Nandita Dasgupta et al.

/ Journal of Pharmacy Research 2012,5(8),4201-4203

Research Article Available online through
ISSN: 0974-6943 http://jprsolutions.info

Antibacterial Activity of Leaf Extract of Mexican Marigold (Tagetes erecta)

Against Different Gram Positive and Gram Negative Bacterial Strains
Nandita Dasgupta*, Shivendu Ranjan, Proud Saha, Rahul Jain, Swati Malhotra and M.A. Arabi Mohamed Saleh.
School of Bioscience and Technology, VIT University, Vellore – 632014, Tamil Nadu, India
Received on:11-05-2012; Revised on: 16-06-2012; Accepted on:28-07-2012

ABSTRACT
The antibacterial effect of Mexican marigold (Tagetes erecta) leaf extract at room temperature against 10 gram positive multidrug resistant bacterial
isolates including Enterococcus faecalis, Enterococcus faecium. Staphylococcus epidermidis, Staphylococcus aureus, Bacillus cereus, Erysipelothrix
rhusiopathiae, Propionibacterium acne, Streptococcus pneumoniae, Streptococcus agalactine, Staphylococcus saprophyticus and 6 gram negative multidrug
resistant bacterial isolates including Escherichia coli, Klebsiella pneumoniae, Pseudomonas auregenosa, Salmonella enteriditis, Acinetobacter baumannii,
Alcaligen faecalis were studied by well diffusion method. The maximum antibacterial effect of Mexican Marigold leaf extract among those micro-organism
was obtained for Acinetobacter baumannii (Acitvity Index = 0.913333333) and Propionibacterium acne (Acitvity Index = 0.906666667) and minimum
was for Streptococcus pneumonia (Activity Index = 0.026666667). The results suggest that species of Mexican marigold i.e. Tagetes erecta has
antibacterial effect against airborne disease causing gram positive and gram negative bacteria and mainly against skin infection causing bacteria, and hence
can be useful in developing drugs for diseases like dermatitis, acne, skin races and also can be developed as antiseptic.

Key words: Tagetes erecta, Leaf extract, Gentamicin, Activity index.

INTRODUCTION
The plant Mexican marigold is classified as: Kingdom: Plantae; Phylum:
Angiosperms; Class: Eudicots; Order: Asterales; Family: Asteraceae;
Subfamily: Asteroideae; Tribe: Tageteae [1]. The tagetes erecta contain 56
species of herbaceous plant in sunflower family. Tagetes genus is originated
in North and South America. Now-a-days it is also cultivated in Asian
countries like India, Bhutan, Nepal, China [2].Use of Tagetes in magical,
religious and medicinal uses have been mentioned in history. Tagetes is used
in the treatment of hiccups [3]. T.erecta is used in garlands to decorate houses
and Idols of God during the festivals in India. Since ancient time parts of this
plant has been used for medicinal purposes and for the skin wash and yellow
dye is used as by Cherokee [4]. Marigold is commonly used in food additives
as a coloring agent and as animal food in fodder (dried flower meal & extract
used as supplement for poultry feed). It is used as an ornamental plant and
also in tannin or dye industry [5]. It is also used in medicine (folklore) and as
a poison for non vertebrates and in plant pest control [6]. Marigold has social
value because of its use in religious purposes. The Tagetes is used to adorn
Idols during festivals in India because of its stability [7]. Tagetes erecta is
used as a food color in the African countries because of its richness in
carotenoid leutin [8]. The species Tagetes lucida is used to prepare sweet
medicinal tea [9]. Tagetes minuta is used to make a popular potato dish called
Ocopa [10]. The essential oil of Tagetes is extracted from its leaves, stems,
stalk and flowers by steam distillation [11]. The main components of this oil
are limonene, ocimene, lagetone and valeric acid [12]. Essential oil of Tagetes
can be attributed to its properties like anti biotic, anti microbial, anti parasitic,
anti septic, anti spasmodic [13].
The essential oil is also used to ease phlegm and catarrh, congestions and in
wound burns, skin diseases, athlete’s foot, colitis and dermatitis [14]. From
scientific studies it is observed that thiophenes (natural phytochemicals that
*Corresponding author.
Nandita Dasgupta
School of Bioscience and Technology,
VIT University, Vellore – 632014,
Tamil Nadu, India
contains sulfur containing ring) may be the active ingredients in Tagetes
species and they have been used against gram negative and gram positive
bacteria in vitro [15]. Tagetes erecta is also used as interplant with cowpea
for control of nematode pests [16]. Tagetes erecta essential oil also has
antioxidant properties [17].
MATERIALS AND METHODS
Microorganisms used
A total of 10 gram positive and 3 gram negative organisms were used in the
study. The isolates of Enterococcus faecalis, Enterococcus faecium.
Staphylococcus epidermidis, Staphylococcus aureus, Bacillus cereus,
Erysipelothrix rhusiopathiae, Propionibacterium acne, Streptococcus
pneumoniae, Streptococcus agalactine, Staphylococcus saprophyticus,
Escherichia coli, Klebsiella pneumoniae, Pseudomonas auregenosa,
Salmonella enteriditis, Acinetobacter baumannii and Alcaligen faecalis were
obtained from the culture collection centre of Institute of Microbial
Technology (IMTECH), Chandigarh, India.
Media Used
The media used in the present investigation was Nutrient Agar media, which
was obtained from Hi – Media Laboratories Ltd., Mumbai, India.
Antibiotic Gentamycin used
The antibiotic standard used in this investigation was Gentamicin G30
susceptibility test discs (30 mg per disc) which were obtained from Hi –
Media Laboratories Ltd., Mumbai, India.
Aqueous stock preparations of Mexican marigold leaf
Fresh Mexican marigold (Tagetes erecta) leaves were collected from nursery
at VIT University, Tamil Nadu, India. 50 gm of surface sterilised Mexican
marigold leaves were gently grinded using mortar and pestle. The crude
extract were taken and centrifuged at 5000 rpm for 10 minutes at 27°C.

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Now the supernatant obtained was the homogenized Mexican marigold leaf
extract which was separated in conical flask. The supernatant extracted was
used as stock for antibacterial activity and stored in the refrigerator.
Re – culturing of micro organisms used or Enrichment of culture
The bacterial cultures were maintained in Peptone Saline Water.
Analyzing Antibacterial Activity of Mexican marigold (Tagetes erecta)
leaf extract using Agar well diffusion assay
100µl of each bacterial strain was used to make lawn culture on sterile
Nutrient Agar plates. 6mm wells were made on the plates using cork borer.
To these well 100µl of Marigold leaves extract was added. 100µl of Mexi-
can marigold leaves extract contained 1.05µg of total dry matter. There
should be sufficient distance between the wells and petriplate wall to avoid
overlapping of zone of inhibition. The plates were incubated in an upright
position at 37°C for 24 hours. The diameter of zone of inhibition was
measured in millimeter and the results were recorded. The inhibition zones
with diameter less than 8mm were not considered as having antibacterial
activity.
SUMMARY AND DISCUSSION
The activity index of Mexican marigold for different gram positive and gram
negative isolated bacterial strain were calculated after antibacterial testing
using agar well diffusion method and antibacterial sensitivity testing with
Gentamicin. The maximum antibacterial effect of Mexican marigold leaf
extract among those micro-organism was obtained for Acinetobacter
baumannii (Acitvity Index = 0.913333333) and Propionibacterium acne
(Acitvity Index = 0.906666667) and minimum was for Streptococcus
pneumoniae (Activity Index = 0.026666667). Reason for minimum activity
index on Streptococcus pneumoniae is its capsulated structure. The results
suggest us that species of marigold i.e. Tagetes erecta has antibacterial
effect against airborne disease causing gram positive and gram negative
bacteria and mainly against skin infection causing bacteria, so it can be
useful in developing drugs for diseases like dermatitis, acne, skin races and
also can be developed as antiseptic.
Table 1: Average Activity index of Mexican marigold (Tagetes erecta)
for 10 gram positive and 6 gram negative isolated bacterial strains.

Antibiotic sensitivity testing Sr. Name of isolated Average zone Average Zone Average
The test microorganisms were also tested for their sensitivity against the No bacterial species of inhibition of inhibition Activity
of Tagetes erecta of Gentamicin Index
antibiotic (Gentamicin 30 mg). Using sterile cotton swabs, the enriched leaf extract (in mm)
cultures were aseptically swabbed on the surface of sterile nutrient agar (in mm)
plates. Using ethanol dipped and flamed forceps, the antibiotic (Gentami-
1. Enterococcus faecalis 1.123333 1.466666667 0.763333333
cin, 30 mg) disc was aseptically placed over the agar plates sufficiently 2. Enterococcus faecium 1.156667 1.466666667 0.783333333
separated from the wells formed to avoid overlapping of inhibition zone. 3. Staphylococcus epidermidis 0.893333 1.143333333 0.776666667
The plates were incubated at 37°C for 24 hours and the diameter of inhibi- 4. Staphylococcus aureus 1.166667 1.466666667 0.79
5. Bacillus cereus 1.15 1.46 0.773333333
tion zones were measured in mm.
6. Erysipelothrix rhusiopathiae 1.196667 1.476666667 0.806666667
7. Propionibacterium acne 1.34 1.46 0.906666667
Measuring Activity Index 8. Streptococcus pneumoniae 0.006667 0.24 0.026666667
Following formula was used to measure Activity Index, 9. Streptococcus agalactine 0.006667 0.25. 0.037533333
10. Staphylococcus saprophyticus 0.006667 0.14 0.047621428
11. Escherichia coli 1.256667 1.49 0.816666667
Activity Index = (Zone of inhibition of extract/Zone of inhibition of antibi- 12. Klebsiella pneumoniae 0.403333 0.893333333 0.443333333
otic). 13. Pseudomonas auregenosa 1.23 1.406666667 0.87
14. Salmonella enteriditis 0.306667 0.666666667 0.453333333
15. Acinetobacter baumannii 1.196667 1.3 0.913333333
Zone of inhibition of stocks against each bacterial species and similarly 16. Alcaligen faecalis 0.983333 1.166666667 0.84
zone of inhibition of Antibiotic (Gentamicin) were measured.

RESULT AND DISCUSSION

Activity Index of Mexican marigold (Tagetes erecta) leaf extract

From the triplicate readings average zone of inhibition for Mexican mari-
gold leaf extract, average zone of inhibition for Gentamicin (30 mg) and
using this activity index of Mexican marigold leaf extract for 10 gram posi-
tive and 6 gram negative isolated bacterial strain were calculated (Table 1).

For gram positive bacterial strains maximum antibacterial effect was found
in Acinetobacter baumannii (Acitvity Index = 0.913333333) and minimum
for Streptococcus pneumonia (Activity Index = 0.026666667). Also Mexi-
can marigold leaf has very less antibacterial effect on Streptococcus
agalactine, Staphylococcus saprophyticus having activity index 0.037533333
and 0.047621428 respectively. For other gram positive bacterial strains
Mexican marigold has activity index between 0.763 and 9.066.

For gram negative bacterial strains maximum antibacterial effect was found
in Acinetobacter baumannii and Pseudomonas auregenosa with the activ-
ity index of 0.913333333 and 0.87 respectively, and minimum for Kleb-
siella pneumoniae (Activity Index = 0.443333333).
Figure 1: Graphical analysis of Activity Index of various isolated gram positive
bacterial strain.

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 Nandita Dasgupta et al. / Journal of Pharmacy Research 2012,5(8),4201-4203
Figure 2: Graphical analysis of Activity Index of various isolated gram negative
bacterial strain.
Figure 3: Zone of inhibition by mexican
marigold on Erysipelothrix rhusiopathiae.
Figure 5: Zone of inhibition by Figure 6: Zone of inhibition by
mexican marigold on Escherichia coli. mexican marigold on Pseudomonas
auregenosa.
Figure 7: Zone of inhibition by
Mexican marigold on Acinetobacter
baumannii.
Figure 10: Zone of inhibition by Figure 11: Zone of inhibition by
Mexican marigold on Streptococcus Mexican marigold on Streptococcus
pneumonia, zero activity index. agalactine, zero activity index.
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Mexican marigold on Alcaligen
faecalis. Source of support: Nil, Conflict of interest: None Declared
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