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Introduction
Bacterial disease can be defined as any form of illness brought about by bacteria. Usually, there are
millions of bacteria living on our body. Bacteria that causes harmful infections are called pathogenic
bacteria (Haines 2013). A few examples of pathogenic bacteria would be Escherichia coli,
Helicobacter pylori, Staphylococcus aureus and Pseudomonas aeruginosa which can cause serious
and life-threatening complications such as bacteremia and toxic shock syndrome (Haines 2013).
Researches and studies are done over many years and this bring about the emergence of
antibiotics. Many fatal and infectious diseases were able to be brought under control, saving
millions of lives. Researchers even thought that synthetic antibiotics might be the solution to cure
infectious disease and they were discouraged in making serious efforts to develop drugs from
simple natural compounds (Thormar 2011).
However, rising in number of resistance to antibiotics is alarming and that new antibacterial
sources are being done now, such as investigation on plants. Infectious diseases cause premature
death, leading the chart with almost 50,000 fatality every day (Mahida and Mohan 2007).
Necessary steps were taken to reduce resistance problem, such as control the consumption of
antibiotics, researches for better understanding of genetic mechanism of resistance and
development of new, synthetic or natural drugs (Nascimento et al. 2000).
Recently, there have been a renewed interest in antibacterial effects of natural compounds
(Thormar 2011). Plant extracts and essential oils were screened for antibacterial activity and bioassay guided fractionation of active extracts were done to isolate the active constituent from the
selected plants (Mohamed et al. 2010).
Scientific
Family
name
Garlic
Allium
Plants part
used
Liliaceae
Bulb
Sativum
Lemon
Citrus limon
Rutaceae
Fruit, juice,
peels and
seeds
Peppermint
Mentha x
Lamiaceae
piperita
Leaves and
stem
Thyme
Thymus
Lamiaceae
vulgaris
Leaves and
flowering
tops
Wormwood
Artemisia
Asteraceae
Whole herb
absinthium
Table 1
Garlic
Lemon
Peppermint
Thyme
Wormwood
Screening methods
There are a few common screening methods used for antibacterial properties of plant extracts.
1. One of the methods used is the disc-diffusion method. This method uses paper discs and are
impregnated with plant extracts and put onto an inoculated agar medium. After incubation, the
diameter around the disc where bacteria growth is stopped or inhibited is measured. The
parameter is known as zone of growth inhibition. There are several disadvantages using this
method such as essential oil unable to diffuse through the agar due to the hydrophobic nature of
the essential oil (Thormar 2011). Another drawback is that this test is qualitative test, as zone of
growth inhibition is only measured in mm, and Minimum Inhibitory concentration cannot be
determined (MichiganStateUniversity 2011).
2. Another method used is the agar- or broth-dilution method. This method offers a better
parameter such as allowing the calculation of Minimum Inhibitory Concentration (MIC). MIC is
simply defined as lowest concentration of the plant drugs that inhibit growth of test bacteria and
more useful than zone of growth inhibition since MIC allows establishment of safe and effective
final concentration in formulated drugs (Thormar 2011).
3. The 96-well plate assays are also used to screen for antibacterial activity and also to determine
the MIC of the drugs. Final concentrations of 100% (undiluted), 50%, 25%, 12.5%, and 6.25% of
serial dilutions of the extracts were prepared using sterile water. Then, bacteria of 108 CFU/mL
were added to each experimental well. The plates were incubated at 37C for 24 hours.
P-iodonitrotetrazolium violet (INT) marker solution was added and presence of antibacterial
activity is indicated by changes of colour from clear to red. MIC is determined by means of digital
images in a computer (Frey and Meyers 2010).
nutrients broth followed by incubation at 37C for 24 hours. Agar dilution method is used to
determine antibacterial activity and MIC. Serial concentrations of thyme extracts were obtained
from 0.02 mgml1 up to 13 mgml1 . Petri plates of BHI containing various concentration of
extracts were inoculated with bacterial strains and spread on solid agar plates. Then, the agar
containing extracts and bacterial strain were incubated for 24 hours. The lowest concentration
extracts that completely inhibit growth of tested bacteria was determined to be the MIC (El-Safey
and Salah 2011).
Antibacterial activity of Wormwood were done to determine the MIC using broth dilution method.
Brain heart infusion (BHI) broth with Tween 80 detergent was used. Bacterial strains were
suspended in the BHI broth to get a final density of 107 cfu ml1 . 40 l Wormwood essential oils
with various dilutions were added to tubes containing the BHI and bacterial strains. The tubes were
then incubated with incubator shaker and the lowest concentration of essential oil used, without
visible growth, was considered the MIC (Taherkhani et al. 2013).
Side effects
Garlic
(WebMD 2014)
Heartburn
Bad breath
Body odor
Lemon
2010)
Peppermint
Heartburn
(Medicinenet 2014)
Uncommon:
Thyme
(Chamberlins.com 2011)
Flushing
Mouth sores
Headache
Wormwood
Dizziness
Heartburn
Muscle weakness
Absinthism
2010)
Digestive disorders
Restlessness
Loss of intellect
Conclusion
The experimental results of garlic extracts are shown in Table 2:
Plant extracts
Zone of Inhibition(mm)
Escherichia coli
Salmonella typhi
Escherichia coli
14.3 0.54
Pseudomonas aeruginosa
18.3 0.72
Staphylococcus aureus
19.3 1.08
Elegalam 2005).
Shigella
13 0.47
Salmonella typhi
15.6 0.56
Table 2
The experimental results of lemon extracts are shown in Table 3:
Plant extracts
Zone of Inhibition(mm)
Escherichia coli
Salmonella parathypi b
Shigella sonnei
Escherichia coli
20
Pseudomonas aeruginosa
19
Staphylococcus aureus
21
Table 3
The experimental results of peppermint extracts are shown in Table 4:
Plant extracts
Zone of Inhibition(mm)
Peppermint oil
Escherichia coli
13
Pseudomonas aeruginosa
12
Salmonella typhi
10.33
Salmonella parathypi b
11
Peppermint juice
Escherichia coli
12.26
Pseudomonas aeruginosa
11.56
Salmonella typhi
9.5
Salmonella parathypi b
Table 4
The experimental results of thyme extracts are shown in Table 5:
Plant extracts
Zone of Inhibition(mm)
Escherichia coli
22.55.5
Staphylococcus aureus
22.71.6
Escherichia coli
MRSA
12
Pseudomonas aeruginosa
Table 5
The experimental results of Wormwood extracts are shown in Table 6:
Plant extracts
Minimal inhibitory
concentration(mg/ml)
Escherichia coli
Staphylococcus aureus
Pseudomonas aeruginosa
Plant extracts
2.5
Zone of inhibition (mm)
Escherichia coli
15
Staphylococcus aureus
32
Pseudomonas aeruginosa
21
Table 6
Based on the results above, all selected plant extracts have shown a significant antibacterial
properties to certain Gram-positive and Gram-negative bacteria. Garlic shows a better antibacterial
properties in the aqueous extracts compared to ethanol extracts, with the highest zone of
inhibition against Gram-positive Staphylococcus aureus. The antibacterial properties of allicin found
in garlic has been shown to have a wide spectrum of antibacterial activity against Gram-negative
and Gram-positive bacteria (Ankri and Mirelman 1999).
In lemon, most of the antibacterial potential lies within the peel, where the zone of inhibition
against the test bacteria showed significant difference compared to lemon juice extracts. The most
susceptible bacterial strains is shown to be Staphylococcus aureus, with the highest zone of
inhibition of 21mm. Lemon peels contain flavonoids, terpenes, ascorbic acid, corydaline alkaloids,
hypericin and other components that contributes to overall antibacterial properties of lemon
extracts (Dhanavade et al. 2011).
The antibacterial properties of peppermint oil and peppermint juice show good inhibition with high
zone of inhibition. From the results, peppermint has shown excellent antibacterial properties in the
extracts of the stem and leaves, E.coli is the most susceptible with slightly higher zone of inhibition
(Saeed et al. 2006).
Thyme essential oil shows a significant inhibitory activity against E.coli and S.aureus with high
degree of zone of inhibition. Thyme aqueous extract showed less activity with lower zone of
inhibition compared to thyme essential oil. The results showed that thyme oil antibacterial
attribute could be due to the presence of Carvacrol and Thymol in Thyme extracts (Cetin et al.
2009).
Wormwood essential oil has shown to have a potent antibacterial effects, with the most
susceptible bacteria being Staphylococcus aureus (MIC: 1 mg/ml and inhibitory diameter 32mm). It
also shows good activity against E.coli and P.aeruginosa with significant inhibitory effects
(Taherkhani et al. 2013).
In conclusion, plant drugs have been proven to show a wide spectrum of activities against bacteria.
With the alarming increase in antibiotic resistant bacteria, plants and herbs potential as
antibacterial should not be overlooked as they can be a good source for potential drug candidate.
(2012 words)
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