight strains. All of them were of Four other strains were found in some female patient
entire medium white colonies. Cells were very short rods. All were samples. Their colonies were large shiny and cells were large,
able to grow on MacConkey agar giving pink colonies. 100% of the compared to bacteria, with very clear nucleus. These strains were
strains were oxidase negative, catalase positive, and indole positive. very similar to yeasts.
 Journal of Applied Pharmaceutical Science 02 (04); 2012: 90-98

Numerical analysis and Identification of the UTI isolates Distribution of uropathogens in collected samples
Characteristic data were transformed into 0/1 codes to be The results indicate that the most predominant
analyzed using SYSTAT program. Cluster analysis was chosen to uropathogen, in the study area, was Escherichia coli as it was
classify members within each group into clusters based on obtained in 43.7% of the isolates followed by Klebsiella
similarity matrix. From each cluster, a representative strain was Pneumonia (14.1%). Both Pseudomonas aeruginosa and Proteus
selected for identification using VITEK® system. mirabilis were represented in 9.4% of the isolates while
Staphylococcus aureus was recorded in 7.8% of the samples.
Gram- positive cocci Enterococcus faecalis and Morganella morganii were represented
The numerical analysis of this group resulted in 2 major in 6.2% of the isolates however, only 3.2% was recorded for
clusters. Strains 1b, 11, 18, 34b, and 41 were grouped in one Pseudomonas fluorescens (Table 2).
cluster at a similarity distance of 98.8. While, strains 8a, 16, 32 and
36b were grouped in another cluster with similarity distance of Table. 2:: Frequency of the uropathogenic isolates .
98.8. Strains 11 and 8a were chosen to represent these clusters, Microorganism Frequency Percentage
respectively, for identification. Escherichia coli 28 43.7%
Klebsiella pneumoniae 9 14.1%
Results of the VITEK® identification of the representative Pseudomonas aeruginosa 6 9.4%
strains showed that strain 8a was found to belong to Enterococcus Proteus mirabilis 6 9.4%
Staphylococcus aureus 5 7.8%
faecalis, and strain 11 was found to belong to Staphylococcus Enterococcus faecalis 4 6.2%
aureus. Morganella morganii 4 6.2%
Pseudomonas fluorescens 2 3.2%

Gram- negative coccobacilli

Results showed two main clusters. Strains 2, 4, 7, 9, 14, Effect of natural oils on bacterial isolates
19, 20, 21, 26, 25, 28, 33, 35, 38, 40, 43, 46 and 48 were grouped Results in figure (5a) show that Dill oil was the most
in one cluster at a similarity distance of 98%, while strains 6a, 8b, effective oil that reduced 61% of E. coli isolates. It also affected
22b, 23a, 24a, 27b, 30b, 34a, 45a, 50a were grouped in another 56% of the Gram- positive cocci, and interestingly, it affected only
cluster with similarity distance of 97%. The whole group was 33% of the Gram- negative bacilli. Generally, dill affected 48% of
homogenous and very similar in all phenotypic characters except in all isolated uropathogens (Fig. 5b).
the sensitivity pattern to antibiotics. 50% of all strains were chosen
for VITEK® system identification, and they were all found to
belong to E. coli.

Gram- negative bacilli

The results showed that members of this group were
classified into two clusters: A was separated at 75% and B was
separated at 83%. Cluster B was sub-divided into two sub-clusters
(C at 74% and D at 73%).
Cluster A was separated at a distance of 75% and
comprised 4 strains (50b, 45b, 30a and 31a). Strains 50b and 31a
were chosen as representative to this cluster for identification and
both found to belong to Morganella morganii.
Cluster C was separated at a distance of 76% and
contained 8 strains, subdivided into 2 sub-clusters (c1 and c2) at a
distance of 72%. Four strains of sub-cluster c1 were chosen for
identification and all found to belong to Pseudomonas aeruginosa,
while the stain chosen from sub-cluster c2 was found to belong to
Pseudomonas fluorescens. These results lead to the conclusion that
all members of cluster C were classified among genus
Pseudomonas.
Cluster D was separated from Cluster B at a distance of
74%. Members of this cluster were also sub-clustered to d1 at 72%
and d2 at 71%. 50% of the members included of each sub-cluster
were submitted for VITEK ® system for identification. Members
of sub-cluster d1 were found to belong to Klebsiella pneumoniae
while members of sub-cluster d2 were found to belong to Proteus
mirabilis. Fig. 5: the effect of natural oil on the uropathogens isolated from the study area.
 Journal of Applied Pharmaceutical Science 02 (04); 2012: 90-98

Results also indicated the effect of Parsley and Celery, DISCUSSION

their stronger effects were against Gram- positive cocci (56%) for
Urinary tract infection (UTI) is an extremely common
both oils, followed by Gram- negative bacilli (48% and 41%,
clinical problem. It is important because it may involve the urethra,
respectively) while their effects on E. coli was much less (29% and
bladder, uterus, and kidney. Because they are not reportable
21%, respectively). Parsley and Celery followed Dill in their
diseases, it is difficult to assess accurately the incidence of UTIs,
general effect. They affected 41% and 34% of the local
not only in Egypt but also in many other countries. This situation is
uropathogens, respectively (Fig. 5a and b).
further complicated by the fact that accurate diagnosis depends on
Thyme showed effects only against Gram- negative bacilli
both the presence of symptoms and positive urine cultures,
and cocco bacilli (37% and 21%, respectively). It had no effect on
although in most outpatient settings this diagnosis is made without
Gram- positive cocci. Its effect generally on all isolates did not
the benefit of culture (Foxman, 2003).
exceed 25%.
55% of tested patients were found to suffer from UTI,
Chamomile was the weakest oil among the tested oils. It
where about 11% of them had renal failure and 16% were from
affected only the Gram- negative while had no effect on Gram-
ICU all had mixed urinary tract infection. These results show that
positive cocci. Generally, it affected only 5% of all isolated
UTI represent a real risk in Egypt, as it is twice higher than those
uropathogens (Fig. 5a and b).
reported by Foxman (2003). Jonathan & Evan (2006) also stated
The inhibition zones with different diameters caused by
that the relation between nosocomial infection as UT mortality in
different oils on plates of different isolates in (Fig.6) reflect the
hospitals remains unclear; however, the highest rates of
antibacterial potency of the used essential oils against different
nosocomial infections are observed in ICUs.
bacterial species infecting urinary tract.
The presence of both pyuria and bacteriuria from a fresh
urine sample is highly indicative for UTI (Watson, 2004). The
study of bacteriology of UTI in 114 patients revealed a high al
failure among patients with bacteriuria was a complicated one
(Kaye, 1972). Another study reported infection rates ranging from
1%–5% after a single brief catheterization to virtually 100% for
patients with indwelling urethral catheters draining into an open
system for longer than 4 days (Fridkin et al., 1997).
According to the current study Gram- negative bacteria
was responsible for 85.9% of UTIs in comparison to Gram-positive
50b: Morganella morganii 36a: Pseudomonas aurginosa bacteria which was 14.1%.. In our study Escherichia coli was the
most predominant uropathogen with 43.7%, followed by Klebsiella
Pneumoniae 14.1%, Pseudomonas auruginosa and Proteus
mirabilis 9.4%, Staphylococcus aureus 7.8%, Morganella
morganii and Enterococcus faecalis 6.2%, and Pseudomonas
fluorescens 3.2%. In a cross sectional study by the University of
Florida, USA of a group of patients, urine specimens were
collected in an emergency department and grew cultures with
greater than 100,000 cfu/ml of single organism on Mackonky and
blood agar. All patients lacking UTI symptoms were excluded.
42: Klebsilla pneumoniae 5: Proteus mirabilis After making all exclusions, 81 patients met the inclusion criteria
of this study. Of these 81 patients 89% had UTI due to Escherichia
coli, 3.7% to Klebsiella, 1.2% to Proteus, 1.2% to Citrobacter,
1.2% to Staphylococcus 1.2% and Enterococcus 3.7% (Mcloughlin
& Joseph 2003).
In this study, there was a significant difference between
the frequency of UTI and gender. The prevalence of UTI was in
female more than in males as after screening 100 (50 males and 50
females) urine samples suspected of having urinary tract infection
that were collected from two areas Alexandria and Kafr Eldwar
using two method of examination urinalysis and urine culture.
8a: Enterococcus faecalis 11: Staphylococcus aureus Only 55 (55%) patients (35 of 63.6% females and 20 of 36.4%
males) gave positive urinary tract infection culture test that is
Fig. 6: Oils sensitivity test of strains 50b, 36a, 42, 5 representing the Gram-
negative bacilli, 8a and 11 representing the Gram- positive cocci where,
meaning the infection spread in females than males. In another
1: Chamomile oil, 2:Celery oil, 3:Dill oil, 4:Parsely oil, and 5: Thyme oil. study, it was seen that significantly higher incidence rate for girls
.
 Journal of Applied Pharmaceutical Science 02 (04); 2012: 90-98

(34.4 episodes per 1000 person years) than for boys (4.4 episodes similar to data previously published for studies conducted
per 1000 person years). Another study also showed that urinary in Canada and other European countries (Zhanel et al., 2008).
tract infections are more common in girls (Brooks & Houston, Often the antibacterial agents in herbs, volatile essential
1977). oils are extracted from plants using steam distillation. These highly
In current microbiological study patients divided into two concentrated oils are often complex mixtures of chemicals
groups; the first group contains 50 patients above 40 years, and the possessing wide-ranging properties. Before modern medicine
second contains 50 patients below 40 years, thirty nine (39%) of started emphasizing chemically synthesized drugs, herbal remedies
first group, and only sixteen (16%) from second group had were the cornerstone of most of the world’s healing traditions and
infection of urinary tract. One study showed increase of women’s even today, are used by 80% of the world’s population who cannot
UTIs between the age groups 20-25 and 41-46 years. The result is afford Western pharmaceutics. As concerns grow about drug side
not surprising since this age span corresponds to a woman’s most effects or bacterial resistance, many are once again turning to
fertile period and to parturition, especially the first delivery, which herbal remedies to treat diverse ailments, including UTIs
is a well-known risk factor for the development of UI, also in (Knobloch et al., 1989). Some studies had shown that Celery Seed
women, a low estrogen level increases the intravaginal pH, oils can help as an herbal remedy today, Celery Seed is most
resulting in the lactobacillus being replaced by a pathogenic agent commonly used as a natural diuretic, as well as a treatment for
(Hagglund et al., 2004). Urinary Tract Infections due to it is anti-bacterial properties
According to the National Institute for Health and Clinical (Maruzzella & Sicurella, 1960). The potent anti-septic action and
Excellence (NIHCE) guidelines, prevention of UTI recurrence mild diuretic effect of the celery comes in handy in the treatment of
includes; relieving constipation and dysfunctional elimination many disorders affecting the human body (Farag, et al., 1989).
syndromes in patient who have had a UTI, encouraging them to Antibacterial activity of various constituents of leaves, flowers and
drink an adequate amount of water. A prolonged course of low- mixtures of Thymus vulgaris extracted with distilled water and
dose antibiotics is effective in reducing the frequency of UTI in 90% ethanol was reported in a study. Alcoholic extracts are more
those with recurrent UTI. Also cranberry (juice or capsule) may efficient on various pathogenic bacteria and mixed extracts have a
decrease the incidence of UTI in those with frequent infections highly antibacterial activity. In general, all extracts in various
(Tangho & Mcaninch, 2004). concentrations with few exceptions are more efficacious on Gram
The data of antibiotic sensitivity pattern has revealed a positive bacteria than on Gram negative bacteria (Goodner et al.,
close relationship with their prophylactic usage. The more 2006). The antibacterial activity of Thmus vulgaris extracts may be
frequently used antibiotics like Penicillin, Erythromycin, due to presence of phenolic constituents (thymol and carvacrol),
Chloramphenicol, and Ampicillin revealed very low levels of which make up a large percentage of the volatile oil (Janssen et al.,
sensitivity (<25%) to all organisms in a group as a whole. The 1987).In general the results agree the results recorded by many
avoidance of prophylactic usage of antibiotics may help surmount workers investigating the used plants ,Celery (Celestin and
this to a certain extent (Harkness, et al., 1975). Heiner, 1993), Dill (Delaques et al, 2002), Thyme (Goodner et al ,
In this study, the antibiotic sensitivity tests showed that 2006) and Camomile (Tayel and El-Taras, 2009).
Enterococcus faecalis and Ps. Aeruginosa were sensitive to Evaluation of Vitek GNI+ and Becton Dickinson
Cefotaxin, Amoxicillin/ Clavulanate, Norfloxacin, Pipracillin/ Microbiology Systems Crystal E/NF identification systems for
Tazobactam, Trimethoprim/ Slfamethoxazol and resistant to identification of members of the family Enterobacteriaceae and
Imipenem and Amikacin. Staphylococcus aureus was resistant to other gram-negative, glucose-fermenting and non-glucose-
all selected antibiotics. But Proteus mirabilis and K. pneumoniae fermenting bacilli. The system is aiming at rapid identification for
were sensitive to Cefotaxin, Amikacin, Amoxicillin/ Clavulanate, which time-consuming supplementary tests are contraindicated
Imipenem, Pipracillin/ Tazobactam and resistant to Trimethoprim/ and/or not often performed in a routine clinical laboratory (Miller,
Slfamethoxazol and Norfloxacin. M. morganii was sensitive to 1999).
Cefotaxin, Amoxicillin/ Clavulanate, Imipenem, Norfloxacin and One major advantage of the VITEK 2 system is its speed
resistant to Amikacin, Trimethoprim/ Slfamethoxazol, Pipracillin/ in reliably identifying gram-negative rods within 3 h. This is
Tazobactam. basically achieved by the more sensitive fluorescence-based
In a study, imipenem demonstrate good activity against technology used in the system. As a broader and more detailed
Enterobacteriaceae, (100% for E coli, 99% for other database has been built by the company and allows a better
Enterobacteriaceae), also piperacillin/tazobactam was the most discrimination between related taxa. However, even the more
potent antibiotic against P. aeruginosa (90% of susceptible strains sensitive fluorescence-based technology used in the ID-GNB card
versus 84% for carbapenems) as reported in other studies (Turner, did not significantly change the outcome of the identifications of
2008). In a nother study, the researcher reported that meropenem, some slowly metabolizing non-fermenting bacteria, which were
imipenem and piperacillin/ tazobactam are very active against categorized as various non-fermenting gram-negative bacilli
Gram-negative bacilli, including Enterobacteriaceae, and the (Stager et al., 1998). Obviously, the VITEK 2 system in
susceptibility data obtained from this multicentre study were conjunction with the ID-GNB card represents an improvement
.
 Journal of Applied Pharmaceutical Science 02 (04); 2012: 90-98

regarding speed compared with the previous VITEK system. In one population-based study of women aged 22-50 years. Scandinavian journal
evaluation, 88.5% of all strains were correctly identified after 3 h, of primary health care 2004;22(2):112.
Harkness, J. L.; Anderston, F. M.; Naomi, D. R-Factor in urinary
whereas in the evaluation of O’Hara et al., applying the previous tract infection. Kidney International, 1975; 8: 130-133
GNI+ card, only 47% of all enteric strains were identified in 3 h or Hindler, F.J.; Munro, S. (2007): Susceptibility testing. In:
less (O’Hara, et al., 1997). Other advantages of the VITEK 2 Clinical Microbiology procedures handbook. 2nd edition, Isenberg, H.D.
(ed.). LSG & Associates Santa Monica, Californnia.
system are the decreased turnaround and hands-on times since the
Janssen, A. M.; Scheffer, J. J. C.; Baerheim, S. A. Antibacterial
system is nearly fully automated. The high degree of automation activity of essential oils. A 1976-1986 literature review. Aspects of the test
may also improve accuracy (Cuziat et al., 1997). Factors affecting methods. Pland Medica, 1987; 53: 395-398
the quality of the identification are the age of the culture (8- to 24- Jonathan, H.C.; Evan, S. Investigation of urinary tract infection.
Curren Pediatrics, 2006;16:248-253.
h cultures are best) and the inoculums but not the age of the Kaye, D. Long term prognosis of Urinary Tract Infection and its
inoculum suspension (Guicherd et al., 2002). management. Saint Louis, (1972),pp.267-278.
Knobloch, K.; Pauli, A.; Iberl, B. Antibacterial and antifungal
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