A Scientific Review Article about Green Tea as an Effective

Antimicrobial for Urinary Tract Infections Caused by Escherichia

coli
Mayramarie L. Flor, Dawn P. Gentolea, Jean-Mae Emmanuel Himagan &
Hanne Christele S. Parcia, Joyzy Faith Pimentel
BACKGROUND: One of the most
common infections caused by
Escherichia coli is Urinary Tract
infection which is also the most
common infection in Filipino
women. With cultural attributes of
people in our country, many
beliefs that Tea made from leaves
of plant can cure these infections
one of this is Green Tea as an
effective antimicrobial treatment
for certain diseases. With the
widely
used
bacteriological
methods, assays and experiments
can prove that Green tea is one of
cheapest fighting armor against
Urinary Tract Infections.
CONTENT: Antimicrobial effect of
Green tea provides effective
curing in Urinary Tract infections
using methods that measures
antimicrobial activity in different
specifications.
Specific
applications are use such as
inhibitory effect of green tea, and
synergistic ability of green tea
and other teas. We discuss this
clinical applications of these
methods in different strains of
E.coli . Concluding remarks and
future application are discussed.
SUMMARY:
Growing
with
discoveries of antibiotic targeting
bacteria, people will always
choose organic and less toxic cure
for diseases like green tea to treat

urinary tract infections. Large

improvements for accuracy and
precision related to the microbial
testing are done to prove the
effectiveness of certain drug. Yet
the easiest and cheapest way is
to invest in organic and non-toxic
substances as a source.
Properties of Green Tea for
Antibacterial Action against
E. coli
Tea is a widely-consumed
drink in the world in terms of
providing leisure and relaxation
given its affordable range of
prices
and
wide
availability
around the world. Aside from
these though, it was later on
suggested to also be beneficial to
the health due to its antihypertensive effects contributing
to reduction of cardiovascular
diseases,
its
anticancer
properties, ultraviolet protection,
bone mineral density increase
and neuro- protection powers, as
well as promotion of oral health
and other physiological functions
(18).
These
benefits
are
specifically
found
from
consumption of green tea. On the
other hand, black tea is known to
have a wild range of beneficial
physiological
and
pharmacological effect, among
these
are,
strengthening
1

capillaries,
anti-inflammatory
effect,
antioxidant,
having
hypocholestrolemic action and
inhibiting the growth of implanted
malignant cell. Green tea is
prepared through an infusion of
different processed leaves of a
variety of an evergreen shrub
Camellia sinensis while the black
tea is prepared in a similar way
except it undergoes fermentation,
an oxidation step (19).
Teas of C. sinensis undergo
different
manufacturing
processes. Green tea is produced
by steaming (Japan) or panning
(China) to prevent catechin
oxidation by polyphenol oxidase2.
Oolong tea is semi-fermented
while black tea is fully fermented.
Human studies suggest that
green tea may contribute to a
reduce the risk of cardiovascular
disease and some forms of
cancer, as well as promote oral
health and other physiological
functions
such
as
antihypertensive
effect,
body
weight control, antibacterial and
antivirasic activity, bone mineral
density
increase,
antifibrotic
properties and neuroprotective
powers. Green tea contains
mainly flavanols or catechins of
epigallocatechin gallate (EGCG),
epigallocatechin
(EGC),
epicatechin gallate (ECG), and
epicatechin (EC) (28).
Urinary tract infection (UTI)
is the second most common type
of infection found in any organ
system, and the most common
type of nosocomial infection. The
strains of Escherichia coli that

infect the urinary tract are

categorized as uropathogenic E.
coli. The antibacterial agents that
have been traditionally used to
treat
UTIs
(Beta-lactams,
fluoroquinolones,
trimethoprim
sulfamethoxazole, nitrofurantoin,
etc.) are becoming less effective
due to the growing worldwide
problem
of
antibacterial
resistance. Cephalosporin, which
is a useful drug against UTI, is
unfortunately the same cause of
various complex side effects and
to a massive extent, even death
(20). To combat the emergence
and spread of multi drug resistant
pathogens, there is a growing
need
to
explore
newer
antimicrobial
compounds
particularly from herbal extracts,
which could be incorporated as
adjuvant therapeutic agents in
the treatment of such life
threatening infections (21).
Green
tea
(Camellia
sinensis
leaves)
has
been
consumed since ancient times in
China and has gained popularity
in
several
other
countries
including
India.
Green
tea
extracts
being
a
popular
beverage consumed worldwide,
inexpensive,
and
non-toxic,
serves as a very important a
source of alternative medicine. It
is known to possess preventive
activity against cardiovascular
diseases and some forms of
cancer. Also, it has antioxidant,
antitumor,
anti-inflammatory,
anti-diabetic, immunomodulatory
and antibacterial activities. Green
tea consists of catechins viz.
epicatechin (EC), epicatechin-32

gallate (ECG), epigallocatechin

(EGC), and epigallocatechin-3gallate (EGCG). Other polyphenols
include
flavanols
and
their
glycosides [1]. EGCG, a main
constituent
of
green
tea
polyphenol, has been reported to
have great antimicrobial activity
and also exhibits synergistic
properties with other antibiotics
against Gram-positive and Gramnegative bacteria (2-4)
Recent studies suggest
that green tea may contribute to
a reduction in the risk of
cardiovascular diseases and some
forms of cancer as well as
promotion of oral health and
other physiological functions such
as anti-hypertensive effect, body
weight
control,
ultraviolet
protection, bone mineral density
increase and neuro-protection
power.
Studies
on
the
antibacterial activity showed that
extracts of tea leaves inhibits the
growth of Staphylococcus aureus
and E.coli which were highly
sensitive (5). S.aureus, Vibrio
parahemolyticus,
Clostridium
perfringens,
Bacillus
cereus,
Pleisomonas
shigelloides
and
Aeromonas sobria failed to grow
in tea normally consumed by
Japanese people. Tea components
also inhibit the growth of Vibrio
choleras
01,
Streptococcus
mutans
Shigella
dysenteriae,
E.coli S. salivarius, Bordetella
pertusis
and
phytopathogen
Eriwinia spp and Pseudomonas
spp (6,7,8,9).These antimicrobial
activity of tea may refers to
several chemical components
found in tea like polyphenolic

compounds (these make up some

30% of dry weight) and generally
known as "tannin" which are
chemically different from other
plant
tannin,
the
simplest
compound in this class are the
catechins (found in green and
black tea), inhibits the growth of
many bacterial species, leaves
also contain flavonols, volatile
flavor components (volatile oil),
vitamin C, minerals and alkaloid
(caffeine) (5,6,7) catechins (found
in green and black tea), inhibits
the growth of many bacterial
species, leaves also contain
flavonols,
volatile
flavor
components (volatile oil), vitamin
C, minerals and alkaloid (caffeine)
(5,10,11)
.According
to
the
reports of many researchers,
antibacterial resistance is a
worldwide growing problem (12)
E.coli are facultative anaerobes in
the normal intestinal flora of
human and animal however
pathogenic
strains
of
these
bacteria are an important cause
of bacterial infections, these
strains are the foremost cause of
urinary tract infections as well as
a major cause of neonatal
meningitis,
nosocomial
septicemia and surgical site
infections
(12,13).
Antibiotic
therapy is the gold standard of
treatment, however, long-term
therapy may result in many side
effects and cause selection of
resistant bacteria (14)
Recent
reports showed that the use of
plant extract may have synergic
effect
with
some
antibiotic
against microorganism, Sub-MIC
of black tea extract made S.
dysenteriae more susceptible to
3

chloramphenicol,
gentamicin,
methicillin and nalidixic acid and
green tea have synergistic effect
with
levofloxacin
against
enterohemorrhagic E.coli (15)
Epicatechin gallate (found in
black and green tea ) reduced the
MIC of oxacillin and -lactam in
methicillin
resistant
S.aureus
(MRSA) (16) and enhanced effect
of Japanese tea on inhibitory
activities of antibiotic against
MRSA (17) ,so the aim of this
study is to investigate the
interactions of sub-MICs doses of
black and green tea water
extracts
and
25
kinds
of
antibiotics against two E.coli
strains isolated from local urinary
tract infections.
Experimental Application
Antibacterial Action

as

Effectiveness
of
certain
drug or extract to test for
antimicrobial activity is measured
by
a
Minimum
Inhibitory
Concentration Determination in
which of the green tea extract
was determined by agar dilution
method.
MHA
plates
that
contained various concentrations
of green tea extract (0, 2.5, 3.0,
3.5,
and
4.0mg/mL)
were
prepared. Suspensions of 0.5
McFarland
standard
dilutions
(equal to 1.5 108 cells/mL)
were prepared from bacterial
cultures grown overnight in LB
broth. A 104 dilution of these
suspensions was prepared, and
100 L of dilution per plate was
inoculated using a spread plate

technique (plated concentration

equals 1.5 103 cells).Inoculated
plates were incubated at 37oC for
48h, and on the basis of colony
counts, the MIC was determined
as the lowest concentration of
green tea extract to show an
inhibitory effect on growth of the
bacteria (no colonies detected).
Each strain was tested three
times (one strain was not
susceptible at even 4.0mg/ml);
94% were Susceptible at 3.5
mg/ml; 76% were susceptible at
3.0
mg/ml;
40%,
were
susceptible at 2.5 mg/ml; The
control strains varied; one being
susceptible at 2.5 mg/ml and
the other susceptible at 3.5
mg/ml. Table 1 shows the MIC
results for the 79 strains that
were susceptible at 4.0 mg/ml
(22).

The green tea extract was shown

to have an inhibitory effect on the
growth of E. coli strains isolated
from UTIs. The MIC results can be
adjusted to reflect the EGC
content, using a value of 18%
EGC as the content in total green
tea
polyphenols
(Vuongetal.,
2011). That makes the adjusted
results as follows: 40% of strains
tested were susceptible at a
4

concentration of EGC at 0.45

mg/ml
(450
g/mL);
36%
susceptible at 0.54 mg/ml (540
g/mL); 18% susceptible at 0.63
mg/ml(630 g/mL); and 5%
susceptible at 0.72 mg/ml (720
g/mL). Since all of the strains
tested (99%) were susceptible at
a concentration of 0.72 mg/mL,
this suggests that EGC might be a
good inhibitor of bacterial growth.
Table 1 shows the MIC data for
total green tea extract and for the
EGC component. The results of
this study have shown that green
tea can have an antimicrobial
effect on E. coli bacteria that
cause UTIs. This is the first time
that green tea has been reported
to have this kind of effect. The
data also adds to the current
information on the potential
health benefits of green tea. It is
our hope that these findings will
encourage further studies on the
antimicrobial potential of green
tea and other plant components
(22).
Antimicrobial activity of Aqueous,
Ethanolic,
Methanolic
and
Acetone extracts of Green tea and
Black tea was determined by Agar
well diffusion method. Green tea
The major difference between
Green tea and Black tea is in their
processing. Green tea is prepared
through an infusion of different
processed leaves of a variety of an
evergreen shrub Camellia sinensis
while the black tea is prepared in a
similar way except it undergoes
fermentation, an oxidation step
catalyzed by polyphenol oxidase.

extracts
exhibited
higher
antimicrobial activity than Black
tea extracts. Amongst Green tea
extracts aqueous extract was
found to be most effective while
amongst Black tea
extracts
methanolic extract was most
effective. All tea extracts have
shown significant antibacterial
activity against S.aureus ATCC
25922 with Aqueous extract of
Green tea exhibiting highest
activity. All Green tea extracts
exhibited
significant
activity
against E.coli ATCC 25923 higher
than Black tea extracts. As
compared to Green tea extracts,
Black tea extracts showed much
lower
activity
against
P.aeruginosa
ATCC
27853.
S.aureus was found to be most
susceptible
to
tea
extracts
followed
by
E.coli
and
P.aeruginosa. Green tea and Black
tea
extracts
have
shown
significant antibacterial activity
with former being more effective
than later. In future there is
immense potential of clinical
application
of
polyphenolic
contents of tea extracts as
adjuvant therapeutic agents to
tackle the menace of growing
antibiotic resistance.
Green tea on the other hand is
subjected to steaming after drying
to
inactivate
the
polyphenol
oxidase enzyme. That is why green
teas have significantly higher
Catechin content than Black teas,
which greatly contributes in the
high antimicrobial property of the
tea (24).
5

Results from the study of Mbuthia

(25) on table 2, antimicrobial
activity for green tea extracts
exhibited an inhibition against both
E.coli ATCC 25922 and S. aureus
ATCC25923 at concentration of
0.1mg/ml.
however,
at
lower
concentration of 0.05mg/ml, green
tea extract did not exhibit any
inhibition
against
the
tested
antibiotic
resistant
bacteria.
Furthermore, on increasing the
concentration of green tea extract,
there was no inhibition against
antibiotic resistant E. coli ATCC
25922 and S. aureus ATCC 26923.
This could be attributed to the
inability of higher concentration of
tea extracts to diffuse through the
Mueller Hinton agar medium.

The bacteria strains use in

this study are part of a research
collection of E. coli isolate from
UTI cultures during the year of

Also there was an evaluation

of green tea extracts interaction
with some antibiotics. Green tea
extract from the TRFK 6/8 cultivar,
exhibited an antagonistic effect
against antibiotic resistant E. coli
ATCC 25923 when combined with
gentamicin Figure 1. However, a
combination of green tea extract
with penicillin G exhibited a
synergeistic effect. In addition, it
was also observed that the green
tea extract from TRFK 6/8 cultivar
exhibited a higher inhibition zone
diameters as compared to penicillin
G and its activity was comparable
to that of gentamicin (Table 3). In
conclusion to the study (25), green
and
black
tea
exhibited
antimicrobial properties against
antibiotic resistant E. coli ATCC
25923 and S aureus ATCC 25923.
Green tea extracts did not impair
the antimicrobial properties of
penicillin G but rather it enhanced
its activity in synergistic manner.
The combination of green tea
extract and penicillin G could be
useful in management of emerging
antibiotic resistant bacteria (25).

2016. E. coli isolates are collect

from urine specimens submit of
selected Al- Hussein medical city
patients during one month period
6

throughout November 2015. The

isolates are further processes by
standard methods to identify as
E. coli isolates. These methods
are Gram stain kit, indole test and
methyl- red reagent (IMVic test)
these strains were tested by
using sensitivity test and had
undergone colony counting to
check green tea and lemon
extract effect. The results of
confirmatory tests on the isolates
that were taken from Al Hussein
medical city patients with urinary
tract
infection
for
all
the
properties
that
E.coli
characteristic with were positive.

Figure 2 The effects of the

90%Concentrations
of green tea extracts
After
refreshing
it
on
nutrient
broth
the
fourth
McFarland standard was taken
and cultivate on Mueller Hinton
agar by taking 0.1m of the
specimen and adding different
concentrations of green tea and
green tea with lemon extracts,
the results showed that the best
inhibitory concentration to inhibit
the growth of bacteria were 90%
for the alcoholic green tea with
lemon extract in the same time
the same concentration give the
best inhibitory rate in the green
tea aqueous extract.

To confirm the results the

concentration
of
90%
was
selected and redistributed on
bacterial grown on Mueller Hinton
agar and after incubation the
number of grown colonies were
counted for the aqueous and
alcoholic extract at concentration
of 90% for each of the green tea
and green tea with lemon
extracts, the green tea with
lemon alcoholic extract shows the
best
results
in
elimination
growing bacteria since only 6
colonies where grown comparing
to the control that exceeded 900
colonies. After studying and
measuring the inhibitory effect of
the antibiotic ciprofloxacin on
E.coli the synergistic effects of
antibiotic and the alcoholic and
aqueous
extracts
in
the
concentration of 90% have been
study and measured for the
purpose of confirm or synergy the
effect of each of the antibiotic in
the termination of bacteria and
the extracts as antibacterial and
antioxidant to the side effects
that caused by the antibiotic
during
treatment
Two
mechanisms was adopted in the
result, and both showed the
significant effect of alcoholic
green tea with lemon extract at
concentration of 90% in the
present of antibiotic, and this has
a major role in reducing or
termination of the bad side
effects that comes with antibiotic
when using green tea with lemon
concomitant with antibiotic (27).
7

Concluding Remarks

An emerging threat of
antibiotic
resistance
among
bacterial strains is posing serious
challenges
to
clinicians
everywhere in the world; they are
left with few options leading to
newer interests to explore new
antimicrobial compounds even
from herbal extract products,
which could be seen as effective
against microbes of interest.
Urinary tract infections are not
new in the society and is
undeniably
an
ongoing
challenge in the clinical world in
terms of looking for effective
medicines to lessen burdens
brought by it without imposing
the
threat
of
antimicrobial
resistance. Green tea and Black
tea extracts are known to exhibit
significant antibacterial activity
against pathogens causing UTI,
which led to numerous studies to
support
this
innovation.
In
conclusion, this study like many
other in vitro studies in support of
the same topic conducted had
proved,
reconfirmed
and
consolidated the fact that tea
extracts do have significant
antimicrobial activity at varying
concentrations against different
bacterial pathogens. This study in
addition had also shown that this
antibacterial activity is dependent
not only on the type of the tea
product but also on different
processing techniques involved in
preparing tea extracts which
determines the concentration of
polyphenols particularly catechins
responsible for this activity (24).
Author

Contributions:

All
8

authors confirmed they have

contributed to the intellectual
content of this paper and have
met the following 3 requirements:
(a) significant contributions to the
conception
and
design,
acquisition of data, or analysis
and interpretation of data; (b)
drafting or revising the article for
intellectual content; and (c) final
approval of the published article.

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from
www.ijpab.com

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