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Antibacterial Activity of Leaf Extract of Mexican Marigold (Tagetes erecta) Against Different Gram Positive and Gram Negative Bacterial Strains
 

Antibacterial Activity of Leaf Extract of Mexican Marigold (Tagetes erecta) Against Different Gram Positive and Gram Negative Bacterial Strains

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ABSTRACT ...

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.

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    Antibacterial Activity of Leaf Extract of Mexican Marigold (Tagetes erecta) Against Different Gram Positive and Gram Negative Bacterial Strains Antibacterial Activity of Leaf Extract of Mexican Marigold (Tagetes erecta) Against Different Gram Positive and Gram Negative Bacterial Strains Presentation Transcript

    • Journal of Pharmacy Research Vol.5 Issue 8.August 2012 Nandita Dasgupta et al. / Journal of Pharmacy Research 2012,5(8),4201-4203 4201-4203 Research Article ISSN: 0974-6943 Available online through www.jpronline.info *Corresponding author. Nandita Dasgupta School of Bioscience and Technology, VIT University, Vellore – 632014, Tamil Nadu, India 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 ofTagetes 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 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 saprophyticusand 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. 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 baumanniiand 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.
    • Journal of Pharmacy Research Vol.5 Issue 8.August 2012 Nandita Dasgupta et al. / Journal of Pharmacy Research 2012,5(8),4201-4203 4201-4203 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. 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 (Gentami- cin, 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 37°C for 24 hours and the diameter of inhibi- tion 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 antibi- otic). 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 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 saprophyticushaving 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). 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 Average zone Average Zone Average No bacterial species of inhibition of inhibition Activity of Tagetes erecta of Gentamicin Index leaf extract (in mm) (in mm) 1. Enterococcus faecalis 1.123333 1.466666667 0.763333333 2. Enterococcus faecium 1.156667 1.466666667 0.783333333 3. Staphylococcus epidermidis 0.893333 1.143333333 0.776666667 4. Staphylococcus aureus 1.166667 1.466666667 0.79 5. Bacillus cereus 1.15 1.46 0.773333333 6. Erysipelothrix rhusiopathiae 1.196667 1.476666667 0.806666667 7. Propionibacterium acne 1.34 1.46 0.906666667 8. Streptococcus pneumoniae 0.006667 0.24 0.026666667 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 12. Klebsiella pneumoniae 0.403333 0.893333333 0.443333333 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 16. Alcaligen faecalis 0.983333 1.166666667 0.84 Figure 1: Graphical analysis of Activity Index of various isolated gram positive bacterial strain.
    • Journal of Pharmacy Research Vol.5 Issue 8.August 2012 Nandita Dasgupta et al. / Journal of Pharmacy Research 2012,5(8),4201-4203 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 4: Zone of inhibition by mexican marigold on Propionibacterium acne. Figure 5: Zone of inhibition by mexican marigold on Escherichia coli. Figure 6: Zone of inhibition by 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. 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. REFERENCES 1. Joy, P.P., Thomas, J., Mathew, S., and Skaria, B.P., Ed. Medicinal Plants. Tropical Horticulture Vol. 2. Naya Prokash, Calcutta pub- lisher, 449-632 (2001). 2. Cruz-Ramírez Luís Alfredo et al, Mexican crops of agroalimentary importance, Advances in Agricultural and Food Biotechnology, 35- 53, (2006) 3. M. Abdul Halim et al., The use of plants in traditional health care practice of the shaiji community in southwestern Bangladesh, Jour- nal of Tropical Forest Science 19(3): 168–175 (2007) 4. Nilani Packianathan,Saravanan Karumbayaram, Formulation and Evaluation of Herbal Hair Dye: An Ecofriendly Process, journal of pharmaceutical sciences and research, 2 (10), 648-656, (2010) 5. Richard Cantrill, Lutein from Tagetes erecta for use in foods for particular nutritional uses, The EFSA Journal, 315 (1), 1-12, (2006) 6. Farjana Nikkon et al., Insecticidal Activity of Flower of Tagetes erecta L. against Tribolium castaneum (Herbst), Research Journal of Agriculture and Biological Sciences,5(5),748-753,(2009). 7. Kapil V Patil et al., Stability Analysis in Marigold (Tagetes erecta L.) for Flower Yield and Quality Parameters, Research Journal of Agri- cultural Sciences,2(2),237-240,( 2011). 8. D. Jothi, Extraction of natural dyes from african marigold flower (tagetesereecta l) for textile coloration, AUTEX Research Journal, 8(2), 49-53,(2008) 9. John M. Maisch, Botanical Medicine Monographs and Sundry on an indigenous species of croton, American Journal of Pharmacy, 57(12), 1-7, (1985) 10. Jose Rafael Lovera, Ed. Food Culture in South America, 1st Edn, Vol 1, Greenwood Publishing Group: 137-200, (2005). 11. Ester R. Chamorro et al., Evaluation of tagetes minuta l. essential oils to control varroa destructor (acari: varroidae), The Journal of the Argentine Chemical Society, 98(1), 39-47 (2011). 12. Mohammad Hadi Meshkatalsadat, Chemical characterization of vola- tile components of tagetes minuta l. cultivated in south west of iran by nano scale injection, Digest Journal of Nanomaterials and Biostructures, 5 (1): 101-106,(2010). 13. H. J. D. Dorman, S. G. Deans, Antimicrobial agents from plants: antibacterial activity of plant volatile oils, Journal of Applied Micro- biology, 88(2): 308-316, (2000) 14. Chowdhury M. S. H. et al., Use of plants in healthcare: a traditional ethno-medicinal practice in rural areas of southeastern Bangladesh, International Journal of Biodiversity Science & Management, 5(1): 41-51 (2009) 15. Roberta Piccaglia, Mauro Marotti, Silvia Grandi, Lutein and lutein ester content in different types of Tagetes patula and T. erecta, In- dustrial Crops and Products, 8(1) : 45–51(1998). 16. T.I. Olabiyi, E.E.A. Oyedunmade, Marigold (Tagetes Erecta L.) as interplantwithcowpeaforcontrolofnematodepests,AfricanCrops Science Conference Proceeding, 8(1): 1075-1078, (2007) 17. Rosa Martha Pérez Gutiérrez et al.,, Antioxidant activity of tagetes erecta essential oil, Journal of the Chilean Chemical Society, 51(2): 883-886, (2006). Source of support: Nil, Conflict of interest: None Declared