NewBioWorld A
Journal of Alumni Association of Biotechnology (2023) 5(1):20-23
RESEARCH
ARTICLE
Comparative Evaluation
of In Vitro Antimicrobial Efficacy of different Species of Curcuma
against Human Pathogenic Bacteria
Krishna Kumar Verma1, Shristi Soni1, Pratiksha Pandey1,
Varsha Sahu1, Sumit Kumar Dubey2, Neha Behar2*
1Department of Microbiology, D.L.S.
P.G. College, Bilaspur,
Chhattisgarh, India
2 Department of Biotechnology, D.L.S. P.G. College, Bilaspur, Chhattisgarh, India
*Corresponding Author Email- neha1_biotech@yahoo.com
ARTICLE INFORMATION
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ABSTRACT
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Article history:
Received
27 April 2023
Received in revised form
23 May 2023
Accepted
Keywords:
Antimicrobial;
Extract;
Curcuma;
Human
pathogenic; bacteria;
Disk-diffusion
method;
Herbal
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Herbal medicines have been commonly used for the treatment
and prevention of disease, health promotion and for the enhancement of the
life span. The Curcuma species
viz., C. Roxb., C. longa Linn and C. amada are used as medicine in traditional
therapeutic practices. The remedy for various diseases including cough,
diabetes and hepatic disorders. The efficiency of turmeric fractions, such as
petroleum ether, chloroform, benzene, methanol and aqueous were evaluated for
their inhibitory effect on clinical and standard strains of pathogenic bacteria.
The methanolic fraction of Curcuma
longa rhizome had high potential to inhibit some pathogenic bacteria
i.e. S. aureus to a greater
degree than other fractions of Curcuma
longa. In our study the results show that the different fractions
(petroleum ether, methanol etc.) of Curcuma
caesia rhizome were more
effective antimicrobial agents than the crude extract of Curcuma longa. The bacteria
isolates include E. coli (MTCC
443), Pseudomonas aeruginosa
(MTCC 424), and Staphylococcus
aureus (MTCC96) Antimicrobial activities were estimated by Disc
diffusion method. The results were pointed as the methanol, acetone and
petroleum ether, extract was more effective on all three pathogenic bacteria.
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Introduction
Medicinal plants represents an excellent source of
antimicrobial agents since ancient times (Verma et al. 2018). In rural areas
many plant materials are used as traditional medicine due to ample availability
and are relatively cheaper than modern medicine (Harit et al. 2013). Plants
generally produce many secondary metabolites (like phenol, alkaloid,
terpenoids, glycosides, etc.) which have significant potential as microbicides,
pesticides, fungicides, and many pharmaceutical drugs. Plant products are still
in use in pharmaceutical preparations (Chattopadhyay et al. 2004).
DOI: 10.52228/NBW-JAAB.2023-5-1-4
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The members of the family Zingiberaceae are found
to be rich source of phytochemical substances. They are rich in curcuminoids and are recognized
for their broad spectrum of biological activities, diversified chemical
structures, physicochemical characteristics as well as functional properties (Kaur
et al., 2018; Khan et al. 2022). Plants belonging to the Zingiberaceae family
are used as a medicine in the traditional system because of their wide spectrum
of biocidal activities (Khan et al. 2022). Many species of Curcuma are
used in traditional medication systems. Many literature have claimed the antifungal,
antibacterial, and inflammatory action of Curcuma longa, Curcuma
caesia, Curcuma aromatica, and Curcuma amada (Naz et al.
2010; Luthra et al. 2001). The rhizome
of these plants has been reported for antimicrobial action since ancient times
(Chopra et al. 1941).
The bacterial strains are significantly involved in
damaging food stuff and causing a variety of diseases in humans. Gram-positive
bacteria such as Staphylococcus aureus and Bacillus subtilis are
mainly responsible for post-operative wound infection, toxic shock syndrome,
and food poisoning. Whereas, Gram-negative bacterium such as Pseudomonas
aeruginosa has been associated with lung, urinary tract, and kidney
infections, and Escherichia coli linked with dysentery (Negi et al. 1999).
P. aeruginosa is the third most common
cause of nosocomial infections with a high mortality rate and E. coli often
causes urinary tract infections, sepsis, and meningitis. Staphylococcus aureus is a major bacterial
human pathogen that causes a wide variety of clinical manifestations. Infections
are common both in community-acquired as well as hospital-acquired settings and
treatment remains challenging to manage due to the emergence of multi-drug
resistant strains such as MRSA (Methicillin-Resistant S. aureus). S.
aureus is found in the environment and is also found in normal human
flora, located on the skin and mucous membranes (most often the nasal area) of
most healthy individuals. S. aureus does not normally cause
infection on healthy skin; however, if it is allowed to enter the bloodstream
or internal tissues, these bacteria may cause a variety of potentially serious
infections. Transmission is typically from direct contact. However, some
infections involve other transmission methods. The
multiple virulence factors, high survival rate, wide environmental spread, and
antibiotic resistance have made these organisms a potential pathogen,
especially to immune-compromised individuals.
The different studies conducted by Singh et al.,
(2015), Prasad et al., (2016), and Sharma et al., (2017) have evaluated the antimicrobial
activity of Curcuma rhizome extract against various pathogenic bacteria,
including Staphylococcus aureus, Escherichia coli, and Pseudomonas
aeruginosa. Their research outcome revealed that the different fractions of
Curcuma rhizome, such as petroleum ether, chloroform, benzene, methanol,
and aqueous fraction, were noted as an effective antimicrobial agent additionally,
they quoted that the methanolic fraction of Curcuma rhizome has been
shown higher potential to inhibit pathogenic bacteria.
The present work aimed to analyze the antimicrobial
effect of different Curcuma (e.g., Curcuma longa, Curcuma
caesia, and Curcuma amada) extracts against human pathogenic
bacterial strains. The findings could be able to support further characterization
of Curcuma as a potent antimicrobial agent to enrich the blend of
traditional and modern medicines.
Materials and Methods
Fresh
rhizome of Curcuma longa (turmeric), Curcuma caesia (Black
turmeric), and Curcuma amada (Mango ginger) were collected from the
herbal garden of D.L.S. P.G. College. The fresh rhizome of these plants was washed
with distilled water to remove soil and other impurities, cut into small pieces
of 0.25 inches, and shaded for 2 days. Further, the rhizome fragments were
dried in a hot air oven at 50 °C for 24 h and were ground into powder and passed
through a sieve (mesh size of 2 mm diameter) and collected the fine powder
which further acted as a sample. The samples were stored at 40C for
further use.
Preparation of Curcuma’s rhizome
extracts: The acetone, ethyl alcohol, methyl alcohol, petroleum
ether, and distilled water extract of powdered samples were prepared using the cold
extraction method. These extracts were then subjected to evaporation to
concentrate and stored at 40C until used.
Assessment of antimicrobial activity
of extracts: The disc diffusion method was used for the detection
of the antimicrobial activity of different extracts. Bacterial cultures of E.
coli (MTCC 443), Pseudomonas aeruginosa (MTCC 424), and Staphylococcus
aureus (MTCC96) were used as test organisms for the assessment of antimicrobial
activities.
The broth of
test bacterial pathogens (at a density of 108 cells ml–1)
was prepared and inoculated into the surface of nutrient agar plates separately.
The discs of 2.0 mm were soaked into Curcuma’s rhizome extracts and placed on
the surface of nutrient agar plates (pre-inoculated with test pathogens), and
incubated for 24 h at 37°C (Harit et al. 2013).
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Curcuma amada (Mango ginger)
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Curcuma caesia (Black turmeric)
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Curcuma longa (Turmeric)
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Fig. 1: Rhizome of C.
longa, C. caesia, and C. amada
Fig. 2: Extracts of C. amada in different solvents.
Results and Discussion
The antibacterial activity of C. longa, C.
caesia, and C. amada was assessed
against one Gram-positive (S. aureus) and two Gram-negative
bacteria (E. coli and P. aeruginosa).
The acetone and distilled water extracts of C.
caesia, have shown maximum antimicrobial activity against E. coli
while ethanol and methanol extract of Curcuma amada have exhibited
maximum antimicrobial action against Pseudomonas aeruginosa and Staphylococcus
aureus respectively (Table 1). The ethanol and methanol extract of C.
amada was effective only against gram-positive bacteria (S.
aureus) whereas the rest extract including petroleum ether, acetone, and distilled
water have not given antimicrobial activity. Kaur et al. (2018) evaluated the antimicrobial potency of
C. amada and C. caesia against S. aureus, Streptococcus
pyogenes, E. coli, and P. aeruginosa and observed that the C.
amada has stronger inhibition capability against both gram-positive and gram-negative
bacteria, and also mentioned that ethanol extract of C. caesia was found
to be an effective antimicrobial agent.
Gupta et
al. (2015) claimed that petroleum ether, benzene, chloroform, methanol, and water extract of C. longa rhizome has been noted with higher
activity against S. aureus between 9 mm, and also similar activity
reported by Negi et al. (1999) while in our study petroleum ether extract of C.
longa has higher activity against E. coli (7.0 mm) than S. aureus (5.5 mm). Adamczak,
et al. (2020) documented that C. longa has significant antimicrobial
action against 6 gram-positive and 9 gram-negative bacterial strains (with
which S. aureus, E. coli, and P.
aeruginosa are included). Our study showed that C.
caesia and C. longa have noteworthy
antimicrobial activity against E. coli,
C. longa and C.
amada have against P. aeruginosa.
Table 1: Antimicrobial Activity of Curcuma
Species
Solvent
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Antimicrobial Activity of Curcuma species (diameter in
mm)
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Escherichia coli
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Pseudomonas aeruginosa
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Staphylococcus aureus
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Curcuma longa
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Curcuma caesia
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Curcuma amada
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Curcuma longa
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Curcuma caesia
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Curcuma amada
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Curcuma longa
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Curcuma caesia
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Curcuma amada
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Ethanol
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5.0
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7.0
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-
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6.5
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-
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7.4
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4.5
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2.2
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5.5
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Methanol
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5.5
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5.5
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-
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6.0
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-
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5.5
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4
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2.3
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5.7
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Petroleum
ether
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7.0
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5.0
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-
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4.7
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4.5
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5.5
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5.5
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-
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-
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Acetone
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2.0
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7.5
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-
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2.5
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3.5
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5.5
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-
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2.5
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-
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Distilled
water
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5.5
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7.5
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-
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4.5
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4.5
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5.5
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-
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3.5
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-
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Conclusion
The present investigation was focused on the different
Curcuma genera including C. caesia, C. longa, and C.
amada to evaluate their antibacterial potential. Different species of Curcuma
were selected as although they belong to same genus but exhibits different
biological and pharmacological attributes. These curcuma rhizomes have diverse
antimicrobial agents that could further be applied for drug development studies.
The phytochemical examinations of these curcuma rhizomes have revealed the
presence of carbohydrates, flavonoids, resins, glycosides, saponins,
phytosterols, and diterpenoids contents. The polyphenols i.e., flavonoids and
phenolic acids, have extensively responsible for antimicrobial potential. The literature
revealed that Curcuma has around 0.40% flavonoid and 0.08% phenol
content that participate in antimicrobial action. Our results
indicated that ethanol extract of C. longa and C. amada exhibited
maximum antibacterial efficacy against P. aeruginosa, whereas aqueous and
acetone extracts of C. caesia displayed maximum activity against E.
coli. The methanolic extract of C.
amada and petroleum ether of C. longa unveiled maximum antimicrobial
activity against S. aureus. The prevention and therapy
of bacterial infections are presently of prime concern in the healthcare
sector, and the inclusion of Curcumin in dietary will help to prevent
pathogenic bacterial infection and purified Curcuma extract in
pharmaceutical preparations.
Conflict of Interest
There is no conflict of interest.
Ethical Compliance Standard
There is no Ethical Compliance Standard.
Acknowledgment
The authors are thankful to D.L.S. P.G. College,
Bilaspur for the well-equipped research facility.
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