7499

PO Box 2345, Beijing 100023, China
www.wjgnet.com
wjg@wjgnet.com
ELSEVIER
World J Gastroenterol 2005;11(47):7499-7507

World Journal of Gastroenterology ISSN 1007-9327
2005 The WJG Press and Elsevier Inc. All rights reserved.
Helicobacter pylori
Bactericidal and anti-adhesive properties of culinary and

medicinal plants against Helicobacter pylori
Rachel OMahony, Huda Al-Khtheeri, Deepaka Weerasekera, Neluka Fernando, Dino Vaira, John Holton,
Christelle Basset
Rachel OMahony, Huda Al-Khtheeri, John Holton, Centre for
Infectious Diseases and International Health, Royal Free and University College London Medical School, London,
United Kingdom
Deepeka Weerasekera, Department of Surgery, University of
Sri Jayewardenapura, Colombo, Sri Lanka
Neluka Fernando, Department of Microbiology, University of
Sri Jayewardenapura, Colombo, Sri Lanka
Dino Vaira, Department of Internal Medicine and Gastroenterology, University of Bologna, Bologna, Italy
Christelle Basset, INSERM 0114, Physiopathologie des Maladies Inflammatoires Intestinales, CHU Lille, France
Supported by the European Union on EC contract QLK2CT-2001-01216 (ADRI)
Correspondence to: Rachel OMahony, Centre for Infectious
Diseases and International Health, Royal Free and University College London Medical School, Windeyer Building, 46 Cleveland
Street, London, W1P 6DB,
United Kingdom.
Telephone: +44-20-76799485
Received: 2005-06-02
Accepted: 2005-04-28
Abstract
AIM: To investigate the bactericidal and anti-adhesive
properties of 25 plants against Helicobacter pylori
(H pylori ).
METHODS: Twenty-five plants were boiled in water to
produce aqueous extracts that simulate the effect of
cooking. The bactericidal activity of the extracts was
assessed by a standard kill-curve with seven strains of
H pylori . The anti-adhesive property was assessed by
the inhibition of binding of four strains of FITC-labeled
H pylori to stomach sections.
RESULTS: Of all the plants tested, eight plants,
including Bengal quince, nightshade, garlic, dill, black
pepper, coriander, fenugreek and black tea, were found
to have no bactericidal effect on any of the isolates.
Columbo weed, long pepper, parsley, tarragon, nutmeg,
yellow-berried nightshade, threadstem carpetweed,
sage and cinnamon had bactericidal activities against
H pylori , but total inhibition of growth was not achieved
in this study. Among the plants that killed H pylori ,
turmeric was the most efficient, followed by cumin,
ginger, chilli, borage, black caraway, oregano and
liquorice. Moreover, extracts of turmeric, borage and
parsley were able to inhibit the adhesion of H pylori
strains to the stomach sections.
CONCLUSION: Several plants that were tested in our

study had bactericidal and/or anti-adhesive effects on
H pylori . Ingestion of the plants with anti-adhesive
properties could therefore provide a potent alternative
therapy for H pylori infection, which overcomes the
problem of resistance associated with current antibiotic
treatment.
2005 The WJG Press and Elsevier Inc. All rights reserved.
Key words: H pylori ; Inhibition; Adhesion; Killing; Plants

OMahony R, Al-Khtheeri H, Weerasekera D, Fernando
N, Vaira D, Holton J, Basset C. Bactericidal and antiadhesive properties of culinary and medicinal plants against
Helicobacter pylori . World J Gastroenterol 2005; 11(47):
7499-7507
http://www.wjgnet.com/1007-9327/11/7499.asp
INTRODUCTION
Over half the human population is colonized by
Helicobacter pylori (H pylori), a Gram-negative, microaerophilic
bacterium. If untreated, infection is usually life-long and
leads to chronic active disease[1]. Although most infected
people are asymptomatic, 5-10% of those infected with
this bacterium develop severe gastroduodenal diseases,
including gastric and duodenal ulcers, gastric lymphomas,
and gastric adenocarcinomas[2].
The current and most effective treatment for peptic
ulcer disease is a triple therapy regimen consisting of
a proton pump inhibitor, such as omeprazole, and two
antibiotics, clarithromycin and either amoxicillin or
metronidazole. However, there is an increase that disturbs
due to the prevalence of antibiotic resistance, which is
high in some areas of the world. Metronidazole resistance
is more common than clarithromycin, the latter particularly
having an adverse effect on the eradication rate. Second
line therapies following failure of one of the initial
regimes include triple and quadruple regimens containing
antibiotics, such as levofloxacin or furazolidone, or even
regimens containing five agents. Although the use of
molecular methods can rapidly detect antibiotic resistance
and host polymorphisms, which may lead to reduced
efficacy of treatment and thus eradication failure[3,4], this is
not a long-term solution to the rising trend of antibiotic
resistance.
7500
ISSN 1007-9327
CN 14-1219/ R
World J Gastroenterol
Resistance to antibiotics is not limited to H pylori and

has been an increasing problem for many years. There is
therefore a constant need for new antimicrobial agents and
novel approaches to treatment, ideally preventing disease,
such as inhibition of adhesion or vaccination[5]. Plants
are known to be the source of phytochemicals which are
beneficial for health and could also prevent diseases[6].
Among these phytochemicals, two are of particular interest
in the case of infectious diseases: antimicrobial and antiadhesive agents.
Numerous studies have been undertaken in order to
find antimicrobial agents from plants against organisms
ranging from viruses to protozoa[7]. The major concern is
the validation in human beings with well-designed clinical
trials, and this has also been true for H pylori infection.
Several in vitro studies have looked at the effect of plant
extracts on H pylori. Anti-microbial effects have been
reported for garlic[8,9], green tea[10], honey[11], thyme[12], some
Iranian plants[13] and the essential oils from several species
of mint[13]. Some of these studies have been validated in
animals and confirmed the potential benefit of using plants
as the source of anti-microbial agents against H pylori.
Although garlic and cinnamon have been tested in human
clinical trials with no significant effect[14], a recent study has
shown that consumption of broccoli sprouts is associated
with the eradication of H pylori in some patients[15], but
more work needs to be done in determining the active
ingredients of broccoli as well as performing studies on a
larger number of patients.
The search for anti-adhesive agents represents the
second alternative to antibiotic therapy, which has received
less interest, although studies in animals and human beings
have proved its potential as a new therapy[16]. Successful
inhibition of adhesion has been shown in vitro with
cranberry juice against H pylori [17] and for the seaweed
Cladosiphon fucoidan [18] . Recently, Shibata et al. [19] have
demonstrated that Cladosiphon fucoidan inhibits adhesion
of H pylori to porcine gastric mucin. By adding the plant
to the drinking water of infected Mongolian gerbils, the
prevalence of animals with infection was shown to be
markedly reduced. Lengsfeld et al. [20] have shown that
adhesion of H pylori to human stomach sections was almost
completely inhibited by pre-incubating H pylori with a fresh
juice preparation of the fruit of the okra plant [Abelmoschus
esculentus (L.) Moench]. Lengsfeld et al. [21] have also
demonstrated that acidic high molecular weight galactans
from blackcurrant seeds could inhibit adhesion of H pylori
to human gastric mucosa tissue sections. Moreover, Lee
et al. [22] have demonstrated the inhibition of H pylori
adhesion by polysaccharide fractions of Panax ginseng and
Artemisia capillaris to a human gastric adenocarcinoma
epithelial cell line.
As mentioned earlier, many plants have been shown to
kill microorganisms but rarely have been studied for their
anti-adhesive properties. We have therefore investigated
both the bactericidal and anti-adhesive properties of
25 plants against H pylori. Sixteen of them have never
been tested before against H pylori and are plants that are
frequently used in cooking as well as in medicine in Sri
December 21, 2005 Volume 11 Number 47
Lanka, Iran, and the Middle East. Moreover, we have used

an in situ adhesion assay[23], with stomach tissues expressing
either the Lewis a (Le a) or Lewis b (Le b) antigen, in order
to determine whether the plant extracts inhibit adhesion
by blocking the major H pylori adhesin BabA (which binds
to Le b) or have an effect on other adhesins.
MATERIALS AND METHODS

H pylori isolates
H pylori NCTC 11637 and six fresh clinical strains from
Italian patients with peptic ulcer disease [obtained by one
of the authors (DV)] were used in the study. All strains
were stored on beads at -80 until use. Bacteria were
grown for 2 d on Columbia agar plates supplemented with
5% horse blood (Oxoid, UK) at 37 under microaerobic
conditions. The number of bacterial cells was determined
by obtaining viable counts of serial dilutions and
measurement of the optical absorbance at 600 nm
(Ultrospec II, LKB, UK) of a suspension of bacterial
cells to prepare a standard curve for H pylori. All studies
thereafter used the same concentration of organism based
upon its absorbance.
Fluorescent labeling of H pylori isolates
Suspensions of 1109 cells/mL were prepared from 1 mL
carbonate buffer (0.15 mol/L NaCl/0.1 mol/L Na2CO3,
pH 9.0). Five microliters of a 10 mg/mL fluorescein
isothiocyanate (FITC, Isomer I, Sigma, UK) solution in
dimethyl sulfoxide (DMSO, Sigma, UK) was added to
each bacterial suspension and then incubated for 1 h with
continuous shaking. The suspensions were then washed
three times with phosphate buffered saline (PBS, 8.0 g/L
NaCl; 1.21 g/L K 2HPO 4; 0.34 g/L KH 2PO 4, pH 7.4)
containing 0.5 g/L Tween 20 (PBST; Sigma, UK), and
resuspended in PBS. Labeling was confirmed by using
flow cytometry (FACSCalibur, Becton Dickinson, UK). All
incubation and washing steps were carried out in the dark
at room temperature.
Plants
Twenty-five plants were obtained from different sources
and are described in Table 1. Extracts from the plants were
prepared as follows: for turmeric, ginger, fenugreek, cumin,
fennel, coriander, and chilli, plants were boiled to 100 in
sterile distilled water (100 mg/mL), filtered through sterile
gauze, neutralized to pH 7.0 and then sterilized. Extracts
from all the other plants were prepared as follows: plants
were boiled in sterile distilled water (100 mg/mL) for 10
min, allowed to cool and filtered through sterile filter paper
(Grade 1, Whatman, UK). Fresh garlic, black peppercorns
and cinnamon sticks were finely chopped before being
boiled. All extracts were stored in the dark at -20 until
use. Before the experiments of adhesion, plant extracts
were filtered again (Grade 1 filter paper, Whatman, UK).
Stomach sections
H pylori-negative biopsies of human stomach were
obtained by one of the authors (DV), with the consent of
the Ethics Committee, St Orsola Hospital, Bologna. Two
OMahony R et al. Plants against H pylori
7501
Table 1 Plants used in the study

English name
Sri Lankan name
Scientific name
Part of the plant used Source
Use
Bengal quince
Belly
Aegle marmelos
Root
Sri Lanka
In cooking & medicine
Black caraway (Europe)/Black

Cumin (USA)
Black pepper
Gammiris
Nigella sativa
Seeds
UK, Supermarket
Piper nigrum
Seed berries/fruit
UK, Supermarket
Black tea
Camellia sinensis
Leaves and shoots
Sri Lanka
In cooking
Borago officinalis
Flowers
Iran

Thee
Borage (Starflower)
Chilli
Mirise
Capsicum anunum
Fruit
Sri Lanka
Cinnamon
Curundu
Cinnamomum verum
Bark
UK, Supermarket
Columbo weed
Veneval
Coscinium fenestratum
climbing root
Sri Lanka
In medicine
Coriander
Kottamalli
Coriandrum sativum
Seeds
Sri Lanka
Cumin (small cumin)
Suduru
Cuminum cyminum
Seeds
Sri Lanka
Dilla
Sududuru
Anthum graveolens
Leaves
UK, Supermarket
Fenugreek
Mathe seeds
Trigonella foenum-graecum
Seeds
Sri Lanka
Garlica
Sudulunu
Allium sativum
Bulb
UK, Supermarket
Ginger
E'guru
Zingiber officinale
Rhizome
Sri Lanka
Liquorice
Val'mee
Glycyrrhiza glabra apofosa
Stem
Sri Lanka
Long pepper
Tiphili
Piper longum
Seeds
Sri Lanka
Nightshade
Ela battu
Solanum surattense
Fruit and root
Sri Lanka
In medicine
Nutmeg
Sadikka
Myristica fragans
Kernel
UK, Supermarket
Oreganoa
Oregano
Origanum vulgare
Leaves
UK, Supermarket
Parsleya
Parsley
Petroselinum crispum
Leaves
UK, Supermarket
Sagea
Minchi
Salvia officinalis
Leaves
UK, Supermarket
Tarragona
Artemisia dracunculoses/
dracunculus
Mollugo cerviana
Leaves
UK, Supermarket
Threadstem carpetweed
Tarragon
Path
Paradagam
Seeds
Sri Lanka
In medicine
Turmeric
Kaha
Curcuma longa
Rhizome
Sri Lanka
Yellow-berried nightshade
Katu val batu
Solanum xanthocarpum
Whole plant
Sri Lanka
In medicine
These plants were bought as both fresh and dried material.
sets of formalin-fixed stomach biopsies were used: those

whose epithelial cells expressed the Lewis a blood group
antigen (Le a stomach) and those expressing the Lewis
b blood group antigen (Le b stomach). Five-micrometer
thick sections of the stomach were cut using a Leica
SM2400 rocking microtome. Sections were collected
on polished glass slides coated with Vectabond (Vector
Laboratories, UK). An antigen retrieval step was used by
boiling for 5 min in an 800-W microwave in plastic Coplin
jars containing 15 mL of citrate buffer (pH 6.0, 100 mol/L
sodium citrate tribasic dihydrate, Sigma, UK). The Lewis
phenotype of the tissue was determined by standard
immunohistochemical staining using the DAKO EnVision
Plus HRP kit (DAKO, UK) with anti-Le a and anti-Le b
antibodies generously donated by Dr. J Bara, Hpital StAntoine, Paris, France. Both antibodies were supplied at
a concentration between 1 and 5 g/mL and diluted 1:10
for the anti-Le a antibodies and 1:50 for the anti-Le b
antibodies.
Viable colony count
Bactericidal activity of the plant extracts was determined
by a viable colony count. One hundred microliters of
a suspension of 108 bacteria/mL was added to 900 L
of plant extract for 60 min. The control consisted of
H pylori incubated with sterile distilled water. Serial
10-fold dilutions were made, and 100 L of each dilution
was plated onto 50 g/L horse blood agar (Oxoid, UK).
These were incubated in gas jars under microaerophilic

conditions using CampyPak (Oxoid, UK) for 3 d, and
colonies were counted (colony forming units per milliliter,
CFU/mL). Plant extracts that killed 100% of H pylori
cells at 60 min (i.e. no colonies grew) were further tested
at 0-, 15-, 30-, and 60-min intervals. All experiments were
performed three times. The effectiveness of the plants at
killing H pylori was expressed as percentage inhibition of
colony growth (i.e. percentage of bacteria killed) compared
to the control.
Microscopic examination
In order to validate the inhibition of adhesion by the plant
extracts, any lytic effect of the extracts on the bacterial
cells was first examined. FITC-labelled and non-labelled
bacteria (1108 cells/mL) were centrifuged (9 000 g for
3 min), re-suspended in 500 L of undiluted plant extract
(100 mg/mL) and incubated with continuous shaking for
1 h at room temperature in the dark. After incubation,
bacteria were centrifuged, washed in PBST and resuspended in 500 L sterile distilled water. One drop of
the bacterial suspension was Gram stained. Using a Zeiss
light microscope, the slides were observed for bacterial
lysis. All the plant extracts were examined, except long
pepper and threadstem carpetweed, which were not tested
due to their limited supply. The negative control consisted
of H pylori incubated with sterile distilled water and the
positive control of H pylori incubated with Puregene PCR
7502
ISSN 1007-9327
CN 14-1219/ R
Table 2 Bactericidal activity of plants against Helicobacter pylori, determined using viable colony count. % inhibition of growth indicates the
percentage of bacteria that were killed by the plants, (i.e. the colonies did not grow), compared to the control.
Plant name
Turmeric
NCTC strain
H pylori clinical strains

3
4
6
100
% inhibition of growth
100
100
100
100
100
100
Minimum of time (min)
15
15
15
15
15
15
15
Ginger
100
100
100
100
100
100
100
30
30
30
30
30
30
30
Fenugreek
60
60
60
60
60
60
60
Cumin (small cumin)

Coriander
Chilli
Bengal quince
Columbo weed
100
100
100
100
100
100
100
30
30
30
30
30
30
30
60
60
60
60
60
60
60
100
100
100
100
100
100
100
30
30
30
30
30
30
30
60
60
60
60
60
60
60
99.9
99.9
99.9
99.5
99.8
99.9
99.8
60
60
60
60
60
60
60
99.9
99.7
99.5
99.9
99.9
99.9
99.9
Yellow-berried nightshade
60
60
60
60
60
60
60
Liquorice
100
100
100
100
100
100
100
60
60
60
60
60
60
30
Nightshade
60
60
60
60
60
60
60
99.9
99.9
100
99.3
99.7
99.9
99.9
60
60
60
60
60
60
60
99.9
99.7
99.9
99.9
99.9
99.9
99.9
60
Long pepper
Threadstem carpetweed
60
60
60
60
60
60
Garlic
60
60
60
60
60
60
60
Parsley
Dill
Sage
Oregano
99.6
99.3
99.9
99.6
99.9
99.99
99.8
60
60
60
60
60
60
60
60
60
60
60
60
60
60
99.9
99.9
99.9
99.7
99.5
99.9
99.9
60
60
60
60
60
60
60
100
100
100
100
100
100
100

Tarragon
Nutmeg
Black pepper
Cinnamon
30
30
30
30
60
30
30
96.9
99.4
99.2
99.9
99.9
99.9
99.9
60
60
60
60
60
60
60
95.9
95.6
99.5
99.4
99.2
96.1
98.9
60
60
60
60
60
60
60
60
60
60
60
60
60
60
100
100
100
100
100
99.9
99.9
60
30
30
30
60
60
60
Black caraway (Europe)/ % inhibition of growth

Black cumin (USA)
100
100
100
100
100
100
100
60
60
60
60
60
60
60
Borage (Starflower)
100
100
100
100
100
100
100
60
60
60
60
60
60
60
Black tea
60
60
60
60
60
60
60
cell lysis solution (Flowgen, UK). The lysis experiments

were performed twice.
Adhesion and adhesion-inhibition assay
Tissue sections were incubated for 30 min in a humidified
atmosphere with 200 L of blocking buffer (PBST+20

g/L BSA) per slide. Next, the sections were washed
three times, by placing in a slide rack in a glass trough
containing 350 mL of PBST, on a rotator for 10 min.
Suspensions of FITC-H pylori were centrifuged and re-
7503
Figure 1 Bright-field images showing H pylori lysis experiment. A: negative control

(H pylori + distilled water); B: positive control (H pylori + cell lysis solution); C:
H pylori + 0.05 g/mL turmeric; D: H pylori + 0.05 g/mL borage; E: H pylorii +
0.05 g/mL parsley. Images taken using x100 objective.
suspended in plant extract to give 1108 bacteria/mL.

This was incubated with continuous shaking for 2 h, and
then washed with the blocking buffer. For the controls,
bacteria were re-suspended in sterile distilled water. The
bacterial suspension (200 L) was then added to each
of the stomach sections and incubated in a humidified
atmosphere for 1 h. After incubation, slides were washed
in PBST as previously, and 200 L of propidium iodide
(PI, 5 g/mL, Sigma, UK) was added and each slide was
incubated for 3 min. Slides were then washed twice in
PBST, air-dried and mounted with Vectashield (V-1000,
Vector Laboratories, UK). All incubation and washing
steps were carried out in the dark at room temperature.
Two tissue sections were used for each plant that was
tested.
A l l p l a n t e x t r a c t s we r e i n i t i a l l y s c r e e n e d f o r
inhibition of adhesion properties at a concentration of
50 mg/mL (diluted in sterile distilled water) and incubated
with H pylori strain 11 637 and Le b stomach sections.
For those plants which had an anti-adhesive effect, the
inhibition assay was repeated using Le a and Le b stomach
sections, and the plants were diluted to concentrations of 5,
10, and 50 mg/mL. Because 50 mg/mL was found to give
Figure 2 Confocal images showing inhibition of H pylori adhesion by A: distilled

water (control); B: turmeric (0.05 g/mL); C: borage (0.05 g/mL) and D: parsley (0.05
g/mL); to stomach sections expressing the Lewis b blood group antigen.
a good inhibition of adhesion, for all further inhibition

experiments, all plants were used at this concentration.
Inhibition of adhesion experiments were then performed
on three clinical strains. All experiments were performed
three times.
Quantification of binding
Sections were observed using a Laser Scanning Confocal
Microscope (Zeiss: Axiovert 100 TV) with a 20 Zeiss
LD-ACHR objective. The excitation wavelengths were 488
nm (for FITC) and 568 nm (for PI). We used 522- and
605-nm band-pass filters to acquire FITC emission and PI
emission, respectively. Digital images (512512 pixels) of
the sections were captured using a Biorad Lasersharp 2000
Confocal Laser Scanning System. Images were converted
to TIF files for processing. Two photographs showing
adjacent areas of the tissue were taken for each tissue
section. Using these digital images, the number of adherent
H pylori were quantified as previously reported (Region of
Interest method with standard area method of counting)
using Metamorph Image analysis software (version 4.5r,
Universal Imaging Corporation, USA)[23]. Using the MannWhitney (non-parametric) test, the difference between the
number of adherent cells for each plant was compared
statistically to the controls (0 g/mL plant extract).
RESULTS
Bactericidal properties of plants
Among the 25 plants tested for their bactericidal activity
against H pylori, eight showed no activity after 60 min of
incubation, these were Bengal quince, nightshade, garlic,
dill, black pepper, coriander, fenugreek and tea (data
7504
ISSN 1007-9327
CN 14-1219/ R
100
% Inhibition
75
50
25
Turmeric
Borage
Parsley
Figure 3 Inhibition of H pylori adhesion by turmeric, borage and parsley to

stomach sections expressing either the Lewis a or Lewis b blood group antigen.
Experiments were performed three times using four strains of H pylori. Mean
percentage of inhibition is shown for each plant. Gray bar, Lewis a stomach; Black
bar, Lewis b stomach.
not shown). The other 17 plants had activity (results are

summarized in Table 2). Turmeric was the most efficient in
killing the seven strains within 15 min. Ginger, cumin, and
chilli were able to kill all the strains within 30 min, whereas
liquorice, oregano, black caraway, and borage killed the
strains within 60 min. Cinnamon had a bactericidal activity
against H pylori, although the seven bacterial strains
differed in their sensitivity. Moreover, the bacterial colonies
were very small, reflecting some resistance to cinnamon.
Finally, the following plants showed a bactericidal
activity against H pylori but without achieving a complete
inhibition of growth (i.e., killing all the colonies) within
60 min: columbo weed, yellow-berried nightshade, long
pepper, threadstem carpetweed, sage, tarragon, nutmeg,
and parsley. Small colonies were seen with yellow-berried
nightshade, threadstem carpetweed, and sage.
Morphological studies
None of the 23 plant extracts that were tested caused
lysis of H pylori cells, as the organisms that remained were
spiral and intact like the negative control. Bacterial cells
incubated with Puregene PCR cell lysis solution were lysed
and no intact spiral shaped bacteria could be observed
(Figure 1).
Anti-adhesive properties of plants
Of the 23 plants screened for anti-adhesive properties,
turmeric, borage, and fresh parsley (50 mg/mL) were
found to inhibit adhesion of H pylori 11637 to the stomach
sections expressing the Le b antigen (Figure 2). Further
testing of these three plant extracts using H pylori 11637
and three clinical isolates confirmed that they were able to
inhibit H pylori adhesion to stomach sections expressing
either the Le b antigen or the Le a antigen (Figure 3).
Mean inhibition rates of the four strains to the stomach
sections expressing the Le a antigen were 61.9%, 61.1%,
and 33.9% for turmeric, borage and parsley, respectively.
The differences between the number of adherent
bacteria compared to the controls (0 g/mL) were: highly
significant (P<0.01) for turmeric; not quite significant
(P = 0.07) for borage; and not significant (P>0.05) for
parsley. For sections expressing the Le b antigen, inhibition

rates were 62.3%, 59.5%, and 48.5% for turmeric, borage
and parsley, respectively. The differences between the
number of adherent bacteria compared to the control
(0 g/mL) were: not quite significant (P = 0.07) for
tur meric; significant (P<0.05) for borage; and not
significant (P>0.05) for parsley. There was no significant
difference between the number of adherent bacteria (to
Le a or Le b stomach sections) when comparing each of
the plant-treatments, except for adhesion of bacteria to Le
a sections where there was a highly significant difference
(P<0.001) between turmeric and parsley.
DISCUSSION
Investigations into plant materials as alternative sources of
antimicrobials have become more common over the past
few years, due to the increased rate of development of
antibiotic resistant organisms. New strategies to combat
infection are also being sought and one such strategy is the
use of anti-adhesive molecules, targeting the primary step
of infection - adhesion of the organism to host tissue[5]. In
this study, we therefore examined both the anti-microbial
and anti-adhesive properties of 25 plant extracts against
H pylori, obtained simply by boiling, as would occur during
the normal cooking process.
Of the eight plants that were not able to kill H pylori
after 60 min of incubation (Bengal quince, nightshade,
garlic, dill, black pepper, coriander, fenugreek, and black
tea), two had been tested previously against H pylori.
Several studies have shown that garlic does kill H pylori
in vitro and in vivo [8,9]. It is probably due to the boiling
method of extraction that this effect was not observed
in this study, since boiling has been shown to reduce the
inhibitory activity of garlic against H pylori[24,25]. Fenugreek
sprouts have been shown to have high anti-microbial
activity against H pylori[26]. In the present study, fenugreek
appeared inactive but the seeds were examined rather than
sprouts, which may account for the difference in results.
Green tea catechins have previously been reported to have
antibacterial effect against H pylori, which was confirmed
in Mongolian gerbils[27]. Moreover, Chinese tea has been
shown to reduce H pylori infection in patients[28]. In this
study, black tea was tested, but no bactericidal activity
against H pylori was observed.
Columbo weed, long pepper, parsley, tarragon, nutmeg,
yellow-berried nightshade, threadstem carpetweed, sage,
and cinnamon had bactericidal activity against H pylori, but
total inhibition of growth was not achieved within 60 min.
Moreover, a reduced colony size was observed with some
of them, such as yellow-berried nightshade, threadstem
carpetweed, sage, and cinnamon. Nutmeg[29], tarragon[30],
and cinnamon[31] have previously been reported to inhibit
the growth of H pylori in vitro. When tested in clinical
trials, however, cinnamon was found to be ineffective at
eradicating H pylori[32].
Among the plants that killed H pylori, turmeric was
the most efficient, followed by cumin, ginger, and chilli.
These results confirmed what has been published before
regarding turmeric[33], chilli[34], and ginger[35], but cumin has

not been previously tested. Other plants, such as borage,
black caraway, and oregano, which have never been tested
before against H pylori, were also found to have bactericidal
effects. Finally, the previous reported bactericidal effect of
liquorice has been confirmed[36,37].
As well as looking at their bactericidal activity, all these
plants were also tested for their anti-adhesive effects, using
an in situ adhesion assay. Stomach tissues expressing either
the Le a or Le b antigen were used. This was in order to
determine whether the plant extracts inhibit adhesion by
blocking the major H pylori adhesin BabA, which binds
to Le b, or have an effect on other adhesins. Most studies
have not attempted to determine which adhesin is being
blocked, and this may be important because combinations
of plants targeting several different adhesins may provide
a more potent therapeutic treatment.
Extracts of turmeric, borage, and parsley were able to
inhibit the adhesion of H pylori strains to both Le a and
Le b stomach sections. Turmeric was the most effective at
inhibiting H pylori adhesion to both Le a and Le b stomach
sections, followed by borage and fresh parsley. From these
results, it seems that both turmeric and borage could
inhibit adhesins other than BabA, because the percentage
of inhibition was almost exactly the same for Le a and Le
b stomach sections. Parsley possibly inhibited BabA as well
as other adhesins, because inhibition occurred on Le a and
Le b stomach sections, but was higher for Le b stomach
sections, indicating that it had a greater effect at inhibiting
BabA. The morphological studies demonstrated that the
reduction in adhesion caused by these plant extracts was
due to the true inhibition of adhesion, but not because
the plants caused the bacteria to lyse. Tarragon is the
only plant in this study that has been tested previously
for its anti-adhesive effects, where it was found to inhibit
H pylori adhesion to human gastric adenocarcinoma
cells[22]. However, in the present study, tarragon failed to
inhibit H pylori adhesion to human stomach sections. This
discrepancy is probably due to the differences between
using cell-lines and whole-tissue.
Plants contain multiple organic components including
phenols, quinones, flavones, tannins, terpenoids, and
alkaloids all of which are known to have bactericidal
effects [7]. These substances are also water-soluble and
therefore very likely to be present in the plant extracts
produced in this study, and are likely candidates
responsible for the killing effect of the extracts on H pylori.
Plants also contain many water-soluble proteins, lectins
and carbohydrates which may bind specifically to sugar
residues, polysaccharides, glycoproteins or glycolipids, such
as the adhesins present on the cell surface of H pylori. In
the event of such interactions occurring, the result would
be to block the availability of the adhesin to its receptor
and hence prevent adhesion of the bacterium to the
stomach sections.
Despite the success of many plant materials at killing
organisms, resistance can develop to these too, as shown
in this study. The present work is of importance because
plant extracts were shown to inhibit adhesion of H pylori
7505
to the stomach - an alternative strategy to bactericidal

compounds. Because inhibition of adhesion works on the
principle of sterically blocking bacteria from attaching
to host tissue, the likelihood of resistance developing in
organisms, which occurs when the organism is killed, is
less likely.
Although the majority of individuals colonized by
H pylori are asymptomatic, a proportion of patients develop
peptic ulcers (duodenal and gastric), and an even smaller
proportion develop gastric cancer. Globally, H pylori is the
major cause of gastric cancer and has been classified as a
Class I carcinogen by the WHO. Some plants are known
to have anti-ulcerogenic and anti-cancer effects. Most of
the plants tested here, which have bactericidal and/or
anti-adhesive properties, also have anti-ulcerogenic or
anti-cancer effects, for example, turmeric[38,39], ginger[40,41],
cumin[42,43], borage[44], liquorice[45], and parsley[46]. Chilli has
recently been shown to have detrimental effects on the
gastric mucosa[47]. The anti-cancer and anti-ulcerogenic
effects of ginger, cumin, liquorice, parsley, turmeric, and
borage combined with their bactericidal and anti-adhesive
properties, as shown in this study, suggests that ingestion
of these six plants could have important therapeutic
implications for patients with H pylori-induced peptic
ulcer disease or gastric cancer. As turmeric killed 100%
of organisms within 15 min, it could therefore be a useful
anti-Helicobacter agent in vivo, because, if given orally, it
would be able to kill H pylori despite the short amount of
time that it would remain in the stomach during digestion.
Moreover, these plants could be used in combination with
antibiotics, possibly increasing the success of eradication,
as has been shown in vitro for cranberry juice[48].
Most studies have used plant extracts that are obtained
by chemical processing, for example, ethanol extracts.
In developing countries where antibiotics are less freely
available, such processing methods of plants would
be impractical and/or expensive. This present study is
therefore of importance because it has demonstrated that
several plant extracts are effective against H pylori and are
obtained simply by boiling the plants. Such a method may
provide a treatment that is simple, relatively inexpensive
and could be incorporated into the normal diet of the
patient, which is highly favorable. Herbal treatments are
usually unregulated and the safety of plants consumed is
often unknown. The plants used and shown to be effective
in this study are already commonly consumed, and thus
they are already known to be safe.
Although the cidal and anti-adhesive effects of ginger,
cumin, liquorice, parsley, turmeric, and borage have been
shown in vitro, further studies are needed to be carried
out to investigate their effects in vivo, to see whether the
extracts are able to remain effective despite the harsh
process of digestion.
ACKNOWLEDGMENTS
We are grateful to Martin Avedian for the idea of testing
borage and for its supply and Ms A. Al-Dossary (School
of Hygiene and Tropical Medicine, UK) for providing
black caraway.
7506
ISSN 1007-9327
CN 14-1219/ R
REFERENCES
1
2
4
5
6
7
8
10
11
12
13
14
15
16
17
18
19
20
21
22
Achtman M and Suerbaum S. Helicobacter pylori: Molecular

and Cellular Biology. Horizon scientific, Norfolk 2001; 61
Montecucco C, Rappuoli R. Living dangerously: how Helicobacter pylori survives in the human stomach. Nat Rev Mol Cell
Biol 2001; 2: 457-466
Basset C, Holton J, Gatta L, Ricci C, Bernabucci V, Liuzzi G,
Vaira D. Helicobacter pylori infection: anything new should we
know? Aliment Pharmacol Ther 2004; 20 Suppl 2: 31-41
Megraud F. H pylori antibiotic resistance: prevalence, importance, and advances in testing. Gut 2004; 53: 1374-1384
Basset C, Holton J, OMahony R, Roitt I. Innate immunity and
pathogen-host interaction. Vaccine 2003; 21 Suppl 2: S12-S23
Lampe JW. Health effects of vegetables and fruit: assessing
mechanisms of action in human experimental studies. Am J
Clin Nutr 1999; 70: 475S-490S
Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999; 12: 564-582
Jonkers D, van den Broek E, van Dooren I, Thijs C, Dorant E,
Hageman G, Stobberingh E. Antibacterial effect of garlic and
omeprazole on Helicobacter pylori. J Antimicrob Chemother
1999; 43: 837-839
Iimuro M, Shibata H, Kawamori T, Matsumoto T, Arakawa
T, Sugimura T, Wakabayashi K. Suppressive effects of garlic
extract on Helicobacter pylori-induced gastritis in Mongolian
gerbils. Cancer Lett 2002; 187: 61-68
Matsubara S, Shibata H, Ishikawa F, Yokokura T, Takahashi
M, Sugimura T, Wakabayashi K. Suppression of Helicobacter
pylori-induced gastritis by green tea extract in Mongolian gerbils. Biochem Biophys Res Commun 2003; 310: 715-719
al Somal N, Coley KE, Molan PC, Hancock BM. Susceptibility
of Helicobacter pylori to the antibacterial activity of manuka
honey. J R Soc Med 1994; 87: 9-12
Tabak M, Armon R, Potasman I, Neeman I. In vitro inhibition
of Helicobacter pylori by extracts of thyme. J Appl Bacteriol
1996; 80: 667-672
Nariman F, Eftekhar F, Habibi Z, Falsafi T. Anti-Helicobacter
pylori activities of six Iranian plants. Helicobacter 2004; 9:
146-151
Martin KW, Ernst E. Herbal medicines for treatment of bacterial infections: a review of controlled clinical trials. J Antimicrob
Chemother 2003; 51: 241-246
Galan MV, Kishan AA, Silverman AL. Oral broccoli sprouts
for the treatment of Helicobacter pylori infection: a preliminary report. Dig Dis Sci 2004; 49: 1088-1090
Ofek I, Hasty DL, Sharon N. Anti-adhesion therapy of bacterial diseases: prospects and problems. FEMS Immunol Med
Microbiol 2003; 38: 181-191
Burger O, Ofek I, Tabak M, Weiss EI, Sharon N, Neeman I.
A high molecular mass constituent of cranberry juice inhibits
helicobacter pylori adhesion to human gastric mucus. FEMS
Immunol Med Microbiol 2000; 29: 295-301
Shibata H, KimuraTakagi I, Nagaoka M, Hashimoto S, Sawada H, Ueyama S, Yokokura T. Inhibitory effect of Cladosiphon
fucoidan on the adhesion of Helicobacter pylori to human gastric cells. J Nutr Sci Vitaminol (Tokyo) 1999; 45: 325-336
Shibata H, Iimuro M, Uchiya N, Kawamori T, Nagaoka M,
Ueyama S, Hashimoto S, Yokokura T, Sugimura T, Wakabayashi K. Preventive effects of Cladosiphon fucoidan against
Helicobacter pylori infection in Mongolian gerbils. Helicobacter
2003; 8: 59-65
Lengsfeld C, Titgemeyer F, Faller G, Hensel A. Glycosylated compounds from okra inhibit adhesion of Helicobacter
pylori to human gastric mucosa. J Agric Food Chem 2004; 52:
1495-1503
Lengsfeld C, Deters A, Faller G, Hensel A. High molecular
weight polysaccharides from black currant seeds inhibit adhesion of Helicobacter pylori to human gastric mucosa. Planta
Med 2004; 70: 620-626
Lee JH, Park EK, Uhm CS, Chung MS, Kim KH. Inhibition of
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
Helicobacter pylori adhesion to human gastric adenocarcinoma epithelial cells by acidic polysaccharides from Artemisia
capillaris and Panax ginseng. Planta Med 2004; 70: 615-619
OMahony R, Basset C, Holton J, Vaira D, Roitt I. Comparison
of image analysis software packages in the assessment of adhesion of microorganisms to mucosal epithelium using confocal laser scanning microscopy. J Microbiol Methods 2005; 61:
105-126
Canizares P, Gracia I, Gomez LA, Garcia A, Martin De Argila C,
Boixeda D, de Rafael L. Thermal degradation of allicin in garlic extracts and its implication on the inhibition of the in-vitro
growth of Helicobacter pylori. Biotechnol Prog 2004; 20: 32-37
Canizares P, Gracia I, Gomez LA, Martin de Argila C, Boixeda
D, Garcia A, de Rafael L. Allyl-thiosulfinates, the bacteriostatic
compounds of garlic against Helicobacter pylori. Biotechnol
Prog 2004; 20: 397-401
Randhir R, Lin YT, Shetty K. Phenolics, their antioxidant and
antimicrobial activity in dark germinated fenugreek sprouts
in response to peptide and phytochemical elicitors. Asia Pac J
Clin Nutr 2004; 13: 295-307
Takabayashi F, Harada N, Yamada M, Murohisa B, Oguni I.
Inhibitory effect of green tea catechins in combination with sucralfate on Helicobacter pylori infection in Mongolian gerbils.
J Gastroenterol 2004; 39: 61-63
Yee YK, Koo MW, Szeto ML. Chinese tea consumption and
lower risk of Helicobacter infection. J Gastroenterol Hepatol
2002; 17: 552-555
Bhamarapravati S, Pendland SL, Mahady GB. Extracts of spice
and food plants from Thai traditional medicine inhibit the
growth of the human carcinogen Helicobacter pylori. In Vivo
2003; 17: 541-544
Wang YC, Huang TL. Screening of anti-Helicobacter pylori
herbs deriving from Taiwanese folk medicinal plants. FEMS
Immunol Med Microbiol 2005; 43: 295-300
Tabak M, Armon R, Neeman I. Cinnamon extracts inhibitory effect on Helicobacter pylori. J Ethnopharmacol 1999; 67:
269-277
Nir Y, Potasman I, Stermer E, Tabak M, Neeman I. Controlled
trial of the effect of cinnamon extract on Helicobacter pylori.
Helicobacter 2000; 5: 94-97
Mahady GB, Pendland SL, Yun G, Lu ZZ. Turmeric (Curcuma
longa) and curcumin inhibit the growth of Helicobacter pylori,
a group 1 carcinogen. Anticancer Res 2002; 22: 4179-4181
Jones NL, Shabib S, Sherman PM. Capsaicin as an inhibitor of
the growth of the gastric pathogen Helicobacter pylori. FEMS
Microbiol Lett 1997; 146: 223-227
Mahady GB, Pendland SL, Yun GS, Lu ZZ, Stoia A. Ginger
(Zingiber officinale Roscoe) and the gingerols inhibit the
growth of Cag A+ strains of Helicobacter pylori. Anticancer
Res 2003; 23: 3699-3702
Fukai T, Marumo A, Kaitou K, Kanda T, Terada S, Nomura T.
Anti-Helicobacter pylori flavonoids from licorice extract. Life
Sci 2002; 71: 1449-1463
Krausse R, Bielenberg J, Blaschek W, Ullmann U. In vitro antiHelicobacter pylori activity of Extractum liquiritiae, glycyrrhizin and its metabolites. J Antimicrob Chemother 2004; 54:
243-246
Azuine MA, Bhide SV. Chemopreventive effect of turmeric
against stomach and skin tumors induced by chemical carcinogens in Swiss mice. Nutr Cancer 1992; 17: 77-83
Huang MT, Lou YR, Ma W, Newmark HL, Reuhl KR, Conney
AH. Inhibitory effects of dietary curcumin on forestomach,
duodenal, and colon carcinogenesis in mice. Cancer Res 1994;
54: 5841-5847
Yamahara J, Mochizuki M, Rong HQ, Matsuda H, Fujimura H.
The anti-ulcer effect in rats of ginger constituents. J Ethnopharmacol 1988; 23: 299-304
Yoshikawa M, Yamaguchi S, Kunimi K, Matsuda H, Okuno Y,
Yamahara J, Murakami N. Stomachic principles in ginger. III.
An anti-ulcer principle, 6-gingesulfonic acid, and three monoacyldigalactosylglycerols, gingerglycolipids A, B, and C, from
42
43
44
45
Zingiberis Rhizoma originating in Taiwan. Chem Pharm Bull

(Tokyo) 1994; 42: 1226-1230
Aruna K, Sivaramakrishnan VM. Anticarcinogenic effects of
some Indian plant products. Food Chem Toxicol 1992; 30: 953-956
Gagandeep, Dhanalakshmi S, Mendiz E, Rao AR, Kale RK.
Chemopreventive effects of Cuminum cyminum in chemically
induced forestomach and uterine cervix tumors in murine
model systems. Nutr Cancer 2003; 47: 171-180
Gonzalez CA, Sanz JM, Marcos G, Pita S, Brullet E, Saigi E,
Badia A, Agudo A, Riboli E. Borage consumption as a possible
gastric cancer protective factor. Cancer Epidemiol Biomarkers
Prev 1993; 2: 157-158
Dehpour AR, Zolfaghari ME, Samadian T, Vahedi Y. The
protective effect of liquorice components and their derivatives
against gastric ulcer induced by aspirin in rats. J Pharm Phar-
7507
46
47
48
macol 1994; 46: 148-149

Al-Howiriny T, Al-Sohaibani M, El-Tahir K, Rafatullah S.
Prevention of experimentally-induced gastric ulcers in rats by
an ethanolic extract of Parsley Petroselinum crispum. Am J
Chin Med 2003; 31: 699-711
Lopez-Carrillo L, Lopez-Cervantes M, Robles-Diaz G,
Ramirez-Espitia A, Mohar-Betancourt A, Meneses-Garcia A,
Lopez-Vidal Y, Blair A. Capsaicin consumption, Helicobacter
pylori positivity and gastric cancer in Mexico. Int J Cancer
2003; 106: 277-282
Shmuely H, Burger O, Neeman I, Yahav J, Samra Z, Niv Y,
Sharon N, Weiss E, Athamna A, Tabak M, Ofek I. Susceptibility of Helicobacter pylori isolates to the antiadhesion activity
of a high-molecular-weight constituent of cranberry. Diagn
Microbiol Infect Dis 2004; 50: 231-235
Science Editor Kumar M Language Editor Elsevier HK

7499

Caricato da

Informazioni sul documento

Copyright

Formati disponibili

Condividi questo documento

Condividi o incorpora il documento

Opzioni di condivisione

Hai trovato utile questo documento?

Questo contenuto è inappropriato?

Copyright:

Formati disponibili

7499

Caricato da

Copyright:

Formati disponibili

PO Box 2345, Beijing 100023, China

World J Gastroenterol 2005;11(47):7499-7507

Bactericidal and anti-adhesive properties of culinary and

CONCLUSION: Several plants that were tested in our

Key words: H pylori ; Inhibition; Adhesion; Killing; Plants

Resistance to antibiotics is not limited to H pylori and

December 21, 2005 Volume 11 Number 47

Lanka, Iran, and the Middle East. Moreover, we have used

MATERIALS AND METHODS

OMahony R et al. Plants against H pylori

Table 1 Plants used in the study

Sri Lankan name

Part of the plant used Source

In cooking & medicine

Black caraway (Europe)/Black

In cooking & medicine

In cooking & medicine

Leaves and shoots

In cooking & medicine

In cooking & medicine

In cooking & medicine

Cumin (small cumin)

In cooking & medicine

In cooking & medicine

In cooking & medicine

In cooking & medicine

In cooking & medicine

Glycyrrhiza glabra apofosa

In cooking & medicine

In cooking & medicine

Fruit and root

In cooking & medicine

In cooking & medicine

In cooking & medicine

In cooking & medicine

In cooking & medicine

In cooking & medicine

Katu val batu

These plants were bought as both fresh and dried material.

sets of formalin-fixed stomach biopsies were used: those

These were incubated in gas jars under microaerophilic

December 21, 2005 Volume 11 Number 47

H pylori clinical strains

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Cumin (small cumin)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Minimum of time (min)

Black caraway (Europe)/ % inhibition of growth

Minimum of time (min)