Comparative in Vitro Study of The Antimicrobial Efficacy of Nigella Sativa Seed Oil (Black Seed Oil) Against Selected Microorganisms With Conventional Antibitics

Volume 4, Issue 9, September – 2019 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
Comparative in Vitro Study of the Antimicrobial

Efficacy of Nigella Sativa Seed Oil (Black Seed Oil)
Against Selected Microorganisms with
Conventional Antibitics
A.D.H. Sudesh, D.N. Wanigasekara, E.D.C. Karunarathne
Gampaha Wickramarachchi Ayurveda Institute
University of Kelaniya
Yakkala, Sri Lanka
Abstract:- This study investigates the antimicrobial microorganisms can be used to delay the emergence of
activity of essential oil obtained from Nigella sativa seeds antibiotic resistance and as successful antimicrobials [5].
(Black seed oil). The objectives are to determine the
antimicrobial efficacy of black seed oil against selected Pharmaceutical concerns invest resources for discovery
pathogens such as Pseudomonas aeruginosa (ATCC programs of novel compounds with considerable
27853), Methicillin Resistant Staphylococcus aureus antimicrobial activity [6]. There is a significant trend to
(MRSA), Methicillin sensitive Staphylococcus aureus determine the antimicrobial potential of untapped natural
(MSSA), Staphylococcus pyogenes (ATCC 19615), sources such as plants, animals and microorganisms for a
Escherichia coli (ATCC 25922 and ATCC 35218) and new generation of antimicrobials [7].
clinically isolated Candida albicans and to compare the
ability of microbial growth inhibition with conventional Although there are 300,000 herbal species that present
antibiotics that are used for the test organisms. Bacteria globally, only 15% plant species have been investigated for
were inoculated on Mueller Hinton Agar and Sabouraud their pharmacological potential [8]. Among various plants
Dextrose Agar was used to inoculate C. albicans. Agar that possess medicinal value, Nigella sativa L. (Black cumin
well diffusion technique was used to evaluate or black seed in English) which belongs to the family
antimicrobial activity of the test organisms. As Dimethyl Ranunculaceae has been considered as one of the most
sulfoxide (DMSO) was served as the solvent for black precious herb around the world [9]. Black cumin seed is
seed oil, it was used as the negative control. Among the widely used in traditional medicine for wide range of
test organisms, staphylococci; MRSA, MSSA, S. ailments [10].
pyogenes and C. albicans were sensitive to the oil. Both
two strains of E.coli, P. aeruginosa were resistant to the This study was conducted with the aims of determining
oil while all test organisms were susceptible to the the antimicrobial effect of black seed oil against selected
antibiotics used. The comparative study revealed that all strains of pathogenic bacteria and yeast and to compare the
three staphylococci are more susceptible to black seed oil antimicrobial efficacy with conventional antibiotics that are
than amoxicillin in tested concentration. effective against the tested pathogens.
Keywords:- Nigella Sativa; Black Seed Oil; Antimicrobial II. MATERIALS AND METHODS
Activity; Antibitic Resistance.
A. Sample Preparation
I. INTRODUCTION
 Black seed oil test sample preparation
Emergence of resistance in pathogenic microorganisms Commercially available black seed oil (Manufacturer;
to antibiotics that are used to treat serious infections causes Bio Extracts (Pvt) Ltd, Colombo 3, Sri Lanka) was mixed
considerably increased mortality [1]. After the introduction with DMSO (Dimethyl sulfoxide) to the ratio of (9:1)
of antibiotics, the antibiotic resistance genes of both aseptically.
pathogens and commensals have been widespread
throughout the environments like hospitals, communities  Microbial culture preparation
etc. [2]. In addition, organisms have been acquiring multi- Pseudomonas aeruginosa (ATCC 27853), Methicillin
drug resistance during the past few years [3].Those multi- Resistant Staphylococcus aureus (MRSA), Methicillin
drug resistant bacteria can cause serious, incurable infections sensitive Staphylococcus aureus (MSSA), Staphylococcus
[4]. pyogenes (ATCC 19615), Escherichia coli (ATCC 25922
and ATCC 35218) and clinically isolated Candida albicans
Novel alternative approaches should be introduced to were inoculated in sterilized test tubes containing Nutrient
face the challenge of antibiotic resistance. Development of Broth separately and incubated overnight at 37oC. After
new effective strategies against antibiotic resistant incubation, cultures were adjusted by adding sterilized
IJISRT19SEP1426 www.ijisrt.com 458

ISSN No:-2456-2165
peptone water until turbidity matched that of a McFarland “BSO”, “+” and “-”. 50 µL of black seed oil test sample was
0.5 standard. added to all wells labeled as BSO. Ciprofloxacin (50 μL; 10
mg/mL) was added to the wells that are named as “+” of P.
 Antibiotic solution preparation aeruginosa inoculated agar plates. Amoxicillin (50 μL; 10
mg/mL) was added to the “+” labeled wells of the agar
 Preparation of Amoxicillin plates inoculated with Methicillin Resistant Staphylococcus
Amoxicillin powder was dissolved in sterile distilled aureus (MRSA), Methicillin sensitive Staphylococcus
water aseptically to obtain the concentration of 10 mg/ mL aureus (MSSA), Staphylococcus pyogenes (ATCC 19615),
Escherichia coli (ATCC 25922 and ATCC 35218). “+”
 Preparation of Ciprofloxacin wells of the C. albicans inoculated plates were added with
Ciprofloxacin powder was dissolved in sterile distilled fluconazole (50 μL; 2.5 mg/mL). All the wells named as “-”
water aseptically to obtain the concentration of 10 mg/mL were filled with 50 μL aliquots of DMSO. The test was
triplicated for all microorganisms used.
 Preparation of Fluconazole
Fluconazole powder was dissolved in sterile distilled Agar plates were incubated at 370C for 24 hours.
water aseptically to obtain the concentration of 2.5 mg/mL Antimicrobial efficacy of black seed oil and antibiotics were
evaluated using the inhibition zone diameter around the
 Inoculation of microbial lawns respective wells that indicated as clear zones. Average was
Aliquots (100 µL each) of turbidity adjusted broth taken from the all three diameters of each triplicate.
cultures of Pseudomonas aeruginosa(ATCC 27853),
Methicillin Resistant Staphylococcus aureus (MRSA),
Methicillin sensitive Staphylococcus aureus (MSSA),
Staphylococcus pyogenes (ATCC 19615), Escherichia coli III. RESULTS AND DISCUSSION
(ATCC 25922 and ATCC 35218) were pipetted out onto
Mueller-Hinton agar plates separately and swabbed Bactericidal activity of Black seed oil was assessed for
uniformly over the agar surface using sterile cotton swabs to six strains of bacteria and one strain of yeast. The
obtain lawns. antimicrobial activity and the potency of black seed oil and
antibiotics were quantitatively assessed by the presence or
100 µL aliquot of Candida albicans culture with absence of inhibition zone and zone diameter. According to
adjusted turbidity was pipetted out onto Sabouraud Dextrose the obtained results, black seed oil was effective against all
Agar and inoculated evenly as mentioned above. tested Staphylococci while it was unable to affect the growth
of two E.coli strains and P. aeruginosa. The growth of the
B. Screening antimicrobial activity of black seed oil and yeast was also affected by black seed oil. All strains used
antibiotics were susceptible to the respective antibiotics. DMSO which
Agar well diffusion method was used to determine the was used as the negative control did not exhibit any
antimicrobial efficacy of black seed oil and antibiotics. inhibition against the tested organisms. The results were
Three wells (diameter 8 mm) were made on each inoculated recorded in Table 1.
agar plate using a sterile cork borer. Wells were labeled as
Fig 1:- Sensitivity testing showing clear zones on Mueller Hinton Agar inoculated with MRSA

ISSN No:-2456-2165
Inhibition zone diameter in mm
Organism
Black seed oil Antibiotic
MRSA 42.33±0.58 12.33±0.58
MSSA 37.67±0.58 28.27±0.58
S. pyogenes 38.33±0.58 34.33±0.58
P. aeruginosa Nil 34.33±0.58
E. coli
Nil 36.33±0.58
(ATCC 25922)
E. coli
Nil 36.00±0.58
(ATCC 35218)
C. albicans 21.00±0.00 35.00±0.00
Table 1:- Antimicrobial activity of black seed oil and antibiotics
DMSO (Dimethyl Sulfoxide) was used as the solvent to thymol [13], thymoquinone [14] and thymohydroquinone
dissolve Black seed oil. Therapeutic substances that are [15] that is known to possess antimicrobial effectiveness.
insoluble in water are probably soluble in DMSO and its high
diffusibility [11] helps to penetrate the test sample through IV. CONCLUSION
the agar medium. To check whether there is an effect of
DMSO towards microbial growth, it was used as the negative Black seed oil possesses considerably higher
control which confirmed that it does not affect the growth of antibacterial effect against MRSA, MSSA and S. pyogenes
tested organisms. (ATCC 19615) than the tested antibiotics. It also affects the
growth of clinically isolated C. albicans strain but the
Black seed oil exhibited significant inhibitory action reference antibiotic is more effective. Black seed oil does not
against MRSA compared to the standard amoxicillin. This affect the growth of Escherichia coli (ATCC 25922 and
result indicates that the antimicrobial action of black seed oil ATCC 35218) and Pseudomonas aeruginosa (ATCC 27853).
is not interfered with the antibiotic resistance. It might Although black seed oil is used for typical application in
possess different modes of action regarding inhibition on test traditional medicine, further studies should be carried out to
organism. evaluate the systemic use and safety of use.
Although growth of Methicillin sensitive REFERENCES

staphylococcus aureus (MSSA) was inhibited by both
Amoxicillin and black seed oil, when considering the extent [1]. Heike von Baum, Reinhard Marre, 2005.
of inhibition, black seed oil exhibited higher capability of Antimicrobial resistance of Escherichia coli and
inhibition. therapeutic implications. International Journal of
Medical Microbiology, Issue 295, pp. 503-508.
Clinically isolated Candida albicans strain was found [2]. Levy, S. B., 2005. Antibiotic resistance—the problem
to be sensitive to Black seed oil. However, quantitatively intensifies. Advanced Drug Delivery Reviews, Issue 57,
anticandidal activity of fluconazole was higher than that of p. 446– 1450.
black seed oil. [3]. Meyer KS, Urban C, Eagan JA, et al. Nosocomial
Outbreak of Klebsiella Infection Resistant to Late-
Both two E.coli strains showed no susceptibility to Generation Cephalosporins. Ann Intern Med.
black seed oil. They were sensitive to Amoxicillin exhibiting 1993;119:353–358.
approximately similar zone diameters. [4]. Cohen, M.L., 1992. Epidemiology of drug resistance:
implications for a post—antimicrobial
S. pyogenes showed to be sensitive to both black seed era. Science, 257(5073), pp.1050-1055.
oil and Amoxicillin. Inhibition zone diameter of black seed [5]. Sadiku, E.R., Fasiku, V., Agboola, O., Ibrahim, I.D.,
oil is larger than the diameter of the antibiotic which Daramola, O.O., Olajide, J.L., Kupolati, W.K., Sanni,
indicates higher inhibitory action of black seed oil on S. S. and Mochane, M.J., Overcoming Antimicrobial
pyogenes. Resistance: New Strategies, Expectations: Greater
Hope. Biomedical Journal, 1, p.5.
P. aeruginosa (ATCC 27853) was found to be resistant [6]. Payne, D.J., Gwynn, M.N., Holmes, D.J. and
to black seed oil while it was susceptible to ciprofloxacin. Pompliano, D.L., 2007. Drugs for bad bugs:
confronting the challenges of antibacterial
Almost all antibiotics are secondary metabolites of soil discovery. Nature reviews Drug discovery, 6(1), p.29.
bacteria and fungi [12].Evaluation of the potency of plant [7]. Taylor, P.W., 2013. Alternative natural sources for a
derived products is particularly relevant to the discovery of new generation of antibacterial agents. International
new antimicrobial drugs. journal of antimicrobial agents, 42(3), pp.195-201.
[8]. De Luca, V., Salim, V., Atsumi, S.M. and Yu, F.,
Antimicrobial activity of black seed oil may be due to 2012. Mining the biodiversity of plants: a revolution in
the presence of active phytochemical substances such as the making. Science, 336(6089), pp.1658-1661.

ISSN No:-2456-2165
[9]. Takruri, H.R. and Dameh, M.A., 1998. Study of the
nutritional value of black cumin seeds (Nigella
sativaL). Journal of the Science of Food and
Agriculture, 76(3), pp.404-410.
[10]. Yimer, E.M., Tuem, K.B., Karim, A., Ur-Rehman, N.
and Anwar, F., 2019. Nigella sativa L.(Black Cumin):
A Promising Natural Remedy for Wide Range of
Illnesses. Evidence-Based Complementary and
Alternative Medicine, 2019.
[11]. Brayton, C.F., 1986. Dimethyl sulfoxide (DMSO): a
review. The Cornell Veterinarian, 76(1), pp.61-90.
[12]. Fischbach, M.A. and Walsh, C.T., 2009. Antibiotics
for emerging pathogens. Science, 325(5944), pp.1089-
1093.
[13]. Randhawa, M.A. and Al-Ghamdi, M.S., 2002. A
review of the pharmacotherapeutic effects of Nigella
sativa. Pak J Med Res, 41(2), pp.77-83.
[14]. Kahsai, A.W., 2003. Isolation and characterization of
active ingredients from Nigella sativa for antibacterial
screening.
[15]. El-Fatatry, H.M., 1975. Isolation and structure
assignment of an antimicrobial principle from the
volatile oil of Nigella sativa L. seeds. Die
Pharmazie, 30(2), pp.109-111.
[16]. Karapinar, M. and Aktuǧ, Ş.E., 1987. Inhibition of
foodborne pathogens by thymol, eugenol, menthol and
anethole. International Journal of Food
Microbiology, 4(2), pp.161-166.
[17]. Ahmad, I. and Beg, A.Z., 2001. Antimicrobial and
phytochemical studies on 45 Indian medicinal plants
against multi-drug resistant human pathogens. Journal
of ethnopharmacology, 74(2), pp.113-123.
[18]. Nascimento, G.G., Locatelli, J., Freitas, P.C. and Silva,
G.L., 2000. Antibacterial activity of plant extracts and
phytochemicals on antibiotic-resistant
bacteria. Brazilian journal of microbiology, 31(4),
pp.247-256

Comparative in Vitro Study of The Antimicrobial Efficacy of Nigella Sativa Seed Oil (Black Seed Oil) Against Selected Microorganisms With Conventional Antibitics

Uploaded by

Copyright:

Available Formats

Comparative in Vitro Study of The Antimicrobial Efficacy of Nigella Sativa Seed Oil (Black Seed Oil) Against Selected Microorganisms With Conventional Antibitics

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Comparative in Vitro Study of The Antimicrobial Efficacy of Nigella Sativa Seed Oil (Black Seed Oil) Against Selected Microorganisms With Conventional Antibitics

Uploaded by

Copyright:

Available Formats

Volume 4, Issue 9, September – 2019 International Journal of Innovative Science and Research Technology

Comparative in Vitro Study of the Antimicrobial

IJISRT19SEP1426 www.ijisrt.com 458

IJISRT19SEP1426 www.ijisrt.com 459

Although growth of Methicillin sensitive REFERENCES

IJISRT19SEP1426 www.ijisrt.com 460

IJISRT19SEP1426 www.ijisrt.com 461

You might also like