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In vitro controlling of selected human diarrhea causing bacteria by clove extracts (Syzygium aromaticum L.)

  • Rural Development Administration, Cheonan, Republic of Korea

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Antibacterial activity of clove extracts (Syzygium aromaticum L.) was proven against five diarrhea causing bacteria. This was further confirmed when compared with commonly used three commercial antibiotics (ciprofloxacin, tetracycline and erythromycin) as a positive control. Significant differences (P<0.0001) were observed in the effect of the antimicrobial agents (clove extracts and antibiotics), and in the sensitivities of the bacterial species (P<0.0001) to the antimicrobial agents. Clove extracts had significant (P<0.001) activity with the acetone extract demonstrating highest activity followed by antibiotics and other extracts against tested bacteria. The zone of inhibition of clove extracts was ranged from 7.33 to 12.00 mm whereas in antibiotics, it was 0.00 to 11.67 mm. Of all the bacteria, Salmonella typhimurium was the most susceptible against all of the extracts as well as concentrations of clove, while low MIC (180 mgml-1) and MBC (680 mgml-1) of the extracts were observed against Shigella dysenteriae. Consequently, clove has a significant antidiarrheal activity and it could be used as an effective antibacterial agent, alternative to the use of antibiotics.
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Rahman et al.
Int. J. Biomol. & Biomed.
In vitro
controlling of selected human diarrhea causing bacteria
by clove extracts (
Syzygium aromaticum
M Mostafizur Rahman1, M Atikur Rahman2, M Soriful Islam3, M Firoz Alam1*
1Biotechnology and Microbiology Laboratory, Department of Botany, Rajshahi University, Rajshahi
6205, Bangladesh
2Forage Biotechnology Laboratory, Department of Animal Bioscience, Division of Applied Life Science,
Gyeongsang National University, Jinju 600-701, South Korea
3Department of Molecular Pathology and Innovative Therapies, Anatomy and Cell Biology, Polytechnic
University of Marche, Via Tronto 10/A, 60020 Ancona,
Received: 20 June 2011
Revised: 22 July 2011
Accepted: 22 July 2011
Key words: Diarrhea, Syzygium aromaticum, antibiotics, antibacterial activity.
Antibacterial activity of clove extracts (Syzygium aromaticum L.) was proven against five diarrhea causing bacteria.
This was further confirmed when compared with commonly used three commercial antibiotics (ciprofloxacin,
tetracycline and erythromycin) as a positive control. Significant differences (P<0.0001) were observed in the effect of
the antimicrobial agents (clove extracts and antibiotics), and in the sensitivities of the bacterial species (P<0.0001) to
the antimicrobial agents. Clove extracts had significant (P<0.001) activity with the acetone extract demonstrating
highest activity followed by antibiotics and other extracts against tested bacteria. The zone of inhibition of clove
extracts was ranged from 7.33 to 12.00 mm whereas in antibiotics, it was 0.00 to 11.67 mm. Of all the bacteria,
Salmonella typhimurium was the most susceptible against all of the extracts as well as concentrations of clove, while
low MIC (180 mgml-1) and MBC (680 mgml-1) of the extracts were observed against Shigella dysenteriae.
Consequently, clove has a significant antidiarrheal activity and it could be used as an effective antibacterial agent,
alternative to the use of antibiotics.
*Corresponding Author: M Firoz Alam
International Journal of Biomolecules and Biomedicine (IJBB)
ISSN: 2221-1063 (Print) 2222-503X (Online)
Vol. 1, No.2, p. 17-26, 2011
Rahman et al.
Int. J. Biomol. & Biomed.
In developing countries diarrhea is the most common
causes of morbidity and mortality (Amstrong and
Cohen, 1999) and it caused several million of deaths in
the world annually (Field, 2003). Many bacteria, virus
and protozoa have been isolated from diarrhea
patients, especially Salmonella typhimurium,
Escherichia coli, Shigella dysenteriae, Proteus
mirabilis (Prescott et al., 2005; Eja et al., 2007),
Yersinia enterocolitica (Okwori et al., 2007), Vibrio
cholera (Zuckerman et al., 2007), Campylobacter
jejuni, Clostridium difficile (Prescott et al., 2005) etc.
bacteria are responsible for acute and chronic diarrhea.
Antibiotics are the essential part for combating
harmful bacterial infections in vivo (Kaushik and
Goyel, 2008), but repeated and improper uses of
antibiotics resulting drug-resistant bacteria. To
overcome this problem an alternative therapy is very
much needed and researchers are looking for
developing alternative strategies (Sivam et al., 1997).
The World Health Organization (WHO) has included a
programme for the control of diarrhea, which involves
the use of traditional herbal medicine (Snyder and
Merson, 1982). Various herbs and spices have been
recognized by their medicinal value used as an
alternative antimicrobial agent to antibiotics, and
several plants have been reported to be used in treating
and managing diarrhea diseases (Agunu et al., 2005).
Cloves (Syzygium aromaticum L.) are the aromatic
dried flower buds, and several studies have
demonstrated on potent antibacterial effects of clove
(Cai and Wu, 1996; Bae et al., 1998; Li et al., 2005; Fu
et al., 2007). The present study was undertaken for in
vitro controlling of human diarrhea causing bacteria
by clove extracts using agar disc diffusion assay.
Materials and methods
Collection of plants and Antibiotics
Locally available flower buds of Syzygium aromaticum
L. (Common name: Clove, Family: Myrtaceae) and
three commercial antibiotics namely ciprofloxacin,
tetracycline and erythromycin (Beximco
Pharmaceuticals Ltd., Bangladesh) were used during
antibacterial study.
Collection of bacterial isolates
Five bacteria with the accession number Salmonella
typhimurium BMLRU1021, Escherichia coli
BMLRU1023, Shigella dysenteriae BMLRU1025,
Proteus mirabilis BMLRU1027 and Yersinia
enterocolitica BMLRU1029 were used in this study.
These bacterial strains were isolated and identified
from stool and urine samples (diarrhea associated)
according to Holt et al. (1994) using their respective
standard strain (collected from ICDDRB, Dhaka,
Bangladesh) in Biotechnology and Microbiology
laboratory, Department of Botany, University of
Rajshahi, Bangladesh.
Preparation of crude extracts
Collected clove flower buds were dried for 3 days in
oven under 60oC then crushed into fine powder using
mortar, pestle and electric blender (Nokia, Osaka-
Japan). Ten-gram dried powder of clove was dipped
into 100ml of different organic solvents (methanol,
ethanol and acetone) separately into a conical flask
followed by air tight with rubber corks, and left for 2
days on orbital shaking (IKA Labortechnik KS 250
Basic Orbital Shaker, Staufen, Germany). The well
refined solution was filtrated through Teton cloth and
Whatman No. 1 filter paper in a beaker followed by
evaporation of solvent using water bath (4 holes
analogue, Thermostatic water bath, China) until
formation of semisolid extract. Semi solid extracts
were dissolved into respective solvent and preserved in
airtight screw cap tube at 4°C for further use.
Preparation of antibiotics
Antibiotics solution was prepared as described by
Ekwenye and Elegalam (2005). The commercial
antibiotics ciprofloxacin (500 mg), tetracycline (500
mg) and erythromycin (250 mg) were crushed
manually using mortar and pestle. Ciprofloxacin and
tetracycline were dissolved in 10 ml de-ionized distill
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Int. J. Biomol. & Biomed.
water separately, and erythromycin was dissolved in 10
ml ethanol (95%). The solutions were preserved at 4°C
until further use.
Antibacterial assay
In vitro antibacterial activity of clove extracts as well as
antibiotics were tested against five studied bacteria
using agar disc diffusion method (Parekh and Chanda,
2007). Under aseptic conditions, sterilized Whatman
no. 1 filter paper discs (6 mm in diameter) were
impregnated with 10 μl of different solvent extracts
(200, 400, and 600 mgml-1) as well as antibiotics
(0.02, 0.04 and 0.06 mgml-1) followed by air-drying
and placed on seeded nutrient agar plates. 30 µl of
bacterial suspension (108 cfu ml-1) was used for
preparing seeded nutrient agar plates. Negative
controls were prepared using respective solvents. The
Petri-plates were incubated at 37°C for 24h. After
incubation, antibacterial activity was determined by
measuring the zone of inhibition in millimeter scale
against the studied bacteria. Each assay was carried
out in triplicate.
Determination of minimum inhibitory concentration
(MIC) and minimum bactericidal concentration
MIC and MBC of plant extracts were determined
according to Doughari et al. (2007). For MIC
determination, 0.5 ml of varying concentrations of the
extracts (150, 180, 200, 220, 250, 280, 300, 320, 350,
380, 400, 420, 450, 480, 500, 520, 550, 580, 600, 620
and 650 mgml-1) were added with nutrient broth (2 ml)
in test tubes, then a loop-full of the test bacteria (108
cfu ml-1) was introduced. A tube containing nutrient
broth was seeded only with the test bacteria, as
described above, to serve as control. The culture tubes
were incubated at 37oC for 24 h. After incubation, the
tubes were examined for microbial growth by
observing for turbidity.
To determine the MBC, for each set of test tubes in the
MIC determination, a loopful of broth was collected
from those tubes that did not show any growth and
inoculated onto sterile nutrient agar by streaking. All
the plates were then incubated at 37oC for 24 h. After
incubation the concentration at which no visible
growth was seen, noted as MBC.
Statistical analysis
Statistical analysis (ANOVA) was performed using
software SPSS (version 10.0; SPSS Inc., Chicago IL,
USA) and MSTAT (version 2.10; Russell, D. Freed,
Michigan State University, USA) and expressed as
mean ± SEM. Least Significant Difference (LSD) test
was used to speculate further if there was a significant
difference within three clove extracts, three antibiotics,
various concentrations, studied bacteria and
interaction effect between them. P values 0.05 were
considered as significant.
The results reveal that studied three concentrations of
clove extract as well as antibiotics have effective
activity against all tested bacteria (Table-1). In clove
extracts, the zone of inhibition was ranged from 0.00
to 12.00 mm. For 200 mgml-1, it was ranged from 0.00
to 7.83 mm, 0.00 to 9.67 mm for 400 mgml-1, and 7.33
to 12.00 mm for 600 mgml-1. In three type of extracts,
acetone gave the best results (10.00 to 12.00 mm)
followed by methanol (7.50 to 9.83 mm) and ethanol
(7.33 to 9.33 mm) at highest concentration (600 mgml-
1). In case of antibiotics, the zone of inhibition was
ranged from 0.00 to 11.67 mm. For 0.02 mgml-1 it was
ranged from 0.00 to 8.67 mm, 0.00 to 9.67 mm for
0.04 mgml-1, and 0.00 to 11.67 mm for 0.06 mgml-1. In
three type of antibiotics, ciprofloxacin gave best results
(10.5 to 11.67 mm) followed by tetracycline (0.00 to
11.00 mm), while erythromycin did not show any
activity. Statistical results of antibacterial activity of
three clove extracts showed significant differences
(P<0.0001) in efficacy among the bacterial strains (S),
concentrations (C) of clove extracts (E), type of extracts
as well as their interaction cases- S×C, S×E, C×E and
S×C×E (Table 2).
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Int. J. Biomol. & Biomed.
Table 1. Antibacterial activities of three solvents extract of S. aromaticum L. and three antibiotics.
S. typhimurium
Y. enterocolitica
Clove extracts
7.33 ± 0.33
7.33 ± 0.33
7.83 ± 0.441
7.50 ± 0.29
7.00 ± 0.00
7.67 ± 0.33
8.33 ± 0.33
7.50 ± 0.50
9.33 ± 0.33
9.33 ± 0.33
9.67 ± 0.33
8.17 ± 0.44
9.33 ± 0.33
9.33 ± 0.33
7.50 ± 0.29
7.33 ± 0.33
9.33 ± 0.33
8.00 ± 0.58
9.83 ± 0.33
7.67 ± 0.66
7.50 ± 0.29
7.50 ± 0.29
9.33 ± 0.33
11.17 ± 0.17
12.00 ± 0.58
11.17 ± 0.44
10.00 ± 0.58
11.33 ± 0.33
8.67 ± 0.33
7.33 ± 0.33
7.33 ± 0.33
7.33 ± 0.33
7.17 ± 0.33
9.67 ± 0.33
9.00 ± 0.58
9.50 ± 0.29
8.33 ± 0.33
9.33 ± 0.33
8.67 ± 0.33
11.00 ± 0.58
11.67 ± 0.33
11.33 ± 0.33
10.50 ± 0.29
10.67± 0.33
11.00 ± 0.58
ET = Ethanol extract; ME = Methanol extract; AC = Acetone extract; CP = Ciprofloxacin; TC = Tetracycline; ER = Erythromycin. *Data were
representing mean zone of inhibition (mm) ± SEM of three replicates.
According to the LSD test results (Table 3), means of
strain, no significant differences were observed among
E. coli, S. dysenteriae and P. mirabilis. But S.
typhimurium and Y. enterocolitica were significantly
different from others and between themselves. Highest
mean value was found against S. typhimurium (6.926)
and decreasing order of sensitivity of selected species
of bacteria against three extracts was Y. enterocolitica
(5.889) > E. coli (4.87) > S. dysenteriae (4.833) > P.
mirabilis (4.741). In case of mean values of
concentration, increasing the concentration level for
extracts had a significant (P<0.05) inhibitory effect on
all test bacteria. The inhibition area that found is larger
as concentration of extracts is increased and highest
for 600 mgml-1 (9.267) followed by 400 mgml-1 (5.089)
and 200 mgml-1 (2.00). The mean value of extracts
shown acetone (8.767) was significantly different from
methanol (3.844) and ethanol (3.744), while no
differences ware found between methanol and ethanol.
Table 2. Statistical results (ANOVA) of antibacterial
activity of three clove extract.
Source of
Sum of
F Value
Strains (S)
Extracts (E)
Like clove extracts, antibacterial activity of three
antibiotics showed significant differences (P<0.0001)
among the bacterial strains (S), concentrations (C) of
antibiotics, type of antibiotics (A) as well as for
interaction items- S×C, S×A, C×A and S×C×A (Table-
4). Mean separation (Table-5) for antibacterial activity
of antibiotics shows that the mean of strain E. coli
(5.296) in the top and significantly different from
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Int. J. Biomol. & Biomed.
others. Rest of strains shows no differences among
themselves. Here also concentrations of antibiotic were
different from each other. All antibiotics were
significantly different from each other and highest for
ciprofloxacin (9.267) followed by tetracycline (1.311)
and erythromycin (0.00).
Table 3. Analysis of mean data of the antibacterial
activity of three clove extracts.
Growth inhibition diameter (mm)
S. typhimurium
6.926 A
E. coli
4.87 C
S. dysenteriae
4.833 C
P. mirabilis
4.741 C
Y. enterocolitica
5.889 B
200 mgml-1
2.00 C
400 mgml-1
5.089 B
600 mgml-1
9.267 A
3.744 B
3.844 B
8.767 A
Means followed by different letter(s) down the column are
significantly different at P<0.05. Data values are means of
three replicates.
Table 4. Statistical analysis of antibacterial activity of
three antibiotics.
Source of
Sum of
F Value
Strains (S)
Antibiotics (A)
The MIC and MBC results of clove extracts are
presented in Table 6. The results reveal that MIC
values were ranged from 180 (S. dysenteriae) to 620
mgml-1 (E. coli and S. dysenteriae). For ethanol
extract, it was ranged from 400 (P. mirabilis) to 620
mgml-1 (E. coli and S. dysenteriae), 400 (S.
typhimurium) to 600 mgml-1 (E. coli, S. dysenteriae
and P. mirabilis) for methanol, and 180 (S.
dysenteriae) to 400 mgml-1 (P. mirabilis) for acetone.
Table 5. Analysis of mean data of the antibacterial
activity of three antibiotics.
Growth inhibition diameter (mm)
S. typhimurium
3.259 B
E. coli
5.296 A
S. dysenteria
3.13 B
P. mirabilis
2.907 B
Y. enterocolitica
3.037 B
0.02 mgml-1
2.533 C
0.04 mgml-1
3.633 B
0.06 mgml-1
4.431 A
9.267 A
1.311 B
0.00 0 C
Means followed by different letter(s) down the column are
significantly different at P<0.05. Data values are means of three
In three types of extract, acetone extract gave lowest
MIC value (180 mgml-1) against S. dysenteriae
followed by methanol and ethanol (400 mgml-1). In
case of MBC values, it was ranged from 220 to 680
mgml-1 under the same strain (S. dysenteriae). For
ethanol extract, it was ranged from 450 (S.
typhimurium and P. mirabilis) to 680 mgml-1 (S.
dysenteriae), 450 (S. typhimurium) to 650 mgml-1 (E.
coli, S. dysenteriae and P. mirabilis) for methanol and
220 (S. dysenteriae) to 450 mgml-1 (P. mirabilis) for
acetone. In three types of extracts, acetone extract gave
lowest MBC value (220 mgml-1) against S. dysenteriae
followed by methanol and ethanol (450 mgml-1).
Although, the primary purpose of spices is to impart
flavor and piquancy to food, the medicinal,
antimicrobial and antioxidant properties of spices have
also been exploited (Souza et al., 2005). Cloves are
antimutagenic (Miyazawa and Hisama, 2003), anti-
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Int. J. Biomol. & Biomed.
inflammatory (Kim et al., 1998), antioxidant (Chaieb et
al., 2007a), antiulcerogenic (Bae et al., 1998; Li et al.,
2005), antithrombotic (Srivastava and Malhotra, 1991)
and antiparasitic (Yang et al., 2003). On the basis of
this information, antibacterial activities of clove
extracts were evaluated for their antidiarrheal
Table 6. Minimum inhibitory concentration (MIC)
and Minimum bactericidal concentration (MBC) of
three solvent extracts of S. aromaticum L. against
studied bacterial strains.
Concentrations (mgml-1)
ET = Ethanol extract; ME = Methanol extract; AC = Acetone extract
The antimicrobial activity has been attributed to the
presence of some active constituents in the extracts.
Clove contains a high eugenol (70-90%) content (de
Guzman and Siemonsma, 1999) which is an
antibacterial compound having wide spectra of
antimicrobial effects against enterobacteria (Burt and
Reinders, 2003; Nanasombat and Lohasupthawee,
2005; Chaieb et al., 2007b). The results of this study
exemplifies that clove extracts have potential source of
antidiarrheal properties because extracts and their
concentrations have significant influence on the
growth of diarrhea causing bacteria and also it has
superior antibacterial activity than antibiotics. Several
investigators conducted related investigation and
recommend clove extracts as a source of antibacterial
agent (Nascimento et al., 2000; Saeed and Tariq,
2008). It has also been reported that, clove oil potently
inhibited the growth of different Gram negative
bacteria (Saeed and Tariq, 2008; Lopez et al., 2005).
Clove extracts had a high activity against E. coli (12.00
mm) and previous studies have documented that E.
coli are known to be multi-drug resistant bacteria
(Saeed et al., 2007; Singh et al., 2002). The results
were also in accordance with those reported by many
investigators (Mandee et al., 2003; Smith-Palmer et
al., 2001; Dorman and Deans, 2000; Hammer et al.,
1999; De et al., 1999). All the tested bacteria, which
were resistant to erythromycin and tetracycline, but
significantly inhabited by clove extracts. Salman et al.
(2008) also found comparable results. In this
experiment, extracts showed different degrees of
growth inhibition depending upon the bacterial
strains. These variations were found because strains
are genetically different from each other, and this is
probably due to the differences in chemical
composition and structure of the cell wall of both types
of microorganisms (Kaushik and Goyel, 2008),
microbial growth, exposure of micro- organisms to
plant oil, the solubility of oil or oil components and the
use and quantity of an emulsifier (Bansod and Rai,
2008). Increasing of the concentrations level of
extracts had a significant (P<0.05) inhibitory effect on
all studied bacteria. Similarly Tylor et al. (2001)
reported that active compounds may be present in
insufficient quantities in the crude extracts to show
activity with the dose levels employed and lack of
activity can thus only be proven by using large dose
(Farnsworth, 1993). Extracts prepared in acetone
extract gave better activity than that of other extracts,
and it could be better solubility of active components
in acetone. It has been reported that different
phytoconstituents have different degrees of solubility
in different types of solvents depending on their
polarity (El-mahmood and Ameh, 2007). This
observation clearly indicates that the polarity of
antimicrobial compounds make them more readily
extracted by acetone solvent, and using organic solvent
does not negatively affect their bioactivity against
bacterial species (Kaushik and Goyel, 2008). Of all the
bacteria, S. typhimurium was the most susceptible
against all of the extracts and concentrations of clove
while P. mirabilis was the most resistance bacteria. In
Rahman et al.
Int. J. Biomol. & Biomed.
this study the low MIC and MBC values observed for S.
dysenteriae is a good indication of high efficacy against
this bacteria and high MIC and MBC values are
indication of low activity (Doughari et al., 2007). In all
cases, three clove extracts consistently displayed
superior potency when compared with antibiotics,
while extracts are a mixture of various plant
constituents and antibiotic is a refined and purified
product (El-Mahmood and Doughari, 2008).
Comparing among the three extracts with positive
control, acetone extract was found most effective for
antibacterial activity and the degree of antibacterial
property of three extracts can be put in the following
order: acetone > methanol > ethanol.
The results of this study revealed that although crude
extracts of clove are not purified but their activity was
very effective against all tested bacteria, and these
extracts could be used as an effective antimicrobial
agent, alternative to the use of antibiotics.
The authors would like to thank the Department of
Pathology, Rajshahi Medical College Hospital,
Rajshahi, Bangladesh, for collecting samples.
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... Syzygium aromaticum is evergreen plant belonging to Myrtaceae family. The essential oil of Syzygium aromaticum i.e., eugenol, exhibits broad spectrum activities Lactobacillus, E. coli and S. enterica [101,102]. It also controls the spoilage food borne bacteria such as S. aureus, P. aeruginosa and E. coli [103]. ...
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Background: Antibiotic resistance is a global problem that presents significant risk to human health. Driven by selective pressure of antimicrobial agents, spontaneous mutation, recombination and horizontal gene transfer events, inappropriate antibiotic prescribing and use outside healthcare settings has increased their impact on healthcare system. Increasing risk for human health lead us to study resistance development mechanisms, associated factors that increase dissemination of resistance genes along with information of imperative measures necessary to curtail the growing menace. Methods: In this article, we emphasized on the state of knowledge regarding imprudent use of antibiotics that act as promoters of resistance development. For this, literature based search for articles and entries related to antimicrobial resistance was done. With ample of data available, selected was performed for the epidemiological and clinical based study to curtail the facts present in these data sets so as to get accurate and important information. Results: Resistance mediated by different determinants such as TEM, SHV, OXA and CTX-M, methods of mobilization that increase spread across species and as such failure to available treatment regimens was studied. Addition to detection methods, information of the inhibitors and natural substance useful in mitigating the effect of multidrug resistance was included to strategies the policies and plans for restricting their spread. Conclusion: As intervention to this growing problem, modified use of antimicrobial agents, employment of different formulations of herbs along with public health interventions in restricting antibiotic use, are believed to be of great help in restricting their dissemination and as such spread to non-pathogenic bacterial isolates.
... Clove (Eugenia caryophyllata) which is represented by the immature un-opened flower buds of the clove tree, has been widely used as a spice, and in popular medicine for various therapeutic applications (15), as remedy for indigestion, atherosclerosis (16), diarrhea (17), ringworm (18), athlete's foot (19) and other fungal infections, especially in Asian countries. Its main constituent is represented by eugenol (4-Allyl-2-methoxyphenol) accounting from 60% to 90% (20,21). ...
Clove essential oil was applied in postharvest trials on peanuts, beans, apricot kernels, and lentils stored in jute bags, and on wheat, maize, rice, and rape kept in silos, at ambient temperature (20°C). Eugenol, accounting for 85.6% of clove essential oil, has been used to assess clove essential oil residues after treatments in dry seeds. Two trials at different concentration and storage time were carried out. Immediately after treatment eugenol residues were under the instrumental limit of detection (LOD, 0.5 μg/Kg), except for wheat, peanuts, and rice (0.03, 0.10, 0.24, 0.23 mg/kg, respectively), and increased till a maximum after 2 weeks, except peanuts and apricot kernel which peaked after 30 days (12.69, and 0.47 mg/Kg, respectively). Sensory analysis showed that ventilation of the seeds allowed to decrease eugenol residue values under the flavor perception capacity of the in house panel (0.10 mg/kg), thus not affecting the organoleptic characteristics of the seeds.
... Antimicrobial activity of Syzygium aromaticum was also studied by Rahman et al., (2011) on five bacterial strains with DIZ values range between 7.33 and 7.83 mm. They have reported high antibacterial activity in acetone extracts rather than ethanolic extracts. ...
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In the present study 200 urine samples were collected from both male and female patients suffering with urinary tract infections (UTI). A total of 75 bacterial cultures were isolated belonging to 5 species: Escherichia coli (44%); Klebsiella pneumoniae (25.33%); Pseudomonas aeruginosa (20%); Enterobacter faecalis (6.66%) and Proteus mirabilis (4%). Six plants (Coriander sativum, Syzygium aromaticum, Cinnamomum cassia, Zingiber officinale, Terminalia chebula and Azadirachta indica) and their parts (leaves, bark, flower, rhizome and fruit) were used to evaluate their antibacterial potential. Aqueous, methanolic and ethanolic extracts were used for studying antibacterial activity by agar well diffusion assay and Minimum inhibitory concentration method. Among the three extracts used, highest antibacterial activity was recorded with ethanolic extracts of Cinnamomum cassia on E. coli and least against K. pneumoniae with diameter of inhibition zones (DIZ) of 21.33 ± 0.57 and 15.66 ± 0.57 mm respectively. Preliminary phytochemical analysis of the plant parts revealed the presence of active compounds such as phenolics, tannins, alkaloids and flavonoids. The results obtained in this study clearly demonstrated higher and broad spectrum antibacterial activity of selected plant extracts on all five UTI isolates compared with ten standard antibiotics used for treating UTI.
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Aims: This study investigated the antimicrobial effect ofaqueous and ethanol extracts of the flower buds of Eugenia caryophyllata (Myrtaeae) against a wide range of bacteria and yeasts cells isolated clinically from patients. Methodology: The agar diffusion method was used to establish theantimicrobial activity and the zones of inhibition caused by the extracts.The antimicrobial effects of 16% and 32% aqueous and ethanol extracts of Eugenia caryophyllata were investigated against 111 pathogenic bacteria and yeasts cells. The microbes used consisted of 11 Proteus mirabilis, 20 Salmonella typhi, 15 Pseudomonas aeruginosa, 18 Escherichia coli, 19 Staphylococcus aureus,12 Klebsiella pneumoniae, and 16Candida albicansspecies. Results: The ethanol extracts inhibited the growth of all the microbes employed in the study with inhibition zones ranging from 8.00±0.00 mm to 24.00±0.00 mm. The aqueous extracts however exhibited different degrees of antimicrobial activity with zones of inhibition ranging from 6.00±0.00 mm to 13.33±0.29 mm. Conclusion: Our study concludes that the aqueous and ethanol extracts of the flower buds of Eugenia caryophyllata have relatively good antimicrobial activity againsta wide range of medically important pathogenic bacteria and Candida albicans in vitro.
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Phytochemical screening of the leaves and roots of Cassia alata (Linn) revealed the presence of some bioactive components, which have been linked to antimicrobial properties. The effects of water, methanol and chloroform extracts on some pathogenic Staphylococcus aureus, Escherichia coli, Streptococcus pyogenes, Pseudomonas aeruginosa and Proteus mirabilis showed that the plant parts can be used to treat infections caused by these bacteria. S. aureus, S. pyogenes and P. mirabilis were more susceptible, while E. coli and P. aeruginosa were less sensitive. The effectiveness of the crude extracts were enhanced at elevated temperatures and at near neutrality pH values, which attests to its use in traditional medicine to treat skin, urinary tract and gastrointestinal infections. The aqueous extract was less effective than the organic solvents, thus suggesting the inability of the traditional medicine practitioners to exhaustively extract all the bioactive components with water. The MICs and MBCs of the extracts against the test bacteria were high and correlate with sensitivity test results. The effectiveness of the extracts were less than the conventional antibiotic, metronidazole.
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The bioactive compounds of root extracts of Carica papaya L. were extracted, using water and organic solvents, and were investigated for antibacterial activity against some pathogenic bacteria using the cup plate agar diffusion method. The aqueous extracts did not show significant activity, but the organic extracts had significant activity with the methanol extracts demonstrating the highest activity against the test bacteria. The extracts demonstrated higher activities against all the gram-negative bacteria than the gram-positive bacteria tested, with the highest activity (14 mm zone of inhibition) demonstrated against Salmonella typhi. Increase in temperature enhanced the activity of the extracts, while alkaline pH decreased the activity. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the extracts ranged between 50-200 mg/ml. Preliminary phytochemical analyses showed that the extracts contain alkaloids, tannins, saponins, glycosides and phenols. Carica papaya may be used for the treatment of gastroenteritis, uretritis, otitis media, typhoid fever and wound infections.
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Oils extracted from fifteen medicinal plants were screened for their activity against A. fumigatus and A. niger by disc diffusion method. Minimum inhibitory concentrations (MICs) of oils (%v/v) against Aspergillus fumigatus and Aspergillus niger done by agar dilution method and minimum inhibitory concentration (MIC) and minimum cidal concentration (MCCs) data (%v/v) obtained by the broth micro dilution method. The results showed that the maximum antimycotic activity was demonstrated by oils of Cymbopogon martini, Eucalyptus globulus and Cinnamomum zylenicum as compared to control, followed by Cymbopogon citratus which showed activity similar to control (miconazole nitrate). The oils of Mentha spicata, Azadirachta indica, Eugenia caryophyllata, Withania somnifera and Zingiber officinale exhibited moderate activity. The oils of Cuminum cyminum, Allium sativum, Ocimum sanctum, Trachyspermum copticum, Foeniculum vulgare and Elettaria cardamomum demonstrated comparatively low activity against A. niger and A. fumigatus as compared to control. Mixed oils showed maximum activity as compared to standard. These results support the plant oils can be used to cure mycotic infections and plant oils may have role as pharmaceutical and preservatives. Key words: Essential oil Antifungal activity Minimum inhibitory concentration Aspergillus
The medicinal plant Parkia biglobosa (Jacq.) was screened for the phytochemical components and antibacterial activity against Escherichia coli , Staphylococcus aureus , Klebsiella pneumoniae and Pseudomonas aeruginosa which are associated with urinary tract infections. Aqueous solutions are more potent than methanolic solutions and activity is concentration dependent. P. aeruginosa is least susceptible than the other organisms.
A crude MeOH extract of Syzygium aromaticum (clove) exhibited preferential growth-inhibitory activity against Gram-negative anaerobic periodontal oral pathogens, including Porphyromonas gingivalis and Prevotella intermedia. By means of bioassay-directed chromatographic fractionation, eight active compounds were isolated from this extract and were identified as 5,7-dihydroxy-2-methylchromone 8-C-beta-D-glucopyranoside, biflorin, kaempferol, rhamnocitrin, myricetin, gallic acid, ellagic acid, and oleanolic acid, based on spectroscopic evidence. The antibacterial activity of these pure compounds was determined against Streptococcus mutans, Actinomyces viscosus, P. gingivalis, and P. intermedia. The flavones, kaempferol and myricetin, demonstrated potent growth-inhibitory activity against the periodontal pathogens P. gingivalis and P. intermedia.
FULL TEXT available free from The antimicrobial activity of plant oils and extracts has been recognized for many years. However, few investigations have compared large numbers of oils and extracts using methods that are directly comparable. In the present study, 52 plant oils and extracts were investigated for activity against Acinetobacter baumanii, Aeromonas veronii biogroup sobria, Candida albicans, Enterococcus faecalis, Escherichia col, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica serotype typhimurium, Serratia marcescens and Staphylococcus aureus, using an agar dilution method. Lemongrass, oregano and bay inhibited all organisms at concentrations of < or = 2.0% (v/v). Six oils did not inhibit any organisms at the highest concentration, which was 2.0% (v/v) oil for apricot kernel, evening primrose, macadamia, pumpkin, sage and sweet almond. Variable activity was recorded for the remaining oils. Twenty of the plant oils and extracts were investigated, using a broth microdilution method, for activity against C. albicans, Staph. aureus and E. coli. The lowest minimum inhibitory concentrations were 0.03% (v/v) thyme oil against C. albicans and E. coli and 0.008% (v/v) vetiver oil against Staph. aureus. These results support the notion that plant essential oils and extracts may have a role as pharmaceuticals and preservatives.
The volatile oils of black pepper [Piper nigrum L. (Piperaceae)], clove [Syzygium aromaticum (L.) Merr. & Perry (Myrtaceae)], geranium [Pelargonium graveolens L'Herit (Geraniaceae)], nutmeg [Myristica fragrans Houtt. (Myristicaceae), oregano [Origanum vulgare ssp. hirtum (Link) Letsw. (Lamiaceae)] and thyme [Thymus vulgaris L. (Lamiaceae)] were assessed for antibacterial activity against 25 different genera of bacteria. These included animal and plant pathogens, food poisoning and spoilage bacteria. The volatile oils exhibited considerable inhibitory effects against all the organisms under test while their major components demonstrated various degrees of growth inhibition.
To quantify the antibacterial properties of five essential oils (EO) on a non-toxigenic strain of Escherichia coli O157:H7 in the presence and absence of a stabilizer and an emulsifier and at three different temperatures. Five EOs known to exhibit antibacterial properties were screened by disc diffusion assay and the most active were selected for further study in microdilution colorimetric assays. Oregano (Origanum vulgare) and thyme (Thymus vulgaris; light and red varieties) EO had the strongest bacteriostatic and bactericidal properties, followed by bay (Pimenta racemosa) and clove bud (Eugenia caryophyllata synonym: Syzygium aromaticum) EO. Oregano oil was colicidal at 625 microl l(-1) at 10, 20 and 37 degrees C. The addition of 0.05% (w/v) agar as stabilizer reinforced the antibacterial properties, particularly at 10 degrees C, whereas 0.25% (w/v) lecithin reduced antibacterial activity. Scanning electron micrographs showed extensive morphological changes to treated cells. Oregano and thyme EO possess significant in vitro colicidal and colistatic properties, which are exhibited in a broad temperature range and substantially improved by the addition of agar as stabilizer. Bay and clove bud EO are less active. Lecithin diminished antibacterial properties. The bactericidal concentration of oregano EO irreversibly damaged E. coli O157:H7 cells within 1 min. Oregano and light thyme EO, particularly when enhanced by agar stabilizer, may be effective in reducing the number or preventing the growth of E. coli O157:H7 in foods.
Worldwide, diarrhea claims several million lives annually, mostly those of infants. Poverty, crowding, and contaminated water supplies all contribute. Almost all of these deaths could have been prevented with adequate fluid replacement. Although its incidence is much lower in the more affluent nations, diarrhea remains one of the two most common reasons for visits to pediatric emergency departments and is also common among the institutionalized elderly. Chronic diarrheas, while less common, often present diagnostic dilemmas and can be difficult to manage. This article will begin by reviewing the relevant physiology; then, with that as context, it will consider pathophysiology, both general aspects and specific diarrheal syndromes. Finally, rationales for management will be briefly considered. The review is amply referenced with some of the references listed at the end of the article and some at the JCI website (