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Multidiscip. Sci. J. (2024) 6:e2024026
Received: March 10, 2023 | Accepted: July 4, 2023
RESEARCH ARTICLE
Published Online: August 24, 2023
https://doi.org/10.31893/multiscience.2024026
Antioxidant and antimicrobial activities of Ocimum
basilicum var. thyrsiflora against some oral
microbes
Akankshya Sahua | Gayatree Nayaka| Sanat Kumar Bhuyanb | Ruchi Bhuyanc |
Dattatreya Karc | Ananya Kuanara
aCentre for Biotechnology, Siksha 'O' Anusandhan University, Kalinga Nagar, Ghatikia, Bhubaneswar, Odisha, India .
bInstitute of Dental Sciences, Siksha 'O' Anusand han University, Bhubaneswar, Odisha, India.
cDepartment of Medical Research, Health Science, IMS & SUM Hospital, Siksha 'O' Anusandhan University, Bhubaneswar, Odisha, India.
1. Introduction
Ocimum basilicum var. thyrsiflora (Lamiaceae) is commonly known as Thai basil. It has narrow, lightly serrated, shiny
green leaves that smell sweet and anise-like in aroma and have a hint of licorice and a little spiciness.
(https://en.wikipedia.org/wiki/Thai_basil) It is most popular in China, Japan, Turkey, and Iran and is also found in South and
Central America, tropical Asia, and Africa (Purushothaman et al 2018). The genus Ocimum is particularly well known for its
antioxidant properties (Chanwitheesuk et al 2005). It has long been used to treat anxieties, coughs, the common cold,
headaches, fevers, diabetes, migraines, neuropathic relief, heart disease, gastrointestinal diseases, insect bites, cramps,
sinuses, and several neurological diseases as an anti-inflammatory and antidepressant (Bora et al 2011). Moreover, the plant's
flowering tops and leaves are apparent as a galactagogue, carminative, stomachic, and anti-inflammatory in traditional
medicine (Sajjadi 2006). Extracts of basil from the roots, stems, flowers, leaves, and seeds have been used in a variety of medical
treatments.
Natural antioxidants derived from plants have some advantages over synthetic antioxidants, such as their affordability,
ease of use, and minimal to nonexistent negative effects. Herbs and spices are now being targeted as key sources of natural
antioxidants as a result of trends toward the preservation of products using natural preservatives (Dessí et al 2001). We recently
reported on the chemical composition and radical-scavenging activities of essential oils from various Indian spices and
medicinal plants (Kar et al 2017). According to a study performed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay for free
radical scavenging, bleaching β-carotene in the linoleic acid system, and inhibiting linoleic acid oxidation, basil essential oil
(Ocimum basilicum L.) showed good antioxidant activity (Hussain et al 2008). The radical-scavenging activities of oils have also
Abstract The accumulation of free radicals is the root cause of many dangerous diseases. Several studies are being
conducted to identify plant-based natural antioxidants and antimicrobial elements. Ocimum basilicum var. thyrsiflora
exhibits anti-inflammatory, antioxidant, antiulcer, antiviral, hypoglycemic, hypolipidemic, antimicrobial, anticancer, wound-
healing, etc. This research aimed to determine the in vitro antioxidant activity of essential oils extracted from the leaves
and seeds of Ocimum basilicum var. thyrsiflora by hydroxyl radical, nitric oxide, 2,2-diphenyl-1-picrylhydrazyl (DPPH),
hydroxyl radical, and azino-bis (3-ethylbenthiazoline-6-sulphonic acid (ABTS) assays. The antibacterial activity of essential
oils extracted from the leaves and seeds of Ocimum basilicum var. thyrsiflora was evaluated by the disc diffusion method
against E. coli, P. aeruginosa, Acinetobacter, S. epidermis, K. pneumoniae, S. aureus, E. faecalis, and P. mirabilis. According
to the findings, the hydroxyl radical (HO•) scavenging assay method revealed more potent antioxidant activity in Ocimum
basilicum var. thyrsiflora essential oils than the other methods. The hydroxyl radical scavenging assay was found to have
IC50 values of 115.20 ± 6.45 and 128.35 ± 6.20 for seeds and leaves, respectively. Ocimum basilicum var. thyrsiflora essential
oils exhibited a strong antibacterial effect against all the tested microorganisms. The highest antibacterial activity was
measured in the essential oil extracted from seeds against P. aeruginosa (20.06±0.30) at a concentration of 50% essential
oil, and the lowest activity was observed in the essential oil extracted from leaves against P. mirabilis (7.46±0.45) at a
concentration of 12.5% essential oil. As a result, it can be a potent natural source of antioxidants to treat many stress- and
anxiety-related diseases and would be a better alternative for the development of new antimicrobial medications to treat
a variety of infectious ailments caused by microbes.
Keywords: Ocimum basilicum var. thyrsiflora, essential oil, antioxidant activity, scavenging activity, free radicals
antibacterial activity
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been studied using various assays given the traditional uses of the herbs, which suggest that they may possess antioxidant
properties.
The rising prevalence of severe opportunistic fungal and bacterial infections is a major issue. Determining new types of
natural compounds that might be useful against bacteria and fungi is therefore extremely important. Plant extracts are part of
ongoing research to discover novel compounds with the potential to combat multiple drug-resistant bacteria. Most novel
antibiotics coming to market today are derived from natural or semisynthetic sources, with approximately 20% of plants found
in the world having undergone pharmacological or biological testing (Mothana et al 2005).
During the last three years, plants with antibacterial activity have received much interest in the search for novel
therapeutic agents. Current research has discovered that sweet basil extracts contain antibacterial effects against Escherichia
coli and Staphylococcus aureus, antifungal effects against Rhizopus solani and Aspergillus niger, and antiviral effects against a
few strains (Nguefack et al 2004, Maisuthisakul et al 2008). There is no published information on the antioxidant and
antibacterial activities of oils derived from various parts of Ocimum basilicum var. thyrsiflora. Therefore, this research aimed
to determine the antioxidant and antimicrobial activities of various plant parts of Ocimum basilicum var. thyrsiflora..
2. Materials and Methods
2.1. Plant material
The plants Ocimum basilicum var. thyrsiflora (Figure 1) were collected from the Medicinal Plants Knowledge Center
(MPKC), Bhubaneswar, Odisha, India, in July 2021. Dr. Pratap Chandra Panda, Senior Scientist, Centre for Biotechnology, Siksha
‘O’ Anusandhan, Bhubaneswar, India, authenticated the plant. A voucher specimen of the plant (No. 1970/CBT) was deposited
in the Centre for Biotechnology, Siksha ‘O’ Anusandhan (Deemed to be University), Bhubaneswar.
Figure 1 (A) Whole plant, (B) Leaf, (C) Seeds of Ocimum basilicum var. thyrsiflora.
2.2. Extraction of essential oils
To remove dust, the freshly collected samples were first washed with tap water, followed by distilled water. The seeds
and leaves were then air-dried in the shade at room temperature. The air-dried leaves and seeds were crushed and pulverized
into a coarse powder in a mortar and pestle. Then, 200 g of coarse powder material was hydrodistilled using a Clevenger
apparatus for 6 hours to achieve an oil yield. By letting the oil air-dry on anhydrous Na2SO4, moisture traces were eliminated.
The oil was then collected in Eppendorf tubes and stored at 4 °C.
2.3. Antioxidant Activity
Several in vitro experiments, including DPPH, ABTS, and nitric oxide free radical scavenging assays, were used to evaluate
the antioxidant activity of oils. All assays were conducted in triplicate, and average values were used.
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2.4. Chemicals
The following chemicals were purchased from Merck India Ltd.: 2, DPPH, sulphanilic acid, sodium nitroprusside, trichloro
acetic acid, thiobarbituric acid, methanol, naphthyl ethylene diamine dihydrochloride, dimethyl sulphoxide, ferric chloride,
hydrogen peroxide, potassium persulphate, ascorbic acid, and butylated hydroxytoluene (BHT).
2.5. Diphenyl-1-picrylhydrazyl (DPPH) assay
Brand-Willium et al.’s (1995) protocol was used to carry out the DPPH assay. This was done by preparing a 0.1 M DPPH
solution in methanol and setting its absorbance at 515 nm to 0.95. Then, 100 L of the sample was mixed with 1 mL of the
DPPH solution, and the combined solution was incubated at 37 °C for 30 minutes. Methanol served as the control chemical.
At 515 nm, the absorbance was noted after 30 minutes. Ascorbic acid served as the standard. To determine the IC50 and
DPPH scavenging activity, the formula below was employed.
DPPH scavenging activity (in %) = [(Absorbance of control - Absorbance of sample)/Absorbance of control] × 100
2.6. Nitric oxide (NO) scavenging activity
Alkaline dimethyl sulfoxide (DMSO) was used to measure the nitric oxide scavenging activity (Koleva et al 2002). Dry
DMSO was allowed to remain in contact with solid potassium superoxide for at least 24 hours. A 200 mL filtrate was then added
to 2.8 mL of an aqueous solution containing 10 mM EDTA, 56 mM nitroblue tetrazolium, and 10 mM potassium phosphate
buffer at pH 7.4. At 540 nm, absorbances were measured after adding test solutions at various concentrations (5-100 g/mL)
and compared to the control.
2.7. Azino-bis (3-ethylbenthiazoline-6-sulphonic acid (ABTS) scavenging activity
The effectiveness of ABTS scavenging activity was analyzed using the Re et al (1999) procedure. To produce a dark ABTS
working solution, 2.45 mM potassium oxidopersulphate solution and 7 mM ABTS solution were mixed. The mixture
solution was then stored in complete darkness for 12 to 16 hours. After being diluted with 50% methanol, the solution's
absorbance at 734 nm was set to 0.7 (± 0.02). One milliliter of the ABTS working solution was added to 100 L of the sample,
and the absorbance was recorded after 1 and 6 minutes. The following formula was used to calculate the ABTS scavenging
activity:
ABTS scavenging activity (in %) = [(Absorbance of control - Absorbance of sample)/Absorbance of control] × 100.
2.8. Hydroxyl radical (HO•) scavenging assay
Using the approach developed by Halliwell and Gutteridge (1981), this activity was determined. First, 100 mL of extract
solution, 200 mL of premixed ferric chloride (100 mM) solution (1:1; v/v), 500 mL of 2-deoxyribose (2.8 mM) in phosphate
buffer (50 mM, pH 7.4), 100 mL of H2O2 (200 mM), and 200 mL of the reaction mixture (100 mL) were mixed. Then, 100 mL of
ascorbate (300 mM) was added to the reaction mixture, which was incubated for 1 hour at 37 °C. Furthermore, 1 mL of TCA
solution (2.8% w/v aqueous) and 1 mL of TBA solution (1% w/v in 50 mM NaOH) were added to the reaction mixture. The
reaction mixture was then heated in a water bath for 15 minutes and allowed to cool. Using the following formula, the hydroxyl
radical scavenging activity at 532 nm was determined.
Hydroxyl radical scavenging activity = (1- Absorbance of sample/Absorbance of control) × 100.
2.9. Test microorganisms
Some periodontal bacteria were identified, and the test bacterial strains were obtained from the Department of
Microbiology, College of Basic Science and Humanities, OUAT, Bhubaneswar. The test bacterial strains were Escherichia coli,
Pseudomonas aeruginosa, Acinetobacter, Staphylococcus epidermidis, Klebsiella pneumoniae, Staphylococcus aureus,
Enterococcus faecalis, and Proteus mirabilis.
2.10. Evaluation of antimicrobial activity
The disc diffusion method, developed by Standard Kirby Bauer, was employed to evaluate the antibacterial effect.
Standard inoculums were prepared by dipping 1-2 colonies into liquid NB and shaking them for 3 hours. After 3 hours, a sterile
spreader was used to spread the liquid bacterial culture onto Mueller-Hinton agar (MHA) plates. Three sterile discs with a 6
mm diameter were placed on each agar plate with bacteria. The discs were impregnated with DMSO-dissolved oils at test
concentrations of 12.5%, 25%, and 50%. After 24 hours of incubation at 37 °C, the zones of inhibition around the discs were
measured and reported in mm. Each bacterium was screened in triplicate, with amoxicillin serving as the positive control and
DMSO solvent serving as the negative control.
2.11. Statistical analysis
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The analyses were carried out in triplicate, and the mean and standard error of the mean (SEM) of the observations
were calculated. Tukey's test (p< 0.05) was used to assess the variations in the means of the IC50 for different samples following
the completion of an analysis of variance (ANOVA).
3. Results
3.1. Antioxidant activity
Various analytical assays were used to evaluate the plant's in vitro antioxidant activities. In comparison to standard
ascorbic acid and BHT, all antioxidant assays significantly support the antioxidant potential of plants. Table 1 and Figure 2
provide the IC50 values for the hydrogen peroxide radical, ABTS radical, nitric oxide radical, and DPPH radical scavenging
activities of essential oils extracted from leaves and seeds.
Table 1 Free radical scavenging activity of Ocimum basilicum var. thyrsiflora essential oil.
Essential
Oil/Standards
IC50 value ± SEM (µg/mL) * by methods
DPPH
Nitric oxide
ABTS
H2O2
Ascorbic acid
55.4 ± 20.12a
331.34 ± 0.08a
39.00 ± 0.15a
183.66 ± 12.89d
BHT
0.18 ± 0.00b
75.00 ± 0.00b
202.35 ± 0.10a
78.33 ± 7.07b
Seeds
185.33 ± 2.08c
515.05 ± 8.25d
220.58 ± 0.81b
115.20 ± 6.45c
Leaves
190.25 ± 2.00b
565.25 ± 8.10d
228.67 ± 0.49b
128.35 ± 6.20b
*Average of determinations made in triplicate. The letters (a-d) in the mean ± SEM indicate significance (p < 0.05).
Figure 2 Free radical scavenging activity of Ocimum basilicum var. thyrsiflora essential oil.
3.2. In vitro antioxidant activity of essential oil extracted from leaves
The essential oil extracted from leaves exhibited a strong hydroxyl radical scavenging effect with an IC50 value of 128.35
± 6.20 µg/mL, which is significantly lower than the standard value of ascorbic acid. At 190.25 ± 2.00 µg/mL for DPPH, 565.25 ±
8.10 µg/mL for nitric oxide radical, and 228.67 ± 0.49 µg/mL for ABTS. The IC50 value of the essential oil extract was higher than
that of the ascorbic acid and BHT standards. The results show that the essential oil extracted from the leaves has a
moderate antioxidant effect.
3.3. In vitro antioxidant activity of essential oil extracted from seeds
The essential oil from seeds has an IC50 value of 115.20 ± 6.45 µg/mL, which is significantly lower than the standard value
for ascorbic acid and indicates a strong hydroxyl radical scavenging effect. The IC50 value of seed oil was higher than the ascorbic
acid and BHT standards, at 185.33 ± 2.08 µg/mL for DPPH, 515.05 ± 8.25 µg/mL for nitric oxide radical, and 220.58 ± 0.81 µg/mL
for ABTS scavenging effect. The outcomes revealed moderate antioxidant activity for the essential oil derived from seeds.
3.4. Antimicrobial activity
The antimicrobial effect of the essential oils of Ocimum basilicum var. thyrsiflora was tested against eight pathogenic
bacteria, as shown in Table 2 and Figure 3. Ocimum basilicum var. thyrsiflora essential oils exhibited a strong antibacterial
effect against all the tested microorganisms. The antimicrobial activity was evaluated by measuring the zone of inhibition.
0
100
200
300
400
500
600
D PP H N IT R IC O XI D E A BT S H 2O 2
IC50 VALUES ΜG/ML
Ascorbic acid BHT Leaves Seed
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Figure 3 Antibacterial activity of different concentrations of essential oils extracted from leaves and seeds of Ocimum basilicum var.
thyrsiflora.
Table 2 Antibacterial activity of different concentrations of essential oils extracted from leaves and seeds of Ocimum basilicum var. thyrsiflora.
Oil concentration of
samples
Diameter of zone of inhibition (mm ±SD)
Name of the microorganisms
E.coli
P.aeruginosa
Acinetobacter
S.epidermis
K.pneumoniae
S.aureus
E.faecalis
P.mirabilis
DMSO (NC)
-
-
-
-
-
-
-
-
Leaves (12.5%)
9.01±0.5
13.5±0.5
11.10±0.65
10.23±0.91
10.63±0.55
8±0.5
11.53±0.50
7.46±0.45
Seeds (12.5%)
10.08±0.6
14.00±0.8
12.09±0.23
10.45±0.86
11.06±0.43
9±0.4
11.89±0.60
9.01±0.23
Leaves (25%)
10.76±0.36
14.68±0.2
12.20±0.36
10.76±0.25
11.6±0.52
9±0.75
12.5±0.5
9.1±0.36
Seeds (25%)
11.23±0.42
16.03±0.23
13.87±0.67
12.38±0.75
12.89±0.25
10.87±0.35
13.76±0.2
10.08±0.24
Leaves (50%)
11.3±0.6
18.33±0.32
13.08±0.21
13.11±0.56
12.43±0.42
11.08±0.8
13.76±0.3
10.34±0.85
Seeds (50%)
13.5±0.5
20.06±0.30
15.93±0.40
15.16±0.47
14.76±0.92
13.6±0.6
15.5±0.5
12.66±0.56
Amoxicillin (10 μg) (PC)
25.63±0.09
28.07±0.56
20.8 7±1.02
21.45±1.22
19.23±0.89
18.90±0.66
19.65±0.8
17.76±1.05
*Values are displayed as the mean zone of inhibition (mm) ± standard deviation of three replicates.
3.5. Antibacterial activity of essential oils of seeds of Ocimum basilicum var. thyrsiflora
The antibacterial activity of different concentrations of essential oils of Ocimum basilicum var. thyrsiflora seeds was
determined. The maximum antibacterial activity was observed in the essential oil extracted from seeds against P. aeruginosa
(20.06±0.30) at a concentration of 50% essential oil, and the minimum activity was noted against S. aureus (9.00±0.4) at a
concentration of 12.5% essential oil. Essential oils extracted from seeds showed antibacterial activity against all the tested
bacteria. The 50% essential oil extracted from seeds exhibited lower antibacterial activity against P. mirabilis (12.66±0.56).
Twenty-five percent essential oils exhibited the maximum antibacterial effect against P. aeruginosa (13.87±0.67) and the
minimum against P. mirabilis (10.08±0.24), while 12.5% showed the strongest antibacterial effect against P. aeruginosa
(14.00±0.8).
3.6. Antibacterial activity of essential oils of leaves of Ocimum basilicum var. thyrsiflora
The antibacterial activity of different concentrations of essential oils of Ocimum basilicum var. thyrsiflora leaves was
studied. The strongest antibacterial effect was evaluated in the essential oil extracted from leaves against P. aeruginosa
(18.33±0.32) at a concentration of 50% of the essential oil and the lowest effect was evaluated against P. mirabilis (7.46±0.45)
at a concentration of 12.5% of the essential oil. Essential oils extracted from leaves showed antibacterial activity against all
tested bacteria. The 50% essential oil extracted from seeds exhibited lower antibacterial activity against P. mirabilis
(10.34±0.85). Twenty-five percent essential oils exhibited the maximum antibacterial effect against P. aeruginosa (14.68±0.2)
and minimum antibacterial effect against S. aureus (9.00±0.75), while 12.5% showed the maximum antibacterial activity against
P. aeruginosa (13.5±0.5).
4. Discussion
Free radicals have a significant effect on pathogenic manifestations. Phytoconstituents derived from plants play a
major role in shielding the antioxidant defense system (Umamaheswari et al 2008). Several methods have been used to
0
5
10
15
20
25
30
35
DMSO NC Leaves 12.50% Leaves 25% Leaves 50%
Seeds 12.50% Seeds 25% Seeds 50%
Amoxicillin 10 μg
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investigate the plant’s antioxidant properties (Kil et al 2009), and it has been found that phytoconstituents are the
best synthetic alternatives (Vongtau et al 2005, Oluyemi et al 2007).
Ocimum basilicum var. thyrsiflora is well recognized for its essential oil and the presence of active phytoconstituents
such as alkaloids, carbohydrates, steroids, proteins, glycosides, tannins, flavonoids, terpenoids, and phenolic compounds
(Lyczko et al 2020). All of these compounds have potent antioxidant properties that actively scavenge the radicals responsible
for lipid peroxidation (Chanwitheesuk et al 2005). According to pharmacological studies, these constituents exhibit
antimicrobial, antioxidant (Bora et al 2011), antiviral (Chiang et al 2005), anti-inflammatory, hypolipidemic (Amrani et al 2009),
anti-platelet aggregation, anti-carcinogenic, antiulcerogenic, and antithrombotic activities. Phenolic compounds' antioxidant
properties are mostly related to their (Beric et al 2008) redox activities, and the antioxidant properties of most of these
phytochemicals have been linked to lower cancer mortality rates in a diversity of human populations (Oyas 2013).
DPPH, a purple bleaching solution high in free radicals, is commonly used to assess a plant's electron-donating ability
(Nunes et al 2012, Kar et al 2017).
The plant has a significant ability to quench ABTS radicals because of its significant ABTS radical quenching ability.
Therefore, it may be used to alleviate radical-related stress (Sahreen et al 2010).
The harmful reactive oxygen radical hydrogen peroxide (H2O2) damages cells when it is converted into the hydroxyl
radical, which can lead to DNA mutations and lipid peroxidation (Gülçın et al 2003). According to current research, essential oil
extracts can quench this radical. This is possible because of the phenolic compounds that help to convert H2O2 to H2O.
Pripdeevech et al (2010) reported that the essential oil of Ocimum basilicum var. thyrsiflora exhibits a potent scavenging
ability for DPPH radicals with an IC50 value of 98.33±2.08 µg/mL. According to Oonsivilai et al (2013), Ocimum basilicum var.
thyrsiflora ethanol extract had the highest antioxidant activity by FRAP assay at a value of 0.0186 ± 0.00 mmol Fe2+/g. Naidu et
al (2016) reported that the scavenging activity of DPPH radicals increased with increasing concentration, i.e., 73.75%, with an
IC50 value of 22 µg/mL. Ocimum basilicum var. thyrsiflora essential oil had 68% linalool and exhibited the highest antioxidant
activity. As a result, it is suggested that plant leaves contain significant antioxidant activity and therefore could be utilized as a
significant source of antioxidants.
According to Adam and Omer (2015), Ocimum basilicum leaf extract had the highest antibacterial activity against E. coli
and P. aeruginosa (7.8 mm inhibition zone) at the lowest concentration of 6.25 mg/ml and the lowest antimicrobial activity
against S. aureus (4.4 mm inhibition zone). Astuti (2016) reported that basil essential oil exhibited antibacterial activity against
S. mutants with an IC90 value of 0.23%. In an experiment, Kaya et al (2008) found that the methanol extracts of Ocimum
basilicum exhibited an antimicrobial effect against S. aureus (15 mm), Shigella specie (13 mm), P. aeruginosa (13 mm), E. coli
RSHI (14 mm), and E. coli ATCC 25922 (13 mm). There was no difference between the acetone and chloroform extracts. Reports
revealed that Ocimum basilicum var. thyrsiflora exhibited antibacterial activity against ampicillin-resistant E. coli and S. aureus
with MICs of 12.5 μL/mL and 6.25 μL/mL, respectively (Avetisyan et al 2017). The report revealed that ethanol, methanol, and
chloroform were used for the extraction of antifungal and antibacterial compounds from different parts of the medicinal plants
(Kar et al 2018a, b, c).
5. Conclusions
In this study, we found that the essential oils of Ocimum basilicum var. thyrsiflora showed potent antioxidant and
antibacterial activities. It showed a stronger antioxidant effect when tested using the hydroxyl radical (HO•) scavenging assay
technique than other methods. It could be utilized as a widely available source of naturally occurring antioxidants and an
excellent source of food supplements. Antioxidants found in natural products play a key role in preventing the harmful effects
of free radicals. In addition, antioxidants derived from plants are less expensive and safer than synthetic alternatives. This
suggests that the Ocimum basilicum var. thyrsiflora essential oils obtained from the plant's seeds and leaves have antibacterial
properties. Therefore, it would be preferable to isolate antimicrobial compounds from the leaves and seeds of Ocimum
basilicum var. thyrsiflora to develop novel antimicrobial therapeutics to treat various infectious ailments caused by microbes.
From this study, we concluded that due to the high antioxidant and antimicrobial activity of Ocimum basilicum var. thyrsiflora,
it might be beneficial to incorporate natural into therapeutic medicines for better implications on human health.
6. List of abbreviations
DPPH - 2-Diphenyl-1-picrylhydrazyl.
ABTS - Azino-bis (3-ethylbenthiazoline-6-sulphonic acid.
MPKC - Medicinal Plants Knowledge Center.
BHT - Butylated hydroxytoluene.
DMSO - Dimethyl sulfoxide.
ANOVA - Analysis of variance.
SEM - Standard error of mean.
SD - Standard Deviation.
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NC - Negative Control.
PC - Positive Control.
MIC - Minimum inhibitory concentration.
Acknowledgment
The authors are grateful to Prof. (Dr.) Sanghamitra Nayak, Head, Centre of Biotechnology, Prof. (Dr.) Sudam Chandra Si,
Dean, Prof. (Dr.) Manoj Ranjan Nayak, President, Siksha ‘O’ Anusandhan University, Bhubaneswar, India, for providing all
facilities.
Ethical considerations
Not applicable.
Conflict of Interest
The authors declare that they have no conflict of interest. The authors Akankshya Sahu and Gayatree Nayak contributed equally
to this work.
Funding
This research did not receive any financial support.
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