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Second International Virtual Conference of Biotechnology Research Center (IVCBRC-2021)
Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
21
Study the effect of Indian premium (Saussurea costus) extract and sesame oil on the
sensitivity of local Acinetobacter baumannii isolates causing infection
Khalid Najim Abdullah Laith Muslah – Najeeb Ezeddin Albayyer
Biology Department / College of science / University of Anbar- Iraq
Corresponding author: Khalid.khaleafea57@gmail.com
Received: 10 /Aug. /2021 , Accepted: 24 /Aug. /2021
Abstract
The current investigation was intended to assess the viability of sesame oil got from Heet city and
Indian premium (saussurea costus) affectability testing against Acinetobacter baumanii isolates
detached from disease destination like urine, sputum, C.S.F, wound swab, and burn swab. The results
indicated that sesame oil activated the gentamycin antibiotic activity from 3.08 mm to 14.7500 mm
and also the Indian premium made the same thing the activation was from 5.4167 mm to 9.3333 mm.
while the addition of sesame oil to Imipenem anti biotic there was inhibition in the activity of
Imipenem from 21.33 mm to 20.083mm and when we added the Indian premium there was activation
to the Imipenem antibiotic was from 22.1667 mm to 23.4167mm. Finally the results of cefotaxime after
adding the sesame oil we noticed that there is decrease in the activity of cefotaxime from 3.500 mm to
1.000 mm and we also noticed that the addition of Indian premium to the antibiotic there was
decrease in the activity from 3.9167 mm to 3.333 mm.
Therefore ower explanation that there is activation to some antibiotics there is also inhibition to the
other by adding different plant extract.
Key Words: Saussurea costus, sesame oil , Acinetobacter baumannii.
Introduction
Acinetobacter is variety Gram-negative with
the widest Gammaproteobacterial class position.
Species of Acinetobacter are not motile and
oxidase – negative and usually Nitrate –negative
(1) patients are Acinetobacter genus a vital
wellspring of disease in crippled patients in clinic
,specifically species Species of Acinetobacter
baumanii of the family Acinetobacter are
rigorously vigorous, non-fermentative ,Gram
negative bacilli . Acinetobacter is frequently
separated from Nosocomial conditions and is
mostly used in concentrated examination units, in
which normal A.baumanii is both incoherent and
pandemic, are the continuous reasons for
Nosocomial pneumonia especially late-start
ventilatory-related pneumonia. (2) The spread of
medication safe microorganism is quite possibly
the most genuine dangers to effective treatment
of microbial sickness(3).Circumstantially the
most recent decade has likewise seen expanding
escalated concentrates on separates and organic
dynamic mixtures disengaged from plant species
utilized for characteristic treatments or home
grown medication (4),(5) for more than millennia
,regular plants have been viewed as an important
wellspring of therapeutic specialists with
demonstrated capability of treating irresistible
illness and with lesser results contrasted with
engineered drug specialists .Sesame has a place
with the family-pedaliaceae and variety –
sesamum . The sort comprises of around 36
species and 19 of which are native to Africa.
Sesame plant is accepted to have started from
Africa (6). In societal medicines in Africa and
Asia, sesame is rumored. All plant parts are
precious. However, the decoction of the leaves in
south-western Nigeria is used for treating injured
and expelled skin, eye and fomentation Mixed
warm water leaf is used for washing stimulated
mouth films. The two leaves and roots were
decoction for chicken box and measles and were
used in Taenia capitis as a hair cleaner (7).
Sesame seed oil, known to the man since the start
of its development, has been used as recovering
oil and privately burnt as a permanent food in
Nigeria notably in southwest and central belt
areas.The sesame oil consists of mainly four fatty
Second International Virtual Conference of Biotechnology Research Center (IVCBRC-2021)
Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
22
acids, while other fatty acids appear in very small
amounts. Due to the fact that sesame seeds are
rich in UFAs, as well as vitamin and minerals
such as calcium, magnesium and phosphorus, its
oil has health benefits. Therefore, sesame oil has
been used as medicine or for pharmaceutical
purposes (8).Sesame oil contains Vitamin E and
several important antioxidants such as sesaminol
and sesamolinol that are believed to promote the
integrity of body-tissues in the presence of
oxidizing compounds (9).Saussurea costus, a
premium known in secret as Kuth in Pakistan and
India has the overpowering flavor of the Western
Himalayas. The species is confined to a small
complice of the Himalayas and develops at a
height of 2600-4000 meters on moist slopes
(10),(11). The species has evolved during the
1920's and 30's in Himac hil Pradesh and
Uttarakhand (12). The species has long been
employed in the area's conventional clinical
assessment procedures. S. costus is the most
industrially suitable species among the Saussurea
kinds. Costus has a powerful, sweet scent of
severe flavor and is used as an antiseptic, in
bronchial asthma management, in particular
vagotonic asthma. Courses of action for this
species are also provided an explanation for the
correction of various contaminations and
circumstances. The oil extracted from the roots is
known as Costus Oil, which is utilized for the
preparation of high quality perfumes and hair oil.
In addition, Costus Oil is predicted to be strong in
illness therapy (13). The Saussura costus
constituents are a total 35 aroma compounds
representing about 92.81% of the total
composition were identified. Aldehyde like (7Z,
10Z, 13Z)-7, 10, 13-hexadecaterinal (25.5%) was
found as a major compound including other
ketones like dehydrocostus lactone (16.7%),
alcohols like elemol (5.84%), γ-costol (1.80%),
vulgarol B (3.14%), valerenol (4.20%), and
terpinen-4-ol (1.60%), etc. Esters and acids were
found to be completely absent in our samples.
Materials and Methods
Sample collection:
The samples of sesame oil collected for the study
was from Heet city. It was extracted by different
farmers of Heet from the locally grown sesame
seeds and the sample of Indian premium
(Saussurea costus) was brought from the Herb
shop in Al-khaldia like powder form.
Sesame is a warm –season annual crop .sesame is
a drought tolerant but doesn't need irrigation
when young farmers know when to pick sesame
seeds harvest occurs 90-150 days from planting
out the crop must be harvested .When the plant is
mature the leaves and stem of sesame plant
change from green to yellow to red.The leaves
start to drop from the plant .If its planted in early
june for example the plant will begin dropping
leaves and drying out in early October it’s not
ready to pick through it takes a while for the
green to disappear from upper seed capsules this
is referred as drying down.After the plant is dried
completely the capsule that cotaines the seed will
automatically explode and shoot out the seeds
and then its harvested then the seeds are squeeze
out and will get sesame oil.
We identify the sesame plant as an annual plant
growing 50 to 100 cm tall with opposite leaves 4
to14 cm with an entire margin ,they are
lanceolate, to 5 cm broad at the base of the plant,
narrowing to just 1 cm broad on the flowering
stem .
Collection of isolates:
Isolates from various departments of the Al-
ramadi education hospital were taken and these
isolates thered from patients following history,
then sampled from a variety of samples, using
procedures that were discussed in relation to
kinds such as urine.,Burn swab ,Wound swab,
sputum and C.S.F . from all specimen only 12
The Acinetobacter baumannii has been
discovered by utilizing isolates Vitek 2. System.
Identification:
According to the specifications of the
manufacturer, recognizable evidence with the
VITEK 2 framework was provided via IDGNB
cards. 41 tests, 18 sugar osmosis tests, 18 sugar
aging tests, 2 decarboxylase tests, and 3 other
tests are included on 64-well plastic ID-GNB
cards (for urease, use of malonate, and
tryptophane deaminase). The cards with vacuum
Second International Virtual Conference of Biotechnology Research Center (IVCBRC-2021)
Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
23
device are vaccinated with the suspension of the
biological entity of 0,5 McFarland prepared for
18 to 20 hours on Macconkey agar plate
(bioMérieux). VITEK 2 peruser inoculator
module. Fluorescence is estimated each 15 min,
and the aftereffects of recognizable proof are
resolved after 3 hr.
Culture media and inoculums preparation:
After we identified all the isolates we prepared
special media used in study as the following:
1-first media was prepared by solving 1.3g of
nutrient broth powder in 90ml of DW and then
we add 10 ml Indian premium extract which was
prepared by solving 100 g of Indian premium in
400 ml boiled and hot DW and left for 24 h and
then we take 10ml of the extract after
centrifugation .And then put it in autoclave for
sterilization. The method of plant extraction is
done by adding 100gm of Indian premium in
powdered form to 400 ml of hot boiled water then
left for 24 hr. in room temperature and after we
centrifuge the hole solution and then that 10ml of
the leached solution and added to 90 ml nutrient
broth.
2- second media is prepared by solving 1.3g of
nutrient broth powder in 90ml DW and then we
add 10 ml of sesame oil which was prepared by
diluting 3ml of sesame oil in 7ml of DMSO after
diluting the sesame oil it was added to 90ml of
nutrient broth and them autoclaved for
sterilization .
After we finish the peppermint of the media its
left to cool up and inoculated by the bacterial
isolates which was identified as Acinetobacter
baumannii by vitek 2 .The isolates were done by
solving a colony from the agar media in 5ml
normal saline until we get a suitable Mcfarland
which was 0.5 measured by a density meter .we
inoculate both of nutrient broth which contains
Indian premium and that contains the sesame oil.
They are left in overnight incubation we will see
turbidity in the broth because of the bacterial
growth.
Agar diffusion method:
We use this method in order to see the inhibition
zone of the antibiotic to the bacteria and it's done
by the following way.
We prepare 100ml of Muller Hinton agar and
poured in to Petri dishes and left until it gets
solidified .After solidification we put a swab in
the broth media that contains growth until if
touches the surface of the media and spread it on
the surface the Muller Hinton agar until we get a
homogenized spread and then we spread three
which were Imipenim (IPM),gentamycin (GN)
and cefotaxim (CTX) anti biotics that used for
study at suitable distances on the Muller Hinton
agar and when finish we incubate the Petri dishes
for 24 hours at 37 C°.
Measurement of zone inhibition:
When the incubation time is finished we will see
inhibition zones on the surface of the media
caused by the effect of the antibiotic on the
bacteria the zone is measured from the edge to
the cross edge of the zone.
Results and Discussion
In the present study ,the antimicrobial activity
was evaluated against Acinetobacter baumannii
species .in the first sesame oil added to
Gentamycin antibiotic were applied on the
bacterial isolate has more effect on the bacterial
isolate than the addition of Indian premium as
seen in table 1 , figure 1 .
Second International Virtual Conference of Biotechnology Research Center (IVCBRC-2021)
Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
24
Table1: The antimicrobial activity of sesame oil after addition to Gentamycin
Medias
N
Mean ± Std. Error
Seasame
12
B
10.83±2.99
Gent+Sesam
12
B
14.75±2.00
Gent CN10
12
A
3.08±1.40
Indian prem
12
AB
6.16±2.27
Genta+Ind prem
12
B
9.33±2.23
Genta CN10
12
AB
5.41±1.59
Total
72
B
8.26± 0.96
P 0.004
Figure 1: The antimicrobial activity of sesame after addition to Geantamycin antibiotic
While in table 2, figure 2 we see that there is no
antimicrobial actitivity of IPM after the addition of sesame oil nor did the Indian premium addition
with IPM.
0
2
4
6
8
10
12
14
16
Seasam
e
Gent+Se
sam
Gent
CN10
Indian
prem
Genta+I
nd prem
Genta
CN10
Mean 10.8333 14.75 3.0833 6.1667 9.3333 5.4167
mm
Second International Virtual Conference of Biotechnology Research Center (IVCBRC-2021)
Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
25
Table 2: shows that there is no antimicrobial activation of antibiotic to bacterial isolates after addition to sesame
and Indian premium
Media
N
Mean ± Std. Error
Sesame oil
12
A
6.41 ± 1.79
IPM+Sesame
12
B
20.08 +± 4.20
IPM
12
B
21.33 ± 3.95
Indian prem
12
A
2.66 ± 1.13
IPM+Ind prem
12
B
23.41 ± 5.09
IPM
12
B
22.16 ± 4.00
Total
72
16.01 ± 1.73
P=0.000
Figure 2: shows the activation of IMP antibiotic after adding Indian premium
In table three we noticed after we done the
experiment that there is no activation of antibiotic
activity when we add the Cefotaxim with sesame
extract and with Indian premium extract as it’s
seen in the table 3 and also in figure 3.
0
5
10
15
20
25
IPM IPM+In
d
Indian IPM IPM+Se
same
Sesame
Series1 22.1667 23.4167 2.6667 21.3333 20.0833 6.4167
mm
Second International Virtual Conference of Biotechnology Research Center (IVCBRC-2021)
Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
26
Table 3: shows the inhibition of antibiotic activity after adding Indian premium and sesame oil
Medias
N
Mean ± Std. Error
Sesame
12
1.00 ± 0.00
CTX30 + Ses
12
1.00 ± 0.00
CTX 30
12
3.50 ±1.68
Indian prem
12
1.00 ± 0.00
CTX 30+Ind prem
12
3.91 ± 1.55
CTX 30
12
3.33 ± 1.59
Total
72
2.2917 ± 0.47
P 0.182
Figure 3: shows inhibition activity of antibiotic when adding Indian premium extract and sesame oil
The over use of antibiotic in the treatment of
infection and the appearance of multi- drug
resistant bacterial strains because of mutations
(which means resistant to two or more
antibiotics).Which leads to find a research
towards the study of anti-microbial agents from
natural sources such as sesame oil and Indian
premium. Sesame oil showed activation with
gentamycin antibiotic and also did the Indian
premium. While the addition of sesame oil to
imipenem antibiotic did not make activation with
antibiotic but when we added indian premium
there was an activity with antibiotic. However,
the addition of sesame oil and Indian premium to
the cefotaxime, there was inhibition in the
activity of the cefotaxime.
0
0.5
1
1.5
2
2.5
3
3.5
4
Sesame CTX30 +
Ses
CTX 30 Indian
prem
CTX
30+Ind
prem
CTX 30
Mean 1 1 3.5 1 3.9167 3.3333
mm
Second International Virtual Conference of Biotechnology Research Center (IVCBRC-2021)
Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
27
Conclusion
The current investigation means the counter
microbial part of the expansion of sesame oil to
Gentamycin and anti-toxin which builds the
hindrance movement of the Gentamycin
medication to the microorganisms. Sesame oil
may be considered as the hotspot for improving
the new broad-based antibacterial strategy. These
outcomes backing and conviction that the
expansion of sesame oil to the anti-infection may
have numerous drug jobs. In similar study by
Oloma et al. (14) who observed that sesame oil
did not show any activity against s.aureus and
klebsiella spp.and his result also revealed that the
sesame oil were active against fungi but other oil
didn’t show any activities in these organism .the
results of Oloma didn’t agree with our result in
which we find that there was activity of sesame
oil on the fungi but not on bacteria. Munir et al.
(15) in Nigeria found that methanolic and
ethanolic extracts have broad spectrum
antimicrobial effect against all the tested
pathogenic micro-organisms except
Streptococcus pneumoniae and Staphylo-coccus
aureus respectively, while the aqueous extract
exhibited inhibitory activity on S. aureus, S.
pneumoniae and Candida albicans.The study by
Munir et al., agree with the results that we find
that there was activity on bacteria when we use
the sesame oil.
The sesame sanani oil and dwaini oil have more
activity against A. niger and A. flavus. These
results do not agree with Anand et al.,(16) who
found antibacterial activity of sesame oil against
bacteria. S. mutans and Lactobacillus acidophilus
were found to be moderate to sensitive to the
sesame oil. This study agrees with the results that
we find that sesame was activated against
bacteria but the sesame oil was not active on
(fungi) and this did not agree with our finding.
Acknowledgement:
I might want to thank the university of Anbar,
collage of science and Biology departmant for
their extraordinary acquired assistance by the
postgraduate labs .Additionally thanks reached
out to both Mr. Mohamed Aljabri and Mr.
Hameed Mahmud for supporting this work.
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Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
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Second International Virtual Conference of Biotechnology Research Center (IVCBRC-2021)
Journal of Biotechnology Research Center
Vol.15 No.2 P-ISSN: 1815-1140
Published Online: (December) 2021 https://doi.org/10.24126/jobrc.2021.15.2.608 E-ISSN: 2708-1370
29
Saussurea costus)
Acinetobacter baumanii
Corresponding author: Khalid.khaleafea57@gmail.com
Acinetobacter baumanii
Acinetobacter baumanii