Antibacterial Activity of Plant Extracts PDF
Antibacterial Activity of Plant Extracts PDF
Antibacterial Activity of Plant Extracts PDF
ISSN 1517-8382
ABSTRACT
The antimicrobial activity of plant extracts and phytochemicals was evaluated with antibiotic susceptible and
resistant microorganisms. In addition, the possible synergistic effects when associated with antibiotics were
studied. Extracts from the following plants were utilized: Achillea millifolium (yarrow), Caryophyllus
aromaticus (clove), Melissa offficinalis (lemon-balm), Ocimun basilucum (basil), Psidium guajava (guava),
Punica granatum (pomegranate), Rosmarinus officinalis (rosemary), Salvia officinalis (sage), Syzygyum
joabolanum (jambolan) and Thymus vulgaris (thyme). The phytochemicals benzoic acid, cinnamic acid,
eugenol and farnesol were also utilized. The highest antimicrobial potentials were observed for the extracts
of Caryophyllus aromaticus and Syzygyum joabolanum, which inhibited 64.2 and 57.1% of the tested
microorganisms, respectively, with higher activity against antibiotic-resistant bacteria (83.3%). Sage and
yarrow extracts did not present any antimicrobial activity. Association of antibiotics and plant extracts showed
synergistic antibacterial activity against antibiotic-resistant bacteria. The results obtained with Pseudomonas
aeruginosa was particularly interesting, since it was inhibited by clove, jambolan, pomegranate and thyme
extracts. This inhibition was observed with the individual extracts and when they were used in lower
concentrations with ineffective antibiotics.
Key words: plant extracts activity; medicinal plants; antimicrobial activity
INTRODUCTION
Even though pharmacological industries have produced a
number of new antibiotics in the last three decades, resistance
to these drugs by microorganisms has increased. In general,
bacteria have the genetic ability to transmit and acquire
resistance to drugs, which are utilized as therapeutic agents (12).
Such a fact is cause for concern, because of the number of
patients in hospitals who have suppressed immunity, and due to
new bacterial strains, which are multi-resistant. Consequently,
new infections can occur in hospitals resulting in high mortality.
From 1980 to 1990, Montelli and Levy (27) documented a
high incidence of resistant microorganisms in clinical
microbiology in Brazil. This fact has also been verified in other
clinics around all over world.
* Corresponding author. Mailing address: Faculdade de Cincias da Sade, Universidade Metodista de Piracicaba, Caixa Postal 68, CEP 13400-911,
Piracicaba, SP. Fax: (+5519) 433-2909. E-mail: [email protected]
247
248
Plant extracts
The methodologies of Harbone (17) and Wagner et al. (37)
were used to process the ethanolic extracts (1:1) from the ten
plants of interest. Seven of them (basil, clove, lemon balm,
rosemary, sage, thyme and yarrow) had essential oils as the main
active ingredient, while the other three (guava, jambolan and
pomegranate) had high contents of tannin. With the exception
of clove, all of the plants with essential oils were cultivated
from seeds in the Greenhouse of Medicinal Plants at the
Methodist University of Piracicaba, SP, Brazil. The seeds were
acquired at Semex, SP, Brazil. Clove was obtained from
medicinal plants market of So Paulo city. The selected plants
with tannins were obtained in the Horticulture Department at
the University of Agriculture Luiz de Queiroz/USP,
Piracicaba, SP, Brazil.
The following lists the plant of interest, plant parts used for
extractions, and compounds obtained.
Thyme (Thymus vulgaris L., Lamiaceae) - [stripped and
dried leaves and flowers]. Chemical constituents: essential
oils (mainly thymol and carvacrol), flavonoids, tannins and
triterpenes (8).
Rosemary (Rosmarinus officinalis L., Lamiaceae) - [leaf].
Chemical constituents: flavonoids, phenolic acids (caffeic,
chorogenic and rosmarinic) and essencial oils (camphor and
cineole) and diterpenes (carnosol) (30)
Lemon balm (Melissa officinalis L., Lamiaceae) - [leaf].
Chemical constituents: essencial oils (containing citral and
citronellal monoterpenes), flavonoids and rosmarinic,
caffeic and chlorogenic acids (8).
Sage (Salvia officinalis L., Lamiaceae) - [leaf]. Chemical
constituents: rosmarinic, caffeic, chlorogenic acids;
carnosol, flavonoids, essential oils (mainly thuyone and
cineole). (30).
Basil (Ocimum basilicum L., Lamiaceae) - [leaf]. Chemical
constituents: essential oils (linalol, estragol and eugenol);
tannins and flavonoids (8).
Yarrow (Achillea millefolium L.,Asteraceae) - [flowerheads]. Chemical constituents: flavonoids, tannins,
comarins, proazulene. (8,11).
Clove (Syzygium aromaticum (L.) Merr. et Perry =
Caryophyllus aromaticus L.) - [dried buds]. Chemical
249
Table 1. Antimicrobial activity caused by plant extracts (hydro-alcoholic fraction) through agar diffusion method.
microorg.
thyme
rosemary
clove
jambolan
1
2
3
4
5
6
7
8
9
10
11
12
13
14
+
+
+
+
-
+
+
-
+
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
lemonbalm pomegranate
+
+
+
-
guava
sage
basil
yarrow
+
+
-
+
-
+
+
-
eugenol
farnesol
benzoic acid
cinnamic acid
1
2
3
4
5
6
7
8
9
10
11
12
13
14
+
+
+
+
-
+
+
-
+
+
+
+
+
250
susceptible microorganisms
resistant microorganisms
90
80
% microorganisms
70
60
50
40
30
20
10
0
thyme
rosemary
clove
jambolan
lemon-balm
pomegran.
guava
sage
basil
yarrow
plant extracts
Figure 1 - Antimicrobial activity from plant extracts against susceptible (6) and resistant (8) antibiotic microorganisms
susceptible microorganisms
resistant microorganisms
90
80
% microorganisms
70
60
50
40
30
20
10
0
eugenol
farnesol
benzoic acid
cinnamic acid
phytochemicals
Figure 2 - Antimicrobial activity from phytochemicals against susceptible (6) and resistant (8) antibiotic microorganisms
251
Table 3. Minimal Inhibitory Concentration (MIC) of plant extracts and phytochemicals against antibiotic resistant bacteria.
Bacteria
P. aeruginosa (11)
K. pneumoniae (7)
K. pneumoniae (8)
Shigella spp (9)
Proteus spp (10)
S. aureus (14)
E. aerogenes (12)
252
thyme
jambolan
70
50
70
300
400
50
100
100
250
20
eugenol
20
10
10
10
Figure 3 - Minimal Inhibitory Concentration (MIC) of plant extracts/phytochemicals against antibiotic resistant bacteria
tested, even those that did not show any activity by themselves.
In the case of pomegranate and clove, the synergetic effect was
noted with the extracts in the concentration of 50 and 10 g/
mL, respectively. Moreover, this effect was also observed for
K. pneumoniae (8) when 20 g/mL of clove extract was
combined to ampicillin. Furthermore, the growth of Proteus
spp.(10) was inhibited when either clove extract (10 g/mL) or
eugenol (5 g/mL) was combined to tetracycline. Synergistic
effects resulting from the combination of antibiotics with
extracts were documented in the literature (28). They studied
the association of anacardic acid and totarol with methicillin
to inhibit strains of S. aureus resistant to methicillin (MRSA).
No synergetic effect was observed when different
concentrations of extracts from lemon balm, clove and eugenol
were combined with ampicillin to inhibit the growth of K.
pneumoniae (7) and E. aerogenes (12). Only the association
of thyme (20 g/mL) with ampicillin was able to cause such an
effect.
The use of plants to heal diseases, including infectious one,
has been extensively applied by people. Data from the literature
as well as our results reveal the great potential of plants for
therapeutic treatment, in spite of the fact that they have not
been completely investigated. Therefore, more studies need to
253
Table 4. Effect of the association of plant extracts/phytochemicals and antibiotics on resistant bacteria. (The concentration of the
antibiotics was 50mg/mL).
Bacteria
Association
5
P. aeruginosa (11)
K. pneumoniae (7)
K. pneumoniae (8)
Proteus (10)
E. aerogenes (12)
J
J + TT
J + AP
J + CO
T
T + TT
T + AP
T + CO
P
P + TT
P + AP
P + CO
C
C + TT
C + AP
C + CO
C
C + TT
C + AP
C + CO
E
E + TT
E + AP
E + CO
J
J + TT
J + AP
T
T + TT
T + AP
20
+
+
+
+
+
+
+
+
+
L
L + AP
C
C + AP
E
E + AP
C
C + AP
E
E + AP
10
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
400
+
+
+
+
+
+
+
+
+
+
+
+
+
+
254
250
+
+
+
+
+
+
ACKNOWLEDGEMENTS
3.
RESUMO
Atividade de extratos vegetais e
fitofrmacos sobre bactrias resistentes a
antibiticos
5.
6.
7.
8.
26.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
27.
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