Nandita Dasgupta et al.

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

Research Article ISSN: 0974-6943

Available online through www.jpronline.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, athletes foot, colitis and dermatitis [14]. From scientific studies it is observed that thiophenes (natural phytochemicals that

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 27C.

*Corresponding author.
Nandita Dasgupta School of Bioscience and Technology, VIT University, Vellore 632014, Tamil Nadu, India

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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 100l 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 100l of Marigold leaves extract was added. 100l of Mexican marigold leaves extract contained 1.05g 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 37C 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. Antibiotic sensitivity testing The test microorganisms were also tested for their sensitivity against the antibiotic (Gentamicin 30 mg). Using sterile cotton swabs, the enriched cultures were aseptically swabbed on the surface of sterile nutrient agar plates. Using ethanol dipped and flamed forceps, the antibiotic (Gentamicin, 30 mg) disc was aseptically placed over the agar plates sufficiently separated from the wells formed to avoid overlapping of inhibition zone. The plates were incubated at 37C for 24 hours and the diameter of inhibition zones were measured in mm. Measuring Activity Index Following formula was used to measure Activity Index, Activity Index = (Zone of inhibition of extract/Zone of inhibition of antibiotic). Zone of inhibition of stocks against each bacterial species and similarly 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 marigold leaf extract, average zone of inhibition for Gentamicin (30 mg) and using this activity index of Mexican marigold leaf extract for 10 gram positive 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 Mexican 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 activity index of 0.913333333 and 0.87 respectively, and minimum for Klebsiella pneumoniae (Activity Index = 0.443333333).
Figure 1: Graphical analysis of Activity Index of various isolated gram positive bacterial strain.

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.
Sr. Name of isolated No bacterial species Average zone of inhibition of Tagetes erecta leaf extract (in mm) 1.123333 1.156667 0.893333 1.166667 1.15 1.196667 1.34 0.006667 0.006667 0.006667 1.256667 0.403333 1.23 0.306667 1.196667 0.983333 Average Zone of inhibition of Gentamicin (in mm) Average Activity Index

1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16.

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 Alcaligen faecalis

1.466666667 1.466666667 1.143333333 1.466666667 1.46 1.476666667 1.46 0.24 0.25. 0.14 1.49 0.893333333 1.406666667 0.666666667 1.3 1.166666667

0.763333333 0.783333333 0.776666667 0.79 0.773333333 0.806666667 0.906666667 0.026666667 0.037533333 0.047621428 0.816666667 0.443333333 0.87 0.453333333 0.913333333 0.84

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Figure 10: Zone of inhibition by Mexican marigold on Streptococcus pneumonia, zero activity index.

Figure 11: Zone of inhibition by Mexican marigold on Streptococcus agalactine, zero activity index.

Figure 2: Graphical analysis of Activity Index of various isolated gram negative bacterial strain.

Figure 3: Zone of inhibition by mexican Figure 4: Zone of inhibition by mexican marigold on Erysipelothrix rhusiopathiae. marigold on Propionibacterium acne.

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 9: Zone of inhibition by Mexican marigold on Alcaligen faecalis.

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Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.5 Issue 8.August 2012

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